IV ... MYCOTAXON 120

MY COTAXON

VOLUME ONE HUNDRED TWENTY — TABLE OF CONTENTS

COVER SECTION

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RESEARCH ARTICLES

Glomus trufemii (Glomeromycetes), a new sporocarpic species

from Brazilian sand dunes Bruno Tomio Goto, Jomar Gomes Jardim,

Gladstone Alves da Silva, Eduardo Furrazola, Yamir Torres-Arias & Fritz Oehl

Two new species of Pluteus section Celluloderma from the

Dominican Republic Alfredo Justo, Eliseo Battistin & Claudio Angelini

The lichen genus Chapsa (Graphidaceae) in India

Santosh Joshi, Dalip K. Upreti & Sanjeeva Nayaka

A new species of Inonotus (Hymenochaetaceae) and Trametes cingulata

(Polyporaceae) newly recorded from Brazil

Maira Cortellini Abrahao & Adriana de Mello Gugliotta

Manoharachariella indica sp. nov. from the Western Ghats, India

Kunhiraman C. Rajeshkumar & Sanjay K. Singh

Russula jilinensis sp. nov. from northeast China

Guo-Jie Li, Sai-Fei Li, Xing-Zhong Liu & Hua-An Wen

Acrogenospora hainanensis sp. nov. and new records of microfungi

from southern China

Jian Ma, Li-Guo Ma, Yi-Dong Zhang, Ji-Wen Xia & Xiu-Guo Zhang

A new species of Nectria on Populus from China

Zhao-Qing Zeng & Wen-Ying Zhuang

A new species of the lichen genus Phlyctis from Maharashtra, India

Gayatri Chitale & Urmila Makhija

Two new freshwater species of Annulatascaceae from China

Dian-Ming Hu, Lei Cai, Ali Hassan Bahkali & Kevin D. Hyde

New records of little-known species of Carbomyces

(Carbomycetaceae, Ascomycota) Gabriel Moreno, Marcos Lizarraga,

Martin Esqueda, Ricardo Galan & Pablo Alvarado

Cantharellus zangii, a new subalpine basidiomycete from

southwestern China

Xiao-Fei Tian, Bart Buyck, Shi-Cheng Shao, Pei-Gui Liu & Yan Fang

Seven lichen species new to Poland

99.

Martin Kukwa, Anna Lubek, Rafat Szymczyk & Anna Zalewska 105

APRIL-JUNE 2012... V

Dissoconium proteae newly recorded from China Rong Zhang,

Yongna Mao, Lu Hao, Hongcai Chen, Guangyu Sun & Mark L. Gleason 119

The fungal collection of the Jaiellonian University Herbarium (KRA),

Krakow, Poland Anna Maria Ociepa, Szymon Zubek & Piotr Mleczko 127

Mycena pseudoinclinata, new to Italy

Alfonso La Rosa, Alessandro Saitta, Riccardo Compagno, & Giuseppe Venturella133

Rossbeevera yunnanensis (Boletaceae, Boletales), a new

sequestrate species from southern China

Takamichi Orihara, Matthew E. Smith, Zai-Wei Ge & Nitaro Maekawa 139

Octaviania violascens: a new sequestrate bolete

from Thailand Rattaket Choeyklin, Thitiya Boonpratuang,

Sujinda Sommai & Sayanh Somrithipol 149

New records of cercosporoid hyphomycetes from Iran

Mahdi Pirnia, Rasoul Zare, Hamid R. Zamanizadeh & Akbar Khodaparast 157

Phaeocollybia nigripes (Agaricomycetes), a new species from Brazil

Victor R.M. Coimbra, Tatiana B. Gibertoni & Felipe Wartchow 171

Peziza michelii and its ectomycorrhizae with Alnus nitida

(Betulaceae) from Pakistan T. Ashraf, M. Hanif & A.N. Khalid 181

Erysiphe javanica sp. nov., a new tropical powdery mildew

from Indonesia Jamjan Meeboon, Iman Hidayat & Susumu Takamatsu 189

Pisolithus: A new species from southeast Asia and a new combination

C. Phosri, M.P. Martin, N. Suwannasai, P. Sihanonth & R. Watling 195

Terriera simplex, a new species of Rhytismatales from China

Xiao-Ming Gao, Chun-Tao Zheng & Ying-Ren Lin 209

The Leptogium juressianum complex in southeastern Brazil

Marcos J. Kitaura & Marcelo P. Marcelli 215

Coriolopsis psila comb. nov. (Agaricomycetes) and two new

Coriolopsis records for Brazil Georgea S. Nogueira—Melo,

Priscila S. de Medeiros, Allyne C. Gomes-Silva, Leif Ryvarden, Helen M.P. Sotao

& Tatiana B. Gibertoni 223

A new species of Postia (Basidiomycota) from Northeast China

Bao-Kai Cui & Hai-Jiao Li 231

Three new Caeoma species on Rosa spp. from Pakistan

N.S. Afshan, A.N. Khalid & A.R. Niazi 239

Nomenclatural status and morphological notes on

Tubifera applanata sp. nov. (Myxomycetes) D.V. Leontyev & K.A. Fefelov 247

Paralepistopsis gen. nov. and Paralepista

(Basidiomycota, Agaricales) Alfredo Vizzini & Enrico Ercole 253

Three new species of Septobasidium (Septobasidiaceae) from Hainan Province

in China Suzhen Chen & Lin Guo 269

Characterization of the causal agent of poplar anthracnose occurring

in the Beijing region Zheng Li, Cheng Ming Tian & Ying Mei Liang 277

VI... MYCOTAXON 120

Sympodioplanus yunnanensis, a new aquatic species from

submerged decaying leaves Guang-Zhu Yang, Kai-Ping Lu, Yue Yang,

Li-Bo Ma, Min Qiao, KeQing Zhang & Ze-Fen Yu

Phaeotrichoconis crotalariae, endophytic on Vitis labrusca

Thais Emanuelle Feijé de Lima, José Luiz Bezerra

& Maria Auxiliadora de Queiroz Cavalcanti

287

291

Studies on Wrightoporia from China 1. A new species from Hunan Province,

South China Jia-jia Chen & Hai-You Yu

A new species of Bipolaris from Iran Abdollah Ahmadpour,

Zeinab Heidarian, Maryam Donyadoost-Chelan,

Mohammad Javan-Nikkhah & Takao Tsukiboshi

Notes on Xylophallus xylogenus (Phallaceae, Agaricomycetes)

based on Brazilian specimens

Larissa Trierveiler-Pereira & Rosa Mara Borges da Silveira

Plectania seaveri sp. nov. (Ascomycota, Pezizales), a new discomycete

from Bermuda Matteo Carbone, Carlo Agnello & Scott LaGreca

New species of Entoloma (Basidiomycetes, Agaricales) from

Kerala State, India C.K. Pradeep, K.B. Vrinda, Shibu P. Varghese & TJ. Baroni

A new species of Infundichalara from pine litter Ondiej Koukol

Clavulicium hallenbergii, a new corticioid species from India

Avneet P. Singh, Jaspreet Kaur & G.S. Dhingra

Vararia longicystidiata sp. nov. (Agaricomycetes) from India

Samita, S.K. Sanyal, G.S. Dhingra & Avneet P. Singh

A new cystidiate variety of Omphalina pyxidata (Basidiomycota,

Tricholomatoid clade) from Italy Alfredo Vizzini,

Mariano Curti, Marco Contu & Enrico Ercole

Four new records of Aspergillus sect. Usti

from Shandong Province, China Long Wang

Systematics of the Gomphales: the genus Gomphus sensu stricto

Admir J. Giachini, Carla M. Camelini,

Marcio J. Rossi, Claudio R.ES. Soares & James M. Trappe

Myxomycetes from China 15: Arcyria galericulata sp. nov.

Bo Zhang, Tian-Hao Li, Qi Wang & Yu Li

New records of Puccinia species on Poaceae from Pakistan

N.S. Afshan, A.N. Khalid & A.R. Niazi

Lectotypification and characterization of the natural phenotype of

Fusarium bactridioides Keith A. Seifert & Tom Grafenhan

Lectotypification of Crepidotus variabilis var. subsphaerosporus

295

301

309

S17

331

343

353

357

361

373

385

401

407

415

Sona Jancovicova & Shaun R. Pennycook 423

A new species of Conidiobolus (Ancylistaceae) from Anhui, China

Yong Nie, Cui-Zhu Yu, Xiao-Yong Liu & Bo Huang

427

APRIL-JUNE 2012...

First record of the sequestrate fungus Neosecotium macrosporum

(Agaricales, Lepiotaceae) for Mexico Marcos Lizarraga,

Martin Esqueda, Mario Vargas-Luna & Gabriel Moreno

New combinations in Lactifluus. 3. L. subgenera Lactifluus and Piperati

A. Verbeken, K. Van de Putte & E. De Crop

Exobasidium ferrugineae sp. nov., associated with hypertrophied flowers

of Lyonia ferruginea in the southeastern USA

Aaron H. Kennedy, Nisse A. Goldberg & Andrew M. Minnis

Passalora aseptata, a new cercosporoid fungus from

northeastern Uttar Pradesh, India Raghvendra Singh,

Balmukund Chaurasia, Kalawati Shukla & Parmatma Prasad Upadhyaya

First records of Craterium aureonucleatum, Perichaena quadrata,

and Physarum mutabile in Italy Riccardo Compagno,

Alfonso La Rosa, & Giuseppe Venturella

Tuber microsphaerosporum and Paradoxa sinensis spp. nov.

Li Fan, Jin-Zhong Cao & Yu Li

Entomophthoromycota: a new phylum and reclassification

for entomophthoroid fungi Richard A. Humber

Moniliophthora aurantiaca sp. nov., a Polynesian species

occurring in littoral forests Bradley R. Kropp & Steven Albee-Scott

ABSTRACTS OF NEW MYCOBIOTAS ONLINE

Checklist of the Argentinean Agaricales 2.

Coprinaceae and Strophariaceae N. Niveiro & E. Alberté

Checklist of the Argentinean Agaricales 3.

Bolbitiaceae and Crepidotaceae N. Niveiro & E. Alberté

Preliminary checklist of the macromycetes from Collestrada

forest ecosystems in Perugia (Italy) Paola Angelini, Giancarlo Bistocchi,

Andrea Arcangeli & Roberto Venanzoni

New, rare, and noteworthy lichens in the Pollino National Park

(Basilicata, southern Italy)

D. Puntillo, M. Puntillo, G. Potenza & S. Fascetti

Saprobic fungi on wood an litter of Alnus alnobetula in the Swiss Alps

Beatrice Senn-Irlet, Rolf Miirner, Elia Martini, Nicolas Kiiffer,

Romano de Marchi & Guido Bieriice

NOMENCLATURE

Nomenclatural novelties proposed in volume 120

VII

437

443

451

461

465

471

477

493

505

506

507

vu ... MYCOTAXON 120

ERRATA FROM PREVIOUS VOLUMES

VOLUME 119

p. 97, lines 37-38 for: unpigmented lower two cells. read: pigmented basal cell.

p. 411, line 15 for: Korea Forest Institute read: Korea Forest Research Institute

PUBLICATION DATE FOR VOLUME ONE HUNDRED NINETEEN

MYCOTAXON for JANUARY-MARCH, VOLUME 119 (I-x1I + 1-511)

was issued on April 11, 2012

APRIL-JUNE 2012... IX

REVIEWERS — VOLUME ONE HUNDRED TWENTY

The Editors express their appreciation to the following individuals who

have, prior to acceptance for publication, reviewed one or more of the

papers prepared for this volume.

Reinhard Agerer

M. Catherine Aime

Joe Ammirati

Vladimir Antonin

Takayuki Aoki

André Aptroot

Mohammad Bahram

Richard Baird

Hans-Otto Baral

Timothy J. Baroni

Reinhard Berndt

Franco Umbertomaria

Bersan

Janusz Blaszkowski

Wolfgang von Brackel

Uwe Braun

Peter Buchanan

Lori M. Carris

Rafael FE. Castafieda Ruiz

Michael A. Castellano

Efren Cazares Gonzalez

Pedro Crous

Yu-Cheng Dai

Kanad Das

Cvetomir M. Denchev

Stephanos Diamandis

Maria Martha Dios

Pradeep K. Divakar

Uno Eliasson

John A. Elix

Francisco C.O. Freire

Walter Gams

Laura Guzman-Davalos

Nils Hallenberg

Shuanghui He

Rosanne Healy

José R. Hernandez

M. Luciana

Hernandez Caffot

Adriana Inés Hladki

Kentaro Hosaka

Peter Jeffries

Mikael Jeppson

Alfredo Justo

Gintaras Kantvilas

Taiga Kasuya

Bryce Kendrick

Paul M. Kirk

Roland Kirschner

Henning Knudsen

Richard P. Korf

Hanns Kreisel

Carlos Lado

Teresa Lebel

Bernardo E. Lechner

Xiao- Yong Liu

Guozhong Lit

Quan Li

Robert Licking

Eric H.C. McKenzie

Alistair McTaggart

Nelson Menolli, Jr.

Andrew M. Minnis

David W. Minter

David W. Mitchell

Pierre-Arthur Moreau

Gabriel Moreno

Jurga Motiejunaite

Edna Dora M.

Newman Luz

Lorelei L. Norvell

Eduardo R. Nouhra

Clark L. Ovrebo

Giovanni Pacioni

Omar Paino Perdomo

Ka-Lai Pang

Shaun R. Pennycook

Ronald H. Petersen

Stephen W. Peterson

Alan J.L. Phillips

Marcin Piatek

Donald H. Pfister

Huzefa Raja

Scott A. Redhead

Gerardo Lucio Robledo

Andrea Irene Romero

Amy Y. Rossman

James Scott

Mark R.D. Seaward

Keith A. Seifert

Hyeon-Dong Shin

Ewald Sieverding

Rosa Mara Bourges da

Silveira

Takuya Sakoda

B.M. Sharma

Hyeon-Dong Shin

Joost A. Stalpers

Brett Summerell

Guangyu Sun

Michal TomSovsky

Mauro Tretiach

Ricardo Valenzuela

Nicolas Van Vooren

Carlos G. Vélez

Else. C. Vellinga

Annemieke Verbeken

Rytas Vilgalys

Yu-Lian Wei

Anthony J. S. Whalley

Zhu-Liang Yang

Ming Ye

Hai-Sheng Yuan

Xiu-Guo Zhang

Zhongyi Zhang

x ... MYCOTAXON 120

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APRIL-JUNE 2012... XI

FROM THE EDITORS

MYCOTAXON, DELAYS, AND 2012 — In 1973 Dick Korf and Gregoire Hennebert noted

that taxonomic papers were unnecessarily delayed by traditional blind peer review and

press processing. They developed Mycotaxon to eliminate these frustrating bottlenecks

by having authors select their own expert reviewers (thus directing the review pace)

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We still strive to process manuscripts and publish as speedily as possible. Unfortunately

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We feel that MycoTaxon remains the most comprehensive journal for fungal taxonomy

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Warm regards,

LorELEI NoRVELL (Editor-in-Chief) with

SHAUN PENNYCOOK (Nomenclature Editor)

22 September 2012

MY COTAXON

http://dx.doi.org/10.5248/120.1

Volume 120, pp. 1-9 April-June 2012

Glomus trufemii (Glomeromycetes), a new sporocarpic species

from Brazilian sand dunes

BRUNO TOMIO GOTO”, JOMAR GOMES JARDIM', GLADSTONE ALVES DA SILVA’,

EDUARDO FURRAZOLA?, YAMIR TORRES-ARIAS? & FRITZ OEHL‘

"Departamento de Botanica, Ecologia e Zoologia, CB, Universidade Federal do

Rio Grande do Norte, Campus Universitario, 59072-970, Natal, RN, Brazil

*Departamento de Micologia, CCB, Universidade Federal de Pernambuco,

Av. Prof. Nelson Chaves, S/N, CEP 50670-420, Cidade Universitaria, Recife, PE, Brazil

*Instituto de Ecologia e Sistemdatica, IES-CITMA, A.P. 11900, La Habana, Cuba

‘Federal Research Institute Agroscope Reckenholz-Tanikon ART, Organic Farming Systems,

Reckenholzstrasse 191, CH-8046 Ziirich, Switzerland

‘CORRESPONDENCE TO: brunogoto@hotmail.com

ABSTRACT — Glomus trufemii, which forms large aggregates (< 850 x 1200 um) in the

rhizosphere of a herbaceous plant community in NE Brazilian sand dunes, is described as

new. Its subglobose glomerospores measuring 72-92 x 79-105 um have two spore wall layers:

an evanescent hyaline 0.3-0.8 um thick outer layer and a laminate orange brown to dark red

brown 7.4—15.5 um thick inner layer. The pigmentation of the subtending hypha is similar but

often much lighter than that of the laminate spore wall layer. The spore size and color, spore

wall structure, and features of the spore base and subtending hyphae separate this species

from similar yellow-brown to brown spored species like G. badium, G. glomerulatum, and

G. brohultii.

Key worps — morphology, Glomeromycota, Glomerales, restinga

Introduction

The Tulasne brothers described the genus Glomus in 1844 for G. macrocarpum

(the type species according to Clements & Shear 1931). Since then many other

fungi with glomoid spores like those of G. macrocarpum have been named,

so that now Glomus comprises >60 species. For many years Glomus was

considered monophyletic based solely on morphological evidence. Morton

& Redecker (2001), based on morphological and molecular data, erected the

genera Paraglomus (Paraglomeraceae) and Archaeospora (Archaeosporaceae),

whose members also formed glomoid spores. Schwarzott et al. (2001) inferred

from phylogenetic SSU rDNA molecular analyses that Glomus is non-

2 ... Goto & al.

monophyletic. Schiifler et al. (2001) subsequently erected a new phylum,

Glomeromycota, in which Glomus was represented by three phylogenetically

different groups: A, B, and C. Oehl & Sieverding (2004) transferred glomoid

species with two spore walls (with the inner wall serving as a germinal wall)

to their new genus Pacispora. Thereafter Walker & Schiifler (2004) transferred

Glomus spurcum C.M. Pfeiff. et al. to the newly established genus Diversispora

and order Diversisporales.

To date, glomoid glomerospores can be found in species representing

Archaeosporales, Diversisporales, Glomerales, and Paraglomerales, making

their identification difficult in the absence of molecular data. Morphological

identification has been facilitated recently in a thorough review of glomoid

spore-forming species (Oehl et al. 2011), which introduced helpful suggestions

based on molecular data associated with morphological differences among the

major phylogenetic clades in the Glomeromycota. Our description of G. trufemii

is based exclusively on the new key morphological criteria, and the species fits

the Oehl’s et al. (2011) revised concept of Glomus.

Material & methods

Study area and sites

Samples were collected in the Parque Estadual das Dunas de Natal “Jornalista Luiz Maria

Alves,’ the first Conservation Unit created in the Municipality of Natal, Rio Grande do

Norte State and currently one of the largest urban conservation areas with dune vegetation

in Brazil. The site is located at 5°46'S 35°12’W with soils analyzed as pH (H,O) =

5.5-5.75, P = 2-4 mg kg", 2,86 g dm? of organic matter. The climate is tropical rainy

(type Am of K6éppen) with a short four-month dry period, with a 25.5 °C mean annual

temperature and 1191 mm mean annual precipitation. In the sand dune ecosystem, the

typical ‘restinga vegetation varies from herbaceous to shrubs and trees (Oliveira-Filho

1993, Oliveira-Filho & Carvalho 1993). The park is composed of mainly represented

by tree species of the Anacardiaceae, Bignoniaceae, Fabaceae, Myrtaceae, and Rubiaceae

(Anacardium occidentale, Caesalpinia echinata, Campomanesia dichotoma, Chamaecrista

ensiformis, Eugenia ligustrina, Guettarda platypoda, Myrcia guianensis, Myrciaria

tenella, Tabebuia roseoalba, Tocoyena sellowiana) and herbaceous species of Poaceae,

Cyperaceae, Asteraceae, Araceae, and Rubiaceae.

Morphological analyses

Glomerospores and sporocarps were separated from the soil samples by sucrose

gradient in plate dishes (Blaszkowski et al. 2006). Sporocarps and glomerospores were

placed in dishes with water and separated under a dissecting microscope. Sporocarps were

fragmented to study spore organization. Glomerospores were mounted on microscope

slides either in water (to check unmodified characteristics of spore wall components and

spore colour; Spain 1990) or permanently in PVLG (Omar et al. 1979) or PVLG with

Melzer's reagent (Koske & Tessier 1983).

Terminology follows Oehletal. (2011) and Furrazolaetal. (2011). Thespore denomination

of Goto & Maia (2006) was used. Zeiss Axioskop compound microscopes with or without

Glomus trufemii sp. nov. (Brazil) ... 3

Nomarski differential interference contrast (DIC) were used for detailed spore observations;

digital images were taken with either an Axiocam camera and AxioVision (v. 3.1 software at

1300 x 1030 dpi) or Canon digital cameras.

Arbuscular mycorrhizal cultures

Bait cultures for the soils from sand dunes (Parque das Dunas de Natal) with spores

and sporocarps were established in the greenhouse of the Universidade Federal do Rio

Grande do Norte with Zea mays L. as host plant, using sterilized soil and 0.5 L pots.

Pure cultures of the new fungus were attempted by inoculating Zea mays seedlings

germinated in autoclaved quartz sand with G. trufemii sporocarps isolated from sand

dunes. After filling 0.5 L pots with sterilized sand dune soil, we formed a small hole in

the center using a glass shaker and discharged spores and sporocarps into the hole using

Pasteur pipettes. Then a Z. mays seedling was placed into the hole, covered with sand,

and grown for one cycle of 3-4 months. The fungus did not produce spores in either bait

cultures grown even for 9-12 months or pure cultures.

Taxonomy

Glomus trufemii B.T. Goto, G.A. Silva & Oehl, sp. nov. FIGs. 1-9

MycoBank MB 561567

Differs from G. brohultii by more regularly shaped subtending hyphae that are pigmented

further below the spore bases and darker colored orange to red brown spores with

slightly thicker walls.

Type: Brazil, Rio Grande do Norte, near Natal, Parque das Dunas de “Natal”, sand dunes

with herbaceous plant community on a convex hillside, 15.Feb.2010, B.T. Goto, in

permanent slide (PVLG) (Holotype: UFRN1482; Isotype: UFRN1483, ZT Myc 15118).

EryMo_oey: trufemii = in honor of Sandra Farto Botelho Trufem, taxonomist, who

long taught many young students and researchers in arbuscular mycorrhizal fungal

taxonomy.

SPOROCARPS formed in aggregates (500-850 x 780-1200 um), orange brown to

dark red brown adherent to living or dead roots with hundreds to thousands of

glomerospores; rarely singly in soils. Melzer’s reaction not observed. Peridium

absent. Sporocarpic hyphae hyaline (3.8—)7.6-10.4(-12.8) um without a septum

and with interwoven arrangements.

GLOMEROSPORES formed terminally on hyphae, subglobose to elliptical

(72-92 x 79-105 um) or rarely globose (72-96 um diam.), orange brown when

young to dark red brown at maturity, perhaps darkening slightly to dark brown

when ageing in soil.

SPORE WALL 7.4-15.5 um thick overall, consisting of two layers (swtl,

SWL2); SwWL1 thin (0.3-0.8 um), hyaline and difficult to observe in mature

spore, but generally visible in the subtending hyphal wall; sw12 thick (7.4-15.5

um), pigmented orange brown to dark red brown, laminated, smooth. Spore

wall layers continuous with subtending hyphal wall layers. The pigmentation of

SWL2 continues into the subtending hyphal wall, but this layer is often lighter

4 ... Goto &al.

Hyphae

™ Septum

yy Aborted

100um

* sw

*® swi1

Glomus trufemii sp. nov. (Brazil) ... 5

in color in the subtending hypha. swil and sw12 do not stain when exposed

to Melzer’s reagent.

SUBTENDING HYPHA (SH) generally present, single, straight or constricted,

cylindrical to sharply curved, light yellow to dark yellow; 5.1-12.7 um diam.

(mean-8.8 tm) at the spore base, the hyphal wall 2.6-5.1 um thick (mean

4.2 um) near the spore base and tapering to approx. 1 um at 15 um from the

base; occlusion formed by introverted sw12 thickening and often through

an additional bridging septum arising from sw1L2, but sometimes the pore

appearing partly open.

ABORTED GLOMEROSPORES frequently observed and forming terminally on

hyphae, generally collapsed or subglobose to elliptical (35-56 x 40-46 um), light

yellow to dark yellow, possibly darkening slightly to light brown when aging in

soils. Spore wall layer (swL2) (<3.8 um) not staining in Melzer’s reagent.

GERMINATION structures unknown.

GLOMEROSPORE DEVELOPMENT was deduced from positively identified

spores in aggregates found in different developmental stages. The hyaline hyphal

wall layer differentiates into a hyaline, evanescent spore wall layer (swL1) and

then a laminate layer (swL2) that becomes more pigmented with increasing

numbers of developing sublaminae. After the glomerospores mature, their pore

is closed by introverted thickening of sw12 and an additional bridging septum

arising from the laminate wall layer.

ARBUSCULAR MYCORRHIZA FORMATION remains unknown.

DISTRIBUTION — So far, the new fungus was detected only in Brazil.

Known in soil from sand dunes mainly harbouring herbaceous species of

Araceae, Asteraceae, Cyperaceae, Poaceae and Rubiaceae (e.g.: Acroceras

zizanioides (Kunth) Dandy, Anthurium affine Schott, Aspilia procumbens

Baker, Centratherum punctatum Cass., Cyperus maritimus Poir., C. meyenianus

Kunth, Mitracarpus eichleri K.Schum., Raddia brasiliensis Bertol., Richardia

grandiflora (Cham. & Schltdl.) Steud.) growing in native, ecologically unstable

restinga ecosystems in Parque das Dunas, Natal (Brazil).

Discussion

Glomus species are very common in sand dunes (Blaszkowski 2010;

Blaszkowski et al. 2009a,b, 2010a,b) and several new species have been

described recently (G. africanum Blaszk. & Kovacs, G. achrum Blaszk. et al.,

Fics. 1-9. Glomus trufemii. 1. Sporocarps in PVLG; note the numerous glomerospores in this

part of sporocarp. 2-3. Fragment of sporocarp with healthy glomerospores and aborted spores.

4, Aborted spores with only one spore wall layer (sw12). 5. Septum formed by sw12. 6. Spores

in different developmental stages. 7. Spore wall layers in mature spore (SwL1&2). 8-9. Spore wall

layers (SwL1&2) continuous with subtending hyphal wall (SW) that form distinct pigmentation.

6 ... Goto & al.

G. bistratum Btaszk. et al., G. indicum Blaszk. et al., G. iranicum Blaszk. et al.,

G. majewskii Btaszk., G. perpusillum Blaszk. & Kovacs). Glomus trufemii is

readily distinguished from previously described sporocarpic Glomus species

by glomerospore size and colour, spore wall structure, (including that at the

spore base), and characters of the subtending hypha. Most of these species form

colourless or pale-coloured spores singly or in loose aggregates (Blaszkowski

et al. 2009a,b; 2010a,b) and thus are easy to distinguish from the sporocarpic

dark-spored G. trufemii. Sporocarps of G. trufemii slightly resemble spore

ageregates of G. aggregatum N.C. Schenck & G.S. Sm. and G. fasciculatum

(Thaxt.) Gerd. & Trappe (Schenck & Smith 1982, Gerdemann & Trappe 1974).

However, the G. fasciculatum sw12 stains in Melzer’s reagent (vs. no reaction

in G. trufemii). Spores of G. aggregatum generally are irregular in shape (vs.

globose to subglobose) and have much thinner spore and subtending hyphal

walls. The spore wall of G. trufemii is one of the thickest among the known

glomeromycotan species with glomoid spores.

Several Glomus spp. form sporocarps without a peridium with similarly

sized and coloured spores similarly distributed in a sporocarp: G. ambisporum

G.S. Sm. & N.C. Schenck, G. atrouva McGee & Pattinson, G. badium Oehl et

al., G boreale (Thaxt.) Trappe & Gerd., G. botryoides RM. Rothwell & Victor,

G. brohultii Sieverd. & R.A. Herrera, G. flavisporum (M. Lange & E.M. Lund)

Trappe & Gerd., G. glomerulatum Sieverd., G. heterosporum G.S. Sm. & N.C.

Schenck, G. invermaium LR. Hall, and G. macrocarpum Tul. & C. Tul. (Oehl et

al. 2011). However, of these only G. badium, G. brohultii, and G. glomerulatum

also form spores with subtending hyphae generally clearly lighter-coloured

than the spore wall (Sieverding 1987, Herrera-Peraza et al. 2003, Oehl et al.

2005, Oehl et al. 2011). In the other species, the wall color of both spore and

subtending hyphae is similar or identical (Oehl et al. 2011).

Glomus glomerulatum produces intercalary yellow brown glomerospores

having 2-4 subtending hyphae (vs only terminal spores with one subtending

hypha in G. trufemii). Moreover, G. trufemii spores are darker (orange-brown

to dark reddish brown) and have a significantly thicker wall.

Glomus badium forms irregularly sized 3-layered spores around an

intrasporocarpic gleba, while G. trufemii generally produces regularly sized

subglobose 2-layered spores in sporocarps lacking glebas.

Glomus trufemii and G. brohultii spores have a similar size and wall structure.

Also, although G. brohultii was not originally described as forming sporocarps

Fics. 10-18. Glomus brohultii. Bi-layered spores (swL1&2). 10. Glomerospores in PVLG; note the

subtending hypha (SH) which is pigmented up to ca. 50 um below the spore base before becoming

subhyaline to hyaline. 11-12. Spores frequently have two subtending hyphae. 13. Mature spore with

sw12 thickened at the spore base. 14. Irregular subtending hypha becoming hyaline in a rather

short distance from the spore base (20-30 um). 16. Hyaline subtending hypha with two layers

(SHL1&2). 17-18. Crushed spores with two spore wall layers (swL1&2).

Glomus trufemii sp. nov. (Brazil) ... 7

8 ... Goto & al.

or aggregates, sporocarps have been observed recently (Oehl, pers. obs.) during

AMF diversity studies in cultivated white and yellow yam sites and adjacent

natural savannas in West Africa (Tchabi et al. 2009), ruling out sporocarp

or aggregate formation as a diagnostic character separating G. trufemii and

G. brohultii. However, G. brohultii spores are paler and lack orange or red

ones— yellow-brown to brown, not orange-brown to dark red-brown — and

its spore walls are generally thinner. Furthermore, in G. trufemii the subtending

hyphal pigmentation extends further below the spore base, both in spores

formed in sporocarps or singly in the soil. Finally, the shape of the subtending

hyphae is much more irregular in G. brohultii (Herrera-Peraza et al. 2003: this

paper, Fics. 10-18).

Acknowledgements

The authors acknowledge, in special, Dr. Janusz Blaszkowski (Department of

Plant Protection, West Pomeranian University of Technology, Szczecin, Poland) and

Dr. Ewald Sieverding (Institute for Plant Production and Agroecology in the Tropics

and Subtropics, University of Hohenheim), for reviewing the manuscript and making

helpful comments and suggestions and appreciate the corrections by Shaun Pennycook,

Nomenclatural Editor, and suggestions by Lorelei L. Norvell, Editor-in-Chief. The study

was supported by the Universidade Federal de Pernambuco which provided a grant to

FE Oehlas ‘visiting professor and Programa de Pés-Graduac¢ao em Sistematica e Evolugao

that invited F. Oehl for a technical visit to Natal, RN.

Literature cited

Blaszkowski J. 2010. Glomus majewskii,a new species of arbuscular mycorrhizal fungi (Glomeromycota).

Polish Botanical Journal 55(2): 265-270.

Blaszkowski J, Renker C, Buscot F. 2006. Glomus drummondii and G. walkeri, two new species of

arbuscular mycorrhizal fungi (Glomeromycota). Mycol. Res. 110: 555-566.

http://dx.doi.org/10.1016/jmycres.2006.02.006

Blaszkowski J, Ryszka P, Oehl F, Koegel S$, Wiemken A, Kovacs GM, Redecker D. 2009a. Glomus

achrum and G. bistratum, two new species of arbuscular mycorrhizal fungi (Glomeromycota)

found in maritime sand dunes. Botany 87: 260-271. http://dx.doi.org/10.1139/B08-138

Blaszkowski J, Kovacs GM, Balazs T. 2009b. Glomus perpusillum, a new arbuscular mycorrhizal

fungus. Mycologia 101(2): 247-255. http://dx.doi.org/10.3852/08-087

Blaszkowski J, Wubet T, Harikumar VS, Ryszka P, Buscot F. 2010a. Glomus indicum, a new arbuscular

mycorrhizal fungus. Botany 88: 132-143. http://dx.doi.org/10.1139/B09-104

Btaszkowski J, Kovacs GM, Balazs T, Orlowska E, Sadravi M, Wubet T, Buscot FE 2010b. Glomus

africanum and G. iranicum, two new species of arbuscular mycorrhizal fungi (Glomeromycota).

Mycologia 102(6): 1450-1462. http://dx.doi.org/10.3852/09-302

Clements FE, Shear CL. 1931. The genera of fungi. Hafner Publishing Co., New York, USA.

Furrazola, E, Torres-Arias Y, Ferrer RL, Herrera RA, Berbara RLL, Goto BT. 2011. Glomus crenatum, a

new ornamented species in the Glomeromycetes from Cuba. Mycotaxon 116: 143-149.

http://dx.doi.org/10.5248/116.143

Gerdemann JW, Trappe JM. 1974. The Endogonaceae in the Pacific Northwest. Mycologia Memoir

No. 5. 76 p.

Glomus trufemii sp. nov. (Brazil) ... 9

Goto BT, Maia LC. 2006. Glomerospores, a new denomination for the spores of Glomeromycota, a

group molecularly distinct from Zygomycota. Mycotaxon 96: 129-132.

Herrera-Peraza RA, Ferrer RL, Sieverding E. 2003. Glomus brohultii: a new species in the arbuscular

mycorrhiza forming Glomerales. Journal of Applied Botany and Food Quality 77: 37-40.

Koske RE, Tessier B. 1983. A convenient, permanent slide mounting medium. Mycol. Soc. Am.

Newsl. 34: 59.

Morton JB, Redecker D. 2001. Two new families of Glomales, Archaeosporaceae and Paraglomeraceae,

with two new genera Archaeospora and Paraglomus, based on concordant molecular and

morphological characters. Mycologia 93(1): 181-195.

Oehl F, Sieverding E. 2004. Pacispora, a new vesicular arbuscular mycorrhizal fungi genus in the

Glomeromycetes. Journal of Applied Botany 78: 72-82.

Oehl F, Redecker D, Sieverding E. 2005. Glomus badium, a new sporocarpic arbuscular mycorrhizal

fungal species from European grasslands of higher soil pH. J. Appl. Bot. Food Qual. 79(1):

38-43.

Oehl F, Silva GA, Goto BT, Sieverding E. 2011. Glomeromycota: three new genera, and glomoid

species reorganized. Mycotaxon 116: 75-120. http://dx.doi.org/10.5248/116.75

Oliveira-Filho AT. 1993. Gradient analysis of an area of coastal vegetation in the state of Paraiba,

Northeastern Brazil. Edinburgh Journal of Botany 50(2): 217-236.

Oliveira-Filho AT, Carvalho DA. 1993. Floristica e fisionomia da vegetacéo no extremo norte do

litoral da Paraiba. Rev. Bras. bot. 16(1): 115-130.

Omar MB, Bolland L, Heather WA. 1979. A permanent mounting medium for fungi. Bulletin of the

British Mycological Society 13: 31-32.

Redecker D, Raab P, Oehl F, Camacho FJ, Courtecuisse R. 2007. A novel clade of sporocarp-forming

species of glomeromycotan fungi in the Diversisporales lineage. Mycological Progress 6: 35-44.

http://dx.doi.org/10.1007/s11557-007-0524-2

Schenck NC, Smith GS. 1982. Additional new and unreported species of mycorrhizal fungi

(Endogonaceae) from Florida. Mycologia 74: 77-92.

Schiifler A, Schwarzott D, Walker C. 2001. A new fungal phylum, the Glomeromycota: phylogeny

and evolution. Mycol. Res. 105: 1413-1421. http://dx.doi.org/10.1017/S0953756201005196

Schwarzott D, Walker C, SchiiSler A. 2001. Glomus, the largest genus of the arbuscular mycorrhizal

fungi (Glomales), is non-monophyletic. Mol. Phylogenet. Evol. 21: 190-197.

Sieverding E. 1987. A VA mycorrhizal fungus, Glomus glomerulatum sp. nov., with two hyphal

attachments and spores formed only in sporocarps. Mycotaxon 29: 73-79.

Spain JL. 1990. Arguments for diagnoses based on unaltered wall structure. Mycotaxon 38: 71-76.

Tchabi A, Burger S, Coyne D, Hountondji FE, Lawouin L, Wiemken A, Oehl F. 2009. Promiscuous

arbuscular mycorrhizal symbiosis of yam (Dioscorea spp.), a key staple crop in West Africa.

Mycorrhiza 19: 375-392. http://dx.doi.org/10.1007/s00572-009-0241-6

Tulasne LR, Tulasne C. 1844. Fungi nonnulli hypogaei, novi v. minus cognito act. Gior. Bot. Ital.

1(2(7-8)): 55-63.

Walker C, Schiifler A. 2004. Nomenclatural clarifications and new taxa in Glomeromycota. Mycol.

Res. 108: 981-982. http://dx.doi.org/10.1017/S0953756204231173

MY COTAXON

http://dx.doi.org/10.5248/120.11

Volume 120, pp. 11-21 April-June 2012

Two new species of Pluteus section Celluloderma

from the Dominican Republic

ALFREDO JUSTO”, ELISEO BATTISTIN* & CLAUDIO ANGELINI?

'Clark University. Biology Department, 950 Main St., Worcester 01610 MA USA

?Natural History Museum, Corso Italia 63, Valdagno 36078 Italy

°Via Tulipifero 9, Porcia 33080 Italy

* CORRESPONDENCE TO: ajusto@clarku.edu

ABSTRACT— Two new species of Pluteus sect. Celluloderma collected in the Dominican

Republic are described based on morphological and molecular (nrITS) characters. Pluteus

crenulatus is characterized by the sulcate crenulate margin of the pileus, clavate cheilo- and

caulocystidia, and absence of pleurocystidia. Pluteus stenotrichus is characterized by the

presence of very narrow and internally septate cells in the pileipellis.

Key worps— biodiversity, Caribbean, phylogeny, Pluteaceae, taxonomy

Introduction

The genus Pluteus Fr. (Pluteaceae, Agaricales) has been the focus of recent

molecular phylogenetic studies (Menolli et al. 2010; Justo et al. 2011a,b),

which essentially support the traditional subdivision of the genus in three

sections (Pluteus, Celluloderma Fayod, and Hispidoderma Fayod) but with

some rearrangements. Sect. Celluloderma still accommodates all species with

non-metuloid hymenial cystidia and a pileipellis arranged as an hymeniderm

or epithelium composed mostly of clavate or spheropedunculate elements

(intermixed or not with elongated cells) but now, based on molecular data, also

includes taxa with a cutis-type pileipellis (previously assigned to sect. Villosi

Schreurs & Vellinga or Hispidoderma) or with distinct partial veil (previously

assigned to Chamaeota (W.G. Sm.) Earle) (Menolli et al. 2010; Justo et al.

2011a,b; Vizzini & Ercole 2011).

Here we describe two new Pluteus species recently collected in the Dominican

Republic. Morphological and molecular (nrITS) data support their inclusion

in sect. Celluloderma. Both taxa are compared with morphologically similar

species and their phylogenetic position is discussed.

12 ... Justo, Battistin & Angelini

Materials & methods

Collections were studied using standard procedures for morphological examination of

Pluteus (e.g., Justo & Castro 2007, Minnis & Sundberg 2010). Descriptive morphological

terms follow Vellinga (1988). The notation [30, 2, 1] indicates that measurements were

made of 30 basidiospores from 2 basidiocarps in 1 collection. Color codes are from

Munsell Soil-Color Charts (Munsell Color 2009). The following abbreviations are used

in the descriptions: avl for average length, avw for average width, Q for the length/width

quotient and avQ for average quotient.

Standard procedures for DNA isolation, PCR, and sequencing were applied (e.g.,

Justo et al. 2011la,b). The “nuclear ribosomal internal transcribed spacers 1 and 2

including the 5.8S region” is abbreviated as nrITS. The final dataset includes sect.

Celluloderma nrITS sequences generated by Justo et al. (2011b), additional sequences

generated by the first author, and sequences currently available in GenBank (mostly

originated by O’Brien et al. 2005, Matheny et al. 2006, Midgley et al. 2007, Malysheva et

al. 2009, Menolli et al. 2010). All accession numbers and sequence geographic origins

are given in Fic. 1. Sequences were aligned using MAFFT version 6 (http://mafft.cbre.

jp/alignment/server/; Katoh & Toh 2008) with the Q-INS-i option, and alignments

were manually corrected using MacClade 4.05 (Maddison & Maddison 2002) before

deposition in TreeBASE (http://purl.org/phylo/treebase/phylows/study/TB2:S12007).

A Maximum Likelihood analysis was run using the RAxML servers with 100 rapid

bootstrap replicates (http://phylobench.vital-it.ch/raxml-bb/index.php; Stamatakis et

al. 2008). Pluteus cervinus (Schaeff.) P. Kumm. and P. petasatus (Fr.) Gillet (both sect.

Pluteus) were used as outgroup taxa.

Results

The phylogenetic position of the new taxa described here is highlighted

in Fic. 1 and discussed in detail under Taxonomy. Sequence identifications

marked with “***” (which require further attention but fall outside the scope

of this paper) are listed here to avoid causing further taxonomic confusion

in GenBank: (i) “Pluteus cf. nanus” (HM562046) and “P. nanus” (jF908611)

probably represent P. cinereofuscus J.E. Lange; (ii) “Pluteus cinereofuscus”

(JF908616) is part of the Pluteus nanus (Pers.) P. Kumm. species complex; (iii)

“Pluteus cervinus” (JF908623) represents a taxon close to Pluteus ephebeus

(Fr.) Gillet; (iv) “Pluteus galeroides”( JF908610, JF908609) probably represents

Pluteus chrysophlebius (Berk. & M.A. Curtis) Sacc.; (v) “Pluteus boudieri”

(jF908627) represents Pluteus phlebophorus (Ditmar) P. Kumm.; (vi) Pluteus

insidiosus (JF908626) falls within collections morphologically assignable to

Pluteus thomsonii (Berk. & Broome) Dennis.

FIGURE 1. Best tree from the Maximum Likelihood analysis of Pluteus sect. Celluloderma. Bootstrap

values 270% are shown on or below the branches. Root length has been reduced to facilitate

graphical representation.

Pluteus spp. nov. (Dominican Republic) ... 13

HQ654907 Pluteus aurantiorugosus var aurantiovelatus Italy

i HQ654908 Pluteus aurantiorugosus var aurantiovelatus Italy

New taxa decribed JF908608 Pluteus aurantiorugosus Italy

1 1 A HM562041 Pluteus aurantiorugosus Spain °

In this article JF908613 Pluteus aurantiorugosus Italy Pluteus aurantiorugosus

100 | HM562121 Pluteus aurantiorugosus Japan

AT ate od, 4 HM562081 Pluteus aurantiorugosus Illinois

%& Mixtini type pileipellis HM562074 Pluteus aurantiorugosus Michigan

72L HM562072 Pluteus aurantiorugosus Massachusetts

9] 98 JF908630 Pluteus romellii Italy

indi HM562062 Plut Ilii_ Spai

Pleurocystidia absent [> [sie Acs nei san

FJ774073 Plut llii Russi ip

or very scarce 75 HM562123 splutsdesronielit Jonah Pluteus romellii

100 HM562078 Pluteus romellii Michigan]

87], HM562183 Pluteus romellii Illinois

964 HM562105 Pluteus romellii Missouri

79,- HM562160 Pluteus aureovenatus Brazil

FJ816663 Pluteus aureovenatus Brazil

100 _}_ Aygs4065 Pluteus aff romellii California Pluteus aureovenatus complex

FJ816667 Pluteus sublaevigatus Brazil

100 HM562173 Pluteus sp. Brazil —————_ —__— Pluteus sp.O

JN603201 Pluteus stenotrichus AJ352 Dominican Republic *

HMS562161 Pluteus sp. Brazil ————————_—— P] teus Sp.

HM562198 Pluteus aff ephebeus England

100] HM562080 Pluteus aff ephebeus France

88] | JF908623 Pluteus cervinus Italy***

199, JF908620 Pluteus ephebeus Italy

76 JF908621 Pluteus ephebeus Italy Pluteus ephebeus complex

HM562044 Pluteus ephebeus Spain

HM562162 Pluteus riberaltensis var conquistensis Brazil

100 HM562091 Pluteus fenzlii Japan ma

100] FJ774082 Pluteus fenzlii Russia Pluteus fenzlii

100 HM562111 Pluteus fenzlii Slovakia

HM562086 Pluteus mammillatus Missouri al

HM562119 Pluteus mammillatus Missouri Pluteus mammillatus

100L M562120 Pluteus mammillatus Florida

98, HM562122 Pluteus podospileus Japan | s

100 HM562049 Pluteus podospileus Spain Pluteus podospileus *

83 HM562196 Pluteus aff podospileus Sweden| Pluteus aff. podospileus *

HM562199 Pluteus seticeps Illinois

100__| HM562192 Pluteus seticeps wisconsin Pluteus seticeps *0O

HM562191 Pluteus seticeps Missouri

100 HM562076 Pluteus eliae Florida H

Nc00008 Plates ae mii | Pluteus eliae / Pluteus sp.

76, FJ816657 Pluteus jamaicensis Brazil : .

100] § FJ816662 Pluteus fuligineovenosus Brazil Pluteus fluminensis com plex

ap FJ816664 Pluteus fluminensis Brazil

FJ816655 Pluteus fluminensis Brazil

100 , HM562146 Pluteus sp. | Pluteus sp.

HM562148 Pluteus sp. Brazil

HM562145 Pluteus sp. Brazil | Pluteus Sp.

HM562046 Pluteus cf. nanus Spain ***

JF908611 Pluteus nanus Italy *** °

B2 HM562108 Pluteus cinereofuscus Portugal Pluteus cinereofuscus

HM562124 Pluteus cinereofuscus Spain

FJ774085 Pluteus eludens Russia

HM562118 Pluteus eludens Madeira Island Pluteus eludens *

76) 9°L HM562185 Pluteus eludens Illinois

HM562056 Pluteus pallescens Spain | Pluteus pallescens

81 HM562115 Pluteus aff cinereofuscus Japan

87L AY969369 Uncultured basidiomycete North eet Pluteus aff. cinereofuscus

HM562201 Pluteus multiformis Spain ° .

88 DQ672275 Uncultured soil basidiomycete austrata| Pluteus multiformis %& /Pluteus Sp.

HM562138 Pluteus phlebophorus Spain

72] HM562137 Pluteus phlebophorus Spain

HM562039 Pluteus phlebophorus Spain

21 JF908627 Pluteus boudieri Italy***

HMS562144 Pluteus phlebophorus Spain Pluteus phlebophorus

HM562184 Pluteus phlebophorus Michigan

87. a HM562112 Pluteus phlebophorus Michigan

“| HEHM562193 Pluteus pallidus Michigan

75 HM562117 Pluteus phlebophorus Japan

0.09 HM562186 Pluteus aff phlebophorus Illinoist Pluteus aff. phlebophorus

HM562116 Pluteus cf eugraptus Japan] Pluteus cf. eugraptus *

JF908616 Pluteus cinereofuscus Italy***

JF908633 Pluteus nanus Italy

_ £4774081 Pluteus nanus Russia Pluteus nanus com plex

HM562216 Pluteus sp VI Illinois

JN603203 Pluteus cf nanus AJ348 Spain

96 80, JF908610 Pluteus galeroides Italy***

75 91/1 jF908609 Pluteus galeroides Italy***

HM562064 Pluteus chrysophlebius Spain “

96 4M562088 Pluteus cheepeniesits seo Pluteus chrysophlebius

74] 'HM562125 Pluteus chrysophlebius Japan

HM562181 Pluteus chrysophlebius Illinois

81) 1 Hms62182 Pluteus chrysophlebius Illinois

64 (90 HM562180 Pluteus chrysophlebius Illinois oe

HM562079 Pluteus rugosidiscus Michiganl Pluteus rugosidiscus

100) HM562066 Pluteus thomsonii Spain

fa0 JF908626 Pluteus insidiosus Italy***

HM562067 Pluteus thomsonii Illinois oe

| )£1774082 Pluteus thomsonii Rusia | Pluteus thomsonii complex %O

100} HMS562053 Pluteus thomsonii Spain

100 JF908607 Pluteus thomsonii Italy

HM562197 Pluteus thomsonii Michigan

FJ816665 Pluteus dominicanus var hyalinus Brazil Pluteus dominicanus var. hyalinus

JN603202 Pluteus crenulatus AJ353 Dominican Republic

HM562143 Pluteus dietrichii Spain 'Pluteus dietrichiiO

HM562038 Pluteus petasatus Spain

100 HM562133 Pluteus cervinus Spain | Outgroup (sect. Pluteus)

14 ... Justo, Battistin & Angelini

Taxonomy

Pluteus crenulatus Justo, Battistin & Angelini, sp. nov. FIG. 2

MycoBAank 563469

Similar to Pluteus tucumanus, also with a sulcate pileus with crenulate margin,

but differing in broader spores, clavate cheilo- and caulocystidia, and absence of

pleurocystidia

Type: Dominican Republic, Prov. Puerto Plata, Sosua, Union, 17 Jan 2011, leg. C.

Angelini, coll. AJ353 (Holotype MICH; GenBank nrITS JN603202. Isotype MCVE

27198).

EryMo_oey: crenulatus refers to the distinctive crenulate pileus margin.

PiLEus 20-40 mm, convex to plano-convex, with a broad umbo, deeply sulcate

in the outer half; surface smooth or minutely granulose at center, granulose

towards the margin; brown, darker at center [7.5YR 3/4, 4/6] and paler towards

margin [7.5YR 5/6, 5/8, 6/6] where it alternates brown and whitish zones, not

hygrophanous, margin distinctly crenulate, white. LAMELLAE crowded, free,

ventricose; < 4mm broad; white when young, later pink with flocculose whitish

edges. STIPE 30-50 x 2-4 mm, cylindrical, slightly broadened at base; white or

white-cream; smooth or pruinose. CONTEXT in stipe and pileus white. SMELL

indistinct. TASTE not recorded. SPORE PRINT not recorded, probably pink.

BASIDIOSPORES [30, 2, 1] 4.5-5.5(-6.0) x 3.9-5.0(-5.5) um, avl x avw = 4.9 x

4.2 um, Q = 1.00-1.26, avQ = 1.17, globose to broadly ellipsoid. BAsip1a 20-35

x 9-12 um, 4-spored, clavate or narrowly utriform. PLEUROCYSTIDIA absent.

LAMELLAR EDGE Sterile, completely covered with cystidia. CHEILOCYSTIDIA

24-73 x 12-34 um, (narrowly) clavate, a few narrowly utriform; colorless;

with thin, smooth walls; crowded, forming a well-developed strip. PILEIPELLIS

a hymeniderm with transitions towards and epithelium; individual elements

29-61 x 14-49 um, mostly clavate or spheropedunculate, a few narrowly

utriform, mucronate, or with basal septum; filled with brown intracellular

pigment, evenly dissolved or more rarely with pigment condensations; with

thin, smooth walls. STrPITIPELLIs a cutis; hyphae 4-13 um wide, cylindrical,

colorless or with brown pigment; with thin, smooth walls. CAULOCYSTIDIA

38-51 x 17-31 um, (narrowly) clavate, some multiseptate; hyaline or with

brown intracellular pigment, some with additional incrusting pigment; with

thin to slightly thickened (less than 1 um), smooth walls; all over stipe surface

in tightly packed clusters. CLAMP CONNECTIONS absent in all tissues.

HABITAT AND DISTRIBUTION—Gregarious, on a fallen log in broad-leaved

forest, January. Known only from the type locality in the Dominican Republic.

ComMENtTS—The deeply sulcate pileus with a crenulate margin is the

most distinctive macroscopic feature of P crenulatus. Microscopically, it is

characterized by the clavate cheilo- and caulocystidia (Fic. 2) and the absence

of pleurocystidia.

Pluteus spp. nov. (Dominican Republic) ... 15

FIGURE 2. Pluteus crenulatus (holotype AJ353). a. Basidiocarps; b. Basidiospores; c. Cheilocystidia;

d. Pileipellis elements; e. Caulocystidia. Scale bars = 10 um.

16 ... Justo, Battistin & Angelini

P. tucumanus is a South American species that also has a sulcate,

appendiculate-crenate pileus (Singer & Digilio 1952; Singer 1956, 1958). The

following observations are based on our examination of its isotype in MICH

(Singer T921 Argentina, Prov. Tucuman, Rio de los Sosas, 18 Jan 1950):

Pluteus tucumanus Singer, Lilloa 25: 269. 1952 [“1951”]. Fic. 3

BASIDIOSPORES [30, 1, 1] 3.9-5.0 x 2.7-4.0 um, avl x avw = 4.5 x 3.6 um,

Q = 1.00-1.34(-1.69), avQ = 1.26, globose to ellipsoid, very rarely oblong.

BasIDIA 25-31 x 7-10 um, 4-spored, narrowly utriform. PLEUROCYSTIDIA

40-53 x 18-22 um, (narrowly) utriform with a distinct pedicel; colorless;

with thin, smooth walls; scarce and scattered, present all over lamellar sides.

LAMELLAR EDGE sterile, completely covered with cystidia. CHEILOCYSTIDIA

29-52 x 12-32 um, (narrowly) utriform or broadly fusiform, a few narrowly

clavate; colorless; with thin, smooth walls; crowded, forming a well-developed

strip. Pileipellis an hymeniderm with transitions towards and epithelium;

individual elements 38-49 x 21-32 um, mostly clavate or spheropedunculate;

filled with brown intracellular pigment, evenly dissolved or more rarely with

pigment condensations; with thin, smooth walls. STIPITIPELLIS a cutis; hyphae

5-13 um wide, cylindrical, colorless or with brown pigment; with thin, smooth

walls. CauLocystTip1a 38-51 x 17-31 um, (narrowly) clavate to (narrowly)

utriform; hyaline or with brown intracellular pigment; with thin, smooth walls;

all over stipe surface, scattered or in clusters. CLAMP-—CONNECTIONS absent in

all tissues.

Although the differences between P crenulatus and P. tucumanus are subtle

(the latter distinguished by narrower spores, presence of pleurocystidia, and

differently shaped cheilo- and caulocystidia), such subtle variations have proved

critical in separating species in sect. Celluloderma (Justo et al. 2011c).

Singer (1958) placed Pluteus fallax Singer and P iguazuensis Singer into

the same morphological group (stirps Tucumanus) as P. tucumanus. We were

unable to examine either species, but based on Singer's (1958) descriptions both

taxa differ from P crenulatus in (among other characters) the non-crenulate

pileus margin and presence of pleurocystidia. Singer also emphasized the

presence of pigment condensations in the pileipellis elements of P. tucumanus

and the absence of such condensations in P fallax and P. iguazuensis. We

observed pigment condensations in both P crenulatus and P. tucumanus, but

the taxonomic significance of this feature needs further study.

Macroscopically, P crenulatus does not resemble any species in sect.

Celluloderma with very rare to absent pleurocystidia — e.g., P diettrichii

Bres., P. insidiosus Vellinga & Schreurs, P. poliocnemis Kihner — which also

differ microscopically in spore and cystidia shape and size (see Kthner &

Romagnesi (1956) and Vellinga (1990) for full descriptions). Pluteus thomsonii

Pluteus spp. nov. (Dominican Republic) ... 17

O09 (Vac

FIGURE 3. Pluteus tucumanus (isotype, Singer T921). a. Basidiospores; b. Pleurocystidia;

c. Cheilocystidia; d. Pileipellis elements; e. Caulocystidia. Scale bars = 10 um.

and P. seticeps (G.F. Atk.) Singer, which sometimes have very rare or absent

pleurocystidia, differ from P crenulatus by the “Mixtini-type’ pileipellis

(see Minnis & Sundberg (2010) and Vellinga (1990) for full descriptions).

Phylogenetically, P crenulatus appears somewhat isolated as sister to all taxa

in sect. Celluloderma except P. diettrichii Bres. (Fic. 1). This placement receives

no statistical support but suggests that P. crenulatus is not closely related to any

of the major lineages presently recognized in sect. Celluloderma (Justo et al.

2011b).

Pluteus stenotrichus Justo, Battistin & Angelini, sp. nov. Fic. 4

MycoBAnk 563470

Characterized by a “Mixtini-type” pileipellis with very narrow, internally septate

cylindrical elements.

Type: Dominican Republic, Prov. Puerto Plata, Sosua, El Castillo, 16 Jan 2011, leg. C.

Angelini, coll. AJ352 (Holotype MICH; GenBank nrITS JN603201. Isotype MCVE

27197).

ETYMOLOGy: stenotrichus (= “with narrow hairs”) refers to the very unusual type of

pileipellis cells.

18 ... Justo, Battistin & Angelini

PILEuS (20-)25-50(-60) mm, convex to plano-convex, with or without a low

broad umbo; surface distinctly rugose-venose at and around center, radially

areolate-rimose in the outer half, exposing the white context; brown, darker at

center [7.5YR 2.5/2, 2.5/3, 3/2, 3/3], paler around center [7.5YR 4/6, 5/6, 5/8]

and much paler at margin [7.5YR 7/4, 7/6, 7/8], not hygrophanous; margin

rimose but not striate or sulcate. LAMELLAE crowded, free, ventricose; < 7 mm

broad; white or cream when young, later pink [5YR 8/3, 8/4], with concolorous

or paler edges. StrPE 30-60 x 4-6 mm, cylindrical, slightly broadened at base;

white or white-cream; smooth or slightly pruinose. CONTEXT in stipe and

pileus white. SMELL indistinct. TASTE not recorded. SPORE PRINT not recorded,

probably pink.

BASIDIOSPORES [30, 2, 1] 4.9-7.0 x 4.5-5.7 um, avl x avw = 5.6 x 5.1 um,

Q = 1.00-1.18, avQ = 1.11, globose to broadly ellipsoid. Bastp1a 25-34 x

7.5-10 um, 4-spored, clavate or narrowly utriform. PLEUROCYSTIDIA 34-83

x 14-32 um, (narrowly) utriform or lageniform, a few broadly conical with

long pedicel; colorless; with thin, smooth walls; relatively common, scattered

over lamellar sides. LAMELLAR EDGE sterile, completely covered with

cystidia. CHEILOCYSTIDIA 25-50(-65) x 14-20(-25) um, (narrowly) clavate,

spheropedunculate, some broadly fusiform, some with basal septum; colorless;

with thin, smooth walls; crowded, forming a well-developed strip. PILEIPELLIS

a hymeniderm with transitions towards and epithelium, composed of three

types of elements: (i) globose, clavate or spheropedunculate 31-76 x 22-60

um; (ii) narrowly clavate or lageniform 53-88 x 15-29 um; (iii) vey narrow

elongated elements, 60-123 x 7-10(-12) um, cylindrical or flexuous, with 0-4

internal septa, usually arising from a basal element < 15 um wide; all elements

filled with brown intracellular pigment, more rarely hyaline; with thin, smooth

walls. STIPITIPELLIS a cutis; hyphae 5-15 um wide, cylindrical, colorless or with

brown pigment; with thin, smooth walls. CauLocystip1a 31-63 x 12-24 um,

(narrowly) clavate, narrowly utriform or conical with basal septum; hyaline or

with brown intracellular pigment; with thin, smooth walls; scattered all over

stipe surface, isolated or in loosely arranged clusters. CLAMP CONNECTIONS

absent in all tissues.

HABITAT AND DISTRIBUTION—Gregarious, apparently terrestrial (among

woody remnants and organic matter) in broad-leaved forest, January. Known

only from the type locality in the Dominican Republic.

COoMMENTS—The very narrow and internally septate elements in the

pileipellis (Fic. 4) are diagnostic for P stenotrichus. Taxa with both clavate-

spheropedunculate and elongated elements in the pileipellis have been

traditionally classified in subsect. Mixtini Singer, but molecular data suggest

that elongated elements in the pileipellis have evolved several times within

sect. Celluloderma, rendering subsect. Mixtini artificial (Justo et al. 2011a,

Pluteus spp. nov. (Dominican Republic) ... 19

FiGURE 4. Pluteus stenotrichus (holotype AJ352). a. Basidiocarps; b. Basidiospores;

c. Pleurocystidia; d. Cheilocystidia; e. Caulocystidia; f. Pileipellis elements - fl clavate

and spheropedunculate, f2 narrowly clavate and lageniform, f3 very narrow and internally

septate. Scale bars = 10 um.

20 ... Justo, Battistin & Angelini

Fig. 1). An extensive literature search and comparison with all species described

with a “Mixtini-type” pileipellis (e.g. Horak 1964, 2008; Horak & Heinemann

1978; Minnis & Sundberg 2010; Orton 1986; Pegler 1977, 1986; Singer 1956,

1958; Vellinga 1990; see also Justo et al. 2011a,b,c for additional taxonomic

references) found none with this type of septate elongated element.

Pluteus stenotrichus is not closely related to any previously sequenced

species with a “Mixtini-type” pileipellis (Fic. 1) and its position in the

romellii/aurantiorugosus group is rather unexpected based on morphology.

Pluteus aurantiorugosus (Trog) Sacc., P. romellii (Britzelm.) Sacc., and taxa

in the species complex around P. aureovenatus Menolli & Capelari all lack

any elongated pileipellis elements and are characterized in part by the bright

yellow or orange-red colors of the pileus and/or stipe. The closest relative of

P. stenotrichus is an undescribed species from Brazil (nrITS HM562173,

collection $P394383) but that taxon differs in the (among other characters)

absence of elongated pileipellis elements and almost total absence of hymenial

cystidia (N. Menolli Jr. & M. Capelari, pers. comm.).

The narrow septate elements in the pileipellis of P crenulatus are reminiscent

of those typically exhibited by some members of sect. Hispidoderma (e.g.,

P. exiguus (Pat.) Sacc.). However the general organization of the pileipellis

differs in those taxa and lacks the well-developed hymeniform layer made up

of clavate or spheropedunculate elements that is typical of P stenotrichus and

the other species with “Mixtini-type” pileipellis in sect. Celluloderma.

Acknowledgments

A. Justo acknowledges financial support from the NSF grant DEB0933081. The

curators of MICH are gratefully acknowledged for sending the collection of Pluteus

tucumanus on loan. Drew Minnis and Nelson Menolli Jr. provided very insightful

comments in the presubmission reviews.

Literature cited

Horak E, Heinemann P. 1978. Flore Illustrée des champignons d'Afrique centrale 6: Pluteus &

Volvariella (compléments). National Botanical Garden of Belgium, Meise

Horak E. 1964. Fungi austroamericani II. Pluteus Fr. Nova Hedwigia 8: 163-199.

Horak E. 2008. Agaricales of New Zealand 1: Pluteaceae-Entolomataceae. Fungi of New Zealand,

vol. 5. Fungal Diversity Research Series 19. 305 p.

Justo A, Castro ML. 2007. Observations in Pluteus section Pluteus in Spain: Two new records for

Europe. Mycotaxon 102: 209-220.

Justo A, Vizzini A, Minnis AM, Menolli Jr N, Capelari M, Rodriguez O, Malysheva E, Contu M,

Ghignone S, Hibbett DS. 201la. Phylogeny of the Pluteaceae (Agaricales, Basidiomycota):

taxonomy and character evolution. Fungal Biology 115: 1-20.

http://dx.doi.org/10.1016/j.funbio.2010.09.012

Justo A, Minnis AM, Ghignone S, Menolli Jr N, Capelari M, Rodriguez O, Malysheva E, Contu

M, Vizzini A. 2011b. Species recognition in Pluteus and Volvopluteus (Pluteaceae, Agaricales):

Pluteus spp. nov. (Dominican Republic) ... 21

morphology, geography and phylogeny. Mycol. Progress 10: 453-479.

http://dx.doi.org/10.1007/s11557-010-0716-z

Justo A, Caballero A, Mufioz G, Minnis AM, Malysheva E. 2011c. Taxonomy of Pluteus eugraptus

and morphologically similar taxa. Mycologia 103: 646-655. http://dx.doi.org/10.3852/10-280

Katoh K, Toh H. 2008. Recent developments in the MAFFT multiple sequence alignment program.

Briefings in Bioinformatics 9: 286-298. http://dx.doi.org/10.1093/bib/bbn013

Kihner R, Romagnesi H. 1956. Espéces nouvelles, critiques ou rares de Volvariacees. Bull. Trimestiel

Soc. Mycol. France 72: 181-249.

Maddison DR, Maddison WP. 2002. MacClade4: analysis of phylogeny and character evolution.

Sinauer Associates, Sunderland, Massachusetts.

Malysheva EF, Malysheva VF, Krasilnikova AA. 2009. Morphological and molecular approaches to

study the genus Pluteus Fr. Mikol. Fitopatol. 43: 216-231.

Matheny PB, Curtis JM, Hofstetter V, Aime MC, Moncalvo J-M, Ge ZW, Yang ZL, Sot JC, Ammirati

J, Baroni TJ, Bougher NL, Hughes KW, Lodge DJ, Kerrigan RW, Seidl MT, Aanen DK, DeNitis

M, Daniele GM, Desjardin DE, Kropp BR, Norvell LL, Parker A, Vellinga EC, Vilgalys R,

Hibbett DS. 2006. Major clades of Agaricales: a multilocus phylogenetic overview. Mycologia

98: 982-995. http://dx.doi.org/10.3852/mycologia.98.6.982

Menolli Jr N, Asai T, Capelari M. 2010. Records and new species of Pluteus from Brazil based on

morphological and molecular data. Mycology 1: 130-153. http://dx.doi.org/10.1080/21501203

.2010.493531

Midgley DJ, Saleeba JA, Stewart MI, Simpson AE, McGee PA. 2007. Molecular diversity of soil

Basidiomycete communities in northern-central New South Wales, Australia. Mycol. Res.

111: 370-378. http://dx.doi.org/10.1016/j.mycres.2007.01.011

Minnis AM, Sundberg WJ. 2010. Pluteus section Celluloderma in the U.S.A. N. Amer. Fungi

5: 1-107. http://dx.doi.org/10.2509/naf2009.005.001

Munsell Color (ed.). 2009. Munsell Soil-Color Charts. Grand Rapids, Michigan.

O’Brien HE, Parrent JL, Jackson JA, Moncalvo JM, Vilgalys R. 2005. Fungal community analysis

by large-scale sequencing of environmental samples. Appl. Environ. Microbiol. 71: 5544-5550.

http://dx.doi.org/10.1128/AEM.71.9.5544-5550.2005

Orton PD. 1986. British fungus flora, agarics and boleti 4: Pluteaceae: Pluteus and Volvariella. Royal

Botanic Garden, Edinburgh.

Pegler DN. 1977. A preliminary Agaric flora of east Africa. Kew Bull., Additional Ser. 6: 1-615.

Pegler DN. 1986. Agaric flora of Sri Lanka. Kew Bull., Additional Ser. 12: 1-519.

Singer R. 1956. Contribution toward a monograph of the genus Pluteus. Transactions of the British

Mycological Society 39: 145-232. http://dx.doi.org/10.1016/S0007-1536 (56)80001-6

Singer R. 1958. Monograph of South American basidiomycetes, especially those of the east slope of

the Andes and Brazil. Lloydia 21: 195-299.

Stamatakis A, Hoover P, Rougemont J. 2008. A rapid bootstrap algorithm for the RAxML Web

servers. Syst. Biol. 75: 758-771. http://dx.doi.org/10.1080/10635150802429642

Vellinga EC. 1988. Glossary. Pp. 54-64, in: C Bas et al. (eds). Flora Agaricina Neerlandica 1.

A.A. Balkema, Rotterdam.

Vellinga EC. 1990. Pluteus Fr. Pp. 31-55, in: C Bas et al. (eds). Flora Agaricina Neerlandica 2.

A.A. Balkema, Rotterdam.

Vizzini A, Ercole E. 2011. A new annulate Pluteus variety from Italy. Mycologia 103: 904-911.

http://dx.doi.org/10.3852/10-382

MY COTAXON

http://dx.doi.org/10.5248/120.23

Volume 120, pp. 23-33 April-June 2012

The lichen genus Chapsa (Graphidaceae) in India

SANTOSH JOSHI, DALIP K. UPRETI* & SANJEEVA NAYAKA

Lichenology Laboratory, CSIR-National Botanical Research Institute

Rana Pratap Marg, Lucknow - 226 001, India

CORRESPONDENCE TO *: upretidk@rediffmail.com

ABSTRACT — This paper provides a detailed account of the 16 species of Chapsa recorded in

India. Chapsa patens and C. platycarpella are new records for the Indian lichen biota.

Key worps — thelotremoid, distribution, identification key, Western Ghats

Introduction

Until recently the thelotremataceous lichen biota was limited in India to

Chroodiscus, Diploschistes, Leptotrema, Myriotrema, Ocellularia, and Thelotrema

(Awasthi 1991, 2000). Based on recent revisions (Frisch et al. 2006, Mangold et al.

2009, Rivas Plata et al. 2010), Singh & Sinha (2010) cited 130 taxa representing 13

genera in the Thelotremataceae Stizenb. for India. However, recent phylogenetic

studies show that taxa previously classified in Thelotremataceae do not form

a separate lineage but are nested within Graphidaceae. The Graphidaceae,

containing the now-synonymous Thelotremataceae (Mangold et al. 2008),

comprises the largest lichen family in India, where it is represented by 428

species and 35 genera.

Massalongo introduced Chapsa in 1860, a genus resurrected recently by

Frisch et al. (2006) to incorporate species earlier classified in Thelotrema,

Chroodiscus, Ocellularia, and Myriotrema. Chapsa is characterized by a thin,

corticolous, endoperidermal to epiperidermal thallus, chroodiscoid ascomata

with a fused to + free proper exciple, and periphysoids (Mangold et al. 2009).

The closely related or similar Acanthotrema, Chroodiscus, and Reimnitzia differ

mainly in spiny paraphysis tips, lack of periphysoids, and isidiate thallus with

columnar crystalline layer, respectively. Moreover, Chroodiscus is typically

foliicolous. Thelotrema and Topeliopsis differ mainly by lacking distinctly

chroodiscoid ascomata. However, several species belonging to the last two

genera require further investigation for proper delimitation and placement.

24 ... Joshi, Upreti & Nayaka

Due to their selective and preferred habitat, most members of thelotremoid

Graphidaceae are restricted to lower altitudes in tropical to subtropical

regions and thus inadequately understood (Hale 1974, 1978, 1981). As a

group, thelotremoid lichens represent a significant component of corticolous

rainforest microlichen biota, with some extending into temperate regimes

(Santesson 1952; Sipman & Harris 1989; Rivas Plata et al. 2008; Purvis et al.

1995). Chapsa is pantropical or subtropical but rarely temperate. Generally,

it occurs below 1000 m and favors shaded microhabitats in rather disturbed

old growth secondary forests (Rivas Plata et al. 2008). Chapsa flourishes in the

southern and eastern regions of India, with some species found at altitudes of

more than 2000 m. The dense moist rain forests of the eastern Himalayas and

the evergreen forests of Western Ghats provide a suitable habitat for its growth.

Most species of Chapsa grow on rough bark, thus indicating a particular type of

forest community predominant in moist humid regions. The species generally

acquire the same colour pattern as the substratum so that in the field many

are difficult to distinguish from bark. The tree bark characteristics appear to

influence the distribution patterns of the taxa in tree communities. Lichens

growing on striate bark concentrate in deep furrows, with their ascomata

emerging from cracks, while in many species the thallus surface mirrors inflated

bark. Most taxa, however, grow on flaking bark with only ascomata protruding

out of the thallus.

The ascomatal features and chemical substances play a vital role in

differentiating the different thelotremoid groups. Based on morphology, the

ascomata may be of the chroodiscoid, fissurinoid, leprocarpoid, platycarpoid,

lamelloid, scabioid, or topeliopsoid types. However, except for C. platycarpa

and C. recurva, all Indian specimens lack secondary compounds.

Sixteen species of Chapsa are currently known from India. The evergreen

forests in the Western Ghats represented by nine species was the most diverse,

with six species known from the tropical rain forests of the eastern Himalayas

and four species known from the Andaman and Nicobar Islands. One report

of Chapsa dilatata (Mull. Arg.) Kalb from Arunachal Pradesh (Dubey et al.

2008, as Thelotrema dilatatum) was based on a misdetermined specimen that

represents a Myriotrema species. All accurately identified Chapsa species

recorded from India are presented here with brief descriptions.

Materials & methods

Material preserved in the herbarium of the National Botanical Research Institute

(LWG) was examined morphologically, anatomically, and chemically. Thin hand-cut

sections of apothecia and thallus mounted in tap water, cotton blue, 5% KOH, and iodine

solution were observed under a compound microscope (LEICA DM 500). Chemical

spot tests and TLC (using solvent system A) were conducted according to Orange et al.

(2001).

Chapsa in India... 25

Taxonomic descriptions

Chapsa alborosella (Nyl.) Frisch, Biblioth. Lichenol. 92: 90, 2006.

The species is characterized by an ecorticate, pale olive green thallus lacking

lichen compounds, round to angular or shortly elongate pale brown apothecia

level with the thallus, 8-spored asci and hyaline, transversely septate, fusiform

to clavate or oblong, I- ascospores.

Chapsa platycarpella differs from the closely related C. alborosella in having a

corticate thallus and a pale to blackish apothecial disc. Patwardhan & Kulkarni

(1977a) and Nagarkar et al. (1988) reported this species from south India as

Ocellularia alborosella. The species is common in tropical evergreen forest of

Kerala in the Western Ghats.

“Chapsa hiata” (Hale), ined.

Known from evergreen forests in the southern part of India, this species is

well characterized by an ecorticate thallus lacking lichen substances, wide and

open apothecia with recurved margins, free proper exciple, 4—8-spored asci,

and small hyaline muriform I- ascospores measuring 14-16 x 4-7um.

Patwardhan et al. (1985) reported it from Karnataka as Thelotrema hiatum.

Similar to Chapsa leprocarpoides, C. velata and C. pseudophlyctis in lacking

thallus compounds, the species differs in having small ascospores. ‘The latter

two species also differ in having a fused proper exciple.

Chapsa indica A. Massal., Atti I. R. Ist. Veneto Sci. Lett. Arti, Ser. 3, 5: 257, 1860.

This taxon is characterised by an ecorticate brownish-white to olive-grey

thallus lacking lichen compounds, rounded to angular or shortly elongated and

slightly branched apothecia that are level with the thallus, 6-8-spored asci, and

hyaline transversely septate oblong-fusiform ascospores.

Patwardhan & Nagarkar (1980) and Nagarkar et al. (1988) first reported the

species from India (as Ocellularia pycnophragmia (Nyl.) Zahlbr.) from tropical

rain forests of Andaman and Nicobar, Assam, and Meghalaya. Similar in having

an ecorticate whitish (-brownish) to olive-grey thallus, C. leprocarpa and C.

pseudophlyctis differ from C. indica in having muriform ascospores.

Chapsa laceratula (Mill. Arg.) Rivas Plata & Licking, Lichenologist 42: 183, 2010.

The species exhibits a pale to dark olive-green to olive brown, corticate

surface lacking secondary metabolites, round to irregular, perithecioid to

apothecioid apothecia with lacerate margins, 1-2(-4)-spored asci, and hyaline,

muriform, fusiform ascospores.

Patwardhan & Kulkarni (1977a) reported Chapsa laceratula (as Thelotrema

laceratulum) from Karnataka, Kerala, Maharashtra, and Tamil Nadu in

southern India. The scabioid apothecia with vertically layered excipulum and

26 ... Joshi, Upreti & Nayaka

<4-spored asci are the major distinguishing characters of C. laceratula, which

morphologically may be confused with Topeliopsis species. It is generally

distributed at lower elevation and growing luxuriantly in evergreen forests of

Western Ghats.

SPECIMENS EXAMINED: INDIA. KARNATAKA: Central Western Ghat, Shimoga district,

3 km before Agumbe Ghat, from Koppa to Agumbe, alt. 649 m, on bark in evergreen

forest, 14.01.2008, H.T. Lumbsch, D.K. Upreti, P.K. Divakar & J. Tandon 19730M/A

(LWG); Uttarkannad, Sharavathi Ghat, near Gersoppa, alt. 718 m, on bark in evergreen

forest, 16.01.2008, H.T. Lumbsch, D.K. Upreti, P.K. Divakar & J. Tandon 19744A-1,

19744B, 1944C-2, 197441, 19745, 19745A-3, 19745C (LWG); Tamit Napvu: Madurai

district, High ways Vattapara (Meghamaali), alt. 1580 m, on fallen twigs, 22.03.1999, S.

Nayaka 92(LWG).

Chapsa leprieurii (Mont.) Frisch, Biblioth. Lichenol. 92: 105, 2006.

The species is characterized by a brown-olive, cartilaginous thallus lacking

secondary compounds, rather small, rounded to slightly elongated or branched

apothecia level with the thallus, 8-spored asci, and transversely septate brown

oblong to ellipsoidal or clavate ascospores.

Chapsa leprieurii has been reported from Karnataka, Kerala, and Tamil

Nadu in southern India (Awasthi 1991, as Thelotrema leprieurii; Singh & Sinha

2010, as T! leprieurii). The brown-spored similar species, C. platycarpa, differs

in producing lichen compounds and a double margin.

Chapsa leprocarpa (Nyl.) Frisch, Biblioth. Lichenol. 92: 108, 2006.

The taxon exhibits a pale to dark olive-grey thallus lacking lichen compounds,

large chroodiscoid apothecia with a lobed and recurved margin, pale brown

to blackish but heavily white pruinose disc, 1-spored asci, and muriform

ascospores.

Awasthi (1991) first recorded C. leprocarpa (as Thelotrema colobicum)

from Andaman and Nicobar Islands. Chapsa leprocarpa inhabits lowland and

submontane regions in the Western Ghats.

SPECIMENS EXAMINED: INDIA. KErAta: Idukki district, .C.R.I campus, Myladumpara,

alt. ca. 1200 m, on bark of tree, 01.03.1984, D.D. Awasthi & G. Awasthi 84.88 (LWG-

LWU); on way Myladumpara to Munnar, Santhampara area, alt. ca. 1200 m, on bark

of tree, 02.03.1984, D.D. Awasthi & G. Awasthi 84.179 (LWG-LWU); Thikkady, Periyar

Tiger Reserve, Sakunthalakadu, on bark, 23.03.2006, B. Haridas 06-009615/B (LWG).

Chapsa leprocarpoides (Hale) Caceres & Liicking, Libri Bot. 22: 52, 2007.

The species is ecorticate, pale olive to fawn coloured, lacking lichen

substances, without compact surface, usually reflecting bark through the

matted thallus surface; it has immersed to semi-immersed, rounded to angular

apothecia with a flesh-coloured pruinose disc and low jagged to lobed margins,

a pale to brownish or hyaline free proper exciple, 8-spored asci, and hyaline

muriform ellipsoidal to oval ascospores.

Chapsa in India ... 27

Chapsa leprocarpoides was collected from evergreen tropical forests of

Karnataka on rough tree barks at lower altitude. The ecorticate “Chapsa hiata”

differs in having rather small ascospores while C. velata and C. pseudophlyctis

have comparatively larger ascospores and a fused proper exciple.

SPECIMENS EXAMINED: INDIA. KARNATAKA: Chikmagalure district, Chamudi Ghat,

Kuvettu, alt. 104 m, on bark in evergreen forest, 15.01.2008, H.T. Lumbsch, D.K. Upreti,

PK. Divakar & J. Tandon 19739R; Uttar Kannad, Sharavati Ghat, near Gersoppa, alt. 718

m, on bark in evergreen forest, 16.01.2008, H.T. Lumbsch, D.K. Upreti, P.K. Divakar &

J. Tandon 19744J (LWG); Shimoga district, Sagar to Talguppa, Ulanahalli, alt. 697 m,

on bark in evergreen forest, 16.01.2008, H.T. Lumbsch, D.K. Upreti, P.K. Divakar & J.

Tandon 19742E, 19742E-1(LWG); Central Western Ghat, Shimoga district, near Jog fall,

Kargal, alt. 644 m, 16.01.2008, H.T. Lumbsch, D.K. Upreti, P.K. Divakar & J. Tandon

19742B-2, 197430-1 (LWG).

Chapsa meghalayensis (Patw. & Nagarkar) Lumbsch & Divakar, Lichenologist 42:

183, 2010.

This taxon is characterized by its ashy-white, ecorticate, granular thallus

lacking lichen compounds, immersed and chroodiscoid apothecia with

recurved margins, lacerate exciple, 2-spored asci, and small muriform brown

ascospores measuring 27-46 x 11-16 um.

First described from Meghalaya by Patwardhan & Nagarkar (1980) as

Leptotrema meghalayense, the species was later transferred to Myriotrema by

Awasthi (1991). Chapsa meghalayensis is endemic to India and collected from

evergreen forests of northeast India.

Chapsa patens (Nyl.) Frisch, Biblioth. Lichenol. 92: 111, 2006. PrP aA

Thallus corticolous, epiperidermal, ecorticate, dark olivaceous grey to

dark grey or blackish, continuous, + cracked due to bark texture, uneven,

bark reflecting through thallus. Prothallus indistinct. Photobiont layer well

developed, up to 50 um thick. Medulla endoperidermal. Apothecia scattered,

sometimes grouped in two, immersed to level with thallus, mostly rounded to

slightly angular or irregular in shape, up to 2.0 mm in diam. Margin raised,

lobed to + continuous, mostly straight to recurved later, brownish to faintly

crystalline pruinose inner side. Disc reddish-brown, exposed, covered by thick

pruina. Proper exciple cupular, pale to hyaline, sometimes brownish, up to 40-

80 um thick laterally. Periphysoides indistinct to 30 um long, inclined towards

epihymenium. Epihymenium 10-15 um high, slightly brownish, granular.

Hymenium, clear, 130-140 um high. Paraphyses entangled, simple, <1.5

um thick, apically thickened, + dendroid branched, adspersed with grayish

granules. Ascus, 1-spored, clavate, 90-130 x 25-40 um. Ascospores hyaline,

densely muriform, with rounded ends, 80-125 x 20-35 um, I-.

CHEMISTRY: K-, PD-, C-; no lichen compound detected in TLC.

ECOLOGY & DISTRIBUTION: The species is a new record to India reported

from tropical rain forests of the Eastern Himalayas where it is common in open,

28 ... Joshi, Upreti & Nayaka

canopied old forests with large tree trunks and generally found at 1700-2500

m. Widely distributed and reported from Africa, Japan, and Sri Lanka.

SPECIMEN EXAMINED: INDIA. WeEsT BENGAL: Eastern Himalaya, Darjeeling district,

Tiger hill, north face of the hill, alt. ca. 2500 m, on bark of tree, 05.03.1967, D.D. Awasthi

& M.R. Agarwal 67.21 (LWG-LWU).

REMARKS: Chapsa leprocarpa, which also lacks lichen substances, differs from

C. patens in having a slightly lower hymenium and smaller ascospores and

grows at lower altitudes (1000-1200 m). The Indian specimen of C. patens

exhibits a dirty grey to dark olivaceous thallus colour hardly distinguished

from the bark, while C. leprocarpa has a pale-green to slightly greyish-green

epiperidermal thallus. However, the hymenium and ascospores of the Indian

sample are intermediate in size and closer to C. patens than C. leprocarpa and

its distribution and thallus and apothecial morphology also agree well with

the diagnostic features of C. patens. Hale (1981) and Matsumoto (2000) both

described C. patens as a stictic acid-producing species that after re-investigation

were shown to lack these substances. The records from Japan and Sri Lanka

should be re-examined (Frisch et al. 2006).

Chapsa platycarpa (Tuck.) Frisch, Biblioth. Lichenol. 92: 113, 2006.

The species is recognized by the dark olive-green to olive-brown or pale

yellowish-brown thallus containing the stictic acid chemosyndrome, a rounded

to slightly irregular apothecioid to chroodiscoid shape in older material,

erumpent apothecia with double margins, 8-spored asci, and transversely

septate ellipsoidal to fusiform brown ascospores.

Earlier placed in the neglected genus Phaeotrema due to its transversely

septate brown ascospores, Chapsa platycarpa was reported from India (Kerala)

as P. platycarpum (Patwardhan & Kulkarni 1977a). The similar C. leprieurii

lacks stictic acid as a major thallus compound.

Chapsa platycarpella (Vain.) Frisch, Biblioth. Lichenol. 92: 118, 2006. PL. 1B

Thallus corticolous, endoperidermal, olive-green to pale-green, continuous,

shiny, compact, cartilaginous, uneven to warty. Prothallus indistinct to

brownish. Medulla white, indistinct, endoperidermal. Phenocortex well

developed, dense, 20-30 um thick. Photobiont layer 20-35 um thick, with

inclusion of calcium oxalate crystals, largely endoperidermal. Apothecia

scattered to aggregate in groups, immersed to level with thallus, round to mostly

angular or irregular in shape, <1.0 mm in diam. Margin thin, fissured to lobed,

recurved, and yellowish to fawn with felty white pruinose to crystalline inner

surface. Disc pale to blackish, distinctly white pruinose. Proper exciple cupular,

pale to hyaline, 6-10 um wide. Periphysoides indistinct to 25 um long, inclined

towards epihymenium. Epihymenium 5-8 um high, granular, unpigmented

Chapsa in India ... 29

to crystalline. Hymenium clear, 55-95 um high. Paraphyses simple, straight,

<2.0 um thick, tips agglutinate with fine greyish to brownish granules. Ascus

8-spored, clavate, 45-90 x 6-9 um. Ascospores, hyaline, transversely 3-5

septate, fusiform with acute end cells, 12-16 x 3-5 um, I-.

CHEMISTRY: K-, PD-, C-; no lichen compounds detected in TLC.

ECOLOGY & DISTRIBUTION: Known to grow on smooth bark of young trees in

lowland rainforests and coastal forests in shade as well as on exposed surfaces.

In India, reported on trees in evergreen tropical forest in Kerala.

SPECIMEN EXAMINED: INDIA. KERALA: Idukki, Adimali Forest Range, Thondi Kappu,

on Myristica beddomei, 15.02.2006, B. Haridas 06-009598(LWG).

REMARKS: Chapsa platycarpella, which is considered a separate species by Frisch

et al. (2006), is otherwise treated as a synonym of C. alborosella. The species

is well characterized and separated from the latter in having a cartilaginous

thallus, thick phenocortex, and loosely aggregated apothecia with blackish

discs covered by bluish-white pruina.

Recently, Rivas Plata et al. (2010) placed the species in synonymy with

C. astroidea (Berk. & Broome) Caceres & Liicking; however following Frisch

(2006) with adequate distinguishing characteristics the species is accepted here

as a new record for India.

Chapsa pseudoexanthismocarpa (Patw. & C.R. Kulk.) Rivas Plata & Liicking,

Lichenologist 42: 183, 2010.

The taxon is characterized by a pale yellowish to greenish brown or pale olive

thallus lacking secondary compounds, apothecia that are rounded to somewhat

irregular and sometimes perithecioid, erumpent, immerged to distinctly

emergent, 4-8-spored asci, and hyaline transversely septate bacilliform-

fusiform ascospores.

Patwardhan & Kulkarni (1977b) described the species from southern

India (Kerala) in Ocellularia. Larger ascospores readily distinguish

C. pseudoexanthismocarpa from C. indica. Other characters include the rough

epiperidermal thallus, lepadinoid apothecia, and lacerate exciple.

Chapsa pseudophlyctis (Ny1.) Frisch, Biblioth. Lichenol. 92: 120, 2006.

The species is characterized by its white to whitish, whitish-grey to pale-grey

thallus lacking secondary compounds, round to angular apothecia with jagged,

often eroded margins having a white crystalline inner surface, 1-6-spored asci,

and hyaline muriform ascospores.

Chapsa pseudophlyctis may sometimes be confused with C. leprocarpa, which

can be distinguished by asci consistently bearing a single larger ascospore.

Jagadeesh Ram & Sinha (2009) first reported C. pseudophlyctis as new to India

from Meghalaya and Sikkim.

30 ... Joshi, Upreti & Nayaka

PLATE. 1. New records of Chapsa from India.

A. Chapsa patens. B. Chapsa platycarpella (note the apothecia).

Scale bars: A = 2mm; B= 1 mm.

Chapsa recurva (G. Salisb.) Frisch, Biblioth. Lichenol. 92: 95, 2006.

Chapsa recurva and C. platycarpa are the only two Chapsa species that

produce stictic acid as a major secondary compound (Rivas Plata et al. 2010).

Nagarkar et al. (1986) reported the species (as Thelotrema recurvum) from

Andaman & Nicobar Islands, but noted the absence of secondary compounds.

Other major distinguishing characters include the corticate whitish grey smooth

thallus, numerous solitary round to elongated semi-emergent chroodiscoid

apothecia, and hyaline muriform ascospores.

Chapsa in India... 31

“Chapsa stellata” (Hale), ined.

The taxon is recognized by the greenish, glaucous, continuous, mostly

epiperidermal thallus lacking lichen substances, dispersed, slightly immersed

chroodiscoid apothecia having a white pruinose disc and recurved margins,

a + fused brown proper exciple, 1-spored asci, and ellipsoidal olivaceous to

brownish muriform ascospores, 88-135 x 22-30 um. Patwardhan & Nagarkar

(1980) reported the species (as Leptotrema stellatum) from Meghalaya in the

eastern Himalayas.

Chapsa velata (Nyl.) Caceres & Licking, Libri Bot. 22: 54, 2007.

The species is characterized by the ashy-white smooth partly hypophloeodal

thallus lacking lichen substances, semi-emergent chroodiscoid apothecia

with erect to recurved margins, fused to free brown proper exciple, hyaline

hymenium, 4-8-spored asci (1-2-spored according to Patwardhan & Nagarkar

1980), and hyaline ellipsoidal muriform ascospores. The species was recorded

from the Andaman Islands (Awasthi 1991, as Thelotrema velatum) and Nagaland

(Patwardhan & Nagarkar 1980, as T. velatum).

Key to Chapsa species recorded from India

Ia. 2kSCOSPOrestirAalisVerselyesep latest. e8ih he Pst t tel Paik has chi doh |e Bloe ttl Le 2

Ib AScosporest(Subs jot TITONIN «042. '3.q.4'¢ sane x poea 4 pean + po iat F paeaen sh sect eens iy

2d HA SCOSPOKES NY AMIS cm, a Mwsere, ai eee AP ae ame Fld a Pt hae NP lo Oa ES LeeLee 3

2D ASCOSPOLES’ DEO WAN: ET Si.e oped e potsate Zeon eh le eR lee eh cde Su eh SLOG CRs ty 6

DT AASCOSPOLES 3s FOCI «ie creey Basecrary Basectery Banu hls Meank ly anohhy Tasneh ty Tatnat strat Casa haab Cisakate 4

Sbt- ASCOSPOres <i AN MINIM. tela re x cosa .a ie 5 hls es wl laie adele 6 at wie Lae + ela oe Sule oa Sale eae S

4a SAscospores (40=)770-120 uti lON SS. s.-..5%, cee oe a eaten atone abe wna C. indica

4b. Ascospores 100-200 um long ...................000- C. pseudoexanthismocarpa

5a. Ascospores 12-16 um long, 4—-6-septate, corticate................ C. platycarpella

5b. Ascospores 17-22 um long, 5-9-septate, ecorticate................. C. alborosella

6a. Stictic acid present, proper exciple free (double margin),

apothecialteise-oreysbeowils $10.29 tt Soh id FLA ETA LAE Le C. platycarpa

6b. Lichen substances absent, proper exciple fused,

dise. pale: brown 10 CrevISh nthe ni dai sebding at feed bata hah did C. leprieurii

PAPASCOSPOFESINV ANI Sot inches Bevcttes Besmartes and, ao Als Mie Al Ware Als Taree idee een a eee 8

7D ASCOSPOFES, DEO WM yc lela + satel ss coe wx + kelses 5 ablahe boat de blots oy Blas om als adele oie 5

BAVA SCOSPOKES: OU MIT LOUS ets irate I cate I pelt bedded on etn te ata heat nc er ne 9

Shi Ascosporesi= GO (Monge: chin peahies pesca ok wha eh ieee ie he de’ ye § 13

9a. Ascospores 12-15 um long, lichen substances absent .................. “C. hiata”

Ob: -Ascosporess-l oyun lores chemastryavariabl ess. tant gi, fact ice died Pct fds Hie ded tga dee ood 10

32 ... Joshi, Upreti & Nayaka

10a, Sticheacid ‘presents cu 25 G0.2. 2400. Vain Gite aae eine yale C. recurva

ONT Le CTE, SUIS EAIICR SAI SOITE cas Ped ais esd Sank Ue ad iG dO cece A cat IE deh 11

lla. Proper exciple free, apothecia often aggregate,

ascospores GOES RALO= 212 WTTe: +. eats! thera ven More woh Save aby baragee hand C. leprocarpoides

11b. Proper exciple fused apothecia solitary, ascospores 30-60 x 12-20 um......... 12

12a. Thallus ashy-white, apothecial disc dark-grey, thinly pruinose

PvE ONES SEITE Cea eau en des che te ha ka Md cA Wak Le C. velata

12b. Thallus pale-olive, apothecial disc pale brown,

distinctly white prUinoSe am a: garg wh he gs gs ed aig Seal de C. pseudophlyctis

13a. Thallus with prosoplectenchymatous cortex, proper exciple layered C. laceratula

13 b wlnalhisecortiGaie. ng OP us. t-ratcas, ke te ee Re we 2S A gE EO eRe ce 14

14a. Ascospores. 2125, pny lone: oe eee es eet mee BM ene BS ane OF matey hoe enone C. patens

146 ASCOSPORES FO PMIONG 1.55 f.t.08 fel EF eee wel! we wee as C. leprocarpa

15a. Ascospores 2746-56 1 U6 filth acts, sctee aces ots eg Gis st oe C. meghalayensis

15b: AScospores:$8= 135562 2—S Oui x sree, stirs gills ies tect tlie ey tetliredey 9 alii “C. stellata”

Acknowledgements

We are grateful to the Director of the CSIR-National Botanical Research Institute,

Lucknow, for providing necessary facilities, and the Ministry of Environment and

Forests, New Delhi, for financial support. The authors are also grateful to Dr. R. Liicking

and Dr. Pradeep. K. Divakar for their valuable comments on manuscript.

Literature cited

Awasthi DD. 1991. A key to microlichens of India, Nepal and Sri Lanka. Bibliotheca Lichenologica

40: 1-337.

Awasthi DD. 2000. Lichenology in Indian subcontinent: a supplement to “A Handbook of Lichens”.

Bishen Singh Mahendra Pal Singh, DehraDun, India.

Dubey U, Upreti DK, Rout J. 2007. Lichen flora of Along town, West Siang district, Arunachal

Pradesh. Phytotaxonomy 7: 21-26.

Frisch A, Kalb K, Grube M. 2006. Contribution towards a new systematic of the lichen family

Thelotremataceae. Bibliotheca Lichenologica 92: 1-556.

Hale ME. 1974. Morden-Smithsonian Expedition to Dominica: the lichens (Thelotremataceae).

Smithsonian Contributions to Botany 16: 1-46.

Hale ME. 1978. A revision of the lichen family Thelotremataceae in Panama. Smithsonian

Contributions to Botany 38: 1-60.

Hale ME. 1981. A revision of the lichen family Thelotremataceae in Sri Lanka. Bulletin of the British

Museum (Natural History), Botany Series 8: 227-332.

Jagadeesh Ram TM, Sinha GP. 2009. New records of lichenized and non-lichenized ascomycetes

from India. Indian Journal of Forestry 32(4): 635-640.

Mangold A, Martin, PM, Licking, R, Lumbsch TH. 2008. Molecular phylogeny suggests synonymy

of Thelotremataceae within Graphidaceae (Ascomycota: Ostropales). Taxon 57: 1-11.

Mangold A, Elix JA, Lumbsch HT. 2009. Thelotremataceae. Flora of Australia 57: 195-420.

Chapsa in India ... 33

Matsumoto T. 2000. Taxonomic studies of the Thelotremataceae (Graphidales, lichenized

Ascomycota) in Japan (1) Genus Thelotrema. J. Hattori Bot. Lab. 88: 1-50.

Nagarkar MB, Sethy PK, Patwardhan PG. 1986. Materials for a lichen flora of the Andaman Islands

— I. Mycotaxon 27: 71-82.

Nagarkar MB, Sethy PK, Patwardhan PG. 1988. Lichen genus Ocellularia (Family Thelotremataceae)

from India. Biovigyanam 14: 24-43.

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society.

Patwardhan PG, Kulkarni CR. 1977a. A contribution to our knowledge of the lichen flora of India

I: Family Thelotremataceae. Kavaka 5: 1-17.

Patwardhan PG, Kulkarni CR. 1977b. Some new taxa of the family Thelotremataceae from Western

Ghats, SW India. Norwegian Journal of Botany 24: 127-131.

Patwardhan PG, Nagarkar MB. 1980. Notes on some lichens from northeast India II: family

Thelotremataceae. Biovigyanam 6: 1-10.

Patwardhan PG, Sethy PK, Nagarkar MB. 1985. A contribution to our knowledge of the lichen

family Thelotremataceae from South India. Biovigyanam 11: 133-140.

Purvis OW, Jorgensen PM, James PW. 1995. The lichen genus Thelotrema Ach. in Europe.

Bibliotheca Lichenologica 58: 335-360.

Rivas Plata E, Licking R, Lumbsch HT. 2008. When family matters: an analysis of Thelotremataceae

(lichenized Ascomycota: Ostropales) as bioindicators of ecological continuity in tropical forests.

Biodiversity and Conservation 17: 1319-1351. http://dx.doi.org/10.1007/s10531-007-9289-9

Rivas Plata E, Licking R, Sipman HJM, Mangold A, Kalb K, Lumbsch HT. 2010. A world-wide

key to the thelotremoid Graphidaceae, excluding the Ocellularia-Myriotrema-Stegobolous clade.

The Lichenologist 42(2): 139-185. http://dx.doi.org/10.1017/S0024282909990491

Santesson R. 1952. Foliicolous lichens I. A revision of the taxonomy of the obligately foliicolous,

lichenized fungi. Symbolae Botanicae Upsalienses 12: 1-590.

Singh KP, Sinha GP. 2010. Indian lichens: An annotated checklist. Botanical Survey of India. Shiva

Offset Press, DehraDun, Uttarakhand.

Sipman HJM, Harris RC. 1989. Lichens. 303-309, in: H Lieth, MJA Were (eds). Tropical Rain

Forest Ecosystems. Amsterdam: Elsevier Science Publishers.

MYCOTAXON

http://dx.doi.org/10.5248/120.35

Volume 120, pp. 35-41 April-June 2012

A new species of Inonotus (Hymenochaetaceae) and

Trametes cingulata (Polyporaceae) newly recorded from Brazil

MaAtrA CORTELLINI ABRAHAO & ADRIANA DE MELLO GUGLIOTTA

Instituto de Botanica, Nucleo de Pesquisa em Micologia

Caixa Postal 68041, CEP 04045-972, Sao Paulo, SP. Brazil

* CORRESPONDENCE TO : mairaabrahao@hotmail.com

ABSTRACT — Two interesting species found during a survey of polypore fungi in northwestern

Sao Paulo State, Southeast Brazil, are described and illustrated. Inonotus multisetifer is

proposed as a new species characterized by resupinate basidiomata with round to angular

pores, 6—9 per mm, acute setal hyphae embedded in trama, subulate hymenial setae, and

globose to subglobose basidiospores. Trametes cingulata constitutes a first record from

Brazil.

KEY worDs — mycodiversity, neotropics, Polyporales, Hymenochaetales

Introduction

The Atlantic Rain Forest, which has 20,000 species of plants of which 6000

are endemic, holds today less than 8% of its original extent in Brazil and has

been rated as one the world’s top five biological hotspots (Mittermeier et al.

1999, SOS Mata Atlantica/INPE 2009).

The state of Sao Paulo is located in southeastern Brazil and has an area of

248,808.8 km’. Mostly inserted in the Atlantic Forest domain (68%), the state

contains remnants of rain forest, Araucaria forest, seasonal semideciduous

forest, transitions between them, and remnants of vegetation of restinga and

mangrove (Kronka 2005, SOS Mata Atlantica/INPE 2009).

Currently, the natural vegetation cover of the state amounts to only 13.94%

of its surface (Kronka 2005, Nalon et al. 2008), concentrated mainly along the

coastal zone, where there is a concentration of natural areas of Integral Protection

(Xavier et al. 2008). In the northwest region, only minor conservation units are

found (Kronka 2005, Xavier et al. 2008), with the majority of the remaining

forest located in private areas where their conservation is key to biodiversity

conservation in the State.

36 ... Abrahao & Gugliotta

The mycodiversity in northwest Sao Paulo State is almost unknown;

currently, only 36 species of basidiomycetes have been cited from this region

(Xavier-Santos 2003, Abrahao et al. 2008, 2009).

As part of a wider project dealing with biodiversity of remnant forest

fragments of the northwest region of Sao Paulo State, this study reports a

new species of Inonotus and a new record of Trametes cingulata from Brazil,

including descriptions, illustrations, and discussions on the species.

Materials & methods

Collections were performed between March 2007 and December 2008 in

Semidecidual Stacional Forest fragments in northwest Sao Paulo State, Brazil.

The material was studied following the classical methods for polypores (Gilbertson

& Ryvarden 1986, Ryvarden 1991, 2004). Micromorphological observations were made

from material mounted in 5% KOH and Melzer’s reagent; measures were made in 5%

KOH in a Leica DM 1000 microscope.

Vouchers are deposited in SP, SJRP, and O (acronyms follow Holmgren & Holmgren

1998).

Nomenclature and classification systems follow those of Kirk et al. (2008),

Centraalbureau voor Schimmelcultures (www.cbs.knaw.nl), and Index Fungorum

(www.indexfungorum.org).

Taxonomy

Inonotus multisetifer Abrahao & Gugliotta, sp. nov. Figs 1-4

MycoBank MB 563746

Differs from Inonotus adnatus in larger basidiomata, slightly narrower setal hyphae,

smaller scant hymenial setae, and larger (8.8-11.2 x 8.8-10 um) globose to subglobose

basidiospores.

Type: BRAZIL. Sado Paulo State: Sdo Jodo de Iracema, SAo Francisco Farm, G5 area

(20°28'25"S 50°17'36"W), 17.IV.2007, M.C. Abrahao 270 (holotype, SP393791; isotype,

SJRP).

ErymMo_ocy: Referring to the abundant setal hyphae.

BASIDIOMA annual, resupinate, up to 8 cm in diameter, ca. 4 mm thick, strongly

adnate, hard and dense when dry. MarGaIn thin to absent. PORE SURFACE

ochraceous to brown, pores round to angular, 6—9 per mm, invisible to the

naked eye; tubes dark brown, up to 4 mm deep. CONTEXT homogeneous, very

thin, virtually absent in parts, cinnamon. HYPHAL SYSTEM monomitic, IKI-;

generative hyphae simple-septate, thin to thick-walled, golden to rusty brown,

(2.5—)3.75 um wide. SETAL HYPHAE abundantly present, dark brown, acute,

thick-walled, 100-200 x 8.75—16.25 um, embedded in trama and not projecting.

HYMENIAL SETAE scant and in some sections totally absent, apparently found

only in the young pores and in the pores mouths, dark brown, subulate, thick-

walled, 33.75—45 x 8.75-10 um. Basrpt1a clavate, hyaline, with four sterigmata,

Inonotus multisetifer sp. nov. (Brazil) ... 37

FiGuRES 1-5 (scale bars = 10 um). Fics. 1-4: Inonotus multisetifer (SP393791, holotype)

— 1, basidiospores; 2, basidia and immature basidiospore; 3, hymenial setae; 4, setal

hyphae. Fic. 5. Trametes cingulata (SP416154) — basidiospores.

22.5—23.75 x 5—6.25 um. Basip1osPorREs globose to subglobose, IKI-, slightly

thick-walled, hyaline to pale yellow, 8.75-11.25 x 8.75-10 um.

SUBSTRATE & DISTRIBUTION —dead hardwood; known only from the type

locality in Brazil.

38 ... Abrahaéo & Gugliotta

ADDITIONAL SPECIMENS EXAMINED —Inonotus adnatus: COSTA RICA. PUNTARENAS:

La Amistad Pacifico, Estacion Progreso, near the station, alt 1180 m, 27.1X.2000,

L. Ryvarden 42795 (0316222-079, isotype). Inonotus micantissimus: BRAZIL. R1o

GRANDE DO SUL STATE: Sao Salvador, III.1944, S.J. Rick 20691 (PACA8878, holotype).

Inonotus pegleri: TANZANIA. EASTERN PROVINCE: KiLosa District, Mikumi

National Park, on dead fallen tree, alt 487 m, 30.IV.1968, D.N. Pegler T725 (K167869,

holotype; 0296, isotype).

ComMENTSs — Inonotus multisetifer is characterized by resupinate basidiomata,

presence of acute setal hyphae in the trama, scant subulate hymenial setae, and

globose to subglobose slightly thick-walled hyaline to pale yellow basidiospores.

This combination of characters relate this species to other Inonotus resupinate

species that also have setal hyphae, hymenial setae, and basidiospores varying

from globose, subglobose to ellipsoid: I. adnatus Ryvarden, I. micantissimus

(Rick) Rajchenb., and I. pegleri Ryvarden. ‘The main diagnostic characteristics

are shown in TABLE 1.

TABLE 1. Key features of Inonotus multisetifer and morphologically related species.

I. multisetifer I. adnatus I. micantissimus I. pegleri

Basidioma <8cm diam. <4cm diam. 10x4cm 12cm

(thickness) (~4 mm) (~3 mm) (<14 mm) (< 15 mm)

Pores/mm 6-9 6-9(-10) 5-6 4-6

Generative hyphae: Thin to thick; Thin to thick; — Slightly thick; Thin to thick;

walls; golden to golden to yellowish to chestnut hyaline to

color rusty brown rusty brown light-brown

Hymenial setae 33.8—-45 x 26.2—-56.2 x 22.5-27.5 X 20-21.2 x

(um) 8.8—10 10-12.5 6.2-11.2 6.2-8.8

Setal hyphae 100-200 x < 150 x 160-300(-400) x 125-520 x

(um) 8.8—16.2 8.8-17.5 8.8-20* 7.5-21.2

Basidiospore shape; (Sub) globose; Globose; (Sub)globose, (apiculate); | Globose;

color hyaline to yellowish hyaline to pale melleous hyaline to

yellowish yellowish

Basidiospore size 8.8-11.2 x 7-8.8 diam. 11.25 x 8.8-10 6-7 um wide

(um) 8.8-10

*(width up to 25 um in KOH)

Inonotus multisetifer is morphologically most closely related to I. adnatus,

a species known only from the type locality in Costa Rica (Ryvarden 2002,

2004, 2005). Inonotus adnatus presents a smaller basidioma, slightly wider

setal hyphae, larger and ventricose hymenial setae, and smaller globose

basidiospores. Hymenial setae are abundant in the type specimen of I. adnatus

but scant to mostly absent in I. multisetifer.

Inonotus multisetifer sp. nov. (Brazil) ... 39

Inonotus micantissimus, which also occurs in Brazil (Rajchenberg 1987,

Baltazar et al. 2010), has larger pores, longer setal hyphae, smaller hymenial

setae, and basidiospores with abundant oily contents.

Inonotus pegleri is easily separated by the combination of larger pores,

smaller hymenial setae, longer setal hyphae, smaller basidiospores, and its

limited distribution in Africa (Ryvarden 2005).

Trametes cingulata Berk., Hooker’s J. Bot. Kew Gard. Misc. 6: 164, 1854 FIG. 5

BASIDIOMA annual, lignicolous, sessile pileate to pseudostipitate, dimidiate,

many basidiomata from one stipe or solitary; pileus semicircular, applanate

and thinner near the margin, with contracted base, soft when fresh and flexible

when dry, 0.7—4.6 x 0.7—3.9 x 0.1—0.8 cm. PILEAR SURFACE in brown and cream

tints, concentrically zonate, glabrous. MarGIN round, concolorous with the

pilear surface, entire, glabrous and sterile. Stipe when present circular, cream

to brown, glabrous, dense, 0.2-1.1 x 0.2-0.9 x 0.5-1.3 cm. PORE SURFACE

poroid, lighter than pilear surface, pores angular to elongated, 7-9 per mm;

tubes stratified, forming up to 4 distinct layers, light-brown, up to 2 mm deep;

dissepiments thin to slightly thick, lacerate. CONTEXT duplex, the upper layer

fibrous, greyish to blackish, thin, at least at the base separated by a thin black

line from the lower part, the latter light-brown, corky, up to 6 mm thick at

the base, negative reaction in KOH. HypHaAt sysTeEM trimitic, IKI-; generative

hyphae hyaline, with clamps, thick-walled, 2.5—-3.75 um wide; skeletal hyphae

yellowish, thick-walled, abundant in the basidioma, golden and solid in the

context, 3.75—6.25 um wide; binding hyphae hyaline to yellow, thick-walled to

semisolid in the context, slightly irregular and moderately branched, frequent,

1.25-3.75 um wide. Cystrip1a absent. Basrp1a clavate, hyaline, thin-walled,

11.25-12.5 x 3.75—5 um. Basip10sPorEs obovoid, hyaline, IKI-, smooth, thin-

walled, 5—6.25 x (2.5—)3.75 um.

SUBSTRATE & DISTRIBUTION — causing white-rot in twigs of dead angiosperm.

Previously cited from Asia, Africa, and Australia (Ryvarden & Johansen 1980,

Young et al. 2004, Gilbert et al. 2008) and Costa Rica and Venezuela in America

(Ryvarden & Iturriaga 2001, Mata et al. 2007).

MATERIAL EXAMINED — BRAZIL. SAO PAULO STATE: Palestina, Boa Vista Farm, P6

area (20°19'16"S 49°30'17”W), 10.XII.2008, A.M. Gugliotta 1410 (SP416154, SJRP); Sao

Joao de Iracema, Sao Francisco Farm, G5 area (20°28'25"S 50°17'36"W), 27.XI.2007,

M.C. Abrahao 319 (SP393683). INDIA. BEHAR: Soane River [BIHAR: Son River], on

dead wood, J. Hooker s.n. (K167868, isotype).

COMMENTS — Macroscopically the species may be recognized by its glabrous

basidioma in brown and cream tints (Ryvarden & Johansen 1980). The angular

to elongated pores, duplex context with a greyish to blackish fibrous upper

AO ... Abrahdo & Gugliotta

layer, separated by a thin black line from the lower light-brown corky part,

and obovoid, hyaline, thin-walled basidiospores support the identification. It

constitutes a new record from Brazil.

Acknowledgments

The authors acknowledge Dr. Leif Ryvarden for identification of Trametes

cingulata (SP393683). We are grateful to FAPESP (first author's fellowship process n°

2006/58786-6 and BIOTA thematic project process n° 2004/04820-3) for financial

support. We also extend our thanks to Gerardo Lucio Robledo and Rosa Mara Borges

da Silveira, who kindly reviewed the manuscript.

Literature cited

Abrahao MC, Gugliotta AM, Silva R, Fujieda RJY, Boscolo M, Gomes E. 2008. Ligninolytic activity

from newly isolated basidiomycete strains and effect of these enzymes on the azo dye orange II

decolourisation. Annals of Microbiology 58: 427-432.

Abrahao MC, Gugliotta AM, Gomes E. 2009. Polipordides (Basidiomycota) em fragmentos de mata

no perimetro urbano de Sao José do Rio Preto, Sao Paulo, Brasil. Revista Brasileira de Botanica

32: 427-440. http://dx.doi.org/10.1590/S0100-84042009000300004

Baltazar JM, Trierveiler-Pereira L, Ryvarden L, Loguercio-Leite C. 2010. Inonotus s.l.

(Hymenochaetales) in the Brazilian herbaria FLOR and SP. Sydowia 62: 1-9.

Gilbert GS, Gorospe J, Ryvarden L. 2008. Host and habitat preferences of polypore fungi in

Micronesian tropical flooded forests. Mycological Research 112: 674-680. http://dx.doi.

org/10.1016/j.mycres.2007.11.009

Gilbertson RL, Ryvarden L. 1986. North American polypores. Synopsis Fungorum 1: 1-433.

Holmgren PK, Holmgren NH. 1998. Index Herbariorum: New York Botanical Garden's Virtual

Herbarium. Available at: http://sweetgum.nybg.org/ih/Holmegren.

Kirk PM, Cannon PF, Minter DW, Stalpers J. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10th ed. Wallingford, CAB International. 771 p.

Kronka FJN. 2005. Inventario florestal da vegetacao natural do Estado de Sao Paulo. Sao Paulo,

Secretaria de Meio Ambiente/Instituto Florestal. 200 p.

Mata M, Ruiz-Boyer A, Carranza J, Ryvarden L. 2007. Nuevos registros de hongos poliporoides

(Basidiomycetes) para Costa Rica. Boletin de la Sociedad Micoldgica de Madrid 31: 123-129.

Mittermeier RA, Myers N, Gil PR, Mittermeier CG. 1999. Hotspots: earth’s biologically richest and

endangered terrestrial ecoregions. Ciudad de Mexico, CEMEX/Agrupacion Sierra Madre. 430

Nalon MA, Mattos IFA, Franco GADC. 2008. Meio fisico e aspectos da fragmentacao vegetal.

15-21, in: Rodrigues RR, Bononi VLR (Orgs.). Diretrizes para a Conservacao e Restauracao da

Biodiversidade no Estado de Sao Paulo. Sao Paulo, Instituto de Botanica.

Rajchenberg M. 1987. Type studies of Polyporaceae (Aphyllophorales) described by J. Rick. Nordic

Journal of Botany 7: 553-568. http://dx.doi.org/10.1111/j.1756-1051.1987.tb02023.x

Ryvarden L. 1991. Genera of polypores. Nomenclature and taxonomy. Synopsis Fungorum 5:

1-363.

Ryvarden L. 2002. Studies in neotropical polypores 17. New neotropical Inonotus species. Synopsis

Fungorum 15: 70-80.

Ryvarden L. 2004. Neotropical polypores. Part 1. Introduction, Ganodermataceae &

Hymenochaetaceae. Synopsis Fungorum 19: 1-227.

Inonotus multisetifer sp. nov. (Brazil) ... 41

Ryvarden L. 2005. The genus Inonotus - a synopsis. Synopsis Fungorum 21: 1-149.

Ryvarden L, Iturriaga T. 2001. Studies in neotropical polypores 9. A critical checklist of poroid

fungi from Venezuela. Mycotaxon78: 393-405.

Ryvarden L, Johansen I. 1980. A preliminary flora of East Africa. Fungiflora: Oslo, 636p.

SOS Mata Atlantica, Instituto Nacional de Pesquisas Espaciais. 2009. Atlas dos remanescentes

florestais da Mata Atlantica, Periodo 2000 a 2005. Available at: http://www.sosma.org.br.

Xavier AF, Bolzani BM, Jordao S. 2008. Unidades de conservacao da natureza no Estado de

Sao Paulo. 23-42, in: Rodrigues RR, Bononi VLR (Orgs.). Diretrizes para a Conservacao e

Restauracao da Biodiversidade no Estado de Sao Paulo. Sao Paulo, Instituto de Botanica.

Xavier-Santos S. 2003. Isolamento, identificagao e perfil enzimatico de fungos decompositores de

madeira da Estagao Ecoldgica do Noroeste Paulista —- Sao José do Rio Preto/Mirassol, SP. 222f.

PhD Thesis (Ciéncias Biolégicas, Microbiologia Aplicada), Universidade Estadual Paulista, Rio

Claro.

Young AM, Fechner NA, Ryvarden L. 2004. A preliminary checklist and introductory notes on the

macrofungi of Lamington National Park. Australasian Mycologist 23: 45-52.

MY COTAXON

http://dx.doi.org/10.5248/120.43

Volume 120, pp. 43-48 April-June 2012

Manoharachariella indica sp. nov. from the Western Ghats, India

KUNHIRAMAN C, RAJESHKUMAR' & SANJAY K. SINGH?

National Facility for Culture Collection of Fungi,

MACS’ Agharkar Research Institute, G.G. Agarkar Road, Pune, India

CORRESPONDENCE TO: ‘rajeshfungi@gmail.com & *singhsksingh@gmail.com

ABsTRACT — Manoharachariella indica is described as a new species from India, differing

from M. lignicola, the only other described species of the genus, by larger conidia, longer

conidiophores, and additional conidiophore septa.

KEY worpDs — anamorphic fungi, dematiaceous hyphomycete, Tamhini Ghats, Maharashtra

Introduction

The Western Ghats, one of the world’s most diverse ‘biodiversity hot

spots, is unique and diverse for its fungal wealth. Studies on microfungi and

exploration of fungal diversity from different microhabitats in the Western

Ghats are routinely conducted by the National Fungal Culture Collection of

India (NFCCI) (Rajeshkumar et al. 2010, 2011a, b; Singh et al. 2009, 2010).

During a September 2011 expedition to explore microfungal diversity in the

Mulshi and adjoining areas we discovered a dematiaceous hyphomycete on a

dead twig from the bamboo dominating the evergreen patch in the valley of the

Tamhini Ghats.

Bagyanarayana et al. (2009) recently established Manoharachariella for

M. lignicola Bagyan. et al. from litter in Andhra Pradesh, India. Monoblastic

integrated conidiogenous cells that produce solitary doliiform obpyriform

dictyoseptate apiculate conidia characterize this dematiaceous hyphomycete

genus. We describe a second species in this genus.

Materials and methods

ISOLATES AND MORPHOLOGY — Conidia were isolated directly from the surface of

a dead twig and observed under a Nikon binocular stereomicroscope (Model SMZ-

1500 with Digi-CAM, Japan). Single conidial cultures were established on 2% potato

dextrose agar plates (PDA; Crous et al. 2009). For morphotaxonomic studies and

photomicrographs, Zeiss (AXIO Imager 2, Germany) and Olympus (Model CX-41,

44 ... Rajeshkumar & Singh

Japan) microscopes were used. Conidia and conidiophores were mounted in lactic acid

cotton blue and measured using an ocular micrometer (and confirmed with software

available with the Zeiss microscope), with 30 observations per structure. Culture colony

characteristics were studied on two different media: 2% malt extract agar (MEA) and

PDA (Crous et al. 2009). Herbarium specimens are deposited in the Ajrekar Mycological

Herbarium (AMH), and the culture NFCCI 2630 was accessioned and preserved at

NFCCI; WDCM-932, Agharkar Research Institute, Pune, India.

Taxonomy

Manoharachariella indica Rajeshkumar & S.K. Singh, sp. nov. PLaTEs 1-3

MycoBank MB 563453

Differs from Manoharachariella lignicola by larger conidia, longer conidiophores, and

additional conidiophore septa.

Type: India, Maharashtra, Tamhini Ghats, on dead twigs, September 2011, K.C.

Rajeshkumar (Holotype, AMH 9438; ex-type culture NFCCI 2630).

EryMoOLoey: indica, referring to the country where this fungus is native.

Colonies effuse, thin, mid to dark brown or blackish brown, mycelium superficial

or semi immersed or immersed, hyphae pale to dark brown, smooth, 4-6 um.

Stroma none. Setae and hypopodia absent. Conidiophores macronematous,

mononematous, straight or flexuous, sparingly branched, arising laterally and

apically from the immersed mycelium, 5—15-septate, smooth, pale to dark brown

76-276 x 6-10.8 um. Conidiogenous cells monoblastic, integrated, terminal or

yw a con Haare —

Fic. 1. Manoharachariella indica (holotype): a. Habit. b. Sticky conidia in false chain. c. Terminal

and lateral conidiogenesis. d—f. Conidiophores and conidia in nature.

Manoharachariella indica sp. nov. (India) ... 45

SO ‘woz

Fig. 2. Manoharachariella indica (holotype): a. Conidial development. b-c. Apiculate conidia.

d,f-g,j-k. Variation in conidial size and shape. e. Conidia with conidiophores. h. Conidia under

phase contrast view. i. Sticky conidia in false chain.

46 ... Rajeshkumar & Singh

Fic. 3. Manoharachariella indica (holotype): a-b. Mature conidiophores. c. Young conidiophores.

d. Hyphal branching and initiation of conidiophores in nature. e. Conidiophores and attached

conidia. f. Conidiophores with young conidia. g. Foot cells of conidiophores. h. Mature conidia

attached with conidiophores, showing multi-polar germination. i. Branched conidiophore. j.

Simple conidiophore.

Manoharachariella indica sp. nov. (India) ... 47

intercalary, pale to mid brown, 4.4-8.6 x 4-7.6 um. Conidia solitary, dry or

sticky, acropleurogenous, simple, doliiform, obpyriform, ellipsoidal or irregular,

apiculate, apiculus rounded or obtuse, smooth, dictyoseptate, longitudinally

10-13-septate, tiered, pale to dark brown or blackish brown, apical and basal

tiers hyaline or subhyaline. 84-110(-120) x 31-44 um; young conidia hyaline

or subhyaline, 56 x 17 um.

Conidia in nature are dry and/or sticky and also form false chains of 2-3

conidia. Conidial germination was observed as being multi-polar in moist

chambers, but also after 12 hours of inoculation on PDA plates.

Colonies on PDA and MEA very slow growing, 1-2 mm diam after 14 days,

pale brown, velutinous; reverse pale brown.

TELEOMORPH: not observed.

Discussion

The monotypic Manoharachariella with its type species M. lignicola

somewhat resembles Septosporium Corda (especially S. rostratum M.B. Ellis,

1961) in having dictyoseptate beaked conidia but differs in the absence of

setae, unbranched conidiophores, and percurrent conidiogenous cells. Two

species of Xenosporium Penz. & Sacc. — X. africanum Piroz. (Deighton

& Pirozynski 1966) and X. boivinii S. Hughes (Hughes 1978)— resemble

Manoharachariella in conidial shape but differ in their monoblastic, acrogenous

or acropleurogenous, obpyriform, apiculate, smooth and dictyoseptate conidia

that are transversely longitudinally septate. In addition, secondary conidia are

absent in Manoharachariella but present in Xenosporium. Manoharachariella,

like Bioconiosporium Bat. & J.L. Bezerra (Ellis 1976), produces solitary

dictyoseptate conidia but the conidia are monoblastic with single apiculus

and lack the two large protuberances that are prominent in Bioconiosporium.

Conidia in both Monodictys S. Hughes (Ellis 1971) and Manoharachariella are

monoblastic, dictyoseptate, and acrogenous but those in Manoharachariella

are apiculate and tiered. Bagyanarayana et al. (2009) noted all these differences

when they established Manoharachariella.

Manoharachariella lignicola is differentiated from M. indica by its smaller

(42.5-50.5 x 25-32 um) 7-9 transversely septate conidia and smaller

conidiophores (< 35 um x 3-4.5 um) with fewer (1-3) septa.

Acknowledgements

We are indebted to Pedro W. Crous (Centraalbureau voor Schimmelcultures, Utrecht,

The Netherlands) and Uwe Braun (Martin-Luther-University, Halle, Germany) for

commenting on this manuscript. Thanks are also due to the Department of Science and

Technology (DST), Government of India, New Delhi for providing financial support for

setting up the National Facility for Culture Collection of Fungi (No. SP/SO/PS-55/2005)

at MACS’ Agharkar Research Institute, Pune, India, and the Director, MACS’ ARI for

providing facility.

48 ... Rajeshkumar & Singh

Literature cited

BagyanarayanaG, Rao NK, KunwarIK.2009. Manoharachariella,anew dematiaceoushyphomycetous

genus from India. Mycotaxon 109: 301-305. http://dx.doi.org/10.5248/109.301

Crous PW, Verkley GJM, Groenewald JZ, Samson RA (eds). 2009. Fungal Biodiversity. CBS

Laboratory Manual Series. Centraalbureau voor Schimmelcultures, Utrecht, Netherlands.

Deighton FC, Pirozynski KA. 1966. Microfungi II. Brooksia and Grallomyces; Acrogenotheca ornata

sp. nov.; the genus Xenosporium. Mycol. Pap. 105. 35 p.

Ellis MB. 1961. Dematiaceous hyphomycetes. III. Mycol. Pap. 79. 23 p.

Ellis MB. 1971. Dematiaceous hyphomycetes. CMI, Kew, UK. 608 p.

Ellis MB. 1976. More dematiaceous hyphomycetes. CMI, Kew, UK. 507 p.

Hughes SJ. 1978. New Zealand fungi. 25. Miscellaneous species. N. Z. J. Bot. 16(3): 311-370.

Rajeshkumar KC, Singh PN, Yadav LS, Swami SV, Singh SK. 2010. Chaetospermum setosum sp. nov.

from the Western Ghats, India. Mycotaxon 113: 397—404. http://dx.doi.org/10.5248/113.397

Rajeshkumar KC, Hepat RP, Gaikwad SB, Singh SK. 201 1a. Pilidiella crousii sp. nov. from northern

Western Ghats, India. Mycotaxon 115: 155-162. http://dx.doi.org/10.5248/115.155

Rajeshkumar KC, Sharma R, Hepat RP, Swami SV, Singh PN, Singh SK. 2011b. Morphology and

molecular studies on Pseudocercospora kamalii sp. nov. a foliar pathogen on Terminalia from

India. Mycotaxon 117: 227-237. http://dx.doi.org/10.5248/117.227

Singh SK, Singh PN, Yadav LS, Hepat RP. 2009. A new species of Gonatophragmium from Western

Ghats, India. Mycotaxon 110: 183-187. http://dx.doi.org/10.5248/110.183

Singh SK, Yadav LS, Singh PN, Sharma R, Rajeshkumar KC. 2010. Anewrecord of Gliocephalotrichum

(Hypocreales) from India. Mycotaxon 114: 163-169. http://dx.doi.org/10.5248/114.161

MY COTAXON

http://dx.doi.org/10.5248/120.49

Volume 120, pp. 49-58 April-June 2012

Russula jilinensis sp. nov. (Russulaceae) from northeast China

Guo-ji£ Li»?, SAt-Fer Li ?, XING-ZHONG Liu! & HUA-AN WEN ™

'State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

No3 'st West Beichen Road, Chaoyang District, Beijing 100101, China

*Graduate University of Chinese Academy of Sciences, Beijing 100049, China

* CORRESPONDENCE TO: wenha@im.ac.cn

ABSTRACT — Russula jilinensis (subg. Coccinula sect. Laetinae), is described from Changbai

Mountains, northeast China. The new species is distinguished by its bright red glabrous pileus

with a cinnamon tinged disc, slightly yellowish context, dark yellow to ocher spore print,

and pileipellis with septate pileocystidia. The morphological characteristics are illustrated in

detail and compared with those of similar species. Identification of R. jilinensis was supported

by the molecular phylogenetic analysis based on the ribosomal DNA internal transcribed

spacer regions (ITS).

Key worps —Russulales, taxonomy, morphology, Basidiomycota

Introduction

Northeast China, including Liaoning, Jilin, and Heilongjiang Provinces

and the eastern part of Inner Mongolia Autonomous Region, covers an area

of 1.236x10° km? (39°-53°30' N 115°-135°E) within the temperate to boreal

continental climate zones. Plant communities range from grassland (eastern

Inner Mongolia) to broadleaf forest (southern Liaoning), while the three main

mountain systems (Great Hinggan, Lesser Hinggan, Changbai) are mostly

covered by coniferous or mixed coniferous—broadleaf forests. The main trees

in northeast China are Pinus pumila, P koraiensis, Larix gmelinii, Betula

platyphylla, Abies nephrolepis, Picea jezoensis, and Quercus mongolica (Jiang et

al. 2003, Xu et al. 2008). In summer and early autumn, these ectomycorrhizal

commensal plants facilitate the fruiting of ectomycorrhizal basidiomycetes,

among which the members of Russula Pers. are very common.

Russula is widely distributed from western Europe to North America in the

northern hemisphere (Romagnesi 1967, Singer 1986, Sarnari 1998, Miller &

Buyck 2002, Bau et al. 2008). Although some new species and varieties have

been reported from southern and southwestern China (Singer 1935, Chiu 1945,

Ying 1983, 1989, Bi & Li 1986, Wen & Ying 2001, Wang et al. 2009), Russula

50 ... Li&al.

species have not been studied systematically elsewhere in China, including the

north. Because of its taxonomic difficulty and the insufficiency of intensive

taxonomic studies in China (Wang et al. 2009), many Russula species reported

from northeastern China are identified using European or American names

and lack voucher support (e.g., Teng 1996, Xie et al. 1986, Li & Bau 2003, Bau

2004). As now in North America (Adamcik & Buyck 2010, 2011; Buyck &

Hofstetter 2011), the lack of endemic systematic studies has probably led to

incorrect estimates of the extent of Russula diversity in northeastern China,.

Although many wood-inhabiting fungi were recently reported from

northeastern China (Dai & Penttila 2006, Yuan et al. 2006, Wei & Dai 2007, Dai

et al. 2008, Xiong et al. 2008, Dai 2010), only a few Russula species have been

reported from this area (Song et al. 2007). In addition, many virgin and natural

forests in this area have not yet been intensively investigated. During a survey

on Russulales in the Changbai Mountains, one interesting Russula was found,

which is described here as a new species.

Materials & methods

Specimens were collected from the Changbai Mountains, Antu County of Jilin

Province, from 2008 to 2010. Macromorphological characteristics of the fresh fruiting

bodies were recorded in field notes. Color names and codes follow Ridgway (1912)

and spore print colors were categorized according to Romagnesi (1967). Collections

were oven-dried at 50-60 °C until their water content was <15% prior to microscopic

examination. Solutions of 10% FeSO, and sulfovanillin (SV) were used for testing the

chemical reaction of dried specimens. Specimens for microscopic examinations were

hand-sectioned and rehydrated in a 5% KOH solution prior to observation under a

Nikon Eclipse 80i microscope. Basidiospores (examined in Melzer’s reagent), basidia,

pleurocystidia, cheilocystidia and elements of the pileipellis and stipitipellis were

measured, with at least 20 elements measured for each character. The abbreviation

(n/m/p) = n spores from m basidiospores of p specimens. Basidiospore dimensions

are shown as (a—) b-c (-d), with the range b-c including 90% of the measured values

and the extreme values shown in parentheses. The spore quotient (Q) = spore length

divided by spore width. Q (in bold) = average quotient value + standard deviation. Spore

ornamentations and apiculi were not included in measurements. Further explanations

of basidiospore data follow Yang (2000). Scanning electron images were also captured

with an FEI Quanta 200 electron microscope. Abbreviations of herbarium names follow

Holmgren et al. (1990). Specimens cited were deposited in the Mycological Herbarium,

Institute of Microbiology, Chinese Academy of Sciences (HMAS).

For phylogenetic studies, genomic DNA was extracted from dried fruiting bodies

briefly crushed with quartz sand in liquid nitrogen. DNA template for amplification

was prepared using a BioTeke plant DNA rapid extraction kit. PCR was conducted with

an Eastwin Dragon Black Vajra thermo cycler using ITS1 (5’-GTAGGTGAACCTGCGG-

3°) and ITS4 (5’-rccTrccGCTTATTGATATGC-3’) primers (Shanghai Sangon, Beijing

TransGen Biotech), EasyTaq DNA polymerase, dNTPs, and PCR buffer to amplify

the ITS1+5.8S+ITS2 region. Amplification of a 50 uL mixture — 35.5 ul ddH,O, 5 ul

Russula jilinensis sp. nov. (China) ... 51

PCR buffer, 4 ul dNTP (2.5 mmol/L), 1 ul DNA template, 2 ul (10 umol/L) each of

two primers, 0.5 ul (Su/ul) Taq DNA polymerase — followed a protocol of 5 min at

94°C (initial denaturation), 35 cycles of 1-min at 94°C (denaturation) + 1 min at 56°C

(annealing) + 1 min at 72°C (extension), and a final 10-min extension at 72°C. The PCR

products were electrophoresed in ethidium bromide-stained agarose gel and sequenced

by SinoGenoMax Co. Ltd, using an ABI 3730XL Analyzer and ABI BigDye 3.1 Cycle

Sequencing Kit. The sequences were deposited in GenBank.

Sequences were aligned and edited in ClustalX (Thompson et al. 1997) and Bioedit

(Hall 1999). Phylogenetic analysis was performed with MEGA version 4.1 Beta (Tamura

et al. 2007), and Minimum Parsimony trees were constructed with the test of inferred

phylogeny a bootstrap of 1000 replications and a random seed (Eck & Dayhoff 1966,

Rzhetsky & Nei 1992). The MP tree was obtained using the Close-Neighbor-Interchange

algorithm with search level in which the initial trees were obtained with the random

addition of sequences (10 replicates). The tree was drawn to scale, with branch lengths

calculated using the average pathway method and in the units of the number of changes

over the whole sequence (Nei & Kumar 2000). All positions containing gaps and missing

data were eliminated from the data set (Complete Deletion option). The bootstrap

consensus tree inferred from 1000 replicates is taken to represent the evolutionary

history of the taxa analyzed. Branches corresponding to partitions reproduced in less

than 50% bootstrap replicates are collapsed. Of the 424 positions in the final dataset,

253 are parsimony informative. The ITS1-5.8S-ITS2 sequences of Albatrellus ovinus

(Schaeff.) Kotl. & Pouzar and Bondarzewia mesenterica (Schaeft.) Kreisel (as B. montana)

were used as outgroups (Wang et al. 2009).

Taxonomy

Russula jilinensis GJ. Li & H.A. Wen, sp. nov. Fics 1-4

MycoBank MB 563682

Differs from Russula integra by shorter basidia and pleurocystidia and smaller spores.

EryMo_Locy: named after the type locality.

Type: CHINA, Jilin Province, Antu County, Erdaobaihe, Changbai Mountains, 42°24'N

128°04'E, alt. 761 m, on ground in mixed coniferous—broadleaf forest dominated by

Pinus koraiensis and Picea, 1 August 2008, M.X. Zhou & Y. Gafforov 08004 (Holotype,

HMAS194253, GenBank GU966632).

PiLEus 6.1-7.3 cm broad, first hemispheric, convex, then more or less convex

to plano-convex, depressed weakly in the center when mature, colors Jasper

Red to Coral Red (x1113'—5’) when juvenile, Pompeian Red to Dragon’s-blood

Red (x1113'/i-5'1) with age, Nopal Red to Brazil Red (13i-5i) when dried; center

Vinaceous-Cinnamon to Pinkish Cinnamon (xx1x13''b—15"'b) when juvenile,

Orange-Cinnamon to Cinnamon (xx1x13'’-15') with age, Mikado Brown

(xx1x13"i) when dried, Nopal Red (13i) in SV, color unchanged in FeSO; viscid

when wet, glabrous, smooth, non-pruinose when dried, pellis peeling 1/2 to

the disc; margin obtuse, not or very slightly striate when old, never cracked.

CONTEXT 1.0-1.5 cm thick from stipe top to pileus center, brittle, white (Lu),

Naples Yellow (xv119'd) when old or dried, color unchanged when bruised;

52 ... Li & al.

Fic.1: Russula jilinensis (Holotype HMAS 194253).

Fic.2: Russula jilinensis (HMAS 262395). Scale bar = 1 cm.

Russula jilinensis sp. nov. (China) ... 53

4/26/2010 WD Mag HV Det Spot

2:42:07 PM.9.6 mm 5000x 12.5 kV ETD 3.0

Fic. 3: Russula jilinensis (Holotype HMAS194253).

Scanning electron microscope (SEM) photo. Basidiospores. Scale bar = 10 um.

taste mild; odor none or indistinct. LAMELLAE 4-7 mm broad, 9-12/cm at

the edge, no lamellulae, decurrent to more or less adnate, not forked, fragile,

Pale Yellow-Orange (11115f) first, Pale Orange-Yellow (11117f) when mature or

dry, color unchanged in FeSO, STIPE clavate, enlarged towards the base, 9.2 x

1.8-2.1 cm, smooth, solid but irregularly hollowing when old, white (L111) when

juvenile, becoming Baryta Yellow, Wax Yellow, or Primuline Yellow (1v21f,

Xv121’, xv119', respectively) towards the base when old or dry, Brazil Red (15i)

in SV, no color change in FeSO,. SPORE PRINT dark yellow, ocher, IIc.

BASIDIOSPORES (80/3/3) (6—)7-8(-8.5) x (5.5-)6-7 um, average 7.5 x 6.3

um, [Q= (1.02) 1.06-1.28 (1.31), Q= 1.18 + 0.06], subglobose to ellipsoid,

without oil droplet, plage indistinct and amyloid; ornamentation amyloid,

mostly isolated, rarely linked by fine lines and not forming a mesh, warts

conic to cylindrical, <0.4-1.0 um tall. Basrp1a (33-)38-44 x 11-13(-14) um,

clavate to subclavate, broadly tapered towards the base, 4-spored, sometimes

2-spored, hyaline in KOH, occasionally containing a large droplet; mature

basidia projecting <15-10 um above the subhymenium; sterigmata 3-5 um.

PLEUROCYSTIDIA (50-)53-56(-63) x (8-)9-10(-11) um, rare, emergent,

fusiform, clavate to subclavate, often with a subacute tip, sometimes witha frayed

small appendage and dense crystal inclusions, projecting <35 um above the

subhymenium, color unchanged in FeSO, dark grey in SV reaction dark grey.

54 ... Li & al.

Fic. 4: Russula jilinensis (Holotype HMAS 194253).

A. Pleurocystidia; B. basidia; C. terminal elements of pileipellis; D. pileocystidia.

Scale bars = 10 um.

CHEILOCYSTIDIA mostly similar to pleurocystidia. PILEIPELLIS two-layered,

100-125 um thick, gelatinous; epicutis an ixotrichoderm, epicuticular hyphae

3-6 um broad, septate, mostly unbranched, erect to suberect, tangled, hyaline

Russula jilinensis sp. nov. (China) ... 55

in KOH, terminal cells thin-walled, cylindrical, with obtuse, undifferentiated,

often tapered ends; pileocystidia 48-64 x 5-7 um, dispersed, numerous in

surface, scattered, clavate, subclavate to cylindrical, with 0-2 septa, contents

crystalline, granulate, slightly turning grey in SV; subcutis hyphae recumbent,

filamentous, interwoven, branched, septate, 2.5-5 um diam., hyaline to pale

yellowish brown in KOH; trama composed of subspherical to angular 13-24

uum diam sphaerocysts and connective hyphae. STIPITIPELLIS composed mostly

of interwoven branched elongated 3-6 um diam hyaline hyphae with inflated

cells; caulocystidia absent. Clamps or laticiferous hyphae absent in all tissues.

Eco.oey: Solitary on soil in mixed coniferous—broadleaf forests, July-

August. Known only from the Changbai Mountains. Edibility: unknown.

ADDITIONAL SPECIMENS EXAMINED: CHINA, JILIN PROVINCE, ANTU COUNTY, Heping

Forest Farm, 43°07'N 128°54’E, alt. 1014 m, in coniferous forest dominated by Pinus,

22 July 2010, L.D. Guo, X. Sun, G.J. Li & L.J. Xie 20100054 (HMAS262395, GenBank

HQ693525); Changbai Mountains Forest Ecological System Research Center, 43°23'N

128°05’E, alt. 811 m, in mixed coniferous—broadleaf forest, 25 July 2010, X. Sun & GJ. Li

20100410 (HMAS262364, GenBank HQ693524).

Discussion

Russula jilinensis is macroscopically characterized by its bright red pileus,

light-yellow lamellae, and a white stipe without red or purple tinge. It shows

superficial affinities with subg. Coccinula sect. Laetinae Romagn., but its

relatively shorter basidia and larger cystidia are more characteristic of subg.

Polychromidia sect. Integrinae Maire (e.g., R. integra f. gigas Romagn.). The R.

jilinensis ITS1-5.8S-ITS2 sequences cluster with R. curtipes FH. Moller & Jul.

Schaff. (Fic. 5), and a blasting with that of R. curtipes (AY061668, Miller &

Buyck 2002) through NCBI databank shows a 93% max identification of 97%

coverage, confirming that R. jilinensis is related to R. integra/curtipes group in

Russula sect. Integrinae.

Russula integra (L.) Fr., which is common in Europe, has been reported

many times from China and neighboring East Siberia (Teng 1964, Romagnesi

1967, Tai 1979, Xie et al. 1986, Ying et al. 1987, Knudsen & Stordal 1992, Mao &

Zhuang 1997, Mao 2000, Bau et al. 2008). However, few specimens of R. integra

have been recorded from northeast China. Russula integra and R. jilinensis

are similar in spore ornamentation and cystidial shape, and both species are

associated with Pinus and Picea in the subalpine zone (Romagnesi 1967, 1985).

However, our microscopic observation followed by intensive comparison of

descriptions of R. integra by Moller & Schaffer (1935) and Romagnesi (1967)

indicates that R. jilinensis is a different species. The latter has shorter basidia

and pleurocystidia and smaller spores than those of R. integra (with spores

8.2-10 x 7-9.2 um, basidia 45-65 x 11-14.5 um, cystidia 82-120 x 8.5-13 um,

in Romagnesi 1967).

56 ... Li&al.

R, jilinensis HQ693525 (HMA S 262395)

— R. jilinensis GU966632 (HMA S 194253)

R, jilinensis HQ693524 (HMA S 262364)

93 p R.aff.curtipes EU248593

R.curtipes AY061668

100 p— R.laricina AY061685

R.cessans AY061730

R.caerulea AY061661

96 R.roseipes AY061716

R.tarci AY061720

R.paludosa AY061703

99 p R.integra AY061683

R.integriformis AY 061684

61 R.xerampelina AY061734

10 R.pascua AY061705

R.nitida AY061696

R.sphagnophila AY061719

R.nauseosa AY061733

98 yR.cuprea AY061667

R.adulterina AY061651

98 R.clariana AY 061664

R.violacea AY061725

99 pR.lepida AY061686

R.aurantiaca AY061658

54 R.consobrina AY061666

R.gracillima AY061678

R.persicina AY061707

R.queletii AYO61711

99 p= R.drimeia AY061672

R.sanguinea AY061718

96 p= R. bicolor GU966633

st bad R. bicolor FJ845435

R.raoultii AF418621

R.aquosa AY061657

oe R.raoultii AY061712

9g [7 R-nana AY061694

TA, R.emetica AY061673

R.betularum AY061729

R.romellii AY0617 14

R.aurea AY061659

100 A.ovinus AY 293165

B.montana AF042646

Fic. 5. One of 13 ITS1-5.8S-ITS2-based phylogenies of Russula jilinensis and related Russula species,

using the Maximum Parsimony method. The percentages of replicate trees in which the associated

taxa clustered together in the bootstrap test (1000 replicates) are shown above the branches.

Another phylogenetically related species is R. curtipes, which shares with

R. jilinensis the short basidia and firm and yellowing context of the R. integra-

curtipes group, but they distinctly differ from each other in pileus color and

habitat: R. curtipes has a dark wine-red, haematite red, to light russet vinaceous

pileus and associates with beech in western Europe with a possible preference

for limestone (Romagnesi 1967), while R. jilinensis has a brighter red pileus and

grows in mixed forests dominated by conifers in efflorescent basalt soils. The

reddish pileus of R. jilinensis distinguishes it from most of the typical violet-

or purple-capped members of R. sect. Integrinae such as R. romellii Maire and

R. carpini R. Girard & Heinem. Another species in R. sect. Integrinae with a

dark-red pileus, R. rubroalba (Singer) Romagn., shares a more or less similar

pileus color and spore ornamentation but is associated with a deciduous forest

and has much longer cystidia than those of R. jilinensis.

Acknowledgments

The authors are grateful to Dr. Y.C. Dai (Institute of Applied Ecology, Chinese

Academy of Sciences) and Dr. Kanad Das (Botanical Survey of India, Sikkim Himalayan

Russula jilinensis sp. nov. (China) ... 57

Regional Centre) for reviewing the manuscript; to Ms. H.M. Li, Ms. A.R. Yan and Dr.

T.Z. Wei (HMAS) for providing the herbarium specimens; to Ms. X.E. Zhu for inking

in line drawings, to Dr. Z.L. Yang (Kunming Institute of Botany, CAS), B.A. Jaffee

(University of California Davis), Dr. S. Pennycook (Landcare Research), Dr. L. Norvell

(Pacific Northwest Mycology Service), and Dr. J.Y. Zhuang for improving the manuscript;

to Dr. L.D. Guo, Dr. X. Sun, Dr. Z. Yong, Mr. M.X. Zhou, Dr. Y. Gafforov, Ms. B. Wu,

Mr. C. Gao, Ms. G.Y. Li, Ms. X.L. Yang, Ms. J.Z. Li, Ms. Q. Qian, Mr. Z.M. Li, Ms. S.F.

Yue, Mr. D. Zhao, Ms. J.X. Yang (Institute of Microbiology, CAS), Ms. L.J. Xie, Dr. C.Y.

Sun and Ms. J. Zhou (National Institute of Occupational Health and Poison Control,

China CDC) for assistance in collecting specimens; to Dr. C.L. Li and Ms. J.N. Liang

(Institute of Microbiology, CAS) in SEM photographing. This project was supported by

the National Natural Science Foundation of China (No. 30770013).

Literature cited

Adamcik S, Buyck B. 2010. Reinstatement of Russula levyana as a good and distinct species of

Russula section Xerampelinae in America. Cryptogamie Mycologie 31(2): 119-135.

Adamcik S, Buyck B. 2011. The species of Russula subsection Xerampelinae described by C.H. Peck

and Miss G.S. Burlingham. Cryptogamie Mycologie 32(1): 63-81.

Bau T. 2004. Mycodiversity in Daqinggou Nature Reserve of Inner Mongolia. Inner Mongolia

Education Press, Hohhot.

Bau T, Li Y, Irina AG, Eugenia MB, Wasiliy AS. 2008. Common wild edible mushroom resource of

Russia. Edible Fungi of China 27(3): 9-13.

Bi ZS, Li TH. 1986. A preliminary note on Russula species from Guangdong, with a new species

and a new variety. Guihaia 6(3): 193-199.

Buyck B, Hofstetter V. 2011. The contribution of tef-1 sequences to species delimitation in the

Cantharellus cibarius complex in the southeastern USA. Fungal Diversity 49:35-46.

http://dx.doi.org/10.1007/s13225-011-0095-z

Chiu WE. 1945. The Russulaceae of Yunnan. Lloydia 8: 31-59.

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 131-343.

http://dx.doi.org/10.1007/s13225-010-0066-9

Dai YC, Penttila R. 2006. Polypore diversity of Fenglin Nature Reserve, northeastern China.

Annales Botanici Fennici 43: 81-96.

Dai YC, Cui BK, Tao WQ. 2008. Phellinus mori sp. nov. (Basidiomycota, Hymenochaetales) from

northern China. Mycotaxon 105: 53-58.

Eck RV, Dayhoff MO. 1966. Atlas of protein sequence and structure. National Biomedical Research

Foundation, Silver Springs, Maryland.

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NTT. Nucleic Acids Symposium Series 41: 95-98.

Holmgren P, Holmgren N, Barnett LC. 1990. Index Herbariorum part I. The Herbaria of the World.

Regnum Veg. 120: 1-693.

Jiang P, Zhao G, Ji Y, Cui GF, Deng HB. 2003. Structure of communities in the northern slope of

Changbai Mountain and its variation along elevation gradients. Chinese Journal of Ecology

22(6): 28-32.

Knudsen H, Stordal J. 1992. Russula. 374-400, in: L Hansen, L Knudsen H (eds). Nordic

macromycetes, vol. 2. Copenhagen.

Li Y, Bau T. 2003. Mushrooms of Changbai Mountains, China. Science Press, Beijing.

Mao XL. 2000. Macrofungi of China. Henan Science and Technology Press, Zhengzhou.

Mao XL, Zhuang JY. 1997. Fungi of Qinling. Chinese Agricultural Science and Technology Press,

Beijing.

58 ... Li&al.

Miller SL, Buyck B. 2002. Molecular phylogeny of the genus Russula in Europe with a comparison

of modern infrageneric classification. Mycol. Res. 106(3): 259-276.

http://dx.doi.org/10.1017/S0953756202005610

Moller FH, Schaffer J. 1935. Le complexe Russula alutacea. Bull. Soc. Mycol. France 51: 105-112.

Nei M, Kumar S. 2000. Molecular evolution and phylogenetics. Oxford University Press, New

York.

Ridgway R. 1912. Color standards and color nomenclature. Robert Ridgway, Washington.

Romagnesi H. 1967. Les Russules d’ Europe et d’ Afrique du Nord. Bordas, Paris.

Romagnesi H. 1985. Les Russules d’ Europe et d’ Afrique du Nord. Reprint with supplement.

J. Cramer, Lehre.

Rzhetsky A, Nei M. 1992. A simple method for estimating and testing minimum evolution trees.

Molecular Biology and Evolution 9:945-967.

Sarnari M. 1998. Monografia illustrata del genere Russula in Europa. Tomo Primo. AMB, Centro

Studi Micologici, Trento.

Singer R. 1935. Supplemente zu meiner Monographie der Gattung Russula. Ann. Mycol. 33:

291 ~ doze

Singer R. 1986. The Agaricales in modern taxonomy. 4th ed. Koeltz Scientific Books, Koenigstein.

Song, B, Li TH, Wu XL, Li JJ, Shen YH, Lin QY. 2007. Known species of Russula from China and

their distribution. Journal of Fungal Research 5(1): 21-42.

Tai FL. 1979. Sylloge fungorum Sinicorum. Science Press, Beijing.

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis

(MEGA) software version 4.0. Molecular Biology and Revolution 24: 1596-1599.

http://dx.doi.org/10.1093/molbev/msm092

Teng SC. 1964. Fungi of China. Science Press, Beijing.

Teng SC. 1996. Fungi of China. Mycotaxon Ltd, New York.

Thompson JD, Gibson TJ, Plewnlak F, Jianmougin F, Higgins DG. 1997. The Clustal X windows

interfaces: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 24: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Wang XH, Yang ZL, Li YC, Knudsen H, Liu PG. 2009. Russula griseocarnosa sp. nov. (Russulaceae,

Russulales), a commercially important edible mushroom in tropical China: mycorrhiza,

phylogenetic position, and taxonomy. Nova Hedwigia 88: 269-282.

http://dx.doi.org/10.1127/0029-5035/2009/0088-0269

Wei YL, Dai YC. 2007. Changbai wood-rotting fungi 15. Henningsomyces leptus sp. nov. Mycotaxon

101: 261-264.

Wen HA, Ying JZ. 2001. Studies on the genus Russula from China II. Two new taxa from Yunnan

and Guizhou. Mycosystema 20(2):153-155.

Xie ZX, Wang Y, Wang B. 1986. Records and maps of the mushrooms in Changbai Mountains. Jilin

Science and Technology Press, Changchun.

Xiong HX, Dai YC, Cui BK. 2008. Perenniporia minor (Basidiomycota, Polyporales), a new polypore

from China. Mycotaxon 105: 59-64.

Xu WD, He XY, Chen W, Liu CF, Zhao GL, Zhou Y. 2008. Ecological division of vegetations in

northeast China. Chinese Journal of Ecology 27(11): 1853-1860.

Yang ZL. 2000. Type studies on agarics described by N. Patouillard (and his co-authors) from

Vietnam. Mycotaxon 75: 431-476.

Ying JZ. 1983. A study on Russula viridi-rubrolimbata sp. nov. and its related species of subsection

Virescentinae. Acta Mycol. Sinica 2(1): 34-37.

Ying JZ. 1989. Studies on the genus Russula Pers. from China 1. New taxa of Russula form China.

Acta. Mycol. Sinica 8(3): 205-209.

Yuan HS, Li J, Huang MY, Dai YC. 2006. Antrodiella stipitata sp. nov. from Heilongjiang Province,

northeast China, and a critical checklist of polypores from the area. Cryptogamie Mycologie

22d 29;

MY COTAXON

http://dx.doi.org/10.5248/120.59

Volume 120, pp. 59-66 April-June 2012

Acrogenospora hainanensis sp. nov. and

new records of microfungi from southern China

JIAN Ma, Li-Guo Ma, Y1-DONG ZHANG, JI-WEN XIA & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT —Acrogenospora hainanensis is described and illustrated from specimens

collected on dead branches in Hainan Province, China. The fungus is characterized by

terminal, monoblastic, percurrently extending conidiogenous cells with solitary, spherical or

subspherical, non-septate and relatively small conidia. Acrogenospora gigantospora, Nakataea

fusispora, and Pseudospiropes costaricensis are newly recorded from China.

KEY worps — anamorphic fungi, taxonomy

Introduction

The forests of southern China have a rich mycota, and mycological

investigations dealing with many new species have been published recently

(Dai & Cui 2006, Cui & Dai 2008, Cui et al. 2009a,b, 2011, Zhang et al. 2009,

2011, Ma et al. 2010). During ongoing mycological surveys in the forests of

Hainan and Guangdong provinces, four interesting anamorphic fungi clearly

related to Acrogenospora, Nakataea, and Pseudospiropes were collected on dead

branches. One of these is an undescribed species of Acrogenospora, while the

other three species are new records for China.

Acrogenospora hainanensis Jian Ma & X.G. Zhang, sp. nov. Fic. 1

MycoBank MB 564117

Differs from Acrogenospora verrucispora in smaller smooth conidia and from A.

sphaerocephala in smaller conidia and more narrow conidial scars.

Type: China, Hainan Province: Tunchang, on dead branches of unidentified broad-

leaved tree, 12 April 2011, J. Ma (Holotype HSAUP H5509; isotype HMAS 146167).

EryMo_oey: refers to the province where the type was found.

Colonies on the natural substratum effuse, black, glistening, hairy. Mycelium

mostly immersed, composed of branched, septate, subhyaline to pale brown,

60 ... Ma & al.

D E

10m

Fic. 1. Acrogenospora hainanensis (holotype). A-C. Conidiophores, conidiogenous cells

with conidia. Note that in C the mature conidium is pushed sidewise following successive

percurrent extension of the conidiogenous cells. D. Conidiophores. E. Conidia.

uioT

ungoT

smooth-walled hyphae. Conidiophores macronematous, mononematous,

single, unbranched, erect, straight or slightly flexuous, septate, smooth,

brown to dark brown, paler towards the apex, 60-80 um long, 2-3.5 um thick.

Conidiogenous cells monoblastic, integrated, terminal, cylindrical, smooth,

pale brown, proliferating percurrently. Conidial secession schizolytic. Conidia

solitary, acrogenous, dry, spherical or subspherical, smooth, brown, non-

septate, 7.5-9.5 x 7-8.5 um, with a truncate base 1.5-2 um wide.

COMMENTS — Acrogenospora, with Monotospora sphaerocephala Berk. &

Broome [= A. sphaerocephala (Berk. & Broome) M.B. Ellis] as the type

species, is characterized by integrated, terminal, monoblastic, percurrently

extending conidiogenous cells on distinct, unbranched conidiophores, and

acrogenous, solitary, non-septate conidia with schizolytic conidial secession.

These characters separate the genus from Domingoella Petr. & Cif. (Petrak

& Ciferri 1932), Brachysporiella Bat. (Batista 1952), and Endophragmiella B.

Sutton (Sutton 1973). Ten species are currently included in Acrogenospora, all

discovered on dead branches, rotten wood, or submerged bamboo (Ellis 1971,

1972, Hughes 1978, Goh et al. 1998, Zhu et al. 2005, Hu et al. 2010). The species

are distinguished primarily on conidial shape, size, and ornamentation (Hughes

1978, Goh et al. 1998, Zhu et al. 2005).

Acrogenospora hainanensis sp. nov. (China) ... 61

Of the known species, Acrogenospora hainanensis is most similar to

A. sphaerocephala (Ellis 1971) and A. verrucispora Hong Zhu et al. (Zhu et al.

2005) in the production of spherical or subspherical conidia. However, the new

species differs from A. verrucispora (verrucose conidia 15-33 x 14-33 um) in

its smaller smooth-walled conidia and from A. sphaerocephala (19-21.5 um

diam., conidial scars 5-7 um wide) in its smaller conidia and narrower conidial

scars.

Acrogenospora gigantospora S. Hughes, New Zealand J. Bot. 16: 314, 1978. Fic. 2

Colonies on the natural substratum effuse, black, glistening, hairy. Mycelium

mostly immersed, composed of branched, septate, pale brown, smooth-walled

hyphae. Conidiophores distinct, single or in groups of 2-4, unbranched, erect,

straight or slightly flexuous, septate, smooth, brown to dark brown, up to 265 um

long, 5.5-8.5 um thick. Conidiogenous cells monoblastic, integrated, terminal,

cylindrical, smooth, pale brown, with multiple percurrent extensions. Conidial

secession schizolytic. Conidia solitary, acrogenous, dry, broadly obovoid to

Fic. 2. Acrogenospora gigantospora. A-C. Conidiophores, conidiogenous cells with

conidia. Note that in A and B the mature conidium is pushed sidewise following successive

percurrent extension of the conidiogenous cells. D. Conidia.

A B C

ind

—

62 ... Ma & al.

subspherical, smooth, dark brown to black, non-septate, 37-42 x 24-30 um,

with a truncate base 5-7.5 um wide.

SPECIMEN EXAMINED: CHINA, GUANGDONG PROVINCE: Chebaling National Nature

Reserve, on dead branches of unidentified broad-leaved tree, 19 Oct 2010, J. Ma (HSAUP

H5402).

COMMENTS - Acrogenospora gigantospora is reported for the first time from

China. Except for its shorter narrower conidiophores, our collection fits well

with the original description of A. gigantospora (Hughes 1978; conidiophores

< 700 x 9-14.5 um). Acrogenospora gigantospora appears to be most closely

related to A. ovalis Goh et al. (Goh et al. 1998) in conidial shape but differs

in having larger conidia and a wider conidial scar. In addition, mature

A. gigantospora conidia are brown to dark brown while those of A. ovalis are

mid orange-brown.

Nakataea fusispora (Matsush.) Matsush., Icon. Microfung. Matsush. Lect.: 100,

1975. FIG. 3

= Vakrabeeja fusispora Matsush., Microfungi Solomon Is. Papua-New Guin.: 66. 1971.

= Pyricularia fusispora (Matsush.) Zucconi, Onofri & Persiani, Micol. Ital. 13(2): 9. 1984.

Colonies on the natural substratum effuse, dark brown, hairy. Mycelium

partly superficial, partly immersed in the substratum, composed of branched,

septate, smooth-walled hyphae. Conidiophores conspicuous, mononematous,

unbranched, straight or flexuous, smooth, septate, brown, paler towards the apex,

64-95 um long, 3.5-4.5 um thick. Conidiogenous cells polyblastic, integrated,

terminal becoming intercalary, sympodially proliferating, denticulate, thin-

walled; denticles subcylindrical, flat and open at the apex, septate at the base.

Conidia solitary, dry, apical and lateral, verruculose, fusiform, acute at the apex,

with a denticle at the base, 3-septate, the cell at each end hyaline or very pale

brown, central cells pale to mid brown; conidia 22-34 x 6-7.5 um.

SPECIMEN EXAMINED: CHINA, HAINAN PROVINCE: tropical forest of Bawangling, on

dead branches of unidentified broad-leaved tree, 11 Dec 2009, J. Ma (HSAUP H5125).

ComMENtTs — Nakataea, established by Hara (1939) with Helminthosporium

sigmoideum Cavara [= N. sigmoidea (Cavara) Hara] as the type species, is

characterized by solitary, apical and lateral conidia seceding rhexolytically

from polyblastic, integrated, terminal becoming intercalary, sympodially

extending conidiogenous cells. The conidia are falcate, sigmoid, fusiform

or obclavate, almost always 3-euseptate, versicolored, end cells paler than

the central cells, with a distinct basal frill (Ellis 1971). Vakrabeeja Subram.

(Subramanian 1956) is based on the same type species as Nakataea and therefore

a superfluous homotypic synonym. Kirk (1983) suggested that Nakataea

should be synonymised with Pyricularia Sacc., because their type species were

morphologically similar and both had Magnaporthe teleomorphs. ITS rRNA

Acrogenospora hainanensis sp. nov. (China) ... 63

D, E

UINQZ

Fig. 3. Nakataea fusispora. A-C. Conidiophores, conidiogenous cells with denticles and

conidia. D. Conidiophore and conidiogenous cell. E. Conidia.

sequence phylogenetic analyses (Bussaban et al. 2005) suggest that N. fusispora

is distinct from Pyricularia species and should be maintained in Nakataea.

Nakataea fusispora is most similar to N. sigmoidea (Hara 1939) in conidial

shape but can be separated from N. sigmoidea (conidia 40-83 x 11-14

um, smooth) by its smaller conidia with verruculose walls. The conidia of

our collection are somewhat wider than cited in the original description

by Matsushima (1971; 4.8-6 um). The conidial length ranges of our and

Matsushima’s material overlap, and other morphological features of our fungus

satisfy the original description of N. fusispora. This fungus is reported for the

first time from China.

Pseudospiropes costaricensis (E.F. Morris) de Hoog & Arx, Kavaka 1:59, 1974

19737] FIG. 4

= Pleurophragmium costaricensis E.F. Morris, Mycologia 64: 893. 1972.

Colonies on natural substratum effuse, brown, hairy. Mycelium partly

superficial, partly immersed in the substratum, composed of branched,

septate, smooth-walled hyphae. Conidiophores distinct, single or in groups,

erect, unbranched, straight or flexuous, smooth, septate, brown to dark brown,

paler toward the apex, 160-290 um long, 6-9 um wide. Conidiogenous cells

holoblastic, polyblastic, indeterminate, terminal becoming intercalary, pale

brown to brown, integrated, always with holoblastic sympodial extensions.

64 ... Ma & al.

Fic. 4. Pseudospiropes costaricensis. A-B. Conidiophores and conidiogenous cells with

conidia. C. Conidiophores and conidiogenous cells. D. Conidia.

Conidiogenous loci enlarged, thickened, protuberant and black, lenticular.

Conidial secession schizolytic. Conidia apical and lateral, solitary, dry, simple,

obclavate or fusiform, pale brown or brown, smooth, 8-11-distoseptate, 40-53

x 11-14 um, apical cell with a hyaline vesicle, basal cell with a truncate base,

2-3.5 um.

SPECIMEN EXAMINED: CHINA, GUANGDONG PROVINCE: Chebaling National Nature

Reserve, on dead branches of unidentified broad-leaved tree, 19 Oct 2010, J. Ma (HSAUP

H5419).

CoMMENTS - Since the establishment of Pseudospiropes M.B. Ellis, based on

Helminthosporium nodosum Wallr. [= P. nodosus (Wallr.) M.B. Ellis], there have

been numerous additions to the genus. However, Sinclair et al. (1997) pointed

out that Pseudospiropes is heterogeneous. Castafieda et al. (2001) reviewed the

genus and proposed three new segregate anamorphic genera —Minimelanolocus

R.F. Castafieda & Heredia, Nigrolentilocus R.F.Castafieda & Heredia, and

Matsushimiella R.F.Castafieda & Heredia— based on the morphology of

conidiogenous loci, conidial septation, and conidial secession. As presently

constituted, Pseudospiropes is characterized by polyblastic, integrated, terminal

becoming intercalary, sympodial, cicatrized conidiogenous cells bearing

broadly enlarged, thickened, often dark and protuberant scars, and solitary,

apical and lateral, distoseptate conidia with schizolytic conidial secession (Ellis

1971, Castaneda et al. 2001). Currently, Pseudospiropes contains 13 valid species

(Ma et al. 2011).

Acrogenospora hainanensis sp. nov. (China) ... 65

Pseudospiropes costaricensis was originally assigned to Pleurophragmium

Costantin and subsequently transferred to Pseudospiropes by de Hoog & Arx

(1974) due to its cicatrized conidiogenous cells with large, often dark and

prominent scars. The conidia of Pseudospiropes costaricensis are morphologically

similar to those of P. hachijoensis Matsush. (Matsushima 1975), P josserandii

(Bertault) Iturr. (Iturriaga & Korf 1990), P nodosus, and P. simplex (Nees &

T. Nees) M.B. Ellis (Ellis 1971). However, P. costaricensis can be easily separated

from P. hachijoensis, P. josserandii, P. nodosus and P. simplex by its conidia, which

have a hyaline vesicle at the apex. Except for having more conidial septa, our

collection overlaps well with the original description of P. costaricensis (Morris

1972; 6-9 septa). This fungus is reported as new to the Chinese mycobiota.

Acknowledgments

The authors express gratitude to Dr Bryce Kendrick and Dr Rafael F. Castafieda-Ruiz

for serving as pre-submission reviewers and for their valuable comments and suggestions.

This project was supported by the National Natural Science Foundation of China (Nos.

31093440, 30499340, 30770015) and the Ministry of Science and Technology of the

People’s Republic of China (Nos. 2006FY120100, 2006FY110500-5).

Literature cited

Batista AC. 1952. Dois novos géneros de fungos imperfeitos. Bol. Secr. Agric. Ind. Com. Est.

Pernambuco 19 (1-2): 106-111.

Bussaban B, Lumyong S, Lumyong P, Seelanan T, Park DC, McKenzie EHC, Hyde KD. 2005.

Molecular and morphological characterization of Pyricularia and allied genera. Mycologia

97(5):1002-1011. http://dx.doi.org/10.3852/mycologia.97.5.1002

Castafieda Ruiz RE, Heredia G, Reyes M, Arias RM, Decock C. 2001. A revision of the genus

Pseudospiropes and some new taxa. Cryptog. Mycol. 22: 3-18.

http://dx.doi.org/10.1016/S0181-1584(01)01057-0

Cui BK, Dai YC. 2008. Wood-rotting fungi in eastern China 2. A new species of Fomitiporia

(Basidiomycota) from Wanmulin nature reserve, Fujian Province. Mycotaxon 105: 343-348.

Cui BK, Dai YC, Bao HY. 2009a. Wood-inhabiting fungi in southern China 3. A new species of

Phellinus (Hymenochaetales) from tropical China. Mycotaxon 110: 125-130.

http://dx.doi.org/10.5248/110.125

Cui BK, Dai YC, Li BD. 2009b. Notes on the genus Rigidoporus (Basidiomycota, Polyporaceae) in

China. Nova Hedwigia 88: 189-197. http://dx.doi.org/10.1127/0029-5035/2009/0088-0189

Cui BK, Du P, Dai YC. 2011. Three new species of Inonotus (Basidiomycota, Hymenochaetaceae)

from China. Mycol. Prog. 10: 107-114. http://dx.doi.org/10.1007/s11557-010-0681-6

Dai YC, Cui BK. 2006. Two new species of Wrightoporia (Basidiomycota, Aphyllophorales) from

southern China. Mycotaxon 96: 199-206.

de Hoog GS, von Arx JA. 1974 [“1973”]. Revision of Scolecobasidium and Pleurophragmium.

Kavaka 1: 55-60.

Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey,

England.

Ellis MB. 1972. Dematiaceous hyphomycetes. XI. Mycol. Pap. 131: 1-25.

Goh TK, Hyde KD, Tsui KM. 1998. The hyphomycete genus Acrogenospora, with two new species

and two new combinations. Mycol. Res. 102(11): 1309-1315.

66 ... Ma & al.

Hara K. 1939. Diseases of the rice plant [Ine no byogai] Ed.2: 185.

Hu DM, Cai L, Chen H, Bahkali AH, Hyde KD. 2010. Four new freshwater fungi associated with

submerged wood from Southwest Asia. Sydowia 62(2): 191-203.

Hughes SJ. 1978. New Zealand Fungi 25. Miscellaneous species. New Zealand J. Bot. 16: 311-370.

Iturriaga T, Korf RP. 1990. A monograph of the discomycete genus Strossmayeria (Leotiaceae), with

comments on its anamorph, Pseudospiropes (Dematiaceae). Mycotaxon 36 (2): 383-454.

Kirk PM. 1983. New or interesting microfungi X. Hyphomycetes on Laurus nobilis leaf litter.

Mycotaxon 18: 259-298.

Ma J, Zhang YD, Ma LG, Ren SC, Zhang XG. 2010. Taxonomic studies of Ellisembia from Hainan,

China. Mycotaxon 114: 417-421. http://dx.doi-org/10.5248/114.417

Ma J, Ma LG, Zhang YD, Castafieda Ruiz RE, Zhang XG. 2011. Pseudospiropes linderae sp. nov.

and notes on Minimelanolocus (both anamorphic Strossmayeria) new to China. Nova Hedwigia

93(3-4): 465-473. http://dx.doi-org/10.1127/0029-5035/2011/0093-0465

Matsushima T. 1971. Microfungi of the Solomon Islands and Papua-New Guinea. Published by the

author, Kobe, Japan.

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Published by the author,

Kobe, Japan.

Morris EF. 1972. Costa Rican hyphomycetes. Mycologia 64(4): 887-896.

http://dx.doi.org/10.2307/3757943

Petrak F, Ciferri R. 1932. Fungi dominicani II. Ann. Mycol. 30(3-4): 149-353.

Sinclair RC, Boshoff S, Eicker A. 1997. Sympodioplanus, a new anamorph genus from South Africa.

Mycotaxon 64: 365-374.

Subramanian CV. 1956. Hyphomycetes - II. J. Indian Bot. Soc. 35(4): 446-494.

Sutton BC. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycol. Pap. 132:

1-143.

Zhang K, Ma J, Wang Y, Zhang XG. 2009. Three new species of Piricaudiopsis from southern China.

Mycologia 101: 417-422. http://dx.doi.org/10.3852/08-147

Zhang YD, Ma J, Wang Y, Ma LG, Castafieda-Ruiz RE, Zhang XG. 2011. New species and record of

Pseudoacrodictys from southern China. Mycol. Prog. 10: 261-265.

http://dx.doi.org/10.1007/s11557-010-0696-z

Zhu H, Cai L, Zhang KQ, Hyde KD. 2005. A new species of Acrogenospora from submerged Bamboo

in Yunnan, China. Mycotaxon 92: 383-386.

MY COTAXON

http://dx.doi.org/10.5248/120.67

Volume 120, pp. 67-73 April-June 2012

A new species of Nectria on Populus from China

ZHAO-QING ZENG*” & WEN- YING ZHUANG"

' State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing

100101, China

? Graduate University of Chinese Academy of Sciences, Beijing 100049, China

CORRESPONDENCE TO *: zhuangwy@im.ac.cn

ABSTRACT - A new species, Nectria zangii, is described on Populus branches from

Donglingshan in western Beijing. The anatomy of perithecia, which become cupulate when

dry, and positive reactions to KOH and lactic acid of the fungus indicates that this species

belongs in the genus Nectria. It is characterized by small, non-septate, allantoid ascospores

and small, subcylindrical to narrowly clavate asci. Sequence analysis of the combined nuclear

ribosomal DNA ITS1-5.8S-ITS2 and partial 6-tubulin gene confirm its taxonomic position

in Nectria as a species new to science.

KEY worps - taxonomy, morphology

Introduction

The genus Nectria (Fr.) Fr. was established in 1849 and typified by

N. cinnabarina (Clements & Shear 1931). About 34 species are currently

accepted in the genus (Rossman et al. 1999, Kirk et al. 2001, Dobbeler 2005a,b,

Marincowitz et al. 2008, Pande 2008). Ten species are known from China (Teng

1963, Tai 1979, Wang et al. 1999, Zhuang & Zhang 2002, Zhang & Zhuang

2003a,b). The genus is characterized by well-developed stromata, subglobose,

globose to ellipsoid, red to dark red, fleshy, soft-textured, uniloculate, warted

perithecia that become cupulate when dry, and associated with coelomycetous

anamorphs (Rossman et al. 1999, Hirooka et al. 2009). Ascospores of Nectria

are variable and usually broadly ellipsoid to long-fusiform, hyaline to yellow-

brown, smooth to striate, non- to multi-septate or muriform. Members of the

genus are typically weak parasites of woody plants, and occur on hardwood

trees and shrubs throughout temperate regions of the Northern Hemisphere

(Samuels et al. 2009, Hirooka et al. 2011).

In this study, a new species of Nectria with small allantoid non-septate

ascospores and subcylindrical narrow asci is described. On the basis of

morphology and sequence analysis of nuclear ribosomal DNA ITS1-5.8S-ITS2

68 ... Zeng & Zhuang

(ITS) and partial B-tubulin gene, its position in Nectria is confirmed, and its

relationship with similar species of the genus is discussed.

Materials & methods

The taxonomic treatments and methods described in Luo & Zhuang (2010) were

generally followed for morphological studies. Water was used as the mounting medium

for microscopic examinations and measurements. Photographs were taken from water

or lactophenol cotton blue mounts with a Canon G5 (Tokyo, Japan) digital camera

connected to a Zeiss Axioskop 2 plus microscope (Géttingen, Germany). For anatomic

studies, longitudinal sections through ascomata were made with a freezing microtome

(YD-1508-II, Yidi Medical Appliance Factory, Jinhua, Zhejiang) at a thickness of ca.

8 um. Continuous measurements of individual structures are based on 30 measurements,

except when otherwise noted. Single-spore isolates were obtained from recent collections.

Colony characters were recorded from cultures on potato dextrose agar (PDA, Gams et

al. 1998). Microscopic descriptions of the anamorphs were taken from cultures after

14 days. Specimens examined are deposited in the Mycological Herbarium, Institute of

Microbiology, Chinese Academy of Sciences (HMAS), and cultures are kept in the State

Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences.

DNA was extracted from mycelium harvested from colonies on PDA after 2 weeks

(Wang & Zhuang 2004). Sequences of ITS and the partial 6-tubulin gene were amplified

with primer pairs ITS5-ITS4 (White et al. 1990) and T1-T222 (O'Donnell & Cigelnik

1997) respectively. The PCR reaction mixture consisted of 2.5 ul 10x PCR buffer, 1.5 ul

MgCl, (25 mM), 1.25 pl each primer (10 uM), 0.5 pl dNTP (10 mM each), 1.25 ul DNA

TABLE 1. Materials used in this study.

SPECIES COLLECTION NO. Hoan DNS Paulin

GENBANK NO. GENBANK NO.

Cosmospora coccinea Rabenh. CBS 114050 HM484537 HM484589

Nectria australiensis Seifert HMAS 83397 GU075855 HM054111

N. berolinensis (Sacc.) Cooke CBS 126112 HM484543 HM484594

N. cinnabarina (Tode) Fr. CBS 125150 HM484684 HM484820

CBS 125158 HM48 4696 HM484830

N. coryli Fuckel CBS 129156 HM484539 HM484596

N. miltina (Mont.) Mont. CBS 121121 HM484547 HM484609

N. nigrescens Cooke CBS 125148 HM484707 HM484806

CBS 129360 HM484711 HM484808

N. pseudocinnabarina Rossman CBS 128673 HM484553 HM484608

N. pseudotrichia (Schwein.) HMAS 183559 HMo54154 HMo54115

Berk. & M.A. Curtis HMAS 183175 HMo054138 HMo54114

N. zangii HMAS 251258 JN9974248 JN997421

holotype, HMAS 251247 JN997425 JN997422

HMAS 188502 JN997426 JN997423

Neonectria ramulariae Wollenw. CBS 151.29 HM054150 HM054124

“Numbers in bold = newly submitted sequences.

Nectria zangii sp. nov. (China) ... 69

template, 0.25 ul Taq polymerase (5 U/ul) and 16.5 ul ddH,O. Reactions were performed

on an ABI 2720 Thermal Cycler (Gene Co. Ltd.) with cycling conditions of denaturation

at 95°C for 5 min, followed by 35 cycles of denaturation at 94°C for 30 s, annealing at 52°C

(ITS region) and at 55°C (B-tubulin gene) for 30 s and elongation at 72°C for 45 s, with

a final extension step at 72°C for 10 min to complete the reactions. PCR products were

purified with the PCR Product Purification Kit (Biocolor BioScience & Technology Co.)

and sequenced with the ABI BigDye Terminator V3.1 Cycle Sequencing Kit (Applied

Biosystems) on an ABI 3730 XL DNA Sequencer (SinoGenoMax Co. Ltd.). ITS5 and

ITS4 (for ITS), and T1 and Bt2b (for the partial B-tubulin gene) were employed as

sequencing primers (White et al. 1990, Glass & Donaldson 1995, O’Donnell & Cigelnik

1997). Sequences of related species were retrieved from GenBank and are listed in

TABLE l.

All sequences were aligned using ClustalX 1.8 (Thompson et al. 1997), and the

alignments were visually adjusted by BioEdit 7.0.5 (Hall 1999) when necessary. A

partition homogeneity test was performed with 1000 replicates in PAUP 4.0b10

(Swofford 2002) to evaluate statistical congruence between sequence data from ITS and

B-tubulin gene regions. The partition homogeneity test (P = 0.01) suggested that the

individual partitions were congruent (Farris et al. 1995, Cunningham 1997). A neighbor-

joining tree was generated using MEGA 4.10 (Tamura et al. 2007) based on combined

sequences of ITS and partial B-tubulin gene with Cosmospora coccinea and Neonectria

ramulariae as the outgroup taxa. The Kimura 2-parameter was selected as the nucleotide

substitution model, and gaps or missing data were pairwise deleted. Bootstrap analyses

were performed with 1000 replicates to test phylogeny branch support.

Taxonomy

Nectria zangii Z.Q. Zeng & W.Y. Zhuang, sp. nov. FIGuRE 1

MYCOBANK 563722

Differs from Nectria miltina by thinner perithecial walls, narrower asci, smaller

ascospores, and occurrence on twigs of Populus sp.

Type: China. Beijing, Donglingshan, alt. 1150 m, on branches of Populus sp., 20 July

2011, Z.Q. Zeng & H.D. Zheng 7684 (Holotype, HMAS 251247).

Erymo.oey: The specific epithet honors the late Chinese mycologist, Prof. M. Zang.

Ascomata perithecial, gregarious, up to 40 in a group, with a well-developed

stroma that is erumpent through bark, superficial, subglobose to globose,

168-200 um high, 151-203 um diam., becoming cupulate upon drying, red

when fresh and reddish brown when dry, turning dark red in 3% KOH aqueous

solution and orange-red to orange in lactic acid, surface somewhat roughened

when dry; with warts or a irregular covering layer of various thickness, 0-32

uum high, cells angular, 3.5-15 x 2.5-5 um, wall 1-1.5 um thick. Ascomatal wall

16-26 um thick, of two layers; outer layer 10-16.5 um thick, cells angular, 2-7

x 3-6 um, wall 1-1.5 um thick; inner layer 5-10.5 um thick, cells flattened,

9.5-15.5 x 1.5-3.5 um, wall 1-1.5 um thick. Asci subcylindrical to narrowly

clavate, 8-spored, with an apical ring, 28-35 x 2.5-3 um (n = 50), biseriate

70 ... Zeng & Zhuang

100 micron 20 micron

PDA, obverse. c. Colony on PDA, reverse. d. Median section of ascomata. e. Structure of ascoma at

apical portion. f. Asci with ascospores. g. Conidiophores and conidia.

to irregularly biseriate. Ascospores allantoid to rod-shaped, straight to slightly

curved, non-septate, hyaline, smooth, 3.5-5.5(-6) x 0.9-1.2(-1.4) um (n =

50).

In culture, colonies reaching 57 mm in diameter after 7 days on PDA at 25°C

under daylight, aerial mycelium sparse to absent, yellowish, submerged mycelia

forming a root-like or vein-like structure, reverse light orange. Conidiophores

unbranched to sparsely branched, septate, 29-79 x 2-3.2 um, arising from agar

surface throughout the colony, dominating near the margin. Microconidia

allantoid to rod-shaped, not or slightly curved, hyaline, aseptate, 3-6 x 0.8-1 um

(n = 50). Macroconidia and chlamydospores not observed.

ADDITIONAL SPECIMENS EXAMINED: CHINA. BeyinG, Donglingshan, alt. 1150m,

on rotten branch of Populus sp., 20 July 2011, Z.Q. Zeng & H.D. Zheng 7648-7651

(HMAS 188501-188504); 7652 (HMAS 251248); 7653-7654 (HMAS 188505-188506);

7655 (HMAS 251249); 7656 (HMAS 188507); 7657-7660 (HMAS 251250-251253);

Nectria zangii sp. nov. (China) ... 71

7663 (HMAS 188508); 7665-7667 (HMAS 188509-188511); 7668 (HMAS 251254);

7669-7671 (HMAS 188512-188514); 7672 (HMAS 251255); 7673-7680 (HMAS

188515-188522); 7681 (HMAS 251256); 7682 (HMAS 188523); 7683 (HMAS 251257);

7685-7694 (HMAS 188524-188533); 7695 (HMAS 251258); 7696-7698 (HMAS

188534-188536).

Comments - Morphologically, the perithecial anatomy, perithecia becoming

cupulate when dry, and positive reaction to KOH and to lactic acid indicates

the new species’ position in Nectria. Among the existing species of the genus,

N. zangii is most similar to N. miltina in shape of asci and ascospores. Nectria

miltina, however, differs in having a thicker perithecial wall (25-35 um thick),

wider asci (28-36 x 3.5-5 um), larger ascospores (5.5-7 x 1.5-2 um), and an

entirely different host. Nectria miltina occurs on leaves of the monocotyledonous

plant family Agavaceae (Rossman et al. 1999) instead of on dicotyledonous

Populus sp.

Sequence analysis of the combined ITS and partial B-tubulin gene from

N. zangii and eight other Nectria species with various ascospore shapes and

septation confirms taxonomic position of the new species (Fic. 2). Our results

support two major distantly related clades within the sampled Nectria species.

Nectria zangii, N. berolinensis, N. miltina, N. coryli, and N. pseudocinnabarina

group together with high bootstrap support (99%). The latter four species were

reported having sporodochial anamorphs (Rossman et al. 1999). The other

81, Nectria zangii HMAS 188502

100 |LN. zangii HMAS 251247

94 N. zangii HMAS 251258

88 N. berolinensis CBS 126112

N. miltina CBS 121121

N. coryli CBS 129156

N. pseudocinnabarina CBS 128673

100 —N. pseudotrichia HMAS 183175

25 N. pseudotrichia HMAS 183559

N. australiensis HMAS 83397

100 ,N. cinnabarina CBS 125158

96 N. cinnabarina CBS 125150

100 N. nigrescens CBS 129360

100 |. nigrescens CBS 125148

Cosmospora coccinea CBS 114050

91 Neonectria ramulariae CBS 151.29

-————_

0.02

Fic. 2. Neighbor-joining tree based on combined sequences of ITS and partial B-tubulin gene

showing the relationships among some Nectria species.

72... Zeng & Zhuang

sampled species, including the type species N. cinnabarina, group together

with 96% bootstrap support (Fic. 2). These results are identical to those by

Hirooka et al. (2011). Nectria berolinensis, shown as sister to N. zangii with 94%

bootstrap support, differs in larger perithecia (250-300 um diam.) and 5-9-

septate larger (16-20 x 7-8 um) muriform elliptical ascospores (Seaver 1909).

Nectria miltina also groups with N. berolinensis and N. zangii with relatively

high support (88%). Thus, both morphology and DNA sequence analysis

support the recognition of N. zangii as a new species.

Acknowledgements

We are very grateful to Dr. A. Y. Rossman and Dr. K. A. Seifert for serving as pre-

submission reviewers, language corrections and valuable suggestions, Dr. L.L. Norvell

for editorial assistance and language corrections, and to Dr. H.-D. Zheng, Dr. J. Luo and

Ms. X. Song for technical help. This work was supported by the National Natural Science

Foundation of China (No. 31070015), and the Knowledge Innovation Program of the

Chinese Academy of Sciences (No. KSCX2-EW-J-6).

Literature cited

Clements FE, Shear CL. 1931. The genera of fungi, 2™ edition. The Wilson Company, New York,

USA.

Cunningham CW. 1997. Can three incongruence tests predict when data should be combined?

Mol. Biol. Evol. 14: 733-740.

Déobbeler P. 2005a. Ascospore diversity of bryophilous Hypocreales and two new hepaticolous

Nectria species. Mycologia 97: 924—934. http://dx.doi.org/10.3852/mycologia.97.4.924

Dobbeler P. 2005b. Three new hypocrealean ascomycetes on bryophytes. Sydowia 57: 179-188.

Farris JS, Kallersjo M, Kluge AG, Bult C. 1995. Testing significance of incongruence. Cladistics 10:

315-319. http://dx.doi.org/10.1006/clad.1994.1021

Gams W, Hoekstra ES, Aptroot A. 1998. CBS course of mycology, 4 edn. Centraalbureau voor

Schimmelcultures, Baarn, Netherlands.

Glass NL, Donaldson GC. 1995. Development of primer sets designed for use with the PCR

to amplify conserved genes from filamentous ascomycetes. Appl. Environ. Microb. 61:

1323-1330.

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NTT. Nucl. Acids Symp. Ser. 41: 95-98.

Hirooka Y, Rossman AY, Chaverri P. 2009. Systematics of the genus Nectria based on a six-gene

phylogeny. Inoculum 60: 22.

Hirooka Y, Rossman AY, Chaverri P. 2011. A morphological and phylogenetic revision of the

Nectria cinnabarina species complex. Stud. Mycol. 68: 35-56.

http://dx.doi.org/10.3114/sim.2011.68.02

Kirk PM, Cannon PE, David JC, Stalpers JA. 2001. Ainsworth & Bisby’s dictionary of the fungi, 9"

edn. CAB International, Wallingford, United Kingdom.

Luo J, Zhuang WY. 2010. Three new species of Neonectria (Nectriaceae, Hypocreales) with notes on

their phylogenetic positions. Mycologia 102: 142-152. http://dx.doi.org/10.3852/08-224

Marincowitz S, Crous PW, Groenewald JZ, Wingfield MJ. 2008. Microfungi occurring on Proteaceae

in the fynbos. CBS Biodiversity series 7: 74.

O’Donnell K, Cigelnik E. 1997. Two divergent intragenomic rDNA ITS2 types within a monophyletic

lineage of the fungus Fusarium are nonorthologous. Mol. Phylogenet. Evol. 7: 103-116.

http://dx.doi.org/10.1006/mpev.1996.0376

Nectria zangii sp. nov. (China) ... 73

Pande A. 2008. Ascomycetes of Peninsular India. Scientific Publishers, Jodhpur, India.

Rossman AY, Samuels GJ, Rogerson CT, Lowen R. 1999. Genera of Bionectriaceae, Hypocreaceae

and Nectriaceae (Hypocreales, Ascomycetes). Stud. Mycol. 42: 1-260.

Samuels GJ, Lu BS, Chaverri P, Candoussau F, Fournier J, Rossman AY. 2009. Cyanonectria, a new

genus for Nectria cyanostoma and its Fusarium anamorph. Mycol. Prog. 8: 49-58.

http://dx.doi.org/10.1007/s11557-008-0577-x

Seaver FJ. 1909. The Hypocreales of North America-II. Mycologia 1: 177-207.

http://dx.doi.org/10.2307/3753513

Swofford DL. 2002. PAUP”: phylogenetic analysis using parsimony (*and other methods). Version

4b10. Sunderland, Massachusetts. Sinauer Associates.

Tai FL. 1979. Sylloge fungorum sinicorum. Academica Sinica, Science Press, Beijing, China (in

Chinese).

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis

(MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596-1599.

http://dx.doi.org/10.1093/molbev/msm092

Teng SC. 1963. Fungi of China. Science Press, Beijing (in Chinese).

Thompson JD, Gibson TJ, Plewniak F, Janmougin F, Higgin DG. 1997. The ClustalX windows

interface: flexible strategies for multiple sequences alignment aided by quality analysis tools.

Nucl. Acids Res. 25: 4876-4883. http://dx.doi.org/10.1093/nar/25.24.4876

Wang L, Zhuang WY. 2004. Designing primer sets for amplification of partial calmodulin genes

from penicillia. Mycosystema 23: 466-473.

Wang YZ, Wu SH, Zhou WN, Zhang DZ, Chen GY, Chen SF, Chen CL, Zeng XX, Liu JH, Xie WR,

Xie HR, Zhong ZX, Jian QY. 1999. List of the fungi in Taiwan. Council of Agriculture, Executive

Yuan, Taipei.

White TJ, Bruns TD, Lee SB, Taylor JW. 1990. Amplification and direct sequencing of fungal

ribosomal RNA genes for phylogenetics. pp. 315-222, in: MA Innis et al. (eds): PCR protocols:

a guide to methods and applications. Academic Press, New York.

Zhang XM, Zhuang WY. 2003a. New Chinese records of Bionectriaceae and Nectriaceae.

Mycosystema 22: 525-530 (in Chinese).

Zhang XM, Zhuang WY. 2003b. Re-examination of Bionectriaceae and Nectriaceae (Hypocreales)

from temperate China on deposit in HMAS. Nova Hedwigia 76: 1-2.

http://dx.doi.org/10.1127/0029-5035/2003/0076-0191

Zhuang WY, Zhang XM. 2002. Re-examination of Bionectriaceae and Nectriaceae (Hypocreales)

from tropical China on deposit in HMAS. Nova Hedwigia 74: 1-2.

http://dx.doi.org/10.1127/0029-5035/2002/0074-0275

MY COTAXON

http://dx.doi.org/10.5248/120.75

Volume 120, pp. 75-79 April-June 2012

A new species of the lichen genus Phlyctis

from Maharashtra, India

GAYATRI CHITALE’ & URMILA MAKHIJA”

Agharkar Research Institute, G.G. Agarkar Road, Pune -411 004, India

CORRESPONDENCE TO: ' gayatri4u@gmail.com & ? uv_makhija@hotmail.com

ABSTRACT—A new species, Phlyctis communis, characterized by 8-spored asci, 7—14(—16)

transversely septate ascospores, and salazinic and norstictic acids, is described from India.

Key worps—Ascomycetes, Ostropales, taxonomy

Introduction

Although taxonomic accounts of several lichen genera from Maharashtra

have been published and many species have been described (Chitale et al. 2008,

2009, 2011, Chitale & Makhija 2008, Dube et al. 2005, Dube & Makhija 2008,

2010), one locally common and abundant interesting species has remained

unpublished. For several years a conspicuously grayish-white unidentified

crustose lichen was known to occur all over Maharashtra, mostly during the

monsoons in the humid high altitude regions. This has now been identified and

is described as a new species of the genus Phlyctis.

The subtropical to temperate lichen genus Phlyctis (Wallr.) Flot., confirmed

by phylogenetic sequence analysis as belonging in the Ostropales (Miadlikowska

et al. 2006), is characterized by a smooth to verrucose crustose thallus,

protococcoid green algal photobiont, sunken or erumpent, round apothecioid

ascomata with indistinct thalline margins, proper exciple present or absent,

unbranched or apically furcate paraphyses, 1-8-spored asci, and ascospores

that are colourless, transversely septate to multicelled-muriform, thin walled,

elongate-ellipsoid, and non-halonate.

Phlyctis species generally contain one or several of the following depsidone

acids: stictic, constictic, norstictic, connorstictic, hypostictic, salazinic psoromic,

neopsoromic and/or protocetraric acids (Galloway & Guzman 1988).

The genus is widely distributed, with 12 species listed by Kirk et al. (2008)

and five others from Great Britain and Ireland (Benfield et al. 2009), China (Ma

76 ... Chitale & Makhija

et al. 2010), India (Joshi et al. 2010), and Australia (Lumbsch et al. 2011) added

since 2008. Of these only five — Phlyctis himalayensis (Nyl.) D.D. Awasthi,

P. nepalensis Rasanen, P. polyphora Stirt., P. karnatakana S. Joshi & Upreti,

and P. subagelaea S. Joshi & Upreti — have been reported from the Indian

subcontinent (Awasthi 2000, Joshi et al. 2010).

In the present study, one new species has been discovered from Maharashtra

that represents the first record of Phlyctis from this area.

Materials & methods

The specimens were examined using a stereomicroscope and a light microscope.

Sections of the thalli and apothecia were stained with Lugol’s iodine solution. All sections

examined were mounted in LPCB (lactophenol with cotton blue). TLC protocols

followed Culberson & Kristinsson (1970) and White & James (1985). All specimens

were observed under UV light (365 nm). The present study is based on the material

preserved in the lichen herbarium of Ajarekar Mycological Herbarium (AMH) and

recent collections by the authors from Maharashtra state.

Taxonomy

Phlyctis communis Chitale & Makhija, sp. nov. Fries 1-4

MycoBank MB 563474

Similis Phlyctidae karnatakanae sed ascosporis 7-14(-16) trans-septatis et acidum

norsticticum et salazinicum continenti differt.

Type: India, Maharashtra, Satara District, Mahabaleshwar, 24.9.1997, U.V. Makhija,

(Holotype, AMH 97.52).

Erymo oey: Latin communis, referring to the plentiful or common occurrence.

THALLUS crustose, corticolous, grayish or greenish-white, spreading over the

bark substratum in 5-12 cm patches, cracked, areolate, matt, sometimes glossy,

white in cracks and 90-120 um thick at the margin; epiphloeodal algal layer

30-35 um thick, green algae single celled, more or less globose, 5-7 um diam.;

hypothallus black.

Ascomarta black, round, oblong, numerous, scattered all over the thallus

and radially arranged in small 1-2(-3.5) mm diam. circles, later grouped and

merging into each other forming patches on the bark of tree trunks, individual

ascomata <1 mm across, immersed in the thallus; disc black, plane to concave,

covered by a white pruina; exciple brownish, composed of loosely interwoven

hyphae in the peripheral region, and blackish-brown at the base; epihymenium

dark brown, 30-33 um thick, KI-; hymenium colourless to light yellow,

72-81 um tall, KI+ blue; paraphyses simple to branched at tips; hypothecium

brownish, 12-15 um thick; asci 8-spored, 90-105 x 18-24 um, entire ascus KI+

blue. Ascospores hyaline, 7-14(-16) transversely septate, very rarely with 1-2

vertical septa, 18-33(-45) x 6-9 um.

Phlyctis communis sp. nov. (India) ... 77

Ficures 1-4. Phlyctis communis (holotype): 1-2, habit; 3, vertical section of the ascocarp;

4, transseptate ascospores. Scale bars: 1-2 = 1 mm; 3-4 = 50 um.

CHEMISTRY— thallus K+ yellow turning red, C-, KC-, P+ yellow; UV-;

norstictic and salazinic acids present.

SELECTED ADDITIONAL SPECIMENS EXAMINED (total specimens = 110, all in AMH)—

INDIA. MAHARASHTRA: KOLHAPUR DISTRICT, Ajra, 6.10.2004, B.C. Behera, N. Verma,

AMH 04.320, U.V. Makhija & G.S. Chitale, AMH 04.389, Amba, 16.10.1974, C.R.

Kulkarni & A.V. Prabhu, AMH 74.1282, on the way to Kumbhi from Gaganbawada,

12.10.2000, B.A. Adawadkar & K.R. Randive, AMH 00.337. Pune District, Amby

valley, 10.2.2006, G.S. Chitale, AMH 06.181; Boma Hills, Khandala, 19.9.1974, M.B.

Nagarkar & C.R. Kulkarni, AMH 74.691, Walwan Dam, 16.9.2002, A.V. Bhosale & G.S.

Chitale, AMH 02.113. Satara District, Mahabaleshwar, 1.11.1973, C.R. Kulkarni,

AMH 73.2938; Panchgani, Tata Holiday Home, 29.9.2003, U.V. Makhija & B.C. Behera,

AMH 03.369. SInpHUDURG District, Ajra to Amboli, 7.12.1974, P.G. Patwardhan &

A.V. Prabhu, AMH 74.2248, 10.10.2000, U.V. Makhija & V.A. Mantri, AMH 00.176.

REMARKS— With respect to external morphology and ascospore size, Phlyctis

communis resembles P. karnatakana, also from India, which differs in fewer

ascospore septa (7 transsepta) and containing only norstictic acid.

78 ... Chitale & Makhija

Phlyctis subuncinata Stirt., also with norstictic acid and transversely septate

ascospores, differs in having a sorediate thallus.

Two other Phlyctis species with transversely septate ascospores —

P. himalayensis from India and P. longifera (Nyl.) DJ. Galloway & Guzman

from New Zealand — differ especially in having larger, 7-septate ascospores.

Phlyctis himalayensis ascospores measure 60-75 x 6-8 um, whereas P longifera

ascospores are 55-86 Xx 5-7 um.

Phlyctis psoromica Elix & Kantvilas, also with transversely septate ascospores,

has only (3-)7-septate ascospores (30-39.3-50(-52) x 4-5.2-6 um) and

contains psoromic acid.

The somewhat similar P polyphora from India shares a whitish thallus and

a pruinose ascomatal disc <1 mm diam. but is distinguished by very large

(60-110 x 7.5-9.5 um) muriform ascospores.

Phlyctis communis is very common and has been collected from semi-

evergreen to dry deciduous forests with relative humidity between 15-90%

where thalli grow in huge patches on the bark of Ficus benghalensis and

Casuarina equisetifolia where it frequently associates with Arthothelium

awasthii Patw. & Makhija.

Acknowledgments

We are thankful to the Ministry of Environment and Forests, Government of India,

New Delhi, for their financial support. The authors are grateful to G. Kantvilas and

A. Aptroot, for reviewing the manuscript, and suggestions to improve the quality of

the manuscript. I also thank Mr. Subhash Gaikwad, for his technical help in preparing

illustrations.

Literature cited

Awasthi DD. 2000. Lichenology in Indian subcontinent. Dehra Dun (India), Bishen Singh

Mahendra Pal Singh.1-123 p.

Benfield B, James PW, Hitch CJB. 2009. Phlyctis. 695-696,in: CW Smith et al. (eds). The lichens of

Great Britain & Ireland, 2nd edn. British Lichen Society, London.

Chitale G, Makhija U. 2008. A new species of the lichen genus Brigantiaea from India. Mycotaxon

104: 409-413.

Chitale G, Dube A, Makhija U. 2008. The lichen genus Physcia and allied genera from Maharashtra,

India. Geophytology 37: 13-21.

Chitale G, Makhija U, Sharma B. 2009. New combinations and new species in the lichen genera

Hemithecium and Pallidogramme. Mycotaxon 108: 83-92.

Chitale G, Makhija U, Sharma B. 2011. Additional species of Graphis from Maharashtra, India.

Mycotaxon 115: 469-480. http://dx.doi.org/10.5248/115.469

Culberson CF, Kristinsson H. 1970. A standardized method for the identification of lichen products.

J. Chromatogr., 46: 85-93. http://dx.doi.org/10.1016/S0021-9673(00)83967-9

Dube A, Makhija U. 2008. A new species of Parmeliella (family Pannariaceae) from India.

Lichenologist 40: 209-212. doi:10.1017/S0024282908007470

Phlyctis communis sp. nov. (India) ... 79

Dube A, Makhija U. 2010. Occurrence of four additional non-hairy species of Leptogium from

Maharashtra, India. Lichenologist, 42(6): 701-710.

http://dx.doi.org/10.1017/S00242829 10000332

Dube A, Chitale C, Makhija U. 2005. The lichen genera Dirinaria and Pyxine (family Physciaceae)

from Maharashtra, India. Phytotaxonomy 5: 83-86.

Galloway DJ, Guzman G. 1988. A new species of Phlyctis from Chile. Lichenologist 20: 393-399.

doi:10.1017/S0024282988000507

Joshi S, Upreti DK, Mishra GK, Divakar PK. 2010. Two new species of the lichen genus Phlyctis in

India. The Bryologist 113(4): 724-727. http://dx.doi.org/10.1639/0007-2745-113.4.724

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th Edition. CABI

Bioscience: CAB International. 771 p.

Lumbsch HT, et al. 2011: One hundred new species of lichenized fungi: a signature of undiscovered

global diversity. Phytotaxa 18: 1-127.

Ma R, Li MH, Wang HY, Zhao ZT. 2010. A new species of Phlyctis (Phlyctidaceae) from China

Mycotaxon 114: 362-366. http://dx.doi.org/10.5248/114.361

Miadlikowska J, et al. 2006. New insights into classification and evolution of the Lecanoromycetes

(Pezizomycotina, Ascomycota) from phylogenetic analyses of three ribosomal RNA- and two

protein-coding genes. Mycologia 98: 1088-1103.

http://dx.doi.org/10.3852/mycologia.98.6.1088

White FJ, James PW 1985. A new guide to microchemical techniques for the identification of lichen

substances. Bulletin British Lichen Society. 57(Suppl.): 1-41.

MY COTAXON

http://dx.doi.org/10.5248/120.81

Volume 120, pp. 81-88 April-June 2012

Two new freshwater species of Annulatascaceae from China

D1An-MinG Hu”, Ler Car", ALI HASSAN BAHKALIP & KEVIN D. HyDE??*#5"

'State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

?International Fungal Research & Development Centre, The Research Institute of Resource Insects,

Chinese Academy of Forestry, Bailongsi, Kunming 650224, China

*Botany and Microbiology Department, College of Science, King Saud University,

Riyadh, Saudi Arabia

‘School of Science, Mae Fah Luang University, Chiang Rai, Thailand

*Institute of Excellence for Fungal Research, Mae Fah Luang University, Chiang Rai, Thailand

* CORRESPONDENCE TO: mrcailei@gmail.com, kdhyde3@gmail.com

ABSTRACT — Two new species belonging to Annulatascaceae are described and illustrated

on submerged wood from a freshwater stream. Annulatascus menglensis, differentiated from

other Annulatascus species by its hyaline ascomatal neck, is also characterized by cylindrical

asci with a large bipartite refractive J- apical ring and hyaline fusiform ascospores without

sheath or appendages. Aqualignicola vaginata, which is distinguished from A. hyalina in

producing a gelatinous sheath surrounding the unicellular hyaline ascospores, is further

characterized by its membranous ascomata with lanceolate setae that surround the neck, long

cylindrical asci with a relatively large apical ring.

KEY worDs —aquatic, Ascomycota, fungi, taxonomy

Introduction

Freshwater fungi are an important ecological group because of their

indispensable function as decomposers in freshwater ecosystems (Wong et al.

1998a); they also have potential application in producing various and useful

bioactive secondary metabolites (Bucher et al. 2004, Zhang et al. 2008, Hosoe

et al. 2010, Dong et al. 2011). Freshwater fungi are however, relatively poorly

investigated comparing to their terrestrial counterpart. Until recently, freshwater

fungi had been reported mainly from Europe, North America, and Southeast

Asia (Cai et al. 2006b, Raja et al. 2009). Although China has a high biodiversity

of fungi and abundant freshwater resources, investigation of freshwater fungi

has been mostly restricted to Hong Kong and Yunnan Province.

Annulatascaceae S.W. Wong et al. comprises many freshwater genera

(Vijaykrishna et al. 2006). The family is characterized morphologically by dark

82 ... Hu &al.

brown or black ascomata with ostiolate necks, long cylindrical unitunicate

asci with a relatively massive, refractive apical ring, and uniseriate, hyaline

to brown, fusiform or ellipsoidal ascospores (Wong et al. 1998b, Ho & Hyde

2000). Seventy-five species representing 21 genera are currently placed in

Annulatascaceae (Kirk et al. 2008).

Our research group has focused on investigating the diversity of freshwater

fungi in China for over ten years (Hyde et al. 2000, Luo et al. 2004, Cai et al.

2005, 2008, Hu et al. 2007, 2010a, b). During our continuing investigation, two

undescribed ascomycetes belonging to the Annulatascaceae were collected on

submerged wood and are described and illustrated in this paper.

Materials & methods

D.M. Hu, who has investigated freshwater ascomycetes on submerged woody

material in the southern provinces of China (including Guizhou, Jiangxi and Yunnan

provinces) since 2008, has collected 875 samples from streams, lakes, ditches, and ponds.

After return to the laboratory, the samples were incubated following Cai et al. (2006a).

Samples were examined for fungal fruiting bodies under a dissecting microscope

(Leica MZ16A) during incubation. Observations and photographs were prepared from

materials mounted in water under a compound microscope (Nikon E800). Single spore

isolation as outlined by Choi et al. (1999) was attempted but unsuccessful. Herbarium

specimens of the fungal taxa described in this study are deposited in the International

Fungi Research and Development Centre (IFRDC).

Taxonomy

Annulatascus menglensis D.M. Hu, L. Cai & K.D. Hyde, sp. nov. PLATE 1

MycoBank MB 563810

Differs from other Annulatascus species by its hyaline neck.

Type: China, Yunnan Province, Mengla, Wudaoban Stream, 21°24'N 101°36’E, alt: 660

m, 3 April 2009, D.M. Hu (Holotype, IFRDC 023-002).

EryMoLoecy: menglensis refers to the collecting site, Mengla, a county in southern

China.

AscoMaTA 180-200 high, 120-150 um in diam., subglobose to ellipsoidal,

superficial, solitary, ostiolate, black, coriaceous. NECK 150-340 x 40-60

um, cylindrical, hyaline, membranous, periphysate. PERIDIUM composed of

compressed cells, outer layers dark brown, inner layers hyaline. PARAPHYSES

ca. 5-6 um wide, broad at the base and tapering towards the apex, hyaline,

septate, smooth-walled, simple, longer than asci. Asc1 115-163 x 10-13 um,

8-spored, cylindrical, unitunicate, persistent, pedicellate, with a large bipartite,

refractive, J- apical ring, ca. 3 um high, 4 um in diam. Ascosporss 21-25 x

8-10 um (mean = 23.8 x 8.5 um, n = 30), uniseriate, overlapping, fusiform, with

acute ends, hyaline, aseptate, guttulate, smooth, thin-walled, without sheaths or

appendages.

Annulatascaceae spp. nov. (China) ... 83

PiaTE 1. Annulatascus menglensis (from holotype). a. Ascoma on submerged wood. b. Squash

mount of an ascoma. c. Peridium comprising angular cells in surface view. d. Paraphyses. e-f. Asci.

g. Upper part of an ascus showing the apical ring. h-k. Ascospores. Scale bars: a = 100 um, b = 50

um, c-d = 5 um, e-f = 20 um, g-k = 5 um.

84 ... Hu &al.

HABITAT & DISTRIBUTION— Saprobic on submerged wood in fresh water

in China.

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE: Mengla, Wudaoban

Stream, 21°24'N 101°36’E, alt. 660 m, 3 April 2009, D.M. Hu (IFRDSC 002-017, IFRDSC

002-018).

COMMENTS — Annulatascus menglensis has black, coriaceous ascomata and

cylindrical asci with a large bipartite, refractive, J- apical ring. It fits well within

the generic concept of Annulatascus (Hyde 1992). Thus far, 14 species have been

included in the genus (Tsui et al. 2002, Barbosa et al. 2008, Abdel-Wahab et al.

2011). The new species resembles A. velatisporus K.D. Hyde (Hyde 1992) in

having fusiform and aseptate ascospores. However, A. velatisporus differs from

A. menglensis in having larger ascospores (26-42 x 9-12 um) with a gelatinous

sheath. Annulatascus menglensis shares similar ascospore dimensions with

A. fusiformis K.D. Hyde & S.W. Wong (Hyde & Wong 2000), A. joannae K.M.

Tsui et al. (Tsui et al. 2002), A. citriosporus J. Frohl. & K.D. Hyde (Frohlich &

Hyde 2000), and A. apiculatus FR. Barbosa & Gusmao (Barbosa et al. 2008)

but lacks their black necks and mucilaginous sheaths and/or appendages.

Annulatascus lacteus K.M. Tsui et al. (Tsui et al. 2002), which also has aseptate

ascospores without sheath or appendage, differs from A. menglensis in having

a milky ascomata.

Aqualignicola vaginata D.M. Hu, L. Cai & K.D. Hyde, sp. nov. PLATE 2

MycoBank MB 563811

Differs from Aqualignicola hyalina in having a hyaline ascomatal neck and a gelatinous

sheath surrounding the ascospores.

Type: China, Yunnan Province, Mengla, Wudaoban Stream, 21°32'N E101°29’E, alt. 620

m, 2 April 2009, D.M. Hu (Holotype, IFRDC 021-043).

ETyMOLOoGy: vaginata (= sheath) refers to the mucilaginous sheath surrounding the

ascospore.

ASCOMATA 100-140 um diam, globose, membranous, brown to black-brown,

submerged, solitary. NecK 40-60 um diam, 200-350 um long, cylindrical,

hyaline, periphysate, covered with setae; setae 15-35 x 4-5 um, lanceolate,

solid, surrounding the neck. PERrpr1uM 7-14 um thick, in surface view of

textura angularis, composed of 5 layers of light-brown, pseudoparenchymatous

cells. PARAPHYSES 4-6 tm in diam, 85-100 long, septate, flexuose, hypha-like,

numerous, tapering towards the apex, embedded in a gelatinous matrix. AscI

145-156 x 5.5-6.5 um, 8-spored, cylindrical, pedicellate, unitunicate, apically

truncate, with a large, J-, refractive, apical ring. Ascospores 11-15 x 5-6

um (mean = 13.4 x 5.3 um, n = 50), uniseriate, ellipsoidal-fusiform, hyaline,

aseptate, thin-walled, multiguttulate, with a fan-shaped, hyaline mucilaginous

sheath surrounding one end of the ascospores.

Annulatascaceae spp. nov. (China) ... 85

PLATE 2. Aqualignicola vaginata (from holotype). a. Ascoma on submerged wood. b. Section of an

ascoma. c. Peridium. d. Section through a neck. e. Hair on neck. f. Paraphyses. g. Ascus. h. Upper

part of an ascus, show the apical ring. i-n. Ascospores (arrows show the mucilaginous sheath and

the multiple guttules). Scale bars: a = 100 um, b = 30 um, c = 5 um, d = 30 um, e, f= 5 um, g = 20

um, h-n = 5 um.

86 ... Hu & al.

HABITAT & DISTRIBUTION — Saprobic on submerged wood in fresh water

in China.

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE: Mengla, Wudaoban

Stream, 21°32'N 101°29’E, alt. 620 m, 2 April 2009, D.M. Hu (IFRDSC 004-003, IFRDSC

004-004, IFRDSC 004-005).

COMMENTS — Previously monotypic, Aqualignicola V.M. Ranghoo et al. is

characterized by membranous ascomata with a neck surrounded by lanceolate

setae, long cylindrical asci with a relatively large apical ring, and hyaline

unicellular ascospores (Ranghoo et al. 2001). The new species fits well with the

generic concept of Aqualignicola but differs from the type species, A. hyalina,

in having a hyaline ascomatal neck and a mucilaginous sheath surrounding the

ascospore. Furthermore, the ascospores and asci of A. vaginata are narrower

than those of A. hyalina (ascospores: 14-15 x 6.25-7.5 um; asci 9-12 um diam.).

Aqualignicola vaginata is also similar to species of Aquaticola W.H. Ho et al.

(Ho et al. 1999) and Longicollum Zelski et al. (Zelski et al. 2011a). However, the

species of the latter two genera have glabrous ascomata and lack the lanceolate

setae surrounding the neck found in A. vaginata. Aqualignicola vaginata can

be compared to the newly described Chaetorostrum quincemilense Zelski et al.

(Zelski et al. 2011b) in that both taxa have a hyaline neck covered with setae;

however the ascospores of A. vaginata are aseptate and much smaller than

those of C. quincemilense (1-3-septate; 30-38 x 10-12 um).

Acknowledgments

Funds for research were provided by a major project of the National Natural Science

Foundation of China, No. 31093440. The Global Research Network for Fungal Biology

and King Saud University are thanked for support.

Literature cited

Abdel-Wahab MA, Abdel-Aziz FA, Mohamed SS, Abdel-Aziz AE. 2011. Annulatascus nilensis

sp. nov., a new freshwater ascomycete from the River Nile, Egypt. IMA Fungus 2: 1-6.

http://dx.doi.org/10.5598/imafungus.2011.02.01.01

Barbosa FR, Gusmao LFP, Raja HA, Shearer CA. 2008. Annulatascus apiculatus sp. nov., a new

freshwater ascomycete from the semi-arid Caatinga Biome of Brazil. Mycotaxon 106:

403-407.

Bucher VVC, Pointing SB, Hyde KD, Reddy CA. 2004. Production of wood decay enzymes, loss of

mass, and lignin solubilization in wood by diverse tropical freshwater fungi. Microbial Ecology

48: 331-337. http://dx.doi.org/10.1007/s00248-003-0132-x

Cai L, Zhang KQ, Hyde KD. 2005. Ascoyunnania aquatica gen. et sp nov., a freshwater fungus

collected from China and its microcylic conidiation. Fungal Diversity 18: 1-8.

Cai L, Ji KF, Hyde KD. 2006a. Variation between freshwater and terrestrial fungal communities on

decaying bamboo culms. Antonie Van Leeuwenhoek 89: 293-301.

http://dx.doi.org/10.1007/s10482-005-9030-1

Cai L, Hyde KD, Tsui CKM. 2006b. Genera of freshwater fungi. Fungal Diversity Research Series

18: 1-261.

Annulatascaceae spp. nov. (China) ... 87

Cai L, Guo XY, Hyde KD. 2008. Morphological and molecular characterisation ofa new anamorphic

genus Cheirosporium, from freshwater in China. Persoonia 20: 53-58.

http://dx.doi.org/10.3767/003158508X314732

Choi YW, Hyde KD, Ho WH. 1999. Single spore isolation of fungi. Fungal Diversity 3: 29-38.

Dong JY, Shen KZ, Sun R. 2011. Recent progress in the research on bioactive substances from

freshwater fungi. Mycosystema 30: 206-217.

Frohlich J, Hyde KD (eds). 2000. Palm microfungi. Fungal Diversity Press, Hong Kong.

Ho WH, Hyde KD. 2000. A new family of freshwater ascomycetes. Fungal Diversity 4: 21-36.

Ho W, Tsui CKM, Hodgkiss IJ, Hyde KD. 1999. Aquaticola, a new genus of Annulatascaceae from

freshwater habitats. Fungal Diversity 3: 87-97.

Hosoe T, Gloer JB, Wicklow DT, Raja HA, Shearer CA. 2010. New nonadride analogues from a

freshwater isolate of an undescribed fungus belonging to the order Pleosporales. Heterocycles

81: http://dx.doi.org/10.3987/COM-10-12009

Hu DM, Zhu H, Cai L, Hyde KD, Zhang KQ. 2007. Sirothecium triseriale, a new chirosporous

anamorphic species from China. Cryptogamie Mycologie 28: 311-314.

Hu DM, Cai L, Chen H, Bahkali AH, Hyde KD. 2010a. Four new freshwater fungi associated with

submerged wood from Southwest Asia. Sydowia 62: 191-203.

Hu DM, Cai L, Chen H, Bahkali AH, Hyde KD. 2010b. Fungal diversity on submerged wood

in a tropical stream and an artificial lake. Biodiversity and Conservation 19: 3799-3808.

http://dx.doi.org/10.1007/s10531-010-9927-5

Hyde KD. 1992. Tropical Australian freshwater fungi IH. Annulatascus velatispora gen. et sp. nov.

Annulatascus bipolaris sp. nov. and Nais aquatica sp. nov. Australian Systematic Botany 5:

117-124. http://dx.doi.org/10.1071/SB9920117

Hyde KD, Wong SW. 2000. Annulatascus fusiformis sp. nov., a new freshwater ascomycete from the

Philippines. Mycologia 92: 553-557. http://dx.doi.org/10.2307/3761515

Hyde KD, Ho WH, Jones EBG, Tsui CKM, Wong WSW. 2000. Torrentispora fibrosa gen. sp.

nov. (Annulatascaceae) from freshwater habitats. Mycological Research 104: 1399-1403.

http://dx.doi.org/10.1017/S0953756200002781

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi, 10" edition. Cromwell

Press, Trowbridge. 771 p.

Luo J, Yin JE, Cai L, Zhang KQ, Hyde KD. 2004. Freshwater fungi in Lake Dianchi, a heavily polluted

lake in Yunnan, China. Fungal Diversity 16: 93-112.

Raja HA, Schmit JP, Shearer CA. 2009. Latitudinal, habitat and substrate distribution patterns of

freshwater ascomycetes in the Florida Peninsula. Biodiversity and Conservation 18: 419-455.

http://dx.doi.org/10.1007/s10531-008-9500-7

Ranghoo VM, Tsui CKM, Hyde KD. 2001. Brunneosporella aquatica gen. et sp. nov., Aqualignicola

hyalina gen. et sp. nov., Jobellisia viridifusca sp. nov. and Porosphaerellopsis bipolaris sp. nov.

(ascomycetes) from submerged wood in freshwater habitats. Mycological Research 105:

625-633. http://dx.doi.org/10.1017/S0953756201003793

Tsui CKM, Ranghoo VM, Hodgkiss IJ, Hyde KD. 2002. Three new species of Annulatascus

(Ascomycetes) from Hong Kong freshwater habitats. Mycoscience 43: 383-389.

http://dx.doi.org/10.1007/s102670200056

Vijaykrishna D, Jeewon R, Hyde KD. 2006. Molecular taxonomy, origins and evolution of freshwater

ascomycetes. Fungal Diversity 23: 351-390.

Wong MKM, Goh TK, Hodgkiss IJ, Hyde KD, Ranghoo VM, Tsui CKM, Ho WH, Wong WSW,

Yuen TK. 1998a. Role of fungi in freshwater ecosystems. Biodiversity and Conservation 7:

1187-1206. http://dx.doi.org/10.1023/A:10088837 16975

88 ... Hu &al.

Wong SW, Hyde KD, Jones EBB. 1998b. Annulatascaceae, a new ascomycete family from the tropics.

Systema Ascomycetum 16: 17-25.

Zhang KQ, Wang L, Dong JY, Song HC, Shen KZ, Wang LM, Sun R, Wang CR, Li GH, Li L. 2008.

Screening and isolation of antibacterial activities of the fermentative extracts of freshwater

fungi from Yunnan Province, China. Annals of Microbiology 58: 579-584.

http://dx.doi.org/10.1007/BF03175561

Zelski SE, Raja HA, Miller AN, Barbosa FR, Gusmao LFP, Shearer CA. 2011la. Longicollum

biappendiculatum gen. et sp. nov., a new freshwater ascomycete from the Neotropics.

Mycosphere 2: 539-545.

Zelski SE, Raja HA, Miller AN, Shearer CA. 2011b. Chaetorostrum quincemilensis, gen. et sp. nov.,

a new freshwater ascomycete and its Taeniolella-like anamorph from Peru. Mycosphere 2:

593-600.

MY COTAXON

http://dx.doi.org/10.5248/120.89

Volume 120, pp. 89-98 April-June 2012

New records of little-known species of Carbomyces

(Carbomycetaceae, Ascomycota)

GABRIEL MORENO”™*, MARCOS LIZARRAGA’*, MARTIN ESQUEDA3,

RICARDO GALAN’ & PABLO ALVARADO’

' Dpto. de Biologia Vegetal, Facultad de Biologia, Universidad de Alcala,

Alcala de Henares, Madrid 28871 Spain

? Dpto. de Ciencias Quimico-Biologicas, Programa de Biologia, Universidad Auténoma de Ciudad

Juarez, Anillo Envolvente Pronaf y Estocolmo s/n,

Ciudad Juarez, Chihuahua 32300 México

* Centro de Investigacion en Alimentacion y Desarrollo, A.C.,

Apartado Postal 1735, Hermosillo, Sonora, C.P. 83000 México

* CORRESPONDENCE TO: gabriel.moreno@uah.es

ABSTRACT— ‘The genus Carbomyces is reported for the first time for the Mexican mycobiota.

Carbomyces emergens, C. gilbertsonii, and C. longii were collected in the Chihuahuan desert,

Mexico. Carbomyces emergens was more frequently collected and more widely distributed,

while C. gilbertsonii and C. longii each have a restricted distribution and represent second

records worldwide. The phenology, chorology, and macro-/microscopic characters (including

spore ornamentation by SEM) are outlined for all species.

Key worps— Pezizales, taxonomy, truffles

Introduction

Several collections of hypogeous and semi-hypogeous species have been

recorded in recent studies of gasteroid and secotioid fungi in Chihuahua State,

including the central Chihuahuan desert, Mexico (Lizarraga et al. 2010; Moreno

et al. 2010). Fungi reported here were found on the soil surface and wrongly

considered in situ to represent basidiomycetes. However, careful examination

revealed that these fungi represent two different ascomycete species in the

genus Carbomyces Gilkey.

Hypogeous ascomycetes, which are widely distributed, occur in different

forest types (e.g., coniferous dominated by Pinaceae, angiospermous with

Betulaceae, Salicaceae, Fagaceae, and Myrtaceae). Some even associate with

herbaceous plants and shrubs, such as those in the Cistaceae. Cistaceous plants

found in arid areas of the Mediterranean basin, southern Europe, northern

90 ... Moreno & al.

Africa, and the Middle East play an important role in plant conservation by

protecting sensitive areas from erosion from heavy rain and human activity. In

these areas, cistaceous plants can form mycorrhizal associations with several

different fungal genera such as Choiromyces, Picoa, Terfezia, Tirmania, and

Tuber.

However, in America, Cistaceae does not occur and arid lands are commonly

dominated by Cactaceae, Fouquieriaceae, Mimosaceae, and Zygophyllaceae.

Carbomyces was described from deserts of New Mexico and California (USA)

for C. emergens and C. longii by Gilkey (1954), who published macro- and

microscopical drawings of both species (Gilkey 1955).

Trappe & Weber (2001) later described in detail those two species and

transferred a third species, C. gilbertsonii, into Carbomyces from the puffball

basidiomycete genus Abstoma G. Cunn. where it had previously been misplaced

(Zeller 1944). These truffles appear endemic to North American deserts,

occurring in the Chihuahuan, Sonoran, and Mohave deserts from New Mexico

to southern California, where they appear from September through April.

Carbomyces, which is adapted to extreme xeric conditions, differs from other

hypogeous ascomycetes in globose emergent ascomata, a two-layered peridium,

and a powdery gleba at maturity resulting from the disintegrated asci (a feature

also shared with Elaphomyces spp. and Ruhlandiella berolinensis Henn.).

Most specimens have been found ex situ since they are easily wind-blown

when mature (lying dry and loose) or eaten and transported (when immature?)

by rodent mycophagists (Zak & Whitford 1986), so that fungal populations

can be easily intermixed and subsequently confused. Nevertheless, in the

Chihuahuan desert (Mexico), we found pure (i.e. non-intermixed) populations

in situ that appeared macroscopically very similar, to other known species based

on spore ornamentation (smooth vs. rough) and shape (ellipsoid vs. globose).

Gilkey (1954) placed Carbomyces in the Terfeziaceae, but Trappe (1971)

erected the monotypic Carbomycetaceae for the genus, a taxonomic position

later accepted by several authors (Trappe 1979, Trappe & Weber 2001, Lumbsch

& Huhndorf 2007). Nevertheless, the true relationship between Carbomyces and

other ascomycetes was unknown until recently (Hansen & Pfister 2006). The

putatively non-mycorrhizal genus Carbomyces is thought to be ectomycorrhizal

(ECM), although no ECM hosts have been identified in its natural habitat

(Trappe 1971; Zak & Whitford 1986; Trappe & Weber 2001).

In the present workall three Carbomyces species —C. emergens, C. gilbertsonii,

C. longii— are reported from Mexico for the first time and redescribed from

recently collected specimens.

Materials & methods

The ascocarps were studied in situ following general methods (Cifuentes et al.

1986, Moreno & Manjén 2010). Dried material was studied at the lab using KOH 5%,

Carbomyces in Mexico ... 91

Melzer’s reagent (MLZ), Cotton blue (CB), Congo red 1%, and Hoyer’s medium for

microscopical observations. A Nikon Eclipse 80i phase-contrast microscope was used

for spore measurements (on dead dried material) under oil immersion including surface

structures such as spines or warts (on 25 spores).

Spore ornamentation was also studied under a scanning electron microscope Zeiss

DSM 950. Critical point drying technique was performed on the samples prior to

mounting, following Moreno et al. (1995).

The collections are kept in the Herbarium UAC] of the “Departamento de Ciencias

Basicas, Universidad Autonoma de Ciudad Juarez,’ Mexico, with duplicate sets in

Herbarium AH of the Universidad de Alcala de Henares and the macromycetes

collection of the Centro de Estudios Superiores del Estado de Sonora (CESUES).

High-quality DNA extract could be obtained from only one fruiting body of

Carbomyces emergens (AH 39344), probably due to the extreme climatic conditions and

high temperatures over long periods to which the other samples were naturally exposed.

DNA extraction and PCR protocols followed Checa et al. (2012). ITS (JQ023163) and

28S nLSU (JQ023164) regions were sequenced, deposited in GenBank, and aligned

with their closest BLAST matches from sequences deposited by Ferdman et al. (2005),

Hansen et al. (2001, 2005), Trappe et al. (2010), and others.

Taxonomy

Carbomyces emergens Gilkey, N. Amer. Fl., Ser. 2, 1: 27 (1954) FIGS 1-3, 11-13

Ascomata subglobose, ovoid to pyriform, 1.2-3 x 1.5-4.4 cm, scattered or

occasionally in small groups. PERIDIUM coriaceous, two-layered: outer layer

very thin, membranous, initially whitish but finally yellow-brownish to dirty

greyish, firmly attached, cracking by fissures or polygonal plates; inner layer

ochraceous to dark brown, 0.2-0.6 mm thick. GLEBA multilocular, although

apparently seeming to be solid, dark brown with sterile, lighter veins forming

a fragile reticule; spore mass pulverulent, dark with olivaceous or vinaceous

tints. Asci globose to subglobose or ovoid, 8-spored, 40-55 x 30-5 um, with

thin brownish walls disintegrating at maturity. Ascospores hyaline to light

yellowish, globose, inamyloid, 9-14 um diam., having a thick wall (1.2-1.5 um)

finely punctate when fully mature (as seen at 400x under LM, feebly verrucose

under the SEM), but appearing smooth when immature.

SPECIMENS EXAMINED: MEXICO, Cuinvuanua: Municipality of Juarez, in arid land:

20.IV.2005 R. Silva (UACJ 06); km 30 road Ciudad Juarez to Casas Grandes, next to

Larrea tridentata Coville, 3.VIII.2008 M. Lizarraga, H. Pelayo (UAC] 1166 in AH 39344;

GenBank Acc. Nos.—ITS: JQ023163, LSU: JQ023164); 31°23’20.21”N 106°23°32.94” W,

alt. 1312 m, 27.[I1.2010, M. Lizarraga, M. Vargas, D. Lopez (UACJ 1555); 8.V.2010

M. Lizarraga, D. Lopez, F. Félix, M. Vargas (UACJ 1556); 15.1X.2010 (UACJ 1684);

6.X1.2010 M. Vargas, M. Lizarraga (UACJ 1687); 31°22’47.44”°N 106°24°3.89"W, alt.

1319 m, 22.X.2010 (UACJ 1688; 30.X.2010 (UACJ 1700); 6.XI.2010 (UACJ 1689);

31°20°13.47°N 106°23°32.95"W, alt. 1334 m, 15.V.2010 M. Vargas, F. Félix, M. Lizarraga,

D. Lopez (UACJ 1686); 15.1X.2010 M. Vargas, M. Lizarraga (UAC] 1685); 10.IV.2011

(UACJ 1683); 31°19°39.98”N 106°30°7.43”W, alt. 1280 m, 23.V.2010 M. Lizarraga, D.

92 ... Moreno & al.

Lépez, M. Vargas (UAC] 1557); 2.X.2010 M. Lizarraga, M. Vargas, D. Saenz (UAC]

1628). Municipality of Ojinaga, La Mula, 29°14’24.0”N 104°26°09.0"W, alt. 1380 m,

24.V.2009 M. Lizarraga, C. Salazar, D. Saenz, D. Lopez, A. Gutierrez, E. Hernandez (HM

22 in CESUES 7550); 24.X.2009 M. Lizarraga, A. Sanchez, A. Gutierrez, C. Salazar, D.

Lopez (HM 171 in CESUES 7551).

REMARKS— Carbomyces emergens has been repeatedly described (e.g., Trappe

& Weber 2001) as having smooth or minutely roughened ascospores, but our

studies show they are clearly warted at or above 400x. This may be because

fully mature ascomata had not been found until now. The species could be

macroscopically confused with C. longii and C. gilbertsonii, which share a

similar habitat, but the species are easily differentiated by shape and spore

ornamentation. Small sporocarps of C. emergens could easily be misidentified

in situ as species of the basidiomycete genus Arachnion Schwein. (Agaricaceae)

but in Arachnion the gleba lacks ascal remnants and contains a capillitium, and

small (< 6 um diam.) verrucose spores.

Zak & Whitford (1986) described the hypogeous development of Carbomyces

emergens and reported the ascomata’s apparent ingestion by rodents after

collecting a few dried sporocarps in Northern Chihuahuan desert. Despite

firm evidence, they suggested that fruiting probably occurs during late winter

(January-March). Our collections UACJ 06 and UACJ 1557 are assumed

to be immature, while the remaining specimens show an evident mature

development.

Trappe & Weber (2001) regard C. emergens as the most common and

widely distributed Carbomyces species, although reported only from American

Southwest —Arizona, California, and New Mexico. Here we report it for the

first time in Mexico as an apparently common species of the central Chihuahuan

desert.

Carbomyces gilbertsonii N.S. Weber & Trappe, Harvard Pap. Bot. 6: 212 (2001)

[nom. nov.] FIGs 4-7, 14-16

= Abstoma longii Zeller, Mycologia 36: 628 (1944) [non Carbomyces longii Gilkey]

ASCOMATA immersed and finally superficial, globose to subglobose,

laterally compressed to irregular, 2-3 cm diam. PERIDIUM two-layered, mostly

persistent, ferruginous at maturity. GLEBA solid, pulverulent, brown to dark

brown. Ascr subglobose, 8-spored, 40-65 um diam., with thin light brown walls

disintegrating at maturity. Ascosporgs hyaline to brownish with age, globose

to subglobose, inamyloid, 20-25 um, ornamented with spine-like elements up

to 2 um long (LM). Under SEM such “spines” consist in small clumps of slender

apically fused baculae, forming pyramids with hollow bases.

SPECIMENS EXAMINED: MEXICO, CurHuaunvua: Municipality of Juarez, 31°22’47.44"N

106°24°3.89" W, alt. 1319 m, in arid soil near to Larrea tridentata, Prosopis sp. and

Opuntia sp., 23.X.2010 M. Lizarraga, M. Vargas, (UACJ 1559, duplicate in AH 39346);

Carbomyces in Mexico ... 93

Fics. 1-3: Carbomyces emergens AH 39344: 1. Ascocarps. 2. Asci with spores. 3. Spores (SEM).

Fics. 4-7 Carbomyces gilbertsonii AH 39347: 4. Ascocarps. 5. Asci with spores. 6. Free spores. 7.

Spores (SEM). Fics. 8-10 Carbomyces longii AH 39345: 8. Ascocarps. 9. Free spores. 10. Spore

(SEM). Scale bars: 1, 4, 8 = 1 cm; 2, 5, 6, 9 = 10 um; 3, 10 = 5 um; 7 = 10 um.

6.XI.2010, (UACJ 1561 in AH 39347); (UACJ 1691); 31°23°20.21”N 106°23’32.94°W, alt.

1312 m, (UACJ 1690); 31°22’21.46”N 106°33’59.26”W, alt. 1225 m, 26.III.2011 (UACJ

1689).

REMARKS— Carbomyces gilbertsonii could be macro- and microscopically

confused with C. longii, but small, ellipsoid spores (17-19 x 12-13 um)

distinguish the latter species. Old Scleroderma cepa specimens could be

confused with C. gilbertsonii specimens without ascal remains, but the smaller

(9-12 um) spores of S. cepa spores are diagnostic. It is noteworthy that

94 ... Moreno & al.

Fics. 11-13: Carbomyces emergens AH 39344: spores (SEM. Fics. 14-16 Carbomyces gilbertsonii

AH 39347: 14-15. Spores (SEM); 16. Spore ornamentation (detail, SEM). Fics. 17-19 Carbomyces

longii AH 39345: 17-18. Spores under SEM. 19. Spore ornamentation (detail, SEM). Scale bar:

11-13, 16 = 2 um; 14-15, 17-18 = 5 um; 19 = 1 um.

C. gilbertsonii was previously known only from the type locality (New Mexico,

U.S.A.); our collections add Chihuahua (Mexico) to its distribution.

Carbomyces longii Gilkey, N. Amer. Fl., Ser. 2, 1:27 (1954) FIGs 8-10, 17-19

Ascomarta subglobose, laterally compressed, pyriform to irregular, 2-3 cm

diam., 2.5-3 x 1.5-2 cm. PERIDIUM two-layered, persistent, outer layer white to

greyish, later light brown to dark brown with reddish tinges at maturity. GLEBA

pulverulent, dark brown to brown, becoming olivaceous. Asci evanescent, only

a few fragments of brownish, disintegrated walls were seen. ASCosPoREs light

yellowish, ellipsoid, 17-19 x 12-13 um, inamyloid, ornamented with regularly

Carbomyces in Mexico ... 95

distributed spines < 1 um long (LM). Under SEM, the spines are scattered,

usually with bent apices.

SPECIMENSEXAMINED: MEXICO, CHIHUAHUA: Municipality of Ahumada, 31°07'32.0”N

106°29°48.0"W, alt. 1280 m, in soil next to Larrea tridentata, 24.V1I.2009 M. Lizarraga,

C. Salazar, D. Saenz, D. Lopez, A. Gutierrez, E. Hernandez (UAC] 1699). Municipality of

Juarez, 31°23°20.21”N 106°23°32.94"W, alt. 1312 m, 27.III.2010 M. Lizarraga, M. Vargas,

D. Lopez (UACJ 1557 in AH 39017); 3.V.2010 (UACJ 1698); 15.V.2010 M. Lizarraga,

D. Lopez, M. Vargas, EF. Félix (UACJ 1558); 15.1X.2010 M. Lizarraga, M. Vargas (UAC]

1697); 22.X.2010 (UAC] 1696); 6.XI.2010 (UAC] 1694); 31°22’47.44”N 106°24°3.89" W,

alt. 1319 m, 30.X.2010 (UACJ 1562, duplicate in AH 39345); 6.X1.2010 (UACJ 1693);

31°22’21.46"N 106°33’59.26”W, alt. 1225 m, 26.11.2011 (UACJ 1695).

REMARKS— Since C. longii asci are fully evanescent, mature sporocarps could

be confused with Arachniopsis albicans Long (Agaricaceae), but the latter is

distinguished by small (3-5 x 3-4 um) oval feebly verrucose basidiospores and

a hyaline capillitium. Co-occurring with C. gilbertsonii, C. longii was previously

known only from the type locality (New Mexico, U.S.A.). Our report from

Chihuahua, Mexico, represents the second world record.

Key to Carbomyces species in the Chihuahuan desert, Mexico

1 Spores globose and smooth to subsmooth (LM) or feebly warted (SEM) ....C. emergens

NPR PoC anG PETY Fe crac lh ees AM chai ah Aaa a chai OM eh e ohdr at eed a dee gM lec, eats eee 2

2-Spores-ellipsoidsscatiered:Spities, 5 2 fern, Horne, hess le lene ele ng geen a C. longii

2’ Spores globose to subglobose, spines in small clumps forming pyramids

ee ee tee PRR PMs Rieko Rumen PRLS PR Uh Wire ota C. gilbertsonii

Molecular results

28S nLSU alignment comprised 39 sequences with 215 variable sites among

563 bases, 133 of them parsimony-informative. Bayesian and parsimony

analyses were generally consistent with each other (Fig. 20) and supported the

three main groups already recognized in Lzessoe & Hansen (2007). The first

group comprises Kalaharituber, Iodowynnea, Hydnotryopsis, Sarcosphaera,

and several Peziza spp. The second group includes Mattirolomyces, Elderia,

and a different set of Peziza spp. The third group includes Terfezia, Tirmania,

Mycoclelandia, Ulurua, Eremiomyces, Ruhlandiella, Hapsidomyces, Plicaria, and

a third set of Peziza spp. The analyses show Carbomyces as somewhat related

to Kalaharituber (Pezizaceae) in the first group (Fig. 20), partially confirmed

by BLAST of the only ITS sequence obtained, which linked C. emergens to

Terfezia and Kalaharituber (84% and 83% identity in 68% and 65% coverage,

respectively), in accordance with data obtained by Karen Hansen (pers.

comm.).

96 ... Moreno & al.

AY544677 Ascobolus carbonarius

AF335148 Peziza luteoloflavida

AF335159 Peziza retrocurvata

AF335156 Peziza phyllogena

AY789328 Peziza phyllogena

gg 7 AY500533 Hapsidomyces venezuelensis

100 L_ AY500553 Plicaria anthracina

400 JN032129 Eremiomyces magnisporus

100 AF435823 Eremiomyces echinulatus

AF335175 Ruhlandiella berolinensis

“a GQ231750 Ulurua nonparaphysata

100 GQ231747 Mycoclelandia bulundari

79 GQ231743 Mycoclelandia arenacea

109 es AF335135 Peziza depressa

100 AF335132 Peziza badia

PL AF335147 Peziza limnaea

95 AF 499448 Terfezia boudieri

100 AF435824 Terfezia claveryi

94 AF335177 Tirmania nivea

100 L__ AF335178 Tirmania pinoyi

AY500548 Peziza lobulata

AF335165 Peziza subviolacea

GQ231753 Mattirolomyces austroafricanus

ae HQ660382 Mattirolomyces spinosus

400 7 GQ231734 Elderia arenivaga

100 L GQ231737 Elderia arenivaga

71 4100 AF335146 Peziza howsei

Po 100 L AF335140 Peziza emileia

99 AF335158 Peziza proteana

100 AY500545 Peziza exogelatinosa

ats AY500552 Peziza vesiculosa

100 a AF335154 Peziza nivalis

100 AF335145 Peziza fimeti

AF335174 Pfistera pyrophila

UACJ 1166 Carbomyces emergens

AY232726 Kalaharituber pfeilii

AF335118 lodowynnea auriformis

400 AF335115 Hydnotryopsis setchellii

100 NG 027607 Sarcosphaera crassa

ob AY500550 Peziza obtuspiculata

100 | AF335157 Peziza polaripapulata 0.05

Carbomyces in Mexico ... 97

Acknowledgements

The authors thank CONABIO (GTO016 project) for the financial support of this

study. One of the authors (M. Lizarraga) extends his gratitude to the collectors and the

“Vicerrectorado de Investigacién e Innovacién” of the Universidad de Alcala for their

assistance in obtaining financial support for his research stage during two months at

the Universidad de Alcala. We express our gratitude to Dr. G. Pacioni and Lda. M?.M.

Dios for reviewing the manuscript and adding a number of useful comments. We wish

to express our gratitude to Mr. D.W. Mitchell for his assistance in correcting the English

and to Mr. A. Priego and Mr. J.A. Pérez (Electron Microscopy Service, University of

Alcala) for their invaluable help with the SEM. We also thank L. Monje and A. Pueblas

of the “Gabinete de Dibujo y Fotografia Cientifica” at the Universidad de Alcala for their

invaluable help in the digital preparation of the photographs. We are also grateful to Dr.

J. Rejos, curator of the AH herbarium for providing specimens management. Finally we

would like to thank Dr. Karen Hansen for her useful comments on the results and Aldo

Gutiérrez (CIAD) for formatting the text.

Literature cited

Checa J, Blanco MN, Moreno G, Manjon JL, Pasaban P, Alvarado P. 2012. Amplistroma longicollis,

a new species and its anamorph state described and sequenced from Europe. Mycological

Progress. http://dx.doi.org/10.1007/s11557-011-0776-8

Cifuentes J, Villegas M, Pérez-Ramirez L, Sierra S. 1986. Hongos. 55-64, in: A Lot, F Chiang (eds).

Manual de herbario. Consejo Nacional de la Flora de México, A.C., México.

Ferdman Y, Aviram S, Roth-Bejerano N, Trappe JM. 2005. Phylogenetic studies of Terfezia pfeilii

and Choiromyces echinulatus (Pezizales) support new genera for southern African truffles:

Kalaharituber and Eremiomyces. Mycological Research 109: 237-245.

http://dx.doi.org/10.1017/S0953756204001789

Gilkey HM. 1954. Tuberales. North American Flora 2, 1: 1-36.

Gilkey HM. 1955 [“1954”]. Taxonomic notes on Tuberales. Mycologia 46: 783-793.

Hansen K, Pfister DH. 2006. Systematics of the Pezizomycetes - the Operculate Discomycetes.

Mycologia 98: 1029-1040. http://dx.doi.org/10.3852/mycologia.98.6.1029

Hansen K, Laessge T, Pfister DH. 2001. Phylogenetics of the Pezizaceae, with an emphasis on

Peziza, Mycologia 93: 958-990. http://dx.doi.org/10.2307/3761760

Hansen K, LoBuglio KF, Pfister DH. 2005. Evolutionary relationships of the cup-fungus genus

Peziza and Pezizaceae inferred from multiple nuclear genes: RPB2, B-tubulin, and LSU rDNA.

Molecular Phylogenetics and Evolution 36(1): 1-23.

http://dx.doi.org/10.1016/j.ympev.2005.03.010

Leessoe T, Hansen K. 2007. Truffle trouble: what happened to the Tuberales? Mycological Research

111: 1075-1099. http://dx.doi.org/10.1016/j.mycres.2007.08.004

Lizarraga M, Esqueda M, Gutierrez A, Pifia C, Barredo-Pool F. 2010. El género Disciseda (Agaricales,

Agaricaceae) en la Planicie Central del Desierto Chihuahuense. Rev. Mex. Micol. 32: 17-23.

Fic. 20: 28S nLSU consensus phylogram constructed in MrBayes 3.1 showing the most probable

relationships between Carbomyces emergens and its closest relatives in the Pezizaceae. Numbers close

to nodes represent maximum parsimony bootstrap proportions (upper), and Bayesian posterior

probabilities (lower). Only nodes supported by at least one inference method were annotated.

98 ... Moreno & al.

Lumbsch HT, Huhndorf SM. 2007. Outline of Ascomycota - 2007. Myconet 13: 1-58.

Moreno G, Manjén JL. 2010. Guia de hongos de la Peninsula Ibérica. Ediciones Omega. 1417 p.

MorenoG, Altés A, OchoaC, Wright JE. 1995. Contribution to the study of the family Tulostomataceae

in Baja California, Mexico. I. Mycologia 87: 96-120. http://dx.doi.org/10.2307/3760953

Moreno G, Lizarraga M, Esqueda M, Coronado ML. 2010. Contribution to the study of gasteroid

and secotioid fungi of Chihuahua, Mexico. Mycotaxon 112: 291-315.

http://dx.doi.org/10.5248/112.291

Trappe JM. 1971. A synopsis of the Carbomycetaceae and Terfeziaceae (Tuberales). Transactions of

the British Mycological Society 57: 85-92. http://dx.doi.org/10.1016/S0007-1536(71)80083-9

Trappe JM. 1979. The orders, families, and genera of hypogeous Ascomycotina (truffles and their

relatives). Mycotaxon 9: 297-340.

Trappe JM, Weber NS. 2001. North American desert truffles: the genus Carbomyces (Ascomycota,

Carbomycetaceae). Harvard Papers in Botany 6: 209-214.

Trappe JM, Kovacs GM, Claridge AW. 2010. Comparative taxonomy of desert truffles of the

Australian outback and the African Kalahari. Mycological Progress 9: 131-143.

http://dx.doi.org/10.1007/s11557-009-0612-6

Zak JC, Whitford WG. 1986. The occurrence of a hypogeous ascomycete in the northern

Chihuahuan desert. Mycologia 78: 840-841. http://dx.doi.org/10.2307/3807532

Zeller SM. 1944. Representatives of the Mesophelliaceae in North America. Mycologia 36: 627-637.

http://dx.doi.org/10.2307/3754840

MY COTAXON

http://dx.doi.org/10.5248/120.99

Volume 120, pp. 99-103 April-June 2012

Cantharellus zangii, a new subalpine basidiomycete

from southwestern China

XIAO-FEI TIAN}, BART BuyCK‘, SHI-CHENG SHAO””,

Pei-Guti Liu’ & YAN FANG>

' Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany,

Chinese Academy of Sciences, 132 Lanhei Road, Kunming,650204, China

? Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences,

88 Xuefu Road, Kunming, 650223, China

> Graduate University of Chinese Academy of Sciences,

19(A) Yuquan Road, Beijing, 100049, China

* Muséum national d’histoire naturelle, Département de Systématique et Evolution,

Case Postale n°39. UMR 7205, 57, Rue de Cuvier, Paris, F- 75231, France

° Department of Analysis, Zibo Water Supply, 14 Gonggingtuan East, Zibo, 255000, China

*CORRESPONDENCE TO: pgliu@mail.kib.ac.cn

ABSTRACT — Cantharellus zangii is described and illustrated from subalpine forest (>3000 m)

in northwestern Yunnan, China. The diagnostic characteristics of C. zangii are the small thin-

fleshed fruit bodies with a long tapering fistulose stipe, a bright orange veined hymenophore,

thin-walled clamped hyphae, and large ellipsoid basidiospores. So far it is only known from

the type locality and its nearby area.

KEY worpDs — taxonomy, endemic species, ectomycorrhizal, Cantharellales

Introduction

Cantharellus Adans. ex Fr., the type genus of Cantharellaceae, was

established by Fries (1821) and later typified with C. cibarius Fr. by Earle (1905).

Thirteen species of Cantharellus have previously been reported from China:

C. appalachiensis R.H. Petersen, C. buccinalis Mont., C. cibarius, C. cinereus

(Pers.) Fr., C. cinnabarinus (Schwein.) Schwein., C. friesii Quél., C. lateritius

(Berk.) Singer, C. minor Peck, C. patouillardii Sacc., C. subalbidus A.H. Sm.

& Morse, C. tuberculosporus M. Zang, C. vaginatus S.C. Shao et al., and

C. yunnanensis W.F. Chiu (Teng 1963, Chiou 1973, Zang 1980, Zhuang 2001,

2005, Tian et al. 2009, Shao et al. 2011). However, some of these collections are

probably misidentified and need to be verified microscopically and molecularly,

100 ... Tian & al.

as has been done by Buyck et al. (2011) and Buyck & Hofstetter (2011) for

North America. During our Cantharellus research, we came across one small

species with a very slender stipe in the subalpine mixed forest of Shangri-La

(northwestern Yunnan, China), which we determined as distinct from already

described species in the genus. It is here described as Cantharellus zangii.

Materials & methods

Macro-morphological fruitbody features were recorded and photographed in the

field; color designations and codes follow Kornerup & Wanscher (1961). In basidiospore

measurements, [n/m/p] = n basidiospores measured from m basidiomata of p collections

in 5% KOH solution, with dimensions given as (a—)b-c(—d), where a and d are extremes

and b-c represent the interval range of 90% measured values. Q = basidiospore length/

width ratio; Q. = average Q of all basidiospores + standard deviation. Herbarium

abbreviation HKAS = Herbarium of Cryptogams, Kunming Institute of Botany, Chinese

Academy of Sciences.

Taxonomy

Cantharellus zangii X.F. Tian, P.G. Liu & Buyck, sp. nov. PLATE 1-5

MycoBank MB 563725

Differs from Craterellus tubaeformis by a plano-convex pileus and bright orange

hymenium and from Cantharellus queletii in the fistulose stipe and hymenium with

branching veins.

Type: China, northwestern Yunnan, Shangri La, Bitahai National Natural Reserve,

9.IX.2008, X.F Tian 417 (Holotype, HKAS55791).

ETYMOLOGY: zangii is in honor of Professor Zang Mu (28 Dec. 1930-10 Nov. 2011),

a distinguished Chinese mycologist who founded the Cryptogamic Herbarium in

Kunming Institute of Botany, Academia Sinica (KUN-HKAS), and pioneered the study

of cryptogam diversity in southwestern China.

MACROCHARACTERS — _ BASIDIOMATA small to medium, _ slender,

submembranaceous, campanuliform to infundibuliform. PrLeus 2-3 cm in

diam, at first umbonate then plane or slightly concave; squamules in the middle;

ochre to ochre yellow (4B8-4B6), with tiny cinereous to grey (4B8-4B6);

margin hygrophanous, smooth, with streaks, sometimes undulate, incurved at

first, then straight. Stipe 7-8 x 0.3-0.4 cm, cylindrical to laterally compressed,

sometimes slightly curved, attenuate towards base; entirely fistulose; dark

orange-yellow (4B5-4B6) to bright orange (4A6). LAMELLAE decurrent,

golden; 1.5-2 mm high, 1-2 mm apart, with transverse venations, with short

branched lamellulae near the edge, seldom anastomosing. CONTEXT about 0.2

cm thick in the cap center; dull white (1A2), fibrous. ODoR Osmanthus-like.

TASTE mild.

MICROCHARACTERS — BASIDIOSPORES [85/4/2] (8—)8.5-11 x (4.5-)5-6.5

(-7) um, Q = (1.3-)1.4-2(-2.2), Q. = 1.7 + 0.2, ellipsoid or adaxially slightly

Cantharellus zangii sp. nov. (China) ... 101

FiGurRE 1-5. Cantharellus zangii (HKAS 55791, holotype): 1. Fruit bodies. 2. Hyphal extremities at

the cap surface. 3. Tramal hyphae with clamps. 4. Basidiospores, often with many tiny oil drops. 5.

Hymenium (with lengthening basidioles and basidia).

102 ... Tian & al.

depressed and more or less reniform; thin-walled, hyaline, nearly colorless,

sometimes with tiny oil drops. Basrp1a 75-85 x 6-9 um, slender, clavate,

5(-6)-spored; sterigmata 5-6 um long. CysTip1A absent. LAMELLAR TRAMA

irregular, composed of colorless and loose hyphae, 3-5 um diam. PILEIPELLIS

a layer composed of mostly radially arranged hyphae, slightly brownish, 7-8

um in diam; the terminal cell more or less clavate; CLAmps very distinct and

abundant in all parts.

ECOLOGY & DISTRIBUTION — Single or in groups on the ground in subalpine

(>3000m) mixed forests dominated by Larix potaninii var. macrocarpa

Y.W. Law and Picea likiangensis (Franch.) E. Pritz. So far known only from

Shangri-La subalpine area, northwestern Yunnan, China.

ADDITIONAL SPECIMENS EXAMINED — CHINA. NORTHWESTERN YUNNAN, Shangri-

La: Big Ravine, alt. 3030 m, 16.IX.2007, Y.C. Li 1537 (HKAS 55743); Haba Snowy

Mountains, alt. 3000 m, 30.1X.2007, Feng Bang 182 (HKAS 55824).

Discussion

Based on its almost membranous pileus and veins as well as the hollow

stipe, Cantharellus zangii could be taken for Craterellus tubaeformis (Fr.) Quel.

(Redhead et al. 2002) at first glimpse. However, C. zangii often has an umbrella-

shaped slender fruit body with a centrally plane or protruding pileus, whereas

Cr. tubaeformis is horn-shaped with centrally depressed caps and has a grey or

grey-white hymenium, never bright orange like C. zangii.

The membranous cap and slender stipe of Cantharellus zangii are also similar

to the European C. queletii (Ferry) Corner (Corner 1966), which differs in a

somewhat solid stipe and the absence of branching veins in the hymenium.

Cantharellus zangii also has some striking similarities with a few of the

smaller American species (Petersen & Ryvarden 1971; Feibelman et al. 1996;

Buyck et al. 2010). The yellow C. minor, for instance, shares the exceptional

character of the fistulose stipe and thin-walled, voluminous hyphae in the

pileipellis. However, its overall size is smaller, it also has slightly smaller spores,

and the terminal cells in the pileipellis usually narrow at the extreme tips. The

yellowish brown C. tabernensis Feib. & Cibula is similar in size to C. zangii

with a very similar pileipellis, but the stipe is not hollow and the spores are

much smaller. The same is true for C. appalachiensis, a more brownish and

slightly more robust sister-species of C. tabernensis, possessing an identical

microscopy.

The small size and abundant clamps together with the thin-walled hyphal

extremities in the cap place this species in C. subgenus Parvocantharellus as

defined by Eyssartier & Buyck (2001), where it is further diagnosed by its large

spores and clavate terminal cells in the pileipellis.

Cantharellus zangii sp. nov. (China) ... 103

Acknowledgments

We wish to express our gratitude to Professor R. Petersen (University of Tennessee)

and Professor M. Verbeken (Ghent University) for reviewing the manuscript. Thanks

also given to Prof. Z.Y. Su and Dr. Z.L. Yang (Kunming Institute of Botany, Chinese

Academy of Sciences) and Dr. S. Pennycook (Manaaki Whenua Landcare Research,

New Zealand) for valuable suggestions. This work was supported by the National

Natural Science Foundation of China (No. 30770007 and 30800005), the Joint Funds

from Chinese National Sciences Foundation and Yunnan Province Government (No.

U0836604), Natural Science Foundation of Yunnan (Key project No.2007C0002Z) and

Foundation of Key Laboratory of Biodiversity and Biogeography, Kunming Institute of

Botany, CAS (No.2008004), as well as Yunnan International Collaborative Program of

innovation to strong provinces by Science and Technology (2009AC013).

Literature cited

Buyck B, Hofstetter V. 2011. The contribution of TEF-1 sequences to species delimitation in the

Cantharellus cibarius complex in the southeastern USA. Fungal Diversity 49: 35-46.

http://dx.doi.org/10.1007/s13225-011-0095-z

Buyck B, Lewis DP, Eyssartier G, Hofstetter V. 2010. Cantharellus quercophilus sp. nov. and its

comparison to other small, yellow or brown American chanterelles. Cryptogamie, Mycologie

bl 17—33:

Buyck B, Cruaud C, Couloux A, Hofstetter V. 2011. Cantharellus texensis sp. nov. from Texas,

a southern lookalike of C. cinnabarinus revealed by tef-1 sequence data. Mycologia 103:

1037-1046. http://dx.doi.org/10.3852/10-261

Chiou WE. 1973. Ten new species of Agaricales from Yunnan, China. Acta Microbiologica Sinica

13(2): 129-135.

Corner EJH. 1966. A monograph of cantharelloid fungi. Oxford University Press. London. 255 p.

Earle FS. 1905. The genera of the North American gill fungi. Bulletin of the New York Botanical

Garden 5: 373-383.

Eyssartier G., Buyck B. 2001. Novitates. Note nomenclaturale et systématique sur le genre

Cantharellus. Documents mycologiques 31(121): 55-56.

Feibelman TP, Bennett JW, Cibula WG. 1996. Cantharellus tabernensis: a new species from the

southeastern United States. Mycologia 88: 295-301. http://dx.doi.org/10.2307/3760934

Fries EM. 1821. Systema Mycologicum. Vol.1. Lundae.

Kornerup A, Wanscher JH. 1961. Farver i Farver. Politikens Forlag. Kobenhavn.

Petersen R, Ryvarden L. 1971. Notes on cantharelloid fungi. IV. Two new species of Cantharellus.

Svensk Botanisk Tidskrift 65: 399-405.

Redhead SA, Norvell LL, Danell E, Ryman S. 2002. Proposals to conserve the names Cantharellus

lutescens Fr.: Fr. and C. tubaeformis Fr.: Fr., Basidiomycota with conserved types. Taxon 51:

559-562. http://dx.doi.org/10.2307/1554875

Shao SC, Tian XE, Liu PG. 2011. Cantharellus in southwestern China: a new species and a new

record. Mycotaxon 116: 437-446. http://dx.doi.org/10.5248/116.437

Teng SC. 1963. Fungi of China. Science Press, Beijing, China.

Tian XE, Shao SC, Liu PG. 2009. Two notable species of the genus Cantharellus Adans (Cantharellales,

Basidiomycota) new to China. Edible Fungi of China 28(4):10-11

Zang M. 1980. Some new species of Basidiomycetes from the Xizang autonomous region of China.

Acta Microbiologica Sinica 20: 29-34.

Zhuang WY. 2001. Higher fungi of tropical China. Mycotaxon Ltd., Ithaca, New York.

Zhuang WY. 2005. Fungi of northwestern China. Mycotaxon Ltd., Ithaca, New York.

MY COTAXON

http://dx.doi.org/10.5248/120.105

Volume 120, pp. 105-118 April-June 2012

Seven lichen species new to Poland

MARTIN KuKWA"*, ANNA LUBEK’, RAFAEL SZYMCZYK? & ANNA ZALEWSKA‘*

"Department of Plant Taxonomy and Nature Conservation, University of Gdansk,

Al. Legionéw 9, PL-80-441 Gdansk, Poland

?Jan Kochanowski University, Institute of Biology, Swietokrzyska 15A, PL-15-406 Kielce, Poland

* EKOPROJEKT Environmental Survey Laboratory, Nowica 24, PL-14-405 Wilczeta, Poland

* Department of Botany and Nature Protection, University of Warmia and Mazury in Olsztyn,

pl. Lodzki 1, PL-10-727 Olsztyn, Poland

*CORRESPONDENCE TO: dokmak@ug.edu.pl

ABSTRACT — Biatora pontica, Buellia violaceofusca, Catillaria croatica, Parmelia ernstiae,

Placopsis lambii, Protoparmelia oleagina and Scoliciosporum curvatum are recorded as new

to Poland. Most reported species represent a group of crustose, often sterile lichens where

secondary chemistry plays an important role in the taxonomy. Characteristics of each species,

notes on similar taxa, distribution, and habitat are provided.

KEY worps — lichenized fungi, chemotaxonomy, lichen metabolites

Introduction

Since the publication of the list of lichens and lichenicolous fungi of Poland

(Faltynowicz 2003), numerous species have been added to the lichen biota of

the country, including some new to science (e.g. Flakus 2007, Kukwa & Kubiak

2007, Motiejunaité & Czyzewska 2008, Krzewicka 2009, Kukwa & Flakus 2009,

Lubek 2009, Zhurbenko et al. 2009, Sliwa & Flakus 2011). Most of these taxa

are either lichenicolous fungi or lichens belonging to critical groups requiring

analyses of secondary lichen metabolites.

In this paper, we report seven lichen species not previously reported from

Poland.

Material & methods

The material studied is deposited in BILAS, KTC, OLS, PRA, UGDA, UPS, and herb.

Printzen. Lichen substances were studied by thin-layer chromatography (TLC) using

the methods of Culberson & Kristinsson (1970) and Orange et al. (2001). The examined

localities are presented according to the modified ATPOL grid square system (Cieslinski

& Faltynowicz 1993, Kukwa et al. 2002).

106 ... Kukwa & al.

The species

Biatora pontica Printzen & Tonsberg, Biblioth. Lichenol. 86: 140. 2003.

CHARACTERISTICS — ‘The species is characterized by its thin, crustose and

areolate thallus with punctiform to confluent, yellowish or light green soralia;

areoles in non-sorediate parts reaching < 1.2 mm in diam. Apothecia are

flat to moderately convex, dark greyish-ochre to brownish-grey and 0.5-0.7

mm in diam.; sometimes they are immersed between soralia. Anatomically

apothecia are characterized by the presence of a blue pigment (Pontica-

blue) in the hypothecium, subhymenium, and (rarely) hymenium and

epihymenium; sometimes an additional red pigment (Pontica-red) is produced

in the hypothecium and hymenium. Ascospores are ellipsoid to bacilliform,

11.9-14.8 x 3.4-3.7 um. Biatora pontica produces thiophanic acid (major),

asemone, and an unidentified diagnostic substance called ‘pontica unknown,

sometimes with minor or trace amounts of other xanthones; soralia react C+

orange (Printzen & Tonsberg 2003). In all Polish specimens thiophanic acid,

asemone, and ‘pontica unknown’ were detected. In only one specimen were

apothecia developed, but the fertile material is only the second such record in

Europe (Printzen & Tonsberg 2003).

ECOLOGY & DISTRIBUTION — Previously B. pontica has been known from

scattered localities in Europe (Austria, Italy, Norway, Russia, Slovenia), Asia

(Turkey), and North America (USA) (Printzen & Tonsberg 2003, Santesson

et al. 2004, Urbanavichus 2010). It is an epiphytic lichen growing in forests on

deciduous trees in Europe and in Turkey, but in eastern North America it is

more commonly found on Picea orientalis (Printzen & Tonsberg 2003).

In Poland B. pontica has been recorded in three localities in northern Poland,

being always found on the bark of Carpinus betulus and Fagus sylvatica in the

bottom of shady river or streams valleys.

SPECIMENS EXAMINED — POLAND. DRAwSKIE LAKELAND. Drawienski National

Park, Stara Wegornia range in valley of the Plociczna river, c. 2 km NNE Ostrowite

village, 53°04’42-46"N 15°59'31-41”E, ATPOL grid square Cb-22, on Carpinus betulus,

08.05.2010, M. Kukwa 7948 (UGDA); ELBLAsSKA UPLAND. Valley of Srebrny Potok

stream, forest section No. 342, 54°10'28"N 19°27'55"E, ATPOL grid square Bd-06,

beech forest in the bottom of the valley, on Fagus sylvatica, 05.08.2007, R. Szymczyk

641 (OLS L-691, dupl. in herb. Printzen); valley of Grabianka river, forest section No.

183b, 54°16'42"N 19°31'27"E, ATPOL grid square Ad-96, beech forest, bottom of the

river valley, on Fagus sylvatica, 27.06.2009, R. Szymezyk 657 (OLS L-762, dupl. in herb.

Printzen).

ComMENTs — When fertile, B. pontica can be confused with B. britannica

Printzen et al. (sorediate), B. hypophaea Printzen & Tonsberg (esorediate),

and B. ocelliformis (Nyl.) Arnold (esorediate) —three other members of the

genus with grey apothecia. They can be easily separated chemically as they

produce argopsin (thalli react P+ orange-red) (Printzen et al. 2001, Printzen

Seven lichens new to Poland ... 107

& Tonsberg 2003); in Poland only B. ocelliformis has been recorded so far

(Faltynowicz 2003). Sterile specimens of B. pontica are morphologically very

similar to B. efflorescens (Hedl.) Rasanen, which, however, contains argopsin

(Printzen et al. 2001). Biatora chrysantha (Zahlbr.) Printzen and Mycobilimbia

epixanthoides (Nyl.) Vitik. et al. can also be confused with B. pontica, but they

contain gyrophoric acid (B. chrysantha) or no lichen metabolites are produced

(M. epixanthoides) (Tonsberg 1992, Printzen & Tonsberg 2003, Printzen & Otte

2005).

Biatora pontica can also be confused with Pyrrhospora quernea (Dicks.)

Korb. which is morphologically very similar when sterile (greenish sorediate

thallus). Both taxa contain thiophanic acid as the major secondary metabolite,

but P. quernea lacks the characteristic substance ‘pontica unknown’ (Tonsberg

1992, Printzen & Tonsberg 2003).

Buellia violaceofusca G. Thor & Muhr, Lichenologist 23: 11. 1991.

CHARACTERISTICS — ‘The species is characterized by its thin, usually

endophloeodal, pale grey or almost white thallus producing dark brownish

soralia with a violet tinge (but greenish when abraded). Soralia are maculiform,

scattered, slightly elevated and usually confluent; no apothecia have been

discovered so far. This lichen does not produce secondary metabolites (Thor &

Muhr 1991), but traces of terpenoids from tree bark can be detected by TLC.

ECOLOGY & DISTRIBUTION — Buellia violaceofusca is widespread but with

a scattered distribution (Coppins et al. 2009); so far it has been found only

in Europe in Austria (Poelt 1994), Belgium (Sérusiaux et al. 1999), Estonia

(Thor & Nordin 1998), Great Britain (Coppins et al. 2009), Norway (Gaarder &

Tonsberg 2010), Slovakia (Palice et al. 2006), and Sweden (Thor & Muhr 1991).

The species usually grows on bark of Quercus spp. in different types of forest,

but in Great Britain it has been reported also on Fraxinus excelsior (Coppins et

al. 2009).

In Poland it has been found only at one locality in bark crevices of old Quercus

sp. in a deciduous forest. Most probably the species is much more common in

the country but has been overlooked due to the inconspicuous thalli and the

habit of growing in deep crevices.

SPECIMEN EXAMINED — POLAND. KASZUBSKIE LAKELAND. Trdjmiejski Landscape

Park, Dolina Ewy valley, 54°24'35"N 18°30'20"E, ATPOL grid square Ac-89, oak-

hornbeam forest, on Quercus sp., 04.04.2004, M. Kukwa 3006 (UGDA L-10664).

REFERENCE MATERIAL EXAMINED — SWEDEN. VARMLAND. VISNUM KIL PAR.,

N6t6n nature reserve, Arskagen, near Lake Vaner, 59°04'N 14°01’E, 50 m, on the

northern side of old Quercus robur in deciduous wood, 27.05.1985, L.-E. Muhr 7911

(UPS L-17315 — holotype).

ComMENTS — Buellia violaceofusca is a very characteristic epiphytic species due

to its thin thallus, violet tinged soredia, and the absence of lichen metabolites.

108 ... Kukwa & al.

In Poland it can be confused with the morphologically similar B. griseovirens

(Turner & Borrer ex Sm.) Almb. Both species have dark outermost soredia, but

B. griseovirens has a thicker thallus and produces atranorin and norstictic acid,

with trace of connorstictic acid and sometimes unidentified pigments (in some

specimens only atranorin or only norstictic acid are present) (Thor & Muhr

1991, Tonsberg 1992, Coppins et al. 2009).

Buellia arborea Coppins & Tonsberg also develops pigmented soredia, but

it can be readily separated by the presence of atranorin and placodiolic acid

(Tonsberg 1992, Coppins et al. 2009); so far the species has not been reported

from Poland, but its discovery there is probable.

Catillaria croatica Zahlbr., Ann. Mycol. 4: 487. 1906.

= Lecania croatica (Zahlbr.). Kotlov, Novosti Sist. Nizsh. Rast. 37: 251. 2004.

CHARACTERISTICS — As the only modern description of C. croatica (by

Harris & Lendemer 2010) is based mainly on North American specimens, a

more detailed description is provided below. The thallus of this corticolous

lichen is light green-grey, superficial, well or poorly developed to immersed,

consisting of scattered or almost contiguous areoles. Soralia are numerous,

rounded, flat, convex or weakly capitate, discrete or patchily coalescing to

form a leprose crust. Soredia are in shades of green, but sometimes externally

pale brown pigmented. Apothecia are sessile, flat or weakly convex, with pale

orange-cream or medium brown discs and paler margins. Anatomically, all

structures are colourless except the pale brownish hypothecium. Asci are

8-spored with narrowly fusiform, 0-1-septate ascospores measuring 12-17

x 4.5 um. Pycnidia were not found in Polish material. The species does not

produce lichen substances detectable by TLC.

ECOLOGY & DISTRIBUTION — Rarely reported from Europe: Austria,

Belgium, Croatia, France, Luxembourg, Romania, Slovakia, Slovenia and

Ukraine (Printzen 1995, Mrak et al. 2004, Hafellner et al. 2005, Eichler et al.

2010, Vondrak et al. 2010), and also known from North America (Harris &

Lendemer 2010). The species grows in forests, usually on bark of broad-leaved

trees, rarely also shrubs (e.g. Sambucus sp.).

In Poland, C. croatica has been found in only one locality, a well-preserved,

humid deciduous forest not seriously influenced by forest management, growing

in association with Anisomeridium polypori (Ellis & Everh.) M.E. Barr, Biatora

ocelliformis, Lepraria lobificans Nyl., L. vouauxii (Hue) R.C. Harris, Opegrapha

rufescens Pers., Pertusaria pertusa (L.) Tuck., and Bacidia sp.

SPECIMEN EXAMINED — POLAND. BIELSK PLAIN. Bialowieski National Park, forest

section No. 273, 52°43°32”N, 23°53’27°E, ATPOL grid square Cg-46, on Acer sp.,

27.06.2009, A. Lubek s.n. (KTC).

ComMMENTS — Catillaria croatica is an inconspicuous, easily overlooked,

corticolous lichen with a sorediate thallus lacking lichen metabolites, with small

Seven lichens new to Poland ... 109

pale apothecia and 0-1-septate ascospores. The species is morphologically very

similar to Mycobilimbia epixanthoides: both are sorediate, lack lichen substances,

and produce pale biatorine apothecia. When fertile, the material is much easier

to determine as both species differ in the type of spores: C. croatica produces

0-1-septate ascospores, whereas M. epixanthoides has 3-septate ascospores. In

the sterile state they may be separated based on substrate preference: Catillaria

croatica grows exclusively on the bark of trees, and M. epixanthoides prefers

bryophytes and is rarely corticolous (Hafellner et al. 2005, Harris & Lendemer

2010). The corticolous material of the latter, according to the cited sources,

could be distinguished by irregular young soralia forming larger irregular

patches, while soralia of C. croatica remain discrete and round, even when they

become crowded with age. From our experience, this might not be true in all

cases and some sterile samples may remain undetermined.

Several other sorediate species, which are morphologically similar to

C. croatica (e.g. Biatora efflorescens or B. chrysantha), can be confused with this

lichen, but they are all readily distinguished by their secondary metabolites

(e.g., Printzen 1995).

Catillaria croatica has been recently transferred to Lecania A. Massal. by

Kotlov (2004), but molecular studies have shown that it nests in Bilimbia De

Not. s.l. and is not phylogenetically related to Lecanias. str. (Reese Neesborg et al.

2007). Therefore we prefer to keep the species in the admittedly heterogeneous

genus Catillaria A. Massal.

Parmelia ernstiae Feuerer & A. Thell, Mitt. Inst. Allg. Bot. Hamburg 30-32: 52. 2002.

CHARACTERISTICS — This foliose lichen is characterized by its heavily

pruinose thalli and isidia, small and non-overlapping lobes, commonly

present lobulae, and the production of atranorin (minor amount, together

with chloratranorin), salazinic acid (major amount, with minor amount of

consalazinic acid), lobaric, lichesterinic, protolichesterinic, nephrosteranic,

isonephrosterinic (all minor amounts) and protocetraric (trace amount) acids

(Feuerer & Thell 2002, Thell et al. 2008). Although not distinctly lobulate,

Polish material agrees well with the description, except nephrosteranic,

isonephrosterinic and protocetraric acids were not detected, most probably due

to their low quantity or overlapping with other metabolites in a similar position

on the TLC plates.

ECOLOGY & DISTRIBUTION — Parmelia ernstiae is widely distributed in

Europe, with a few records known from Africa. It has been reported from

Austria, Belgium, Bosnia and Herzegovina, Britain, Bulgaria, Czech Republic,

Denmark, Estonia, France, Germany, Greece, Ireland, Lithuania, Luxembourg,

the Netherlands, Slovenia and Spain (Feuerer & Thell 2002, Sérusiaux et al.

2003, Thell 2003, Molina et al. 2004, Otte 2005, Santesson et al. 2004, Thell

110 ... Kukwa & al.

et al. 2007, Suija et a. 2007, Motiejiinaité et al. 2008, Thell et al. 2008, Berger

et al. 2009, Seaward 2010, Diederich et al. 2011) and in Africa from Algeria

and the Canary Islands (Sérusiaux et al. 2003). It is a typically corticolous

species growing on deciduous or rarely coniferous trees in open situations (e.g.

roadsides, churchyards) and forests (Thell 2003, Santesson et al. 2004, Thell et

al. 2007); an exceptional saxicolous record is also known (Thell et al. 2008).

In Poland it has been found only at one locality in northwestern Poland ona

roadside elm in a well-lit situation. All available material labelled as P. saxatilis

in UGDA has been examined, but as no more specimens of P. ernstiae have

been traced, it probably should be regarded as rare in the country.

SPECIMEN EXAMINED — POLAND. DRAwSKIE LAKELAND. road N of Gtusko village,

53°02'59"N 15°56'34"E, ATPOL grid square Cb-22, on roadside Ulmus sp., 05.05.2010,

M. Kukwa 7793 (UGDA).

CoMMENTS — Based on DNA sequences and different morphology (Feuerer

& Thell 2002) and more recently on the composition of secondary metabolites

(Thell et al. 2008), P ernstiae has been recently segregated from P. saxatilis (L.)

Ach., from which it can be easily separated morphologically by the epruinose

thalli and isidia and chemically by the absence of lobaric and fatty acids (Feuerer

& Thell 2002, Thell et al. 2008).

Parmelia ernstiae can also be confused with another isidiate species,

P. serrana A. Crespo et al. Thalli of both taxa are pruinose, but in P serrana

lobes are larger, rounded, and commonly overlapping and lobaric acid is not

produced (Molina et al. 2004, Thell et al. 2008). Parmelia serrana has not been

found in Poland, although its distribution range (see Thell et al. 2008) suggests

it probably occurs there.

Placopsis lambii Hertel & V. Wirth, Flecht. Baden-Wiirtt. Verbreit.: 511. 1987.

CHARACTERISTICS — ‘The species is characterized by its placodioid thallus

with deeply incised and radiating marginal lobes, shiny upper surface, usually

blackish and more or less rounded soralia, non-lobate cephalodia (absent in

some specimens), and the production of 5-O-methylhiascic and gyrophoric

acids as major substances (Moberg & Carlin 1996, Gilbert & Purvis 2009,

Harrold et al. 2010). The cephalodia were not developed in Polish specimen.

ECOLOGY & DISTRIBUTION — Placopsis lambii is widespread but rather

scattered. In Europe it has been reported from Austria, Belgium, Britain

Finland, France, Germany, Iceland, Ireland, the Netherlands, Norway, Russia,

and Sweden (Wirth 1987, Diederich 1994, Moberg & Carlin 1996, Aptroot et al.

1999, Hafellner & Turk 2001, Santesson et al. 2004, Harrold et al. 2010, Seaward

2010, Diederich et al. 2011). In Africa it has been found in Kenya, Lesotho, and

Tanzania (Moberg & Carlin 1999) and in the Americas in Bolivia, Chile, Costa

Rica, and Ecuador (Galloway 2002, Galloway & Arvidsson 2007). It is also

Seven lichens new to Poland... 111

known from New Zealand (Galloway 2001) and asiatic Russia (eastern Siberia

and Russian Far East; Urbanavichus 2010).

In Poland P. lambii has been recorded only once in northern Poland where it

was found on a pile of stones in a sunny but rather humid habitat near several

ponds. This stand is also noteworthy for other rare and endangered lichens in

Poland, e.g., Rhizocarpon lecanorinum Anders and Xanthoparmelia mougeotii

(Schaer.) Hale.

SPECIMEN EXAMINED — POLAND. KaASZzuBSKIE LAKELAND. Vicinity of Szumles

village, 54°09'33"N 18°13'26"E, ATPOL grid square Bc-07, open area by small ponds,

on stone, 08.05.2008, M. Kukwa 5968, M. Bulinski, J. Zarembska (UGDA L-16515, dupl.

in BILAS and PRA).

ComMENTsS — Placopsis lambii is very similar and morphologically almost

indistinguishable from P gelida (L.) Linds. Placopsis gelida tends to be matte or

shiny only at the lobe tips of the upper surface and has more elongated soralia that

are rarely blackish. However, those are subjective characters, and the chemistry

is a most reliable diagnostic feature; both taxa contain gyrophoric acid, but

P. lambii additionally produces 5-O-methylhiascic acid (Gilbert & Purvis 2009,

Harrold et al. 2010). Due to their similarity one could treat both species as one

variable entity, but molecular studies supported their distinctiveness (Schmitt

et al. 2003).

Some P. lambii morphs might be also mistaken for Trapelia placodioides

Coppins & P. James, which, however, lacks radiating marginal lobes and

cephalodia; T’ placodioides also produces gyrophoricacid, but 5-O-methylhiascic

acid is absent (Gilbert & Purvis 2009, Purvis et al. 2009).

Protoparmelia oleagina (Harm.) Coppins, Lichenologist 24: 368. 1992.

= Lecanora oleagina Harm., Lich. France 5: 1023. 1913.

CHARACTERISTICS — Protoparmelia oleagina is a crustose, often sterile,

epiphytic lichen with a more or less continuous, dull olivaceous brown, scurfy

granular-isidiate thallus and apothecial margin; the apothecial disc is also

olivaceous. Ascospores are fusiform, 9.5-15 x 2-3.5 um (Coppins & Chambers

2009, Brodo & Aptroot 2005). The species produces lobaric acid, but often in

low concentration (Brodo & Aptroot 2005). One Polish specimen was fertile

(Kukwa 7837a), but very few apothecia were developed.

ECOLOGY & DISTRIBUTION — Protoparmelia oleagina occurs only in Europe,

where it has been recorded from Austria, Belgium, Finland, France, Germany,

Great Britain, Italy, the Netherlands, Norway and Sweden (Aptroot et al. 1999,

Scholz 2000, Hafellner & Turk 2001, Santesson et al. 2004, Brodo & Aptroot

2005, Nimis & Martellos 2008, Coppins & Chambers 2009, Diederich et al.

2011).

In Poland P. oleagina has been found on deciduous trees in humid forests at

two localities in western Pomerania (NE Poland).

112 ... Kukwa & al.

SPECIMENS EXAMINED — POLAND. DRAWSKIE LAKELAND. Drawienski National Park,

eastern slopes of Drawa river valley, 2 km NE of Zatom village, Tragankowe Urwisko

range, 53°09'12"N_ 15°51'49-54"E ATPOL grid square Cb-12, deciduous forest, on

Fagus sylvatica, 06.05.2010, M. Kukwa 7837a (UGDA). SLOWINSKIE Coast. Bialogora

nature reserve, forest section No. 22, 54°49'27"N, 17°57'52"E, ATPOL grid square Ac-

36, wet pine-birch forest, on Betula pendula, 22.09.2010, M. Kukwa 8268a, A. Jabtonska,

M. Oset (UGDA L-16660).

ComMENTS — Protoparmelia oleagina is easily distinguished by its corticolous

habitat and more or less continuous, dull olivaceous brown, scurfy granular

and isidiate thallus. Morphologically it is most similar to P. ochrococca (Nyl.)

P.M. Jorg. et al., but the thallus of the latter is not isidiate, consists of subglobose

areoles, and lacks lichen substances (Brodo & Aptroot 2005, Coppins &

Chambers 2009); so far P. ochrococca has not been recorded from Poland.

Chemically and morphologically P oleagina can be confused with

P. hypotremella Herk et al., which is known from several localities in Poland

(see Kubiak et al. 2010). Both taxa contain lobaric acid, but the thallus of

P. hypotremella consists of dispersed round flat granules or microsquamules;

it is also brighter, pale grayish-brown to olive-grey (Brodo & Aptroot 2005,

Coppins & Chambers 2009).

Morphologically, P oleagina resembles small forms of P. badia (Hoftm.)

Hafellner, but the latter is a saxicolous lichen with ellipsoid-fusiform ascospores

with distinctly pointed apices (Coppins & Chambers 2009).

Some epiphytic species of the genus Lecanora Ach. also have dark apothecial

discs (e.g. L. argentata (Ach.) Malme or L. persimilis (Th. Fr.) Arnold), but they

lack brown pigmentation, often produce atranorin or usnic acid in the cortex,

and their ascospores are generally wider (Sliwa 2007, Coppins & Chambers

2009).

Scoliciosporum curvatum Sérus., Nord. J. Bot. 13: 458. 1993.

CHARACTERISTICS — This crustose lichen is characterized by its pale green

to dark grey-green granular thallus, chlorococcoid photobiont with rather large

cells (12—)14—20(-22) um, numerous and very small (up to 0.16 mm diam.),

pale pink, orange to brown apothecia, 2 um wide, simple to furcated or rarely

anastomosing paraphyses, 8-16-spored asci and 1-septate, curved (lunulate)

or slightly sigmoid spores with acute ends, 7-11 x 1.5-3 um. Paraphyses

are abundant only in young apothecia. The species does not produce lichen

substances (Sérusiaux 1993, Edwards et al. 2009).

ECOLOGY & DISTRIBUTION — This species occurs only in Europe, being much

more common in western countries. It has been reported from Austria, Britain,

Czech Republic, Denmark, France, Germany, Hungary, Ireland, Norway,

Slovakia, Romania, Spain, Sweden, Switzerland and Ukraine (Sérusiaux 1993,

Poelt 1994, Boom et al 1995, Palice 1999, Sochting & Alstrup 2002, Clerc 2004,

Seven lichens new to Poland... 113

Santesson et al. 2004, Lisickaé 2005, Edwards et al. 2009, Vondrak et al. 2009,

Seaward 2010, Dymytrova 2011). Dymytrova (2011), citing Nimis & Martellos

(2003), cited S. curvatum from Italy, but Nimis & Martellos (2008) have included

it not in the Italian checklist but only in the iconographic archive (along with

several other species not reported from Italy). The report by Himelbrant (2008)

of S. curvatum from Poland is a misprint (Himelbrant, pers. comm.), making

the specimens cited below the first records for Poland.

Most central European localities of S. curvatum lie in the mountains

(490-1600 m a.s.l.), but those from Poland are in the lowlands. The species is

treated as an endangered species (extremely rare and potentially endangered

or vulnerable) in some countries and has been included in the lichen red lists

of Austria (Turk & Hafellner 1999), Czech Republic (Liska et al. 2008), and

Switzerland (Scheidegger et al. 2002).

Scoliciosporum curvatum is typically a foliicolous lichen growing in sheltered

and humid situations on leaves and (sometimes) twigs of evergreen shrubs

such as Buxus, Camellia, and Rhododendron (Edwards et al. 2009). The species

has also been found on needles (rarely twigs) of spruces, firs and occasionally

pines, typical of southwestern Scandinavia (Santesson et al. 2004) and the

mountains of central Europe (Poelt 1994, Palice 1999, Guttova & Palice 1999,

Lisicka 2005).

In Poland S. curvatum has been found only in two localities in Romincka

Forest in northeast Poland where it was collected from spruce needles inside a

wet spruce forest and in a managed pine forest.

SPECIMENS EXAMINED — POLAND. RoMINCKA Foresst. Puszcza Romincka Forest

Landscape Park, Zytkiejmska Struga nature reserve, forest section No. 63, c. 3.5 km

WSW of Zytkiejmy village, 54°20'51"N 22°37'55"E, ATPOL grid square Af-86, boggy

spruce forest, on needles of spruce, 17.09.2003, A. Zalewska, W. Faltynowicz s.n. (OLS);

forest section No. 184, c. 4.5 km of SW Zytkiejmy village, 54°19'49"N 22°37'37"E,

ATPOL grid square Af-86, pine forest with spruce, on needles of spruce, 19.09.2003, A.

Zalewska, W. Faltynowicz s.n. (OLS).

CoMMENTS — Placement of S. curvatum within Scoliciosporum A. Massal. is

problematic as it is separated from other representatives by its higher number

of ascospores per ascus and different organization of paraphyses (Sérusiaux

1989, 1993). The generic concept appears to be unclear, since several other

species with deviating characters are included, especially S. abietinum

T. Sprib., S. intrusum (Th. Fr.) Hafellner, and S. coniectum Kantvilas & Lumbsch

(Sérusiaux 1993, Hafellner 2004, Spribille et al. 2009, Kantvilas & Lumbsch

2010). Molecular analyses would clarify its circumscription and explain its

relationship with S. curvatum.

Scoliciosporum curvatum is inconspicuous and easily overlooked, as

its thallus may resemble a green algal cover over the substrate. In the field

it can be confused with other species growing in the same habitat, but it is

114... Kukwa & al.

easily recognizable after microscopic examination. In Poland, only two other

species, Fellhanera bouteillei (Desm.) Vézda and F. subtilis (Vézda) Diederich

& Sérus., have been found on needles of Abies alba (F. bouteillei) or Picea abies

(FE. subtilis) (Faltynowicz 2003). The thallus of E subtilis is grey to green with

smaller photobiont cells (5-15 um in diam.), apothecia are whitish and larger

(0.15-0.4 mm in diam.), ascospores are oblong, fusiform and (1-)3-septate,

and it always develops pinkish pycnidia. Fellhanera bouteillei has a verruculose-

rimose thallus, often entirely covered by dull bluish-green farinose soredia.

The apothecia and pycnidia superficially resemble those of F subtilis, but

ascospores are only 1-septate (as in S. curvatum) but also ovoid to oblong-ovoid

(sometimes soleiform) and often constricted at the septa; the F. bouteillei thallus

also contains usnic acid, zeorin, and + asemone (Sérusiaux 1996, Aptroot et al.

2009).

Some S. curvatum specimens might also be mistaken for Fellhanera

viridisorediata Aptroot et al., which has not been found in Poland. It has a

similar green granular thallus, with firstly punctiform, then coalescing, soralia

and (0—)1-septate elongate-ellipsoid ascospores. The species is often sterile, but

then it differs in the presence of roccellic acid (Aptroot et al. 2009).

Scoliciosporum gallurae Vézda & Poelt, recently recorded in Poland

(Kukwa & Kubiak 2007), which may also superficially resemble S. curvatum,

produces gyrophoric acid and is known from twigs in open, nutrient-enriched

situations.

Acknowledgments

We are very grateful to Dr Jurga Motiejunaité (Vilnius) and Prof. Mark R. D. Seaward

(Bradford) for reviewing the manuscript, and very valuable comments on a previous

version of the paper. We would also like to thank Dr Christian Printzen (Frankfurt am

Main) and Dr Zdenék Palice (Prihonice) respectively for confirming our determinations

of Biatora pontica and Placopsis lambii, Dr Anders Nordin (Uppsala) for hospitality

during visits to UPS, and Dr Dmitry E. Himelbrandt (Saint Petersburg) for information

on the distribution of Scoliciosporum curvatum.

Literature cited

Aptroot A, Herk CM van, Sparrius LB, Boom PPG van den. 1999. Checklist van de Nederlandse

lichenen en lichenicole fungi. Buxbaumiella 50: 4-64.

Aptroot A, Edwards BW, Sérusiaux E, Coppins BJ. 2009. Fellhanera Vézda (1986). 398-401, in: CW

Smith et al. (eds). The lichens of Great Britain and Ireland. London, British Lichen Society.

Berger FE, Preiemetzhofer F, Tiirk R. 2009. Atlas der Verbreitung der Flechten in Oberésterreich.

Stapfia 90: 1-320.

Boom PPG van den, Etayo J, Breuss O. 1995. Interesting records of lichens and allied fungi from the

Western Pyrenees (France and Spain). Cryptog., Bryol. Lichénol. 16: 263-283.

Brodo IM, Aptroot A. 2005. Corticolous species of Protoparmelia (lichenized Ascomycotina) in

North America. Canad. J. Bot. 83: 1075-1081. http://dx.doi.org/10.1139/b05-091

Seven lichens new to Poland... 115

Cieslinski S, Faltynowicz W. 1993. Note from editors. 7-8, in: S Cieslinski, W Faltynowicz (eds).

Atlas of the geographical distribution of lichen in Poland. 1. Krakow, W. Szafer Institute of

Botany, Polish Academy of Sciences.

Clerc P. 2004. Les champignons lichénisés de Suisse. Cryptog. Helv. 19: 1-320.

Coppins BJ, Chambers SP. 2009. Protoparmelia M. Choisy (1929). 753-755, in: CW Smith et al.

(eds). The lichens of Great Britain and Ireland. London, British Lichen Society.

Coppins BJ, Scheidegger C, Aptroot A. 2009. Buellia De Not. (1846). 228-238, in: CW Smith et al.

(eds). The lichens of Great Britain and Ireland. London, British Lichen Society.

Culberson CF, Kristinsson H. 1970. A standardized method for the identification of lichen products.

Jour. Chromatogr. 46: 85-93. http://dx.doi.org/10.1016/S0021-9673(00)83967-9

Diederich P. 1994. New or interesting lichenicolous fungi. 4. Clauzadeomyces verrucosus gen. et sp.

nov. (Deuteromycotina). Bull. Soc. Linn. Provence 45: 417-420.

Diederich P, Ertz D, Stapper N, Sérusiaux E, Broeck van den D, Boom van den P, Ries C. 2011. The

lichens and lichenicolous fungi of Belgium, Luxembourg and northern France.

http://www.lichenology.info [viewed online on 1 October 2011].

Dymytrova LV. 2011. Notes on the genus Scoliciosporum (Lecanorales, Ascomycota) in Ukraine.

Polish Bot. J. 56: 61-75.

Edwards BW, James PW, Purvis OW. 2009. Scoliciosporum A. Massal. (1852). 839-841, in: CW

Smith et al. (eds). The lichens of Great Britain and Ireland. London, British Lichen Society.

Eichler M, Cezanne R, Diederich P, Ertz D, Broeck D van den, Boom P van den, Sérusiaux E. 2010.

New or interesting lichens and lichenicolous fungi from Belgium, Luxembourg and northern

France. XIII. Bull. Soc. Naturalistes Luxemb. 111: 33-46.

Faltynowicz W. 2003. The lichens, lichenicolous and allied fungi of Poland. An annotated checklist.

Biodiversity of Poland 6: 1-435. W. Szafer Institute of Botany, Polish Academy of Sciences,

Krakow.

Feuerer T, Thell A. 2002. Parmelia ernstiae - a new macrolichen from Germany. Mitt. Inst. Allg.

Bot. Hamburg 30-32: 49-60.

Flakus A. 2007. Lichenized and lichenicolous fungi from mylonitized areas of the subnival belt in

the Tatra Mountains (Western Carpathians). Ann. Bot. Fenn. 44: 427-449.

Gaarder G, Tonsberg T. 2010. Buellia violaceofusca new to Norway. Graphis Scripta 22: 22.

Galloway DJ. 2001. Additional lichen records from New Zealand 36. Placopsis lambii Hertel &

V. Wirth. Australasian Lichenology 49: 36-38.

Galloway DJ. 2002. Taxonomic notes on the lichen genus Placopsis (Agyriaceae: Ascomycota) in

southern South America, with a key to species. Mitt. Inst. Allg. Bot. Hamburg 30-32: 79-107.

Galloway DJ, Arvidsson L. 2007. Notes on Placopsis (Ascomycota: Trapeliaceae) in Ecuador.

Biblioth. Lichenol. 96: 87-102.

Gilbert OL, Purvis OW. 2009. Placopis (Nyl.) Linds. (1867). 710-711, in: CW Smith et al. (eds). The

lichens of Great Britain and Ireland. London, British Lichen Society.

Guttova AA, Palice Z. 1999. LiSajniky Narodného parku Muranska planina I - Hrdzava dolina.

35-47, in: M Uhrin (ed.).Vyskum a ochrana prirody Muranskej planiny 2. Revuca.

Hafellner J. 2004. Notes on Scoliciosporum intrusum. Fritschiana 49: 29-41.

Hafellner J, Turk R. 2001. Die lichenisertien Pilze Osterreich - eine Checkliste der bisher

nachgewiesenen Arten mit Verbreitungsangaben. Stapfia 76: 3-167.

Hafellner J, Petutschnig W, Taurer-Zeiner C, Mayrhofer H. 2005. Uber einige bemerkenswerte

Flechtenfunde in Karnten, hauptsachlich in den Gurktaler Alpen. Carinthia II 195: 423-440.

Harris RC, Lendemer JC. 2010. A review of Lecania croatica (syn. Catillaria croatica) in North

America. Opuscula Philolichenum 8: 4-49.

116 ... Kukwa & al.

Harrold P, Walkinshaw S, Coppins BJ, Ellis CJ. 2010. Species discrimination and the distribution of

Placopsis gelida and P. lambii in Great Britain. Brit. Lich. Soc. Bull. 107: 44-51.

Himelbrant DE. 2008. Rod Scoliciosporum A. Massal. 96-106, in: MP Andreev et al. (eds).

Handbook of the lichens of Russia. 10. Izdatel’stvo Nauka, St-Petersburg.

Kantvilas G, Lumbsch TH. 2010. A new species and a new record of Australian Scoliciosporum.

Australasian Lichenology 66: 16-23.

Kotlov YV 2004. Preliminary checklist of lichen family Catillariaceae. Novosti Sist. Nizsh. Rast.

37: 234-252.

Krzewicka B. 2009. Some new records of Verrucaria from Beskid Niski Mts. Acta Mycol. 44:

265-273.

Kubiak D, Zaniewski P, Wrzosek M. 2010. Notes on the distribution of Sphinctrina anglica and its

host in Poland. Polish Bot. J. 55: 239-242.

Kukwa M, Flakus A. 2009. New or interesting records of lichenicolous fungi from Poland VII.

Species mainly from Tatra Mountains. Herzogia 22: 191-211.

Kukwa M, Kubiak D. 2007. Six sorediate crustose lichens new to Poland. Mycotaxon 102:

155-164.

Kukwa M, Motiejunaite J, Rutkowski P, Zalewska A. 2002. New or interesting records of lichenicolous

fungi from Poland. Part I. Herzogia 15: 129-139.

Lisicka E. 2005. The lichens of the Tatry Mountains. VEDA, Slovak Academy of Sciences.

Bratislava.

Liska J, Palice Z, Slavikova $. 2008. Checklist and Red List of lichens of the Czech Republic. Preslia

80: 151-182.

Lubek A. 2009. Jamesiella anastomosans, a lichen species new the Poland. Polish Bot. J. 54:

125-127.

Moberg R, Carlin G. 1996. The genus Placopsis (Trapeliaceae) in Norden. Symb. Bot. Upsal. 31(3):

SIIB 25;

Moberg R, Carlin G. 1999. Placopsis lambii, new to Africa. Lichenologist 31(6): 647-648.

Molina MC, Crespo A, Blanco O, Lumbsch HT, Hawksworth DL. 2004. Phylogenetic relationships

and species concepts in Parmelia s. str. (Parmeliaceae) inferred from nuclear ITS rDNA and

B-tubulin sequences. Lichenologist 36: 37-54. http://dx.doi.org/10.1017/S0024282904013933

Motiejunaité J, Czyzewska K. 2008. Additions to the biota of lichens and lichenicolous fungi of

Poland, with a note on Lecania prasinoides in eastern and central Europe. Polish Bot. J. 53:

L5s- 162:

Motiejunaite J, Alstrup V, Randlane T, Himelbrant D, Stoncius D, Hermansson J, Urbanavichus

G, Suija A, Fritz O, Prigodina LukoSiené I, Johansson P. 2008. New or noteworthy lichens,

lichenicolous and allied fungi from Birzai district, Lithuania. Bot. Lith. 14: 29-42.

Mrak T, Mayrhofer H, Batic F. 2004. Contributions to the lichen flora of Slovenia XI. Lichens from

the vicinity of Lake Bohinj (Julian Alps). Herzogia 17: 107-127.

Nimis PL, Martellos S. 2003. Checklist of Italian Lichens 3.0. University of Trieste, Dept. of Biology.

http://dbiodbs. univ.trieste.it/global/italic_ico [viewed online on 9 November 2011].

Nimis PL, Martellos S. 2008. ITALIC - The information system on Italian lichens. Version 4.0.

University of Trieste, Dept. of Biology. http://dbiodbs.univ.trieste.it [viewed online on 9

November 2011].

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

London, British Lichen Society.

Otte V. 2005. Noteworthy lichen records for Bulgaria. Abh. Ber. Naturkundemus. Gorlitz 77:

77-86.

Seven lichens new to Poland... 117

Palice Z. 1999. New and noteworthy records of lichens in the Czech Republic. Preslia 71:

289-336.

Palice Z, Guttova A, Halda JP. 2006. Lichens new for Slovakia collected in the National Park

Muranska planina (W Carpathians). 179-192, in: A Lackovicova et al. (eds), Central European

lichens — diversity and threat. Ithaca, Mycotaxon Ltd.

Poelt J. 1994. Bemerkenswerte Flechten aus Osterreich, insbesondere der Steiermark. Mitt.

Naturwiss. Vereins Steiermark 124: 91-111.

Printzen C. 1995. Die Flechtengattugn Biatora in Europa. Biblioth. Lichenol. 60: 1-275.

Printzen C, Otte V. 2003. Biatora longispora, new to Europe, and a revised key to European and

Macaronesian Biatora species. Graphis Scripta 17: 56-61.

Printzen C, Tonsberg T. 2003. Four new species and three new apothecial pigments of Biatora.

Biblioth. Lichenol. 86: 133-145.

Printzen C, Lumbsch HT, Orange A. 2001. Biatora britannica sp. nov. and the occurrence of Biatora

efflorescens in the British Isles. Lichenologist 33: 181-187.

http://dx.doi.org/10.1006/lich.2000.0319

Purvis OW, Coppins BJ, Woseley PA, Fletcher A. 2009. Trapelia M. Choisy (1929). 904—908, in: CW

Smith et al. (eds). The lichens of Great Britain and Ireland. London, British Lichen Society.

Reese Naesborg R, Ekman S, Tibell L. 2007. Molecular phylogeny of the genus Lecania

(Ramalinaceae, lichenized Ascomycota). Mycol. Res. 111: 581-591.

http://dx.doi.org/10.1016/j.mycres.2007.03.001

Santesson R, Moberg R, Nordin A, Tonsberg T, Vitikainen O. 2004. Lichen-forming and

lichenicolous fungi of Fennoscandia. Uppsala, Museum of Evolution, Uppsala University.

Scheidegger C, Clerc P, Dietrich M, Frei M, Groner U, Keller C, Roth I, Stofer S$, Vust M. 2002.

Rote Liste der gefahrdeten Arten der Schweiz. Baum- und erdbewohnende Flechten. Bern,

Bundesamt fir Umwelt, Wald und Landschaft BUWAL.

Schmitt I, Lumbsch HT, Sechting U. 2003. Phylogeny of the lichen genus Placopsis and its allies

based on Bayesian analyses of nuclear and mitochondrial sequences. Mycologia 95: 827-835.

http://dx.doi.org/10.2307/3762011

Scholz P. 2000. Katalog der Flechten und flechtenbewohnenden Pilze Deutschlands. Schriftenreihe

Vegetationsk. 31: 1-298.

Seaward MRD. 2010. Census catalogue of Irish lichens. 3rd edition. National Museum Northern

Ireland, Belfast.

Sérusiaux E. 1989. Foliicolous lichens: ecological and chorological data. Bot. J. Linn. Soc. 100:

87-96. http://dx.doi.org/10.1111/j.1095-8339.1989.tb01710.x

Sérusiaux E. 1993. New taxa of foliicolous lichens from Western Europe and Macaronesia. Nordic

J. Bot. 13: 447-461. http://dx.doi.org/10.1111/j.1756-1051.1993.tb00080.x

Sérusiaux E. 1996. Foliicolous lichens from Madeira, with the description of a new genus and two

new species and a world-wide key of foliicolous Fellhanera. Lichenologist 28: 197-227.

Sérusiaux E, Diederich P, Brand AM, van den Boom PPG. 1999. New or interesting lichens and

lichenicolous fungi from Belgium and Luxembourg. VIII. Lejeunia, n.s. 162: 1-95.

Sérusiaux E, Diederich P, Ertz D, van den Boom PPG. 2003. New or interesting lichens and

lichenicolous fungi from Belgium, Luxembourg and northern France. IX. Lejeunia, n.s. 173:

1-48.

Sliwa L. 2007. A revision of the Lecanora dispersa complex in North America. Polish Bot. J. 52:

1-70.

Sliwa L, Flakus A. 2011. Lecanora microloba, a new saxicolous species from Poland. Lichenologist

43: 1-6. http://dx.doi.org/10.1017/S0024282910000551

118 ... Kukwa & al.

Sochting U, Alstrup V. 2002. Danish lichen checklist. Ver. 2. Botanical Institute, University of

Copenhagen, Copenhagen. www.bi.ku.dk/lichens/dkchecklist [24. October 2011].

Spribille T, Bjork CR, Ekman S, Elix JA, Goward T, Printzen C, Tonsberg T, Wheeler T. 2009.

Contributions to an epiphytic lichen flora of northwest North America: I. Eight new species

from British Columbia inland rain forests. Bryologist 112: 109-137.

http://dx.doi.org/10.1639/0007-2745-112.1.109

Suija A, Leppik E, Randlane T, Thor G. 2007. New Estonian records. Lichens and lichenicolous

fungi. Folia Cryptog. Estonica 43: 73-76.

Thell A. 2003. Parmelia ernstiae — new to the Nordic lichen flora. Graphis Scripta 14: 10.

Thell A, Hansen ES, Karnefelt I, Feuerer T. 2007. The distribution of Parmelia ernstiae in Denmark.

Biblioth. Lichenol. 96: 299-304.

Thell A, Elix JA, Feuerer T, Hansen ES, Karnefelt EI, Schiller N, Westberg M. 2008. Notes on the

systematics, chemistry and distribution of European Parmelia and Punctelia species (lichenized

ascomycetes). Sauteria 15: 545-559.

Thor G, Muhr L-E. 1991. Buellia violaceofusca, a new lichen from Sweden. Lichenologist 23:

11-13.

Thor G, Nordin A. 1998. 16 lichens new to Estonia. Folia Cryptog. Estonica 32: 123-125.

Tonsberg T. 1992. The sorediate and isidiate, corticolous, crustose lichens in Norway. Sommerfeltia

14: 1-331.

Turk R, Haffelner J. 1999. Rote Liste gefahrdeter Flechten (Lichenes) Osterreichs. 2. Fassung.

187-228, in: H Niklefeld et al. (eds). Rote Listen gefahrdeter Pflanzen Osterreichs. 2 Auflage.

Griine Reihe des Bundesministeriums fiir Umwelt, Jugend und Familie, 10. Graz, Austria

Medien Service.

Urbanavichus G. 2010. A checklist of the lichen flora of Russia. Nauka, Sankt Petersburg.

Vondrak J, Soun J, L6kés L, Khodosovtsev A. 2009. Noteworthy lichen-forming and lichenicolous

fungi from the Bikk Mts, Hungary. Acta Bot. Hung. 51: 217-230.

Vondrak J, Palice Z, Khodosovtsev A, Postoyalkin S. 2010. Additions to the diversity of rare or

overlooked lichens and lichenicolous fungi in Ukrainian Carpathians. Chornomors'k. bot. z.

6: 6-34.

Wirth V. 1987. Die Flechten Baden-Wiirttembergs. Verbreitungsatlas. Eugen Ulmer, GmbH & Co.,

Stuttgart.

Zhurbenko MP, Kukwa M, Oset M. 2009. Roselliniella stereocaulorum (Sordariales, Ascomycota), a

new lichenicolous fungus from the Holarctic. Mycotaxon 109: 323-328.

http://dx.doi.org/10.5248/109.323

MY COTAXON

http://dx.doi.org/10.5248/120.119

Volume 120, pp. 119-125 April-June 2012

Dissoconium proteae newly recorded from China

RONG ZHANG’, YONGNA Mao’, Lu Hao’,

HONGCAI CHEN?, GUANGYU SUN’ & MARK L. GLEASON?

' State Key Laboratory of Crop Stress Biology in Arid Areas and College of Plant Protection

& College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, P. R. China

*Department of Plant Pathology and Microbiology, Iowa State University, Ames, Iowa 50011, U.S.A

* CORRESPONDENCE TO: sgy@nwsuaf.edu.cn

ABSTRACT — We document the first report of Dissoconium proteae from China. This fungus

was found on the cuticle of apple fruit collected from orchards in Shaanxi Province. Its

morphology is compared with other Dissoconium species and a phylogenetic analysis based

on ITS sequence is presented.

Key worps — sooty blotch and flyspeck, taxonomy, internal transcribed spacer, rDNA

Introduction

Sooty blotch and flyspeck (SBFS) is a worldwide disease complex more

than 60 fungi that blemish the cuticle of pome fruits (such as apple) in humid

temperate regions and cause cosmetic damage entailing significant economic

losses (Batzer et al. 2005). The common name “flyspeck” refers to species in the

SBFS complex that appear on fruit surfaces as clusters of black, shiny, round

to oval, flattened sclerotium-like bodies with no mycelial mat (Batzer et al.

2005).

The hyphomycete genus Dissoconium de Hoog et al. has lightly pigmented

conidiophores that produce septate and aseptate conidia in a sympodial manner.

The conidia are hyaline, one- or two-celled, and detach forcibly in pairs (Hoog

et al. 1983).

Currently the genus includes nine species: Dissoconium aciculare de Hoog

et al., D. australiense Crous & Summerell, D. commune Crous & Mansilla,

D. dekkeri de Hoog & Hijwegen, D. eucalypti Crous & Carnegie, D. maliG.Y. Sun

et al., D. musae Arzanlou & Crous, D. proteae, and D. subuliphorum (Matsush.)

R.F. Castaneda (Arzanlou et al. 2008; Crous et al. 1999, 2004, 2007, 2008; Hoog

et al. 1983, 1991; Zhang et al. 2007). Batzer et al. (2005) reported five putative

Dissoconium species within the sooty blotch complex on apple fruit.

120 ... Zhang & al.

Recently, during investigations of SBFS of apple in China, we identified four

fungal isolates that represent the first record of Dissoconium proteae from China

based on ITS sequence analysis and morphological comparison.

Materials & methods

IsOLATION. In September 2008, apples with flyspeck signs were collected from

orchards in Shaanxi Province. The thalli were transferred to a potato dextrose agar slant

(potato 200 g, dextrose 20 g, agar 10 g; PDA) directly from the apple surface, and then

cultured at 23+1°C in darkness (Sun et al. 2003). Colony description of the species was

based on a 1-month-old pure culture. Morphological characteristics are described from

specimens obtained from a cover slip inserted into the surface of the media. Voucher

specimens and cultures are conserved in the Fungal Herbarium of Northwest A&F

University, Yangling, China (HMUABO).

DNA SEQUENCING. Template DNA was extracted from fungal mycelium according

to Li et al. (2011); primer pairs used for ITS amplification and sequencing were ITS1-F

TABLE 1. Sequences used in the phylogenetic analysis

SPECIES GENBANK CODE REFERENCE

Dissoconium proteae (mynbxd10) HM070373 This paper

D. proteae (mynbxd2) HM070374 This paper

D. proteae (mynbxd1.1) HM070375 This paper

D. proteae (mynxc1.1) HM070376 This paper

D. aciculare AY725519 Crous et al. (2004)

AY725521 Crous et al. (2004)

D. australiense EF394854 Crous et al. (2007)

D. eucalypti EF394855 Crous et al. (2007)

D. mali EF627451 Zhang et al. (2007)

D. musae EU514226 Arzanlou et al. (2008)

EU514225 Arzanlou et al. (2008)

D. proteae EU707897 Crous et al. (2008)

Dissoconium sp. AY598874 Batzer et al. (2005)

AY598875 Batzer et al. (2005)

AY598877 Batzer et al. (2005)

FJ425205 Diaz Arias et al. (2010)

Mycosphaerella communis

(Gtisni. Deeorintune) AY725544 Crous et al. (2004)

DQ302950 Crous et al. (2006)

M. lateralis (anam.: D. dekkeri) EU301078 Burgess et al. (2007)

Ramichloridium apiculatum EU041794 Arzanlou et al .(2007)

EU041791 Arzanlou et al .(2007)

R. cerophilum EU041798 Arzanlou et al. (2007)

R. indicum EU041799 Arzanlou et al .(2007)

Ramichloridium sp. FJ425199 Diaz Arias et al. (2010)

Stenella araguata AF362066 Crous et al. (2001)

Dissoconium proteae new to China... 121

(Gardes & Bruns 1993) and ITS4 (White et al. 1990). Amplification was completed with

the following cycling parameters: initial denaturation at 94°C for 3 min followed by 35

cycles of denaturation at 94°C for 30 s, annealing at 52°C for 30 s, and extension at 72°C

for 10 min. The PCR products were sequenced by Organism Technology Co., Shanghai,

China.

The ITS nucleotide sequences generated in this study were added to sequences

downloaded from GenBank (TABLE 1) that had high similarity according to a BLAST

search (National Center for Biotechnology Information’s nucleotide blast program).

Preliminary alignments were performed using CLUSTAL-X (1.83) (Thompson et

al. 1997), imported into BioEdit version 5.0.9.1 (Hall 1999), and manually adjusted.

Phylogenetic analysis of aligned DNA sequences was performed with PAUP version

4.0b10 for 32-bit Microsoft Windows (Swofford 2001). Heuristic searches were

performed with 1000 random sequence additions. Clade stability was evaluated by 1000

bootstrap replications. Other measures for parsimony, including tree length, consistency

index, retention index and rescaled consistency index (CI, RI and RC, respectively),

were also calculated. Stenella araguata was used as the outgroup taxon.

Results: DNA phylogeny

A multiple alignment of the rDNA-ITS was generated with 21 sequences

obtained from GenBank plus the sequences of isolates mynbxd10, mynbxd2,

mynbxd1l.1 and mynxcl.l. An MP tree with 354 length (CI = 0.7232,

RI = 0.8622, RC = 0.6235) was constructed (Fic. 1). Two major clades were

resolved in the MP trees. One of the major clade with 100% bootstrap value

contained three species in Dissoconium, Mycosphaerella, and Ramichloridium.

The other major clade also had a bootstrap value of 100%. Our isolates and

an authentic Dissoconium proteae isolate identified by Crous et al. (2008) fell

within a single clade with 71% bootstrap support.

Taxonomy

Dissoconium proteae Crous, Persoonia 20: 68. 2008. Fic. 2

Mycelium external consisting of branched, septate, smooth, hyaline hyphae,

0.8-2.8 um wide. Conidiophores 9-20 x 2.2-4.7 um, solitary, arising from

hyphae, subcylindrical, subulate, tapering to a bluntly rounded or truncate

apex, straight to gently curved, smooth, hyaline, becoming medium pale brown

with age, aseptate; conidiogenous loci terminal and lateral, visible as slightly

thickened, darkened scars, 0.5 um wide. Conidia 6.3-11.5 x 2.2-4 um, solitary,

hyaline to pale olivaceous, smooth, ellipsoid, non-constricted to slightly

constricted at median septum, apex obtuse, base obconic-truncate, tapering

pronounced at somewhat protruding hilum, unthickened, not darkened, 1 um

wide. Secondary conidia developing adjacent to primary conidia, hyaline to

subhyaline, aseptate, ellipsoid, tapering prominently towards a protruding,

truncate base, 4.3-8 x 1.6-3.8 um; anastomosing with primary conidia after

122 ... Zhang & al.

EU301078 Mycosphaerella lateralis

AY463143 Dissoconium aciculare

90 || 4¥639404. Dissoconium aciculare

89 |' EF394855 Dissoconium eucalypti

AY598874 Dissoconium sp.

97' AY598875 Dissoconium sp.

EU707897 Dissoconium proteae

mynxct.1

mynbxd1.1

mynbxd2

mynbxd10

AY598877 Dissoconium sp.

FJ425205 Dissoconium sp.

EU041799 Ramichloridium indicum

100, £U041794 Ramichloridium apiculatum

EU041791 Ramichloridium apiculatum

100 EF627451 Dissoconium mali

EU514226 Dissoconium musae

98 |! EU514225 Dissoconium musae

EF394854 Dissoconium australiense

DQ302950 Mycosphaerella communis

91! AY725544 Mycosphaerella communis

FJ425119 Ramichloridium sp.

EU041798 Ramichlondium cerophilum

AJ244261 Stenella araguata

10_

Fic. 1 The majority consensus tree (length = 354, CI = 0.7232, RI = 0.8622, RC = 0.6235) derived

from a heuristic search option in PAUP version 4.0b10 for 32-bit Microsoft Windows with 1000

randomizations of sequence input orders and 1000 bootstrap replications using the data set of ITS1,

5.88 and ITS2. Bootstrap values higher than 50% are indicated above or below the tree branches.

active discharge. In some cases the secondary conidia were observed to

germinate.

CULTURAL CHARACTERISTICS: Colonies on PDA spreading, with sparse

aerial mycelium and regular smooth margins; surface white or sienna, with

patches of white and cinnamon; forming clusters of black sclerotia (remaining

infertile) on PDA; reaching 15 mm diam after 1 mo.

CHARACTERISTICS ON THE HOST: On apple peel, the fungus produced dark,

shiny, round to oval, slightly protuberant sclerotium-like bodies (Fic. 2A).

SPECIMENS EXAMINED: China: Shaanxi Province, Xianyang, Bin County, 35°04'N

108°09’E, alt. 1108 m, on Malus xdomestica (Rosaceae), 18 Sep. 2008, Y.N. Mao,

HMUABO (the Fungal Herbarium of Northwest A&F University) 8885, 8877, 8866

(with dried culture), culture mynbxd10, mynbxd2, mynbxd1.1. China: Shaanxi Province,

Xianyang, Xunyi County, 35°13’N 108°33’E, alt. 1300 m, On Malus xdomestica, 18 Sep.

2008, Y.N. Mao, HMUABO 8682 (with dried culture), culture mynxcl.1.

Dissoconium proteae new to China... 123

Fic. 2 Dissoconium proteae isolate mynbxd10. A. Signs on apple peel. B. Colony on PDA

agar after 30 days. C-D. Conidia, conidiogenous cells, and hyphae. E. Conidiogenous cell.

F, Conidium and secondary conidium. Bars: B=10 mm, C-F =10 um.

124 ... Zhang & al.

Discussion

Of the more than 60 fungal species that blemish the surface of apples

worldwide (Diaz Arias et al. 2010), many have been described quite recently.

For example, Li et al. (2010) and Ma et al. (2010) reported three species of

Zygophiala (Z. cryptogama and Z. cylindrica; Z. qianensis) associated with

flyspeck on apples. Our report provides the first evidence that Dissoconium

proteae can cause flyspeck on apple fruit. Crous et al. (2008) described

D. proteae from leaves of Protea sp. (Proteaceae), causing a leaf spot.

Currently there are nine species in Dissoconium. Based on phylogenetic

analysis of the ITS region and morphological characters of the anamorph, we

identified four isolates as Dissoconium proteae. This is the first report of the

species from apple, and a new record for China.

Acknowledgments

This work was supported by National Natural Science Foundation of China

(31170015, 31171797), the 111 Project from Education Ministry of China (B07049), Top

Talent Project of Northwest A&F University and the earmarked fund for Modern Agro-

industry Technology Research System (nycytx-08-04-04).The authors wish to thank

Dr Eric H.C. McKenzie (Landcare Research, Auckland, New Zealand) and Professor

Zhongyi Zhang (College of Plant Protection, Yunnan Agricultural University, Kunming,

Yunnan, China) for reviewing the manuscript.

Literature cited

Arzanlou M, Groenewald JZ, Gams W, Braun U, Shin H-D, Crous PW. 2007. Phylogenetic and

morphotaxonomic revision of Ramichloridium and allied genera. Studies in Mycology 58:

57-93. http://dx.doi.org/10.3114/sim.2007.58.03

Arzanlou M, Groenewald JZ, Fullerton RA, Abeln ECA, Carlier J, Zapater M-F, Buddenhagen IW,

Viljoen A, Crous PW. 2008. Multiple gene genealogies and phenotypic characters differentiate

several novel species of Mycosphaerella and related anamorphs on banana. Persoonia 20: 19-37.

http://dx.doi.org/10.3767/003158508X302212

Batzer JC, Gleason ML, Harrington TC, Tiffany LH. 2005. Expansion of the sooty blotch and flyspeck

complex on apples based on analysis of ribosomal DNA gene sequences and morphology.

Mycologia 97: 1268-1286. http://dx.doi.org/10.3852/mycologia.97.6.1268

Burgess TI, Pegg GS, Barber PA, Carnegie AJ, Hardy GESJ 2007.

http://www.ncbi.nlm.nih.gov/ nucleotide/162568812?report=genbank

Crous PW, Hong L, Wingfield MJ, Wingfield BD, Kang JC. 1999. Uwebraunia and Dissoconium,

two morphologically similar anamorph genera with different teleomorph affinity. Sydowia 51:

155-166.

Crous PW, Kang JC, Braun U. 2001. A phylogenetic redefinition of anamorph genera in

Mycosphaerella based on ITS rDNA sequence and morphology. Mycologia 93: 1081-1101.

Crous PW, Groenewald JZ, Mansilla JP, Hunter GC, Wingfield MJ. 2004. Phylogenetic reassessment

of Mycosphaerella spp. and their anamorphs occurring on Eucalyptus. Studies in Mycology 50:

195-214.

Crous PW, Wingfield MJ, Mansilla JP, Alfenas AC, Groenewald JZ. 2006. Phylogenetic reassessment

of Mycosphaerella spp. and their anamorphs occurring on Eucalyptus. II. Studies in Mycology

55: 99-131. http://dx.doi.org/10.3114/sim.55.1.99

Dissoconium proteae new to China... 125

Crous PW, Summerell BA, Carnegie AJ, Mohammed C, Wingfield MJ, Himaman W, Groenewald

JZ. 2007. Foliicolous Mycosphaerella spp. and their anamorphs on Corymbia and Eucalyptus.

Fungal Diversity 26:143-185.

Crous PW, Summerell BA, Mostert L, Groenewald JZ. 2008. Host specificity and speciation of

Mycosphaerella and Teratosphaeria species associated with leaf spots of Proteaceae. Persoonia

20:59-86. http://dx.doi.org/10.3767/003158508X323949

Diaz Arias MM, Batzer JC, Harrington TC, Wong AW, Bost SC, Cooley DR, Ellis MA, Hartman

JR, Rosenberger DA, Sundin GW, Sutton TB, Travis JW, Wheeler MJ, Yoder KS, Gleason ML.

2010. Diversity and biogeography of sooty blotch and flyspeck fungi on apple in the eastern and

midwestern United States. Phytopathology 100: 345-355.

http://dx.doi.org/10.1094/PHY TO-100-4-0345

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes- application

to the identification of mycorrhizae and rusts. Molecular Ecology 2:113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NT. Nucl Acids Symp Ser. 41: 95-98.

Hoog GS de, Oorschot CAN van, Hijwegen T. 1983. Taxonomy of the Dactylaria complex. II.

Dissoconium gen. nov. and Cordana Preuss. Proceedings van de Koninklijke Nederlandse

Akademie van Wetenschappen, Section C. 86: 197-206.

Hoog GS de, Hijwegen T, Batenburg-van der Vegte WH. 1991. A new species of Dissoconium.

Mycological Research 95: 679-682. http://dx.doi.org/10.1016/S0953-7562(09)80814-9

Li HY, Zhang R, Sun GY, Batzer JC, Gleason ML. 2010. New species and record of Zygophiala on

apple fruit from China. Mycological Progress 9: 245-251.

http://dx.doi.org/10.1007/s11557-009-0633-1

Li HY, Sun GY, Batzer JC, Crous PW, Groenewald JZ, Karakaya A, Gleason ML. 2011. Scleroramularia

gen. nov. associated with sooty blotch and flyspeck of apple and pawpaw from the Northern

Hemisphere. Fungal Diversity 46:53-66. http://dx.doi.org/10.1007/s13225-010-0074-9

Ma YQ, Zhang R, Sun GY, Zhu HX, Tang M, Batzer JC, Gleason ML. 2010. A new species of

Zygophiala associated with the flyspeck complex on apple from China. Mycological Progress 9:

151-155. http://dx.doi.org/10.1007/s11557-009-0635-z

Sun GY, Zhang R, Zhang Z, Zhang M. 2003. Isolation of sooty blotch and flyspeck fungi from apple

surface by picking up the thalli. Acta Phytopathology Sinica 33: 479-480.

Swofford DL. 2001. PAUP*. Phylogenetic analysis using parsimony (*and other methods) version

4.0 beta version. Sinauer Associates, Sunderland, Massachusetts, U.S.A.

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The CLUSTAL-X windows

interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 25: 4876-4882.

White TJ, Bruns TD, Lee SB, Taylor JW. 1990. Analysis of phylogenetic relationships by amplification

and direct sequencing of ribosomal RNA genes. 315-322, in: Innis MA, Gelfand DH, Sninsky

JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic, New York.

Zhang R, Zhang Z, Zhang M, Sun GY, Gleason ML. 2007. A new species of Dissoconium from

China colonizing apples. Mycotaxon 101: 165-172.

MY COTAXON

http://dx.doi.org/10.5248/120.127

Volume 120, pp. 127-132 April-June 2012

The fungal collection of the Jagiellonian University Herbarium

(KRA), Krakow, Poland

ANNA MarIA OCIEPA’', SZYMON ZUBEK? & PIOTR MLECZKO ”

'W. Szafer Institute of Botany, Polish Academy of Sciences &

?Institute of Botany, Jagiellonian University

ul. Lubicz 46, 31-512 Krakow, Poland

“CORRESPONDENCE TO: ubmleczk@cyf-kr.edu.pl

ABSTRACT — ‘The paper presents a short history of the mycological collection of the

Jagiellonian University Herbarium (KRA, Krakow, Poland). The Herbarium holds over 22,000

specimens, including some interesting European and extra-European fungal exsiccata. One

of the most valuable fungal collections (including many type specimens) in the Herbarium is

that containing Javanese fungi gathered by Marian Raciborski at the end of 19" century.

Key worps — fungi of Pieniny Mts., fungi of Gorce Mts., ascomycetes, basidiomycetes,

Peronospora

The history

The origin of the Jagiellonian University Herbarium dates back to 1780,

when the Faculty of Chemistry and Natural History was established, and with it

the Natural History Room (Kohler 1999; Kohler 2001a). The Herbarium, which

was long associated with the Chair of Botany at the Botanical Garden, is now

part of the Institute of Botany of Jagiellonian University.

The first record of fungi being kept in the Herbarium dates from 1812, and

the oldest specimen with a date placed on the label is from 1807 (Fic. 1). In the

past the Herbarium collections served mainly an educational function (Kohler

2001b). Until 1910 each set of specimens, including purchased, exchanged, or

donated exsiccata and collections, was kept separately and a taxonomic criterion

was not applied to the Herbarium’s arrangement. Between 1910 and 1918 the

collections were merged, and all specimens were arranged taxonomically (Kohler

2001a).

After the Second World War the fungal collection was separated from plant

collections and arranged by Barbara Guminska, who in 1950 became the first

curator of the Herbarium’s mycological section (Kohler 2001b) and served

128 ... Ociepa, Zubek & Mleczko

PLATE 1. The oldest specimen in the fungal collection of the Jagiellonian University Herbarium:

“Geastrum hygrometricum Pers. Normandie. 1807. D. Delaroche (?). Habitat in Italia, Helvetia,

Germania umbrosis. 4.”

as curator until her retirement in 1993. The next curator, Katarzyna Turnau

(1993-2005), was succeeded by Piotr Mleczko, who currently holds this post.

Until 2006 the specimens in the mycological collection were not numbered. In

that year all specimens were counted and most of them were assigned numbers.

The collection now contains 20,195 numbered specimens: 5564 Ascomycota,

11,098 Basidiomycota, 2682 anamorphic fungi, and 851 fungus-like organisms

(mostly Peronospora in the Oomycota). In addition it holds also about 900

unnumbered macromycete specimens collected by Barbara Guminska in Pieniny

National Park, Poland.

The collections

One of the most valuable fungal collections is the Javanese collection

of Marian Raciborski. Raciborski’s collection dates back to 1896-1900 and

consists of dried specimens marked with ‘J’ (Java) after the number (Fic. 2)

as well as some samples preserved in ethanol-formaldehyde solution, which

are in the Botanical Garden Museum (Kohler & Zemanek 1989). According

to Wojewoda (1986), Raciborski described 235 new fungal taxa from Java, all

tropical microfungi except for one macromycete. Many genera and species

described by Raciborski are still accepted under his names: e.g. Aldona

KRA fungal collection (Poland) ... 129

woe

= “Tolyposporvum bogorience Rac,

Bull Al Acad, Sq) Cracow'e 41909:349,

1404

determ. Kalman Vanky

A697] 192%

olyposporium bogoriense Racib.— HOLOTYPUS!

' = Sporisorium bogoriense (Racib.) M. Piatek

i Rev. Marcin Piatek :

W. Szafer Institute of Botany PAS, Krakow 26.VII.2005

PLATE 2. A specimen from the Javanese collection of Marian Raciborski: “Ustilago bogoriense Rac.,

Panicum sp., Bogor.’

(A. stella-nigra), Alina (A. jasmini), Anhellia (A. tristis), Balladyna (B. gardeniae),

Farysia (F. javanica), Goplana (G. aporosae, G. micheliae, G. mirabilis), Lambro

(L. insignis), Ordonia (O. orthobasidion), Skierka (S. agallocha, S. canarii), and

Telimena (T: erythrinae) (Index Fungorum 2011).

Other important exsiccata kept in KRA are those of Karl Wilhelm Gottlieb

Leopold Fuckel, Otto Jaap, Gottlob Ludwig Rabenhorst, Heinrich Rehm, Pier

Andrea Saccardo, Trajan Savulescu, Hans Sydow, and Charles Wright (see the

list below), which comprise many syntypes. At present only the exsiccati of

Ascomycota are catalogued, comprising 134 syntypes.

The fungal section also holds many specimens gathered in Poland, including

three holotypes: Exobasidium dubium Racib., Wawelia regia Namysl. (an exotic

species found in the Botanical Garden of the Jagiellonian University, Krakow),

and Geopyxis rehmii Turnau. The most important Polish collections were

130 ... Ociepa, Zubek & Mleczko

contributed by Wladystaw Wojewoda (1932-2010) from Ojcéw National Park

(macromycetes), Katarzyna Turnau from the Gorce Mts. (larger ascomycetes),

and Barbara Guminska from Pieniny National Park. Recently Piotr Mleczko

and co-workers made a significant collection of fungi, mostly from southern

Poland, which is now being incorporated into the Herbarium and will increase

the number of specimens by approximately 1000.

The most important fungal exsiccata and collections in the Herbarium

mycological section are listed below. It should be noted that the Taxonomic

Literature series (Stafleu & Cowan 1976-88; Stafleu & Mennega 1992-2000) and

the monographs by Stevenson (1971) and Pfister (1985) mostly do not mention

the sets stored at Jagiellonian University Herbarium.

The names of fungal collections, dates, and authors follow (where possible)

Stafleu & Cowan (1976-88), Stafleu & Mennega (1992-2000), Pfister (1985), or

Stevenson (1971). Otherwise data are based on exsiccata labels or, with respect to

author life spans, from various sources cited with each collection).

Abbreviations

IF - Index Fungorum (www.indexfungorum.org)

SC-1-7 — Stafleu & Cowan (1976-88), vols. 1-7

Pf — Pfister (1985)

St — Stevenson (1971)

POLISH COLLECTIONS

* Fungi bialowiezenses exsiccati, Wincenty Siemaszko (1887-1943) [Kohler 2002, Pf,

SC-5].

¢ Fungi parasitici Poloniae exsiccati, Marian Raciborski (1863-1917) [Piekietko-

Zemanek 1986, Wojewoda 1986].

¢ Mycotheca polonica, Marian Raciborski and Boleslaw Namystowski (1882-1929)

[Kohler 2002, Wojewoda 1986].

¢ Mycotheca polonica, Jézef Kochman (1903-95) and Boguslaw Salata (1940-99)

[Majewski 1995, Chmiel & Mulenko 1999, Pf].

EUROPEAN COLLECTIONS (EXCEPT POLAND)

* Ascomyceten, Ascomycetes exsiccatae, Heinrich Rehm (1828-1916) [St; Pf; SC-6].

* Cryptogamae Cechoslovenicae exiccatae, Frantisek Smarda (1904-68) [SC-5].

¢ Flora Hungarica, Gabriel Bohus (1914-2005) [IF].

¢ Flora hungarica exsiccata. Sectio botanica Musei Nationalis Hungarici, Budapest.

¢ Flora Lituana Exsiccata Fungi parasitici, Antanas Minkevicius (1900-98) [Botanikos

ir genetikos katedra 2011; Pf].

¢ Flora Romaniae exsiccata, Alexandru Borza (1887-1971) [Pop 1972; Pf; SC-1].

¢ Fungi bavarici exsiccati, Andreas Allescher (1828-1903) and Johann Nepomuk

Schnabl (1853-99) [Pf; SC-1,5].

¢ Fungi bohemici, FrantiSek Bubak (1865-1925) [SC-1].

KRA fungal collection (Poland) ... 131

¢ Fungi Europaei exsiccati Klotzschii herbarii vivi mycologici continuatio. Editio nova.

Series secunda, Gottlob Ludwig Rabenhorst (1806-81) [Pf; St; SC-4].

¢ Fungi rhenani exsiccati, Karl Wilhelm Gottlieb Leopold Fuckel (1821-76) [SC-1].

¢ Fungi Schemnitziensis, Andreas Kmet (1841-1908) [Lizon 1973; Pf].

* Fungi selecti exsiccati, Otto Jaap (1864-1922), Carl Brick (1863-1924) [Pf; SC-2].

* Griby Rossii, Gavril Stepanowich Nevodovskij (1874-1952) and Voldemar Tranzschel

(1868-1942) [Karczmarz & Salata 1982; Pf; SC-6].

¢ Herbarium mycologicum romanicum, Trajan Savulescu (1889-1963) [Pf SC-5].

Klotzschii herbarium vivum mycologicum, Gottlob Ludwig Rabenhorst [Pf St;

SC-4].

Klotzschii herbarium vivum mycologicum, editio novo, Gottlob Ludwig Rabenhorst

[Pf; St; SC-4].

° Kryptogamae exsiccatae editae a Museo Palatino Vindobonensi, Ginter von Beck

(1856-1931), Alexander Zahlbruckner (1860-1938), Karl von Keissler (1872-

1965), Franz Petrak (1886-1973) [Pf; St; SC-1,2,7].

¢ Mycotheca germanica, Paul Sydow (1851-1925), Hans Sydow (1879-1946) [Pf

SC-6].

¢ Mycotheca veneta, Pier Andrea Saccardo (1845-1920) [Pf; SC-4].

Plantae cryptogamicae, quas in Arduenna collegit (Plantae cryptogamicae Arduennae),

Marie-Anne Libert (1782-1865) [Pf; SC-3].

EXTRA-EUROPEAN COLLECTIONS

* Cryptogamae parasiticae in Insula Javae lectae exsiccatae, Marian Raciborski [Pf]

¢ Fungi cubenses wrightiani, Charles Wright (1811-86) [St; SC-7].

¢ Fungi exotici exsiccati, Hans Sydow [St; Pf; SC-6].

* Ohio Fungi, William Ashbrook Kellerman (1850-1908) [St; Pf; SC-2].

Loan requests should be directed to: Institute of Botany Herbarium (Fungal

collection), Jagiellonian University, Kopernika 31, 31-501 Krakow, Poland.

Curator of the mycological collection: Dr. Piotr Mleczko.

The Herbarium web-page is http://www.ib.uj.edu.pl/en/?¢d=zielnik

Acknowledgments

Anna Maria Ociepa would like to thank Professor Adam Zajac for employment

in Herbarium of Jagiellonian University in 2006. The authors wish to thank Professor

Reinhard Agerer and Professor Reinhard Berndt for their very valuable comments on

the manuscript.

Literature cited

Botanikos ir genetikos katedra. 2011. Profesorius Antanas Minkevicius [http://www.bg.gf.

vu.lt/?Personalas:Anks%C4%8 Diau_dirb%C4%99_i%26scaron%3Bkiliausi_katedros_

%C5%BEmon%C4%97s:Profesorius_Antanas_Minkevi%C4%8Dius (viewed online on 4

November 2011)]. (in Lithuanian)

Chmiel MA, Mulenko W. 1999. Professor Boguslaw Salata (1940-1999). Acta Mycologica 34(1):

3-5.

132 ... Ociepa, Zubek & Mleczko

Index Fungorum 2011. [http://www.indexfungorum.org/Names/Names.asp (viewed online 4

November 2011)].

Karczmarz K, Satata B. 1982. Studies and botanical achievements of G.S. Nevodowski (1874-1952).

Annales Universitatis Mariae Curie-Sklodowska, Sectio C, Biologia 37: 223-231. (in Russian,

with English summary)

Kohler P. 1999. History of the Jagiellonian University Herbarium (KRA) (Cracow, Poland) in

1780-1910. Kwartalnik Historii Nauki i Techniki 44(2): 7-60. (in Polish with English

summary)

Kohler P. 2001a. History of the Jagiellonian University Herbarium (KRA) (Cracow, Poland)

in 1910-2000. Kwartalnik Historii Nauki i Techniki 46(4): 77-104. (in Polish with English

summary)

Kohler P. 2001b. History of the Jagiellonian University Herbarium (KRA), Cracow, Poland. Taxon

50: 943-945.

Kohler P. 2002. Botany at the Academic Society of Cracow, Academy of Sciences and Letters and

the Polish Academy of Sciences and Letters (1815-1952). Studia i materialy do dziejéw Polskiej

Akademii Umiejetnosci. Tom II. Polska Akademia Umiejetnosci, Krakow. (in Polish with

English summary)

Kohler PS, Zemanek A. 1989. The Marian Raciborski collection in the Museum of the Jagiellonian

University Botanical Garden. Zeszyty Naukowe UJ 926, Prace Botaniczne 18: 135-148. (in

Polish with English summary)

Lizon P. 1973. Andrej Kmet a mykologicky vyskum Slovenska. Ceska Mykologie 27(3): 177-179.

Majewski T. 1995. Jézef Kochman (1903-1995). Rocznik Towarzystwa Naukowego Warszawskiego

58: 55-57. (in Polish)

Pfister DH. 1985. A bibliographic account of exsiccatae containing fungi. Mycotaxon 23: 1-139.

Piekietko-Zemanek A. 1986. A calendar of life of Marian Raciborski. 9-43, in: J Kornas (ed.):

Marian Raciborski. Studia nad zyciem i dzialalnoscia naukowa. Zeszyty Naukowe UJ, Varia

210. (in Polish with English summary)

Pop E. 1972. In memoriam Elexandre Borza (1887-1971). Vegetatio 25(5-6): 273-277. (in French)

http://dx.doi.org/10.1007/BF02198407

Stafleu FA, Cowan RS. 1976-88. Taxonomic literature. Volume I-VII. Regnum Vegetabile 94-116.

Stafleu FA, Mennega EA. 1992-2000. Taxonomic literature. Supplement I-VI, Regnum Vegetabile

125-137.

Stevenson JA. 1971. An account of fungus exsiccati containing material from the Americas. Beihefte

zur Nova Hedwigia 36: 1-563.

Wojewoda W. 1986. Mycology in the scientific output of Marian Raciborski. 59-78, in: J Kornas

(ed.): Marian Raciborski. Studia nad zyciem i dzialalnoscia naukowa. Zeszyty Naukowe UJ,

Varia 210. (in Polish with English summary)

MY COTAXON

http://dx.doi.org/10.5248/120.133

Volume 120, pp. 133-137 April-June 2012

Mycena pseudoinclinata, new to Italy

ALFONSO LA Rosa, ALESSANDRO SAITTA, RICCARDO COMPAGNO

& GIUSEPPE VENTURELLA*

Dipartimento di Biologia vegetale e Biodiversita, Universita di Palermo,

Via Archirafi 38, I-90123 Palermo, Italy

* CORRESPONDENCE TO: giuseppe. venturella@unipa. it

ABSTRACT - The first report of Mycena pseudoinclinata in Italy is accompanied by notes on

the taxonomy, ecology, and distribution of this uncommon basidiomycete.

Key worps — Mycenaceae, sect. Fragilipedes, basidiomycetes

Introduction

Mycena pseudoinclinata (Mycenaceae Overeem) belongs to sect. Fragilipedes

(Fr.) Quél., which is considered the largest section in Mycena (Pers.) Roussel.

The species was originally described by Smith (1947), who distinguished it from

M. inclinata (Fr.) Quél. by the presence of both pleurocystidia and cheilocystidia

and absence of (or fewer) fibrillose flecks on the stipe. This taxon has a limited

distribution all over the world, particularly in Europe where it has recently been

recorded only in Switzerland and France (Robich 2003). This paper deals with

the first record of M. pseudoinclinata in Italy, which we recently collected in a

reforested area in the neighborhood of the town of Palermo (Sicily).

Materials & methods

Basidiomata were identified while fresh and microscopic features were observed

in H,O using a Leica DMLB microscope; spore measurements were based on

100 observations. Nomenclature follows the Index Fungorum (http://www.

indexfungorum.org/Names/Names.asp). The description of the characteristics of

the species is based on personal observations that were subsequently compared

with the descriptions given by Smith (1947) and Robich (2003). The collection is

stored in the fungal dried reference collection of the Herbarium Mediterraneum

Panormitanum, Palermo (PAL).

134 ... La Rosa & al.

Taxonomy

Mycena pseudoinclinata A.H. Sm., North Amer. Species Mycena: 327. 1947.

Fics. 1B, 2

Pileus 20-30 mm, ash-gray, conical to campanulate, pruinose, surface moist

and glabrous, disc slightly umbonate, margin striate. Lamellae close, white,

notched, margin concolorous to the face of lamellae; lamellulae present. Stipe 70

x 3 mm, fragile, hollow, almost concolorous with the pileus, darker at the base

with a slight whitish felt. Flesh typically with a farinaceous odor and taste.

Basidiospores 8-10 x 6-6.5 um, ellipsoid to broadly ellipsoid, smooth,

amyloid. Basidia, 30-35 x 7.5-8.5 um, tetrasporic, claviform. Cheilocystidia

20-60 x 9-12 um, claviform, sphaerical-pedicellate, with irregular excrescences

of different shapes < 10-15 um in length, well-spaced. Epicutis composed of

cylindrical hyphae, smooth, 2-5 um diam., with terminal elements 50-70 x

2-6 um. Subhymenial layer with sub-globose hyphae, isodiametric, 3-8 um

diam. Hyphae of stipe cylindrical, innermost 8-25 um and outermost 1.5-2.5

um diam., smooth or with sparse excrescences, dextrinoid.

COLLECTION EXAMINED: ITALY. Sicity: Raffo Rosso, neighborhood in Palermo, 423

m, on rotten trunks of Eucalyptus camaldulensis Dehnh. (Myrtaceae), 13 Jan 2011, GPS:

38°10'46"N 13°15'40"E, coll. R. Compagno & A. La Rosa (PAL 960).

ComMENTs — Robich (2003) included mycenas with large basidiomata in sect.

Mycena. Some species that might be confused with others in sect. Filipedes

(Fr.) Quél. or Fragilipedes can be differentiated by their basidioma shape and

cheilocystidial type. Mycenas in sect. Fragilipedes are primarily differentiated

through cheilocystidial and/or pleurocystidial shapes, here the main character

differentiating M. pseudoinclinata is the numerous large irregular protuberances

on the cheilocystidia. Although Smith (1947) referred to pleurocystidia in his

original description of M. pseudoinclinata, Maas Geesteranus (1988a,b) correctly

described this species as lacking them. Robich, who cited the cheilocystidia as

restricted to the lamellar edge (Robich 2003), includes the species in his revised

key to Mycena sect. Fragilipedes of the northern hemisphere.

Smith (1947) first reported M. pseudoinclinata as densely cespitose and

occurring in Tennessee, Pennsylvania, and Michigan (U.S.A.). Bi et al. (1993)

report the species from China on fallen twigs in broad-leaved woods. In Europe

Gerault (2005) reports the species from France and Switzerland in broad-leaved

forests mainly characterized by oaks.

We found that M. pseudoinclinata also grows in Eucalyptus plantations

(Fic. 1A) and confirm its role in decaying wood and its typical cespitose habit,

as demonstrated by our collection of eight connate basidiomata per trunk

(Fic. 1B). The literature notes that M. pseudoinclinata fruits from spring (April)

through autumn; in Sicily we have collected basidiomata as early as January, no

Mycena pseudoinclinata, new to Italy ... 135

Fic. 1: Mycena pseudoinclinata.

A. Habitat in Eucalyptus camaldulensis plantation. B. Basidiomata.

136 ... La Rosa & al.

10 um

O®

“8

d -—— 10m

Fic. 2: Mycena pseudoinclinata (PAL 960(Mic)].

a. Basidiospores. b. Claviform cheilocystidia with irregular excrescences of different shapes.

c. Epicutis with cylindrical hyphae. d. Vesiculose and swollen hyphae in the pileitrama.

Mycena pseudoinclinata, new to Italy ... 137

doubt due to the mild weather conditions on the island. Our report represents

the southernmost limit of its distribution in Europe.

Acknowledgements

The authors wish to thank Dr Vladimir Antonin (Czech Republic) and Dr Stephanos

Diamandis (Greece) for critically reviewing the manuscript. The authors would also like

to thank Dr. Cassandra Funsten (USA) for her help with linguistic revision.

Literature cited

Bi ZS, Zheng GY, Li TH. 1993. The macrofungus flora of China's Guangdong Province. The Chinese

University Press, Shatin, N.T., Hong Kong. 737 p.

Gerault A. 2005. Florule evolutive des Basidiomycotina du Finistére. Heterobasidiomycetes.

Tricholomatales. Vers. 2.1. 205 p. http://projet.aulnaies.free.fr/Florules/TRICHOLOMATALES.

pdf

Maas Geesteranus RA. 1988a. Conspectus of the mycenas of the northern hemisphere - 9. Section

Fragilipedes, species A-G. Proc. Kon. Ned. Akad. Wetensch., Ser. C, 91: 43-83.

Maas Geesteranus RA. 1988b. Conspectus of the mycenas of the northern hemisphere - 9. Section

Fragilipedes, species I-R. Proc. Kon. Ned. Akad. Wetensch., Ser. C, 91: 129-159.

Robich G. 2003. Mycena d’Europa. Associazione Micologica Bresadola, Trento. 728 p.

Robich G. 2006. A revised key to the species of Mycena, section Fragilipedes of Northern hemisphere.

Persoonia 19: 143.

Smith AH. 1947. North American species of Mycena. Ann. Arbor: University of Michigan Press.

London: Geoffrey Cumberlege, Oxford University Press. 521 p.

MY COTAXON

http://dx.doi.org/10.5248/120.139

Volume 120, pp. 139-147 April-June 2012

Rossbeevera yunnanensis (Boletaceae, Boletales),

a new sequestrate species from southern China

TAKAMICHI ORIHARA*”?, MATTHEW E. SMITH3,

ZAI-WEI GE* & NITARO MAEKAWA?

"Kanagawa Prefectural Museum of Natural History,

499 Iriuda, Odawara-shi, Kanagawa 250-0031, Japan

*The United Graduate School of Agricultural Sciences, Tottori University,

4-101 Koyama-cho-minami, Tottori 680-8553, Japan

*Department of Plant Pathology, University of Florida, Gainesville FL 32611-0680, USA

‘Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, China

°Fungus/Mushroom Resource and Research Center, Faculty of Agriculture, Tottori University,

4-101 Koyama-cho-minami, Tottori 680-8553, Japan

* CORRESPONDENCE TO: t_orihara@nh.kanagawa-museum.jp

ABSTRACT — A new sequestrate (truffle-like) species, Rossbeevera yunnanensis, is described

based on a collection from Chuxiong Prefecture, Yunnan Province, China. The species is

morphologically characterized by its large, narrow, fusoid to fusiform basidiospores and

remarkably thin peridium. Maximum likelihood and neighbor joining phylogenies of our

nLSU rDNA dataset indicate that the species constitutes the earliest diverging lineage within

the genus Rossbeevera and has a close phylogenetic relationship to species of Leccinellum.

Morphological and phylogenetic relationships to the other Asian members of Rossbeevera

are discussed.

Key worps — Chamonixia, FTA, hypogeous fungi, Leccinum, Rosbeeva

Introduction

The sequestrate (truffle-like) genus Rossbeevera T. Lebel & Orihara (erroneous

orthographic variant: “Rosbeeva”) was erected to accommodate Australasian

and Asian species of Chamonixia Rolland (Lebel et al. 2012a,b), which are

phylogenetically close to but distinct from Chamonixia sensu stricto within

Boletaceae. The genus is characterized by ellipsoid to fusiform basidiospores

with 3-5 longitudinal ridges, bluish green to cyanescent discoloration of

basidiomata, and a thin cutis-like whitish peridium. Phylogenetically, this genus

is closely related to the epigeous bolete genera Leccinum Gray and Leccinellum

Bresinsky & Manfr. Binder and sequestrate genera Chamonixia and Octaviania

Vittad. (Lebel et al. 2012a, Orihara et al. 2012). Rossbeevera species have

140 ... Orihara & al.

been previously reported from Australia—R. vittatispora (G.W. Beaton et al.)

T. Lebel and R. westraliensis T. Lebel, New Zealand—R. pachydermis (Zeller

& C.W. Dodge) T. Lebel, Southeast Asia—R. mucosa (Petri) T. Lebel, China—

R. bispora (B.C. Zhang & Y.N. Yu) T. Lebel & Orihara, and Japan—R. eucyanea

Orihara and R. griseovelutina Orihara (Lebel et al. 2012a,b).

During a fieldtrip in Yunnan Province, China, two of the authors (Smith & Ge)

collected Rossbeevera basidiomata that could not be ascribed to any currently

known species. Here we propose this material as a new species, Rossbeevera

yunnanensis, based on morphological evidence as well as phylogenetic analyses

of the large subunit of the nuclear ribosomal DNA (nLSU).

Materials & methods

Taxon sampling and macro- and microscopic characterization

Fresh basidiomata were located and collected using a truffle rake. Specimens were

placed in wax paper bags and transported to the laboratory within six hours where they

were photographed and rapidly dried in a closed container with silica drying beads.

The dried specimens are deposited in the Herbarium of Cryptogams, Kunming Institute

of Botany, Chinese Academy of Sciences (KUN, with HKAS accession numbers),

Kanagawa Prefectural Museum of Natural History (KPM), and the University of Florida

Herbarium at the Florida Museum of Natural History (FLAS).

For standard light microscopy and differential interference contrast microscopy,

hand-cut sections of dried specimens were mounted in water, 3% KOH, lacto-glycerol,

or 1% phloxine B aqueous solution. To determine the amyloid reaction, dried material

was stained with Melzer’s reagent. Spore dimensions (length x width range) and their

standard deviations (SD), and length of hilar appendages were measured based on 50

randomly selected spores. The length : width ratio (Q) and hilar appendage: spore length

ratio (HA/S, newly introduced here) are presented to describe a range of the basidiospore

proportion. Measurements include the hilar appendage but not ornamentation or

pedicel. Basidium ranges are given as length range x width.

Molecular methods

DNA was extracted from a dried basidioma using the Indicating FTA Cards

(Whatman International Ltd, Maidstone, England). Internal tissue ca. 1 mm thick was

soaked in 99.5% ethanol for 2 min before DNA extraction. The extraction was done

according to the manufacturer's protocol for plant samples (available at http://www.

whatman.com/References/WGI_1397_PlantPoster_V6.pdf) using FTA Cards, more

convenient and efficient over traditional methods (Dentinger et al. 2010). Furthermore,

we demonstrate here that this technique is useful for dried tissues after a brief ethanol

pretreatment. For PCR amplification and sequencing reactions, we used the primer

pair LROR / LRS (Vilgalys & Hester 1990). For PCR conditions, sequencing and editing

of the obtained sequence, see Lebel et al. (2012a). The nLSU sequence is deposited in

GenBank.

Sequences included in the nLSU dataset were retrieved from GenBank based on the

previous analyses of Rossbeevera and allied genera (Lebel et al. 2012a). Some sequences

Rossbeevera yunnanensis sp. nov. (China) ... 141

of Leccinellum spp., which are shown to be paraphyletic with Rossbeevera, were selected

according to the phylogeny in Lebel et al. (2012a) and were used to root the phylogenetic

tree. Multiple sequence alignment was performed using Clustal X ver. 1.83 (Thompson

et al. 1997) and the data were manually adjusted in SeaView (Galtier et al. 1996). Gaps

were treated as “missing” data for all analyses.

The neighbor-joining (NJ) analysis based on p distance was conducted with Clustal

X ver. 1.83 with 1000 bootstrap replicates and with default settings. Subsequently, the

maximum likelihood (ML) analysis was done by PhyML ver. 3.0 (Guindon et al. 2010)

under GTR + I + G model based on hLRT estimated by MrModeltest 2.3 (Nylander

2004), starting the analysis using a tree estimated by the Nearest Neighbor Interchanges

(NNIs) and setting the number of bootstrap replicates to 1000. The resulting trees were

visualized with FigTree ver. 1.3.1 (Rambaut, available at http://tree.bio.ed.ac.uk/).

Results

The finally aligned nLSU dataset was 837 bp long, including full length

reads for all 19 sequences but excluding the primer regions. Settings of the best

model estimated by hLRT in MrModeltest 2.3 were as follows: base frequencies

(A = 0.2700, C = 0.1941, G = 0.2904, T = 0.2455), proportion of invariable

sites (I = 0.7687), and gamma distribution shape parameter (a = 1.9606).

Maximum likelihood analysis generated one ML tree (In L = -2078.576887;

Fic. 1). Branches that were moderately to strongly supported by ML bootstrap

values (>60%) were also strongly supported in the NJ topology (BS >70%). In

the Rossbeevera clade, R. eucyanea and R. griseovelutina were each supported to

be monophyletic by high BS values but the Australasian species were not well

differentiated from each other, as shown in Lebel et al. (2012a). The Chinese

Rossbeevera sp. MES420 formed a basal branch within the Rossbeevera clade

and was phylogenetically distinct from the other members of Rossbeevera

included in this study.

Taxonomy

Rossbeevera yunnanensis Orihara & M.E. Sm., sp. nov. Fic. 2

MycoBank MB 563649

Differs from Rossbeevera bispora by thinner or partially lacking peridium, fusoid to

fusiform basidiospores, and 2—4-spored basidia.

Type: China, Yunnan Province, ChuXiong Yi Autonomous Prefecture, Mt. Zixi, 19

Sept. 2010, M. E. Smith et Z.-W. Ge, MES420 (holotype - HKAS 70689, isotype - KPM-

NC0017850, GenBank JN979437).

ETyMOLOGy: yunnanensis (Latin) refers to the type locality.

BASIDIOMATA Sparse, ca. 1 cm, subglobose, soft, surface covered with very thin,

minutely felty, whitish to almost translucent peridium gradually turning bluish

gray or black when touched or bruised, the peridium partially absent, exposing

the gleba. GLEBA off-white to beige when immature, reddish brown to blackish

142 ... Orihara & al.

Rossbeevera westraliensis AUS HQ647161

Rossbeevera westraliensis AUS HQ647160

Rossbeevera vittatispora AUS HQ647148

Rossbeevera pachydermis NZ DQ534620

53/ -

pate Rossbeevera pachydermis NZ HQ647157

Rossbeevera vittatispora AUS HQ647150

Rossbeevera vittatispora AUS HQ647159

Rossbeevera griseovelutina JPN HQ693877

100/100 | Rossbeevera griseovelutina JPN HQ693876

Rossbeevera griseovelutina JPN HQ693878

96/97

Rossbeevera eucyanea JPN HQ693880

100/100 ' Rossbeevera eucyanea JPN HQ693879

Rossbeevera yunnanensis CHINA JN979437

Leccinellum albellum AY612811

Leccinellum aff. griseum JPN JN378509

Leccinellum carpini AF139691

Leccinellum rugosiceps AY612813

Leccinellum crocipodium AF139694

a oat ee 100/100 L Leccinellum crocipodium AF454590

0.1 substitutions / site

FiguRE 1. Maximum likelihood (ML) tree of the nLSU dataset of Rossbeevera species with

Leccinellum species selected as the outgroup. ML and Neighbor-Joining (NJ) bootstrap (BS) values

(1000 replicates; only BS >50% are shown) are indicated above or below branches or at nodes as

MLBS/NJBS. Abbreviations: AUS = Australia; NZ = New Zealand; JPN = Japan.

brown at maturity, rubbery, composed of minute, irregular locules, turning

bluish gray in some portions when cut and exposed to air, trama subgelatinous,

somewhat translucent. STIPE-COLUMELLA present but reduced, somewhat

dendroid or as a small basal pad, subgelatinous, translucent. ODOR unknown.

BASIDIOSPORES (14.4—)16.9-23.4(-24) x 6.8-9(-9.5) um, (mean + SD

= 20.1 + 1.60 x 7.8 + 0.61), Q = 2-3.8 (mean = 3.1), symmetric, fusoid to

fusiform, inamyloid, nondextrinoid, colorless at first then becoming reddish

brown at maturity, with 3-4 or rarely 5 smooth, longitudinal ridges up to 2.3

um high (mostly ca. 1 um high) in water, walls 0.7-1.5 um thick, with a large

hilar appendage (2-)2.3-3.8 um long (mean 3.1), HA/S = 0.12-0.22, (mean =

0.16) at the base and a minute, incomplete hollow (> 1 um in diameter) at the

tip. Basrpra 17.5-30 x 6.8-11 um (mean = 23.5 x 8.4 um; n = 15), cylindrical

to cylindro-clavate, colorless to pale yellow-brown to reddish brown, 2-, 3- or

Rossbeevera yunnanensis sp. nov. (China) ... 143

4-spored. HymEenium developed when immature but collapsed at maturity,

colorless, hymenial cystidia not seen; BASIDIOLES cylindrical or clavate to

clavulate. SUBHYMENIUM not developed. TRama of densely or somewhat

loosely interwoven, partly branched, colorless, non-inflated, thin-walled (< 0.8

uum thick), filamentous hyphae 3.5-8 um broad. STIPE-COLUMELLA composed

of loosely interwoven, subgelatinous, thin-walled (< ca. 1 um thick) filamentous

hyphae (2-)5-8(-9.5) um broad. PERIDIUM thin, absent in some portions, up

to 60 um thick in the dried basidioma, yellowish brown under light microscopy,

composed of repent, non-inflated, thin-walled (< 0.8 um thick) filamentous

hyphae 2.5-6 um broad but the hyphae are collapsed in most portions. CLAMP

CONNECTIONS absent in all tissues.

HABITAT AND DISTRIBUTION: Hypogeous in mixed forest dominated by

Pinus yunnanensis Franch., Lithocarpus mairei (Schottky) Rehder, L. dealbatus

(Hook. f. & Thomson ex Miq.) Rehder, and unidentified Quercus spp.; known

only from Yunnan Province, China.

Discussion

Rossbeevera yunnanensis shares several morphological characteristics

with the other Rossbeevera species, including the bluish color change of the

peridium and reddish brown basidiospores with 3-5 unbranched longitudinal

ridges. ‘This new species also has the large, distinctly fusoid to fusiform (Q =

2.0-3.8) basidiospores and remarkably thin peridium characteristic of the

genus. Rossbeevera griseovelutina, which is reported from Japan, has slightly

larger basidiospores (14.4-31.9 x 6.7-10.4 um, mean = 22.2 x 8.7 um) and the

Q value is smaller (Q = 2.0-3.3; Lebel et al. 2012a). In addition, the latter species

has a more fully developed peridiopellis composed of more or less vertically

oriented, partially inflated hyphae. Rossbeevera yunnanensis also differs

morphologically from another known Chinese species, R. bispora, because

R. yunnanensis has a thinner peridium, larger and narrower basidiospores, and

its basidia often bear 4 spores rather than 2 spores characteristic of R. bispora

(Zhang & Yu 1989). We have examined the holotype of R. bispora (GDGM5688)

and have confirmed the above-noted differences. The Southeast Asian species,

R. mucosa is readily distinguished from R. yunnanensis in its ellipsoid to fusoid

basidiospores (13—)15-17 um in diameter with a lower Q value (Q = 1.76-2.05;

Lebel et al. 2012a).

The HA/S ratio, which is newly introduced in this study, could be helpful to

describe spore shapes objectively when spore sizes vary widely within collections

or sporocarps of a particular species. Basidiospores of R. griseovelutina are

somewhat similar to those of R. yunnanensis as discussed above, but the

mean HA/S ratio of R. griseovelutina was significantly higher than that of

R. yunnanensis. However, the ranges overlapped: HA/S of R. griseovelutina =

0.12-0.24, mean 0.18, n = 25 vs. HA/S of R. yunnanensis = 0.12-0.22, mean

144 ... Orihara & al.

= 0.16. Basidiospore morphology is widely used as a key character for the

taxonomy of sequestrate fungi (Castellano et al. 1989). The HA/S difference

is proposed as a new, useful diagnostic character that could be useful in the

taxonomy of Rossbeevera and other sequestrate genera.

In our phylogeny R. yunnanensis formed the earliest diverging lineage

within the Rossbeevera clade with moderate to strong bootstrap support

(ML: 67%; NJ: 91%). Although we have not been able to include sequences of

R. bispora, it is possible that the two Chinese species are closely related within

Rossbeevera, because they share a similar overall morphology, including a cutis-

like peridium and relatively large basidiospores with mostly 3-4 longitudinal

ridges. Sequences from additional genes and specimens of Rossbeevera from

China and the adjacent regions would help to clarify the evolutionary origin of

this sequestrate genus.

Although we have examined only one R. yunnanensis collection, its

morphological characteristics and unique phylogenetic position are sufficient

to discriminate the species from the other members of Rossbeevera. We would

prefer to have additional specimens in order to provide more ecological

information about R. yunnanensis, such as details of its phenology and host tree

preferences. However, since hypogeous fungi are difficult to find and collect, we

felt it was important to describe this new Rossbeevera species to promote further

research on the systematics of Asian sequestrate fungi. Unique sequestrate

fungi have been found in Asia for more than a century (e.g. Corner & Hawker

1953; Smith & Schmull 2011). However, several recent studies highlight the fact

that Asia is likely a hotspot for unique, endemic truffles and probably hosts a

high diversity of undescribed sequestrate fungi (Yang et al. 2006; Desjardin et

al. 2008, 2009; Orihara et al. 2008, 2010, 2012; Kinoshita et al. 2011). Clearly,

more taxonomic work on sequestrate fungi is needed in this region.

Acknowledgments

The study was supported by Research Fellowships for Young Scientists (No. 21-6052)

from the Japan Society for the Promotion of Science and partially by the Grant-in-Aid

for the Global COE Program “Advanced Utilization of Fungus/Mushroom Resources

for Sustainable Society in Harmony with Nature” from the Ministry of Education,

Culture, Sports, Science and Technology of Japan. Financial support for the field trip

FIGURE 2 (right). Rossbeevera yunnanensis (holotype): a. Basidiomata. A reduced columella is

present in the center of the section (arrow); b. Significantly reduced peridium of dried material

(arrow); c. Thick portion of the peridium; d. Mature basidiospores with longitudinal ridges

(arrows) mounted in lacto-glycerol; e. Immature basidiospores viewed from above (arrow);

f. 3- and 4-spored basidia mounted in lacto-glycerol after a pretreatment with 1% phloxine B

aqueous solution; g. 2-spored basidium mounted in lacto-glycerol. Scale bars: a = 1 cm; b = 20 um;

c = 30 um; d-g = 10 um.

Rossbeevera yunnanensis sp. nov. (China) ... 145

146 ... Orihara & al.

was provided by the National Natural Science Foundation of China (No. 30800004)

and the Knowledge Innovation Program of the Chinese Academy of Sciences (No.

KSCX2-EW-J-24). The authors gratefully acknowledge Dr. Kentaro Hosaka (National

Museum of Nature and Science, Ibaraki, Japan) and Ms. Rosanne Healy (University of

Minnesota) for serving as pre-submission reviewers and their helpful comments on the

manuscript.

Literature cited

Castellano M, Trappe JM, Maser Z, Maser C. 1989. Key to spores of the genera of hypogeous

fungi of north temperate forests with special reference to animal mycophagy. Mad River Press,

California.

Corner EJH, Hawker LE. 1953. Hypogeous fungi from Malaya. Transactions of the British

Mycological Society 36: 125-137.

Dentinger BM, Margaritescu S, Moncalvo J-M. 2010. Rapid and reliable high-throughput methods

of DNA extraction for use in barcoding and molecular systematics of mushrooms. Mol Ecol

Resources 10: 628-633. http://dx.doi.org/10.1111/j.1755-0998.2009.02825.x

Desjardin DE, Wilson AW, Binder M. 2008. Durianella, a new gasteroid genus of boletes from

Malaysia. Mycologia 100: 956-961. http://dx.doi.org/10.3852/08-062

Desjardin DE, Binder M, Roekring S, Flegel T. 2009. Spongiforma, a new genus of gastroid boletes

from Thailand. Fungal Diversity 37: 1-8.

Galtier N, Gouy M, Gautier C. 1996. SEAVIEW and PHYLO_WIN: Two graphic tools for sequence

alignment and molecular phylogeny. Systematic Biology 12: 543-548.

http://dx.doi.org/10.1093/bioinformatics/12.6.543

Guindon S, Dufayard J-F, Lefort V, Anisimova M, Hordijk W, Gascuel O. 2010. New algorithms and

methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML

3.0. Systematic Biology 59: 307-321. http://dx.doi.org/10.1093/sysbio/syq010

Kinoshita A, Sasaki H, Nara K. 2011. Phylogeny and diversity of Japanese truffles (Tuber spp.)

inferred from sequences of four nuclear loci. Mycologia 103: 779-794.

http://dx.doi.org/10.3852/10-138

Lebel T, Orihara T, Maekawa N. 2012a. The sequestrate genus Rosbeeva T. Lebel & Orihara gen. nov.

(Boletaceae) from Australasia and Japan: new species and new combinations. Fungal Diversity

52: 49-71. http://dx.doi.org/10.1007/s13225-011-0109-x

Lebel T, Orihara T, Maekawa N. 2012b. Erratum to: The sequestrate genus Rossbeevera T. Lebel &

Orihara gen. nov. (Boletaceae) from Australasia and Japan: new species and new combinations.

Fungal Diversity 52: 73. http://dx.doi.org/10.1007/s13225-011-0118-9

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology

Centre, Uppsala Univ, Uppsala, Sweden.

Orihara T, Kasuya T, Phongpaichit S, Dissara Y. 2008. Radiigera tropica (Geastraceae, Geastrales), a

new species from a tropical rain forest of Thailand. Mycotaxon 105: 111-117.

Orihara T, Sawada F, Ikeda S, Yamato M, Tanaka C, Shimomura N, Hashiya M, Iwase K. 2010.

Taxonomic reconsideration of a sequestrate fungus, Octaviania columellifera, with the proposal

of a new genus, Heliogaster, and its phylogenetic relationships in the Boletales. Mycologia 102:

108-121. http://dx.doi.org/10.3852/08-168

Orihara T, Smith ME, Shimomura N, Iwase K, Maekawa N. 2012. Diversity and systematics of the

sequestrate genus Octaviania in Japan: two new subgenera and eleven new species. Persoonia

28: 85-112. http://dx.doi.org/10.3767/003158512X650121

Rossbeevera yunnanensis sp. nov. (China) ... 147

Smith ME, Schmull M. 2011. Tropical truffles: English translation and critical review of F von

Hohnel’s truffles from Java. Mycological Progress 10: 249-260.

http://dx.doi.org/10.1007/s11557-010-0694-1

Thompson JD, Gibson TJ, Pleweniak F, Jeanmougin F, Higgins DG. 1997. The Clustal X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 25: 4876-4882.

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several Cryptococcus species. The Journal of Bacteriology 172:

4238-4246.

Yang ZL, Trappe JM, Binder M, Sanmee R, Lumyong P, Lumyong S. 2006. The sequestrate genus

Rhodactina (Basidiomycota, Boletales) in northern Thailand. Mycotaxon 96: 133-140.

Zhang B-C, Yu Y-N. 1989. Chamonixia bispora sp. nov. (Boletales) from China. Mycotaxon 35:

277-281.

MY COTAXON

http://dx.doi.org/10.5248/120.149

Volume 120, pp. 149-155 April-June 2012

Octaviania violascens: a new sequestrate bolete from Thailand

RATTAKET CHOEYKLIN™, THITIYA BOONPRATUANG?’,

SUJINDA SOMMAI’ & SAYANH SOMRITHIPOL’

‘Enzyme Technology and Waste Management Research Unit,

Kasetsart Agricultural and Agro-Industrial Product Improvement Institute (KAPI),

Kasetsart University, Bangkok, Thailand

*Mycology Laboratory, Bioresources Technology Unit,

National Center for Genetic Engineering and Biotechnology,

Thailand Science Park, Pathumthani, Thailand

* CORRESPONDENCE TO: rattaketster@gmail.com

ABSTRACT — Octaviania violascens sp. nov. is proposed for a new sequestrate bolete

from Khao Yai National Park, Nakhon Ratchasima Province, Thailand. A comprehensive

description and illustrations are provided.

Key worps — Basidiomycota, truffle, Octavianiaceae

Introduction

While walking along an elephant trail collecting mushrooms in a bamboo

grove in Khao Yai National Park, Nakhon Ratchasima Province, ‘Thailand,

we encountered a rose-colored fungus where recent rain had washed away

the topsoil. The distinctive characteristics of this species, besides the rose

color of its basidiocarps, were that it bruised deep reddish violet, contained

non-glutinous gleba, and had clamp connections in the peridial layer and

basidia containing only two highly ornamented spores. Its micromorphology

clearly placed it in Octaviania and it is the first record of this genus from

Thailand. Octaviania is included in the group of sequestrate basidiomycetes

called truffles. In Thailand 8 species of sequestrate fungi have been reported,

including Arcangeliella rosea (Harkn.) Zeller & C.W. Dodge (Ellingsen 1982),

A. beccarii (Petri) Zeller & C.W. Dodge (Chandrasrikul et al. 2008), Descomyces

cf. albellus (Massee & Rodway) Bougher & Castellano (Dissing 1963, Ellingsen

1982), Mycoamaranthus cambodgensis (Pat.) Trappe, et al. (Chandrasrikul et al.

2007, 2008, Lumyong et al. 2003), Radiigera tropica Orihara & T. Kasuya, and

three other unknown species from Dissing (1963) in the genera Hymenogaster,

150 ... Choeyklin & al.

Leucogaster, and Melanogaster. This paper describes a newly discovered species,

Octaviania violascens, from Thailand.

Materials & methods

COLLECTION SITE: The study site is in a bamboo grove, in a secondary forest near

the 28 km marker post along the road from the northern entrance in Khao Yai National

Park, Nakhon Ratchasima Province, Thailand.

FUNGAL MATERIALS: Dried specimens are deposited in the BIOTEC Bangkok

Herbarium (BBH).

MorpHo ocy: Macromorphological data were recorded from dried specimens using

an Olympus model XZ30 stereomicroscope. Micromorphological data were recorded

from dried specimens using an Olympus model BX51 microscope. Drawings were

made using an Olympus Camera Lucida model U-DA. Measurements and drawings

were made from slide preparations stained with cotton blue, Melzer’s reagent, or 3%

potassium hydroxide. The following abbreviations are used for chemical reactions: IKI

(Melzer’s reagent, with IKI- = non—amyloid and non-dextrinoid; IKI+ = amyloid or

dextrinoid), KOH (3% potassium hydroxide), and CB (cotton blue; CB+ = cyanophilous;

CB(+) = weakly cyanophilous; CB- = acyanophilous). Basidium width was measured

at the broadest part, and basidium length was measured from the apex (sterigmata

excluded) to basal septum. Color terms and notations in parentheses follow Kornerup

and Wanscher (1963). Spore statistics include: x = the arithmetic mean of the spore

length by spore width (+ standard deviation) for n spores measured; Q = the quotient of

length and width in any one spore, indicated as a range of variation in n spores measured;

Qm = the mean of Q—values (+ standard deviation). Ultrastructures were recorded from

dried specimens using a Hitachi S-3400N scanning electron microscope.

Taxonomy

Octaviania violascens Choeyklin, Boonprat. & Somrith., sp. nov. PLATES 1-3

MycoBank MB 563238

Differt a O. purpurea, absque columella et rhizomorpha, loculus rotundatus, spora

verruca angulari, basidia bispora pariete crasso; praesentia fibularum et cellularum

spheropedunculatarum et hypharum laticiferarum.

TYPE — THAILAND. Nakhon Ratchasima, Pak Chong district, Khao Yai National Park, 28

km. marker post (Bambusa spp. forest), 14°30'59.47"N 101°22'08.29"E, 23 August 2010,

coll. Rattaket Choeyklin & Sujinda Sommai, RCK 00093 (Holotype BBH 30342).

EtryMoLocy -— violascens (Latin) = turning violet.

Basidiocarps 9-18 mm tall x 12-22 mm wide (dried specimens), ovoid,

subglobose to turbinate, white to cream colored near base, lumpy or folded on

the upper part; smooth to cracking to form squamules, the squamules reddish

grey to greyish rose (12B2-3), staining reddish violet to deep violet (18C-D8)

where bruised or cut; dull, dry, hard and cartilaginous. Peridium thin, 0.5-0.6

mm diam., white, quickly changing from violet white to pale violet (16A3-4)

when cut. Gleba lacunate, 0.5-1 mm diam., locules filled with powdery brown

Octaviania violascens sp. nov. (Thailand) ... 151

PLaTE 1: Octaviania violascens. 1. Basidiocarps in various shapes; 2. Peridial layer cracking to

form squamules and deep violet stains where bruised; 3. Gleba lacunate, quickly changing to violet

brown when cut, basidiocarps without rhizoid (holotype). Bar = 10 mm.

spores when dried, dark brown when fresh, solid when fresh and with a hollow

central cavity when dried, watery, quickly changing to violet brown (11F8) when

cut; sterile tissue between locules fibrous, white but immediately changing to

reddish violet to deep violet when bruised (18C-D8). Latex absent. Columella

absent. Odor not distinctive.

Basidiospores without spines (11-)12-15(-19) x 11-14(-15) um (x = 13.84

+ 1.53 x 12.44 + 1.16 um, Q = 1.0-1.46, Qm = 1.09 + 0.11, n = 25); with spines

(12-)15-17(-18) x (10-)15-20 um (x = 15.79 + 1.36 x 16.40 + 2.14 pm, Q =

0.67-1.50, Qm = 0.98 + 0.15), orthotropic and heterotropic, symmetric and

asymmetric, globose to subglobose, walls 2.0-2.5 um thick, pale yellowish

brown, golden-brown, brown to dark brown in water and 3% KOH, IKI+

(weakly dextrinoid), CB-, spore ornamentation echinulate to conical spines,

1-2 um tall x 2 um wide at base x 1 um wide at apex, most apical spines acute

152 ... Choeyklin & al.

PLATE 2: Octaviania violascens (holotype). 1. Basidiospore formation; 2. 2-spored basidia; 3. Basidia

shapes; 4. Spheropedunculate cell from peridial context; 5. Hymenium with basidia; 6. Oleiferous

hyphae; 7. Skeletal hyphae. Bar = 10 um.

but some blunted or slightly bent, hyaline in 3% KOH and water, IKI+ (weakly

dextrinoid), CB+; pedicel 6-10 x 3 um, walls 1-1.5 um thick, pale yellowish-

brown. Basidia (12.5-)20-31(-35) x (5-)7.5-15 um, 2-spored, lageniform

to fusoid, walls >1 tm thick, hyaline when young to pale yellowish brown to

brown when mature in water and KOH, IKI-, CB+, basally septate; sterigmata

7-8 um tall x 3 um wide, walls 1 um thick, pale brown in water and 3% KOH,

IKI-, CB+, septa present at the sterigmal base. Peridial context 170-300 um

diam., containing two types of hyphae consisting of oleiferous and binding

hyphae; oleiferous hyphae 3-4 um wide, thin-walled, brown in water and 3%

Octaviania violascens sp. nov. (Thailand) ... 153

PLATE 3: Octaviania violascens (holotype): scanning electron micrographs of spores. 1. Mature

basidiospore, 2. Basidiospore wall, 3. Basidiospores variation, Bar 1-2 = 10 um, 3 = 3 um

KOH, IKI-, CB+; binding hyphae 4-6 um width, thick-walled, walls 1-1.5 um

thick, hyaline in water and 3% KOH, IKI-, CB+, and pinkish when stained

with phloxine. Sphaeropedunculate cells 27.5-30 x 27.5-30 um, embedded

in the peridial context, pedicel 7 x 2 um, ovoid, thick-walled, hyaline, IKI-.

Subhymenium IKI+ (weakly dextrinoid). Tramal plate 25-30 um diam., thin-

walled, hyaline, IKI-. Peridiopellis of repent hyphae, interwoven, thin-walled,

hyaline, IKI-. Sterile tissues that separate the locules are identical to the tramal

plate. Clamp connections present only in the peridial tissue.

154 ... Choeyklin & al.

ECOLOGY & DISTRIBUTION: Solitary to gregarious or in small clusters,

subhypogeous to hypogeous, in sandy soil in a Bambusa spp. (Bambusaceae)

forest, in open secondary forest. Thailand, Nakhon Ratchasima Province, Khao

Yai National Park, 28 km marker post.

Discussion

Octaviania violascens may be diagnosed by its echinulate spores with conical

spines and thick-walled 2-spored basidia. The new species is most similar to

O. purpurea Coker & Couch (Coker & Couch 1928) but differs in the texture

of the gleba, absence of a columella and rhizomorphs, spore shape, colour,

and ornamentation, presence of laticiferous hyphae, and basidia shape and

sterigmata number (TaBLE 1). Spheropedunculate cells in the peridial context

and clamp connections found in O. violascens are absent in O. purpurea

(TABLE 1).

Heliogaster Orihara & K. Iwase also produces an empty-chambered

nonglutinous gleba, but unlike Octaviania its gleba is soft, not rubbery and its

peridium composed of filamentous hyphae (not partly inflated hyphae with

isodiametric cells) and spores with a perisporium. In the absence of DNA

analyses, we retain O. violascens within Octaviania based on morphology.

TABLE 1. Morphological characters in Octaviania violascens and O. purpurea.

MORPHOLOGY O. violascens O. purpurea

Sporocarp texture Rubbery Not reported

Spore ornamentation Echinulate with conical spines Angular warts

Spore shape Globose Subglobose

Spore color Pale yellowish brown to brown Yellowish brown to dark brown

Basidia 2-spored, thick-walled 1-spored, thin-walled

Basidial shape Lageniform to fusoid Jug-shaped

Basidial size ~20-31 x 7.5-15 um 30-44 x 5.5-7.8 um

Clamp connection Present Absent

Spheropedunculate cells Present Absent

Rhizoid & columella Absent Present

Locules Round Labyrinthiform

Acknowledgments

We express appreciation to Michael Castellano and Teresa Lebel for reviewing this

paper. We thank Tim Flegel for reviewing English. We are grateful Ellen Bloch (curator

of New York Botanical Garden) for the loan of Octaviania purpurea Coker & Couch

#780569. This study was funded by BIOTEC, Thailand (TRF/BRT - The biodiversity

Research and Training Program grant #BRT_R650001) for field survey. We appreciate

the Department of National Parks, Wildlife and Plant Conservation, Ministry of Natural

Resources and Environment for permitting collecting inside Khao Yai National Park.

Octaviania violascens sp. nov. (Thailand) ... 155

Literature cited

Chandrasrikul A, Phanichapol D, Boonthavikoon T, Chalermpong A. 2007. Mushrooms in

Thailand. The Royal Institute Press, Thailand. 272 p.

Chandrasrikul A, Suwanarit P, Sangwanit U, Morinaga T, Nishizawa Y, Murakami Y. 2008. Diversity

of mushrooms and macrofungi in Thailand. Kasetsart University, Bangkok. 514 p.

Coker WC, Couch JN. 1928. The gasteromycetes of the eastern United States and Canada. Chapel

Hill The University of North Carolina Press, North Carolina. 508 p.

Dissing H. 1963. Studies in the flora of Thailand 25: discomycetes and gasteromycetes. Dansk

Botanisk Arkiv 23: 117-130.

Ellingsen HJ. 1982. Some gasteromycetes from northern Thailand. Nordic Journal of Botany 2:

283-285. http://dx.doi.org/10.1111/j.1756-1051.1982.tb01190.x

Kornerup A, Wanscher JH. 1963. Methuen handbook of colour. Methuen & Co Ltd press, Great

Britain. 224 p.

Lumyong S, Sanmee R, Lumyong P, Yang ZL, Trappe JM. 2003. Mycoamaranthus cambodgensis

comb. nov., a widely distributed sequestrate basidiomycete from Australia and southeastern

Asia. Mycological Progress 2: 323-325. http://dx.doi.org/10.1007/s11557-006-0069-9

Orihara T, Kasuya T, Phongpaichit S, Dissara Y. 2008. Radiigera tropica (Geastraceae, Geastrales),

a new species from a tropical rain forest of Thailand. Mycotaxon 105: 111-117.

Orihara T, Sawada F, Ikeda S, Yamato M, Tanaka C, Shimomura N, Hashiya M, Iwase K. 2010.

Taxonomic reconsideration of a sequestrate fungus, Octaviania columellifera, with the proposal

of a new genus, Heliogaster, and its phylogenetic relationships in the Boletales. Mycologia 102:

108-121. http://dx.doi.org/10.3852/08-168

MY COTAXON

http://dx.doi.org/10.5248/120.157

Volume 120, pp. 157-169 April-June 2012

New records of cercosporoid hyphomycetes from Iran

MAHDI PIRNIA’, RASOUL ZARE’,

HAMID R. ZAMANIZADEH! & AKBAR KHODAPARAST?

"Science and Research Branch, Islamic Azad University, Tehran, Iran

*Department of Botany, Iranian Research Institute of Plant Protection,

PO. Box 1454, Tehran, 19395, Iran

*Department of Plant Protection, University of Guilan, Rasht, Iran

* CORRESPONDENCE TO: pirnia@ymail.com

ABsTRACT — Eight collections from different localities in the northern provinces of Iran

were obtained during spring-summer 2010-11. Seven species on eight host plants were

identified: Cercospora pantoleuca on Plantago lanceolata, Cercosporella primulae on Primula

macrocalyx, C. virgaureae on Conyza bonariensis, Passalora bondartsevii on Medicago sp.,

Pseudocercospora danaicola on Danae racemosa, P. paraguayensis on Eucalyptus camaldulensis,

and Scolecostigmina confluens on Crataegus melanocarpa and C. pseudomelanocarpa. ‘These

taxa are new records to Iran. Cercosporina danaicola is recombined as Pseudocercospora

danaicola.

Key worps — Cercospora-like fungi, taxonomy, Asia Minor, leaf spot pathogens

Introduction

Cercosporoid fungi include over 30 morphologically similar anamorphic

hyphomycetous genera that are mainly associated with leaf spots on various host

plants such as cereals, vegetables, ornamentals, forest trees, and weeds (Crous

& Braun 2003). Cercospora Fresen. is one of the largest genera in this group.

In his monograph of Cercospora, Chupp (1954) listed over 1800 species names

and accepted 1419 species. He adopted a broad generic concept and mainly

considered the characteristics of conidia (hilum thickness, pigmentation,

number, and arrangement). In a major taxonomic treatment, Deighton (1967,

1973, 1974, 1976, 1979) divided and reclassified many Cercospora species into

several segregate genera, including Cercosporella Sacc., Cercosporidium Earle,

Paracercospora Deighton, Pseudocercospora Speg., Pseudocercosporella Deighton,

and Pseudocercosporidium Deighton. Other morphologically similar fungi are

members of Passalora Fr. and Scolecostigmina U. Braun. Cercospora species are

characterized by acicular hyaline septate conidia with conspicuous hila produced

158 ... Pirnia & al.

on pigmented unbranched septate smooth conidiophores. Cercosporella species

are easily distinguished from Cercospora species by colourless structures with

thickened darkened conidiogenous loci. In Pseudocercospora, conidiogenous

loci and hila are inconspicuous, not thickened, and not darkened. Passalora is

distinguished by pigmented conidiophores and pigmented ellipsoid-fusiform

to obclavate-subcylindrical conidia. Scolecostigmina is diagnosed by thick-

walled verruculose conidiophores with conspicuous annellations in the upper

part. Conidia in this genus are subcylindric-obclavate, smooth to verruculose,

transversely multi-euseptate, and occasionally with a few longitudinal septa.

Crous & Braun (2003) published an annotated checklist for Cercospora and

Passalora names with 5720 taxa and taxonomic re-allocations of numerous

species. Recently Braun & Crous (2007) proposed several new species,

combinations, and names after re-examining the type collections of Cercospora

species and other related genera.

Iranian records of Cercospora and other related genera have been only

poorly studied. Scharif & Ershad (1966) presented a list of fungi on various

host plants including a few Cercospora and Pseudocercospora species. Ershad

(1990, 2000, 2002) reported four Cercospora species from Iran and later (Ershad

2009) corrected 59 Cercospora names previously published from Iran, partly

according to Crous & Braun (2003), thus reducing the number of species to 22

with 14 uncertain species. He did not follow Crous & Braun (2003) in merging

morphologically indistinguishable taxa on various host plants in Cercospora

apii s. lat. Recently Pirnia et al. (2010, 2012a, 2012b) studied Cercospora,

Passalora, Ramularia, and Ramularia-like species in Iran following the species

concepts proposed by Crous & Braun (2003) and Braun (1998). They linked

six host plants with C. apii s. lat. Furthermore, some specimens deposited in

the fungus reference of the Iranian Ministry of Agriculture (Iranian Research

Institute of Plant Protection, Tehran) have been examined and proved to be

morphologically indistinguishable from C. apii.

Materials & methods

Specimens with leaf spot symptoms from different localities in northern Iran

(i.e., Guilan, Mazandaran, Golestan, northern Khorasan provinces) were collected

during spring-summer 2010-11. Microscopic slides were prepared from stromata,

conidiophores, and conidia in 25% lactic acid. Species were identified based on stromata

(presence/absence) and their development, conidia (pigmentation, shape, dimensions),

hila (thickness, darkness), conidiophores (pigmentation, dimensions) and conidial scars

(position, thickness, darkness). Drawings were made using a drawing tube attached to

an Olympus BH-2 microscope.

Results

In our study on cercosporoid hyphomycetes of Iran, we identified 70 taxa on

100 host plants (TABLE 1). Of the 21 taxa that are new records for the mycobiota

Cercosporoids new to Iran...

TABLE 1. Cercosporoid hyphomycetes and hosts identified from Iran

PATHOGEN

*Cercospora acnidae

C. althaeina

*C. apii

C. beticola

*C. bizzozeriana

*C. caricis

C. cheiranthi

C. lactucae-sativae

*C. mercurialis

*C. pantoleuca

*C. peckiana

C. sorghi

C. traversiana

C. violae

C. zebrina

C. zonata

*Cercosporella primulae

*C. virgaureae

Neoovularia ovata

Passalora bolleana

*P. bondartsevii

P. calotropidis

*P. chaetomium

P. circumscissa

P cousiniae

P. dubia

P. graminis

P. microsora

P. personata

P. phaeopappi

P punctum

P. rosae

P rosicola

*P. ziziphi

Pseudocercospora abelmoschi

P. atromarginalis

P. cruenta

*P. danaicola

*P. griseola

*P. heteromalla

P. jujubae

P. kaki

P. neriella

*P. paraguayensis

P. punicae

Hosts

Amaranthus chlorostachys var. chlorostachys

*Gossypium hirsutum

Abutilon theophrasti, Euphorbia heterophylla, Gerbera jamesonii,

Medicago sp., Pelargonium zonale, Petunia hybrida, Solanum

lycopersicum, Vigna sinensis, Zantedeschia aethiopica

Beta vulgaris, B. maritima

Cardaria draba

Carex orbicularis

Cheiranthus cheiri

Lactuca sativa, *L. serriola

Mercurialis annua

Plantago lanceolata

Rumex sanguineus, R. crispus

*Sorghum halepense

Trigonella foenum-graecum

Viola sylvestris, Viola sp.

Medicago sp.

Vicia faba

Primula macrocalyx

Conyza bonariensis

Salvia hypoleuca, S. limbata, S. nemorosa

Ficus carica

* Medicago sp.

Calotropis procera

Euphorbia marschalliana

Cerasus vulgaris, Prunus domestica

Cousinia sp.

*Chenopodium album

*Agropyron sp., *Poa annua, * Stipa sp.

Tilia begoniifolia

Arachis hypogaea

Phaeopappus aucheri

Anethum graveolens, Foeniculum vulgare, Petroselinum sativum

*Rosa persica, Rosa sp.

Rosa sp.

Ziziphus spina-christi

Hibiscus cannabinus

Solanum nigrum

Vigna sinensis

Danae racemosa

Phaseolus vulgaris

Rubus sp.

Ziziphus spina-christi

Diospyros lotus, D. kaki

Nerium oleander

Eucalyptus camaldulensis

Punica granatum

159

160 ... Pirnia & al.

Table 1, concluded

PATHOGEN

P. rubi

*P. salicina

P. salvadorae

P. sphaerellae-eugeniae

P vitis

Ramularia anchusae

R. brunnea

R. cynarae

R. geranii var. geranti

R. grevilleana var. grevilleana

R. heraclei

*R. inaequalis

R. lamii var. lamii

*R. macularis

*R. pratensis var. pratensis

R. rhabdospora

R. rubella

R. rumicis

R. sambucina

R. simplex

R. uredinicola

R. urticae

Ramulariopsis gossypii

*Scolecostigmina confluens

Sirosporium celtidis

Hosts

Rubus sp.

Salix alba

Salvadora persica

Eugenia jambos

Vitis sylvestris, V. vinifera

Anchusa italica, A. ovata

Tussilago farfara

Carthamus oxyacantha, C. tinctorius

Geranium pyrenaicum

Fragaria x ananassa, Potentilla reptans

Heracleum persicum

*Calendula persica

*Mentha piperita, Mentha sp.

Chenopodium album

Rumex crispus, Rumex sp.

Plantago lanceolata

*Rumex conglomeratus

Rumex crispus

Sambucus ebulus

Ranunculus oxyspermus, R. sahendicus

Uredinia of Melampsora sp. on Salix babylonica

Urtica dioica, Urtica urens

Gossypium hirsutum

Crataegus melanocarpa, C. pseudomelanocarpa

Celtis australis

* = new pathogen and host records

of Iran, seven are treated in detail in this paper. All specimens are deposited in

the fungus reference of Iranian Ministry of Agriculture “IRAN” at the Iranian

Research Institute of Plant Protection. Synonyms are listed in Crous & Braun

(2003) and Braun (1995, 1998).

Taxonomy

Cercospora pantoleuca Sacc., Michelia 1: 268 (1878) Fic. 1

Leaf spots circular, numerous, brown to blackish brown with gray center,

1-3 mm in diameter; caespituli amphigenous, mostly epiphyllous, punctiform;

stromata small to fairly prominent, brown, 20-25 um wide; conidiophores

fasciculate, 8-15 stalks, arising through stomata, pale olivaceous-brown, paler

towards the tip, aseptate or with few inconspicuous septa, straight, subcylindrical

to geniculate-sinuous, not branched, smooth, thin, 15-50 x 2.5-5 um; conidial

scars conspicuous, thickened and darkened, terminal and lateral, 1.5-2 um

wide; conidia formed singly, hyaline, acicular, narrowly obclavate, straight to

Cercosporoids new to Iran... 161

Fic. 1. Cercospora pantoleuca on Plantago lanceolata.

A. Conidiophores, B. Conidia, C. Symptoms on leaf (Bar = 50 um).

slightly curved, smooth, thin, 3-7-septate, base truncate, tip obtuse to subacute,

40-90 x 2.5-4 (-5) um; hilum thickened and darkened.

SPECIMEN EXAMINED: IRAN, NORTHERN KHORASAN PROVINCE, Shirvan, on Plantago

lanceolata L., 3 July 2011, B. Bicharanlou (IRAN 15498 F).

Note—This species is characterized by short conidiophores and conidia.

Conidiophores are pale olivaceous-brown to subhyaline, and conidial scars

are conspicuous, terminal, and lateral. These characters distinguish this

species from other Cercospora species. The Iranian specimen examined closely

resembles Braun's (1995) description of this species but slightly differs in having

faintly pigmented conidiophores.

Cercosporella primulae Allesch., Ber. Bayer. Bot. Ges. 2: 18 (1892) Fic. 2

Leaf spots circular to subcircular, yellowish, ochraceous to brown, margin

indefinite, surrounded by a yellowish halo, 2-8 mm in diameter; caespituli

amphigenous, punctiform, whitish; stromata substomatal to intraepidermal,

composed of few swollen hyphal cells, colourless; conidiophores in small

fascicles, 3-5 stalks, rarely solitary, arising through stomata or erumpent through

the cuticle, hyaline, straight to geniculate-sinuous, not branched, smooth, thin-

162 ... Pirnia & al.

A B

Fic. 2. Cercosporella primulae on Primula macrocalyx.

A. Conidiophores, B. Conidia, C. Symptoms on leaf (Bar = 50 um).

walled, 0-1-septate, 20-45 x 2.5-5 um; conidial scars conspicuous, thickened;

conidia formed singly, hyaline, obclavate-acicular to subcylindrical, smooth,

thin, indistinctly multiseptate, 0-7-septate, base truncate to obconicallytruncate,

tip subacute, 20-85 x 3-5 um; hilum slightly thickened and darkened.

SPECIMEN EXAMINED: IRAN, GOLESTAN PROVINCE, Gorgan, on Primula macrocalyx

Bunge, 13 May 2011, M. Pirnia (IRAN 15479 F).

Note—This taxon is characterized by moderately short and_ hyaline

conidiophores and conidia. Because of its hyaline conidiophores and conidia

as well as the structure of the scars, Braun (1995) maintained this species in

Cercosporella.

Cercosporella virgaureae (Thiim.) Allesch., Hedwigia 34: 286 (1895) Fic. 3

Leaf spots subcircular to irregular, at first yellowish brown, later grayish

brown, 2-6 mm in diameter; caespituli hypophyllous, whitish to grayish white,

punctiform to dense; stromata substomatal, composed of some aggregated

swollen hyphal cells, conidiophores fasciculate, 3-8 stalks, arising from

stromata, emerging through stomata, hyaline, below straight, geniculate-sinuous

towards the apex, not branched, 0-1-septate, 25-65 x 4-8 um; conidiogenous

cells integrated, terminal; conidial scars conspicuous, thickened and darkened,

terminal and lateral; conidia formed singly, hyaline, subcylindrical to obclavate,

Cercosporoids new to Iran ... 163

Fic. 3. Cercosporella virgaureae on Conyza bonariensis.

A. Conidiophores, B. Conidia, C. Symptoms on leaf (Bar = 50 um).

straight to slightly curved, smooth, thin, multiseptate, 3-7 septa, base truncate

to obconically truncate, tip mostly obtuse, 40-95 x 4-7 um; hilum distinct.

SPECIMEN EXAMINED: IRAN, GUILAN PROVINCE, Lahijan, on Conyza bonariensis (L.)

Cronquist, 29 Nov. 2010, M.R. Mirzaee (IRAN 15480 F).

Note—Morphology of the specimen examined agrees with the description

provided by Braun (1995). Cercosporella virgaureae is widespread and has a

wide host range on many genera of Asteraceae. This taxon is well characterized

by hyaline to subhyaline structures, long conidiophores and conidia, and

numerous conspicuous conidial scars on conidiophores.

Passalora bondartsevii U. Braun & Melnik, Trudy Bot. Inst. im. V.L. Komarova 20:

43 (1997) Fig. 4

Leaf spots circular to irregular, pale brown, surrounded by a yellowish

halo, 5-10 mm in diameter; caespituli epiphyllous, punctiform; conidiophores

in small to dense fascicles, 7-20 stalks, arising from stromata, brown, erect,

subcylindrical, slightly geniculate-sinuous towards the apex, smooth, thin,

continuous to septate, sometimes constricted at the septa, 40-90 x 4-5 um;

164 ... Pirnia & al.

Fic. 4. Passalora bondartsevii on Medicago sp.

A. Conidiophores, B. Conidia, C. Symptoms on leaf (Bar = 50 um).

conidiogenous cells integrated, terminal; conidial scars moderately thickened

and darkened; conidia formed singly, olivaceous-brown, ellipsoid, obovoid,

smooth to rough, 0-3-septate, base obconically truncate, apex obtuse to rounded,

10-20 x 5-7.5 um; hilum darkened, unthickened to slightly thickened.

SPECIMEN EXAMINED: IRAN, MAZANDARAN PROVINCE, Babolsar, on Medicago sp., 25

June 2011, M. Rafiee (IRAN 15499 F).

Note—This taxon was originally established by Braun & Melnik (1997)

on Onobrychis vicifolia Scop., from Russia and is only known from the type

collection. Passolora bondartsevii is diagnosed by ellipsoid-obovoid to

obclavate-subcylindrical, smooth to faintly rough conidia. In our specimen

the conidia are ellipsoid, obovoid, and mostly rough, but we did not see any

obclavate-subcylindrical conidia. However, other characters agree well with

the description in Braun & Melnik (1997). Therefore, we assign the Iranian

collection tentatively to P bondartsevii even though another host genus is

involved.

Pseudocercospora danaicola (Vienn.-Bourg.) Pirnia & Zare, comb. nov. Fic. 5

MycoBank MB 563391

= Cercosporina danaicola Vienn.-Bourg. Ann. Phytopathol.

2(4): 689 734 (1971 [“1970”]) as “danaecola”.

Leaf spots circular to subcircular, sometimes with spacious blight specially at

the tip of leaves, reddish brown, 10-15 mm in diameter; caespituli amphigenous,

Cercosporoids new to Iran... 165

punctiform; stromata present, globular, well developed, dark brown, 45-60 um

wide; conidiophores in dense fascicles, arising from stromata, pale olivaceous-

brown, subcylindrical, sinuous, rarely once geniculate, not branched, smooth,

thin, aseptate, 8-30 x 2-3 um; conidiogenous loci inconspicuous; conidial

scars unthickened and inconspicuous; conidia formed singly, pale olivaceous,

acicular, straight to slightly curved, smooth, thin, indistinctly 1-5-septate, base

truncate, tip subacute, 65-130 x 2.5-4 um; hilum inconspicuous, unthickened

and not darkened.

SPECIMENS EXAMINED (deposited as “Cercosporina danaecola”): IRAN, GOLESTAN

PROVINCE, Khan-bebin, Shirabad forest, on Danae racemosa Moench, 7 May 1976,

D. Ershad (IRAN 515 F); MAZANDARAN PROVINCE, Nowshahr, on D. racemosa, 19

Aug. 1973, collector unknown (IRAN 516 §, neotype, designated here).

Note—The name Cercosporina danaicola was originally published by Viennot-

Bourgin based on material from Iran (Viennot-Bourgin et al. 1971). Careful

microscopic examination of morphological characteristics showed that the

conidiogenous loci and hila are inconspicuous, unthickened, and not darkened.

Crous & Braun (2003) place species with these morphological features into

Pseudocercospora. Type material could not be traced, but based on the Iranian

Fic. 5. Pseudocercospora danaicola on Danae racemosa.

A. Conidiophores, B. Conidia, C. Symptoms on leaf (Bar = 50 um).

166 ... Pirnia & al.

specimens examined, this species can be transferred to Pseudocercospora. The

collection IRAN 516 is designated as neotype.

Fic. 6. Pseudocercospora paraguayensis on Eucalyptus camaldulensis.

A. Conidiophores, B. Conidia, C. Symptoms on leaf (Bar = 50 um).

Pseudocercospora paraguayensis (Tak. Kobay.) Crous, Mycotaxon 57: 270 (1996)

Fic. 6

Leaf spots irregular, vein-limited, pale brown to dark grayish brown, border

slightly raised, 2-5 mm in diameter, sometimes coalescing into a large blotch;

caespituli amphigenous, mostly hypophyllous, punctiform; stromata present,

well developed, brown, 50-70 um wide; conidiophores densely fasciculate,

compact, subhyaline to pale olivaceous, subcylindrical, aseptate, not branched,

sinuous to rarely once geniculate, straight to curved, rounded at the apex,

10-30 x 2-3 um; conidial scars unthickened and inconspicuous; conidia

formed singly, subhyaline or very pale olivaceous, narrowly obclavate, straight

to curved, indistinctly 2-6-septate, apex subacute or subobtuse, base truncate

to obconically truncate, 22-70 x 2-3 um; hilum inconspicuous, unthickened

and not darkened.

SPECIMEN EXAMINED: IRAN, GOLESTAN PROVINCE, Shastkola forest, on Eucalyptus

camaldulensis Dehnh., 8 Nov. 2010, M. Pirnia & R. Zare (IRAN 15481 F).

Note—Kobayashi introduced Cercospora paraguayensis on Eucalyptus sp. from

Paraguay (see Crous 1998).

Cercosporoids new to Iran... 167

Fic. 7. Scolecostigmina confluens on Crataegus pseudomelanocarpa.

A. Conidiophores, B. Conidia, C. Symptoms on leaf (Bar = 50 um).

Scolecostigmina confluens (Lieneman) U. Braun, New Zealand J. Bot. 37: 325 (1999)

Fic. 7

Leaf spots irregular, dark brown, scattered on leaf surface, but mostly at

the tip and edge of leaves, 5-15 mm in diameter; caespituli amphigenous,

mostly epiphyllous, punctiform; stromata prominent, subglobose, dark brown,

30-65 um wide; conidiophores in dense fascicles, 15-30 stalks, arising from

stromata, dark brown, paler and attenuated towards the apex, 0-1-septate,

cylindrical, straight to curved, sinuous to rarely once geniculate in the upper

part, not branched, verruculose, thick-walled, 25-75 x 4-7 um; conidiogenous

cells integrated, terminal, proliferation percurrent, annellate; conidia solitary,

dark brown, paler and narrower towards the tip, subcylindrical-obclavate,

slightly curved, verrucose, thick-walled, transversely pluriseptate, occasionally

with few longitudinal or oblique septa, base truncate, tip subacute, 75-140 x

4—8 um.

SPECIMENS EXAMINED: IRAN, GOLESTAN PROVINCE, Ghorogh forest park, on Crataegus

pseudomelanocarpa Popov ex Lincz. 14 Sep. 2010, M. Pirnia (IRAN 15472 F); GUILAN

PROVINCE, Rasht, Saravan, on C. melanocarpa M. Bieb., 27 July 2007, A. Khodaparast

(IRAN 15473 F).

Note—The genus Scolecostigmina was originally described as having thick-

walled verruculose conidiophores with conspicuous annellations and conidia

168 ... Pirnia & al.

that are subcylindrical-obclavate, smooth to verruculose, transversely multi-

euseptate conidia, and occasionally with a few longitudinal septa (Braun et al.

1999).

Acknowledgments

The authors are grateful to U. Braun (Martin-Luther-Universitat, Halle, Saale,

Germany) for providing part of his work on cercosporoid hyphomycetes from Russia as

well as his pre-submission review. A.J.L. Phillips (Universidade Nova de Lisboa, Portugal)

is thanked for preliminary review of the manuscript. We also thank S. Pennycook for

improving the text, particularly the nomenclature. A.R. Ghorbanian (Damghan Branch,

Islamic Azad University, Iran) and B. Djavadi (Iranian Research Institute of Plant

Protection, Iran) are thanked for identifications of host plants.

Literature cited

Braun U. 1995. A monograph of Cercosporella, Ramularia and allied genera (phytopathogenic

hyphomycetes), Vol. 1. IHW-Verlag, Eching.

Braun U. 1998. A monograph of Cercosporella, Ramularia and allied genera (phytopathogenic

hyphomycetes), Vol. 2. IHW-Verlag, Eching.

Braun U, Crous PW. 2007. The diversity of cercosporoid hyphomycetes: new species, combinations,

names and nomenclatural clarifications. Fungal Diversity 26: 55-72.

Braun U, Melnik VA. 1997. Cercosporoid fungi from Russia and adjacent countries. Trudy Bot.

Inst. im. V.L. Komarova (St. Petersburg) 20: 1-130.

Braun U, Mouchacca J, McKenzie EHC. 1999. Cercosporoid hyphomycetes from New Caledonia

and some other South Pacific islands. New Zealand Journal of Botany 37: 297-327.

http://dx.doi.org/10.1080/0028825X.1999.9512636

Chupp C. 1954. A monograph of the fungus genus Cercospora. Ithaca, New York. Published by the

author.

Crous PW. 1998. Mycosphaerella spp. and their anamorphs associated with leaf spot diseases of

Eucalyptus. Mycologia Memoirs 21: 1-170.

Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs: 1. Names published in Cercospora

and Passalora. CBS Biodiversity Series 1: 1-569.

Deighton FC. 1967. Studies on Cercospora and allied genera. II. Passalora, Cercosporidium and

some species of Fusicladium on Euphorbia. Mycological Papers 112: 1-80.

Deighton FC. 1973. Studies on Cercospora and allied genera. IV. Cercosporella Sacc.,

Pseudocercosporella gen. nov. and Pseudocercosporidium gen. nov. Mycological Papers 133:

1-62.

Deighton FC. 1974. Studies on Cercospora and allied genera. V. Mycovellosiella Rangel and a new

species of Ramulariopsis. Mycological Papers 137: 1-75.

Deighton FC. 1976. Studies on Cercospora and allied genera. VI. Pseudocercospora Speg., Pantospora

Cif. and Cercoseptoria Petr. Mycological Papers 140: 1-168.

Deighton FC. 1979. Studies on Cercospora and allied genera. VII. New species and redispositions.

Mycological Papers 144: 1-56.

Ershad D. 1990. New records of two Cercospora species for Iran. Iranian Journal of Plant Pathology

26: 41.

Ershad D. 2000. Two new species of mitosporic fungi. Rostaniha 1: 1-9.

Ershad D. 2002. A new Cercospora species from Iran. Rostaniha 3: 47-50.

Cercosporoids new to Iran... 169

Ershad D. 2009. Fungi of Iran. Iranian Research Institute of Plant Protection, Tehran.

Pirnia M, Zare R, Zamanizadeh HR, Khodaparast A. 2010. Contribution to the identification of

Cercospora species in Iran. Rostaniha 11(2): 183-189.

Pirnia M, Zare R, Zamanizadeh HR, Khodaparast A, Javadi Estahbanati AR. 2012a. Contribution

to the identification of the genus Passalora in Iran. Applied Entomology and Phytopathology

80 (1) (in press).

Pirnia M, Zare R, Zamanizadeh HR, Khodaparast A, Javadi Estahbanati AR. 2012b. Taxonomic

study of the genus Ramularia and Ramularia-like genera in Iran. Rostaniha 13(1) (in press).

Scharif G, Ershad D. 1966. A list of fungi on cultivated plants, shrubs and trees of Iran. Ministry of

Agriculture, Plant Pests and Diseases Research Institute, Evin, Tehran.

Viennot-Bourgin G, Ale-Agha N, Ershad D. 1971 [“1970”]. Les champignons parasites de I’Iran.

(Nouvelle contribution). Annales de Phyopathologie 2(4): 689-734.

MY COTAXON

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Volume 120, pp. 171-179 April-June 2012

Phaeocollybia nigripes (Agaricomycetes), a new species from Brazil

VICTOR R.M. COIMBRA”, TATIANA B. GIBERTONI! & FELIPE WARTCHOW?

' Universidade Federal de Pernambuco, Departamento de Micologia/CCB,

Av. Prof. Nelson Chaves, s/n, CEP: 50670-901, Recife, PE, Brazil

? Universidade Federal da Paraiba, Departamento de Departamento de Sistematica e Ecologia, CEP:

58051-970, Jodo Pessoa, PB, Brazil

* CORRESPONDENCE TO: vick_mat_coimbra@yahoo.com. br

ABSTRACT —Phaeocollybia nigripes is described from an Atlantic Forest fragment in

Pernambuco, Northeast Brazil. It is characterized by a stipe that is uniformly slender with a

slightly swollen base and dark grayish brown below to almost black at the apex, brown pileus,

large basidiospores that average 12.4 x 6.6 um, hyphoid to narrowly clavate cheilocystidia,

and abundant clamp connections. The holotype of P megalospora var. megalospora, the

putatively most similar taxon, was examined. A key to the six taxa reported from Brazil is

also presented.

Key worps — Agaricales, Hymenogastraceae, neotropic, taxonomy

Introduction

Phaeocollybia R. Heim was originally described to accommodate taxa

among the brown-spored Agaricales with smooth and moist to viscid pilei and

long pseudorhizas (Heim 1931). Matheny et al. (2006), based on multilocus

phylogenetic analyses, placed the genus as represented by P festiva (Fr.)

R. Heim in the Hymenogastraceae clade, but Kirk et al. (2008), continued to

refer Phaeocollybia to Cortinariaceae. More recently, Petersen et al. (2010), also

using molecular data, treated P festiva and P dissiliens A.H. Sm. & Trappe in the

bolbitioid clade with Bolbitius Fr., Panaeolus (Fr.) Quél., and Conocybe Fayod.

Phaeocollybia has been rarely reported in Brazil. The few taxa known from

this country were all reported from the Amazon region by Singer (1987),

who cited P brasiliensis .A. Aguiar ex Singer, P. elaeophylla Singer, P. flava

I.A. Aguiar ex Singer, P megalospora I1.A. Aguiar ex Singer var. megalospora,

and P. megalospora var. tetraspora Singer.

In this paper, we describe a new species based on a collection from the

Atlantic Forest of Pernambuco, Northeast Brazil, and provide a key to the

Brazilian Phaeocollybia species.

172 ... Coimbra, Gibertoni & Wartchow

Materials & methods

One basidioma was collected in an Atlantic Forest fragment called Ecological

Reserve of Dois Irmaos (8°01'15"S 34°52'30"W), a 387.4 ha area comprising a remnant

of Atlantic Forest and a public zoo in the urban perimeter of the municipality of Recife

(Ribeiro et al. 2007, Souza et al. 2009).

Colors are coded according to Watling (1969). The key to species is based mainly on

data given by Singer (1987). Basidiospore data terminology follows Tulloss et al. (1992)

but based on 25 basidiospores measured from a single basidioma (Wartchow 2009).

Abbreviations include L(W) = basidiospore length (width) average, Q = the length :

width ratio range as determined from all measured basidiospores, and Q = the Q value

averaged from all basidiospores measured.

Scanning electron microscopy (SEM) studies were conducted at the “Laboratorio de

Microscopia Eletrénica’ (DF/UFPE). Sections were removed from dried basidiomata

and mounted directly on aluminum stubs using carbon adhesive tabs. The fragments

were coated with 8-13 nm of gold using a sputter coater and examined with a scanning

electron microscope.

Phaeocollybia nigripes Wartchow & V. Coimbra sp. nov. FIGs. 1-6, 8

MycoBank MB 518815

Ab Phaeocollybia megalospora var. megalospora differt stipe coloribus apice niger versus

fuscus basin, cheilocystidiis tenui clavati vel cylindraceous et catenatus, et pileipellis

hipodermio ex elementis pro ratione voluminosis.

Ho torype: Brazil, Pernambuco, Recife, Ecological Reserve of Dois Irmaos, 10.iii.2009

V.R.M. Coimbra, J.M. Baltazar & L. Trierveiler-Pereira s.n. (URM 82277, holotype).

Erymo oy: Latin for ‘black foot; in reference to the stipe color.

PiLEus 11 x 10 mm, conic, reddish brown to dark brown (“Date brown 24’),

surface smooth, margin slightly short-sulcate and involute. LAMELLAE adnexed,

close, lamellulae present, dark grayish brown (slightly lighter than “Drab 33’)

with dark gray edges (under 10x lens), edges entire. Veil not evident. STIPE 70

x 1.5-2 mm (upper portion apart from pseudorhiza 55 mm long), cylindrical

above a swollen base, central, dark brown (‘Cigar brown 16’), shiny metallic

graphite-black at apex with a vinaceous base under intense light, surface

smooth, hollow. Pseudorhiza thin, spiraling, % of the stipe length.

BASIDIOSPORES (10—)10.8-14.5(-15) x (5.5-)6-7.3(-8) um (L = 12.4 um; W

= 6.6 um; Q = (1.69-)1.77-2.00(-2.29); Q = 1.88), amygdaliform to sometimes

limoniform, thin-walled, surface minutely verrucose, brown. BAsip1a 24-33 x

9-11 um, clavate, thin-walled, with prominent sterigmata, predominantly 2-

sterigmate. PLEUROCYSTIDIA absent. CHEILOCYSTIDIA (13—)16-25 x 5-6 um,

narrowly clavate to hyphoid (located in terminal position of chains of short

celled hyphae), forming dense tufts on the sterile edge, thin-walled, hyaline.

TIBIIFORM DIVERTICULA abundant on pseudorrhizal pellis, 11-32 x 1.8-3 um,

narrowly lanceolate to subcapitate, sometimes with apical droplet, some with

small lateral projections, hyaline, colorless, thin walled; on stipe more scattered

Phaeocollybia nigripes sp. nov. (Brazil) ... 173

Fic. 1. Phaeocollybia nigripes (holotype). Basidioma. Scale bar = 10 mm.

and slightly smaller 11-16 x 2-3 um, hyaline, thin walled. PILEIPELLIs with

periclinal to sometimes anticlinal hyphae of 3-6 um wide, pigments encrusting,

yellowish brown, showing some gelatinized zones; hypodermium somewhat

pseudoparenchymatous, elements 20-27 x 13-18 um, yellowish-brown.

LAMELLAR TRAMA regular and parallel, hyphae ranging 4.5-15.5 um, thin-

walled. CLAMP CONNECTIONS abundant.

ECOLOGY & DISTRIBUTION: Solitary on soil under dicotyledonous trees,

including representatives of the ectomycorrhizal families (Smith & Read 2008)

Caesalpiniaceae [Senna Mill., Chamaecrista (L.) Moench, Dialium L., Hymenaea

L., Sclerolobium Vogel, Zollernia Maximil. & Nees], Nyctaginaceae (Boerhavia

174 ... Coimbra, Gibertoni & Wartchow

Fics. 2-6. Phaeocollybia nigripes (holotype): 2. Basidioma. 3. Basidiospores. 4. Basidia. 5. Hyphal

elements of the lamella edge. 6. Pileipellis and adjacent inflated elements of the hypodermium.

Fic. 7. P. megalospora var. megalospora (holotype): Pileipellis and adjacent hyphae of the

hypodermium. Scale bars: 2 = 10 mm; 3-7 = 10 um.

L., Guapira Aubl.) and Polygonaceae (Coccoloba P. Browne). Known only from

the type locality in the Atlantic Forest of Northeast Brazil.

ADDITIONAL SPECIMEN EXAMINED: Phaeocollybia megalospora var. megalospora—

BRAZIL. Amazonas, road Manaus-Caracarai km 45, 11.vii.1977, R. Singer & I. Araujo,

Singer B 10082 (INPA 102697, holotype!).

REMARKS: The distinctive microscopic features of P nigripes include

basidiospores longer than 11 um, predominantly 2-spored basidia, cylindric-

hyphoid to narrowly clavate cheilocystidia, anda slightly pseudoparenchymatous

hypodermium. The pseudorhiza of our new species was broken during

collection.

Following the classification proposed by Bandala & Montoya (1994),

P. nigripes would be referred to subgen. Fibulophaeocollybia Bandala & Montoya

sect. Subattenuatae Singer based on the presence of clamp connections (most

easily seen at the cheilocystidial bases) and limoniform basidiospores longer

than 7 um (Bandala & Montoya 1994). Norvell & Exeter (2009), who felt that

infrageneric classification is premature in the absence of molecular analyses of

the whole genus, suggested that cheilocystidial morphology might be a better

Phaeocollybia nigripes sp. nov. (Brazil) ... 175

» -

\ =

isk “6; BEE

13kU ~6, BEB Zhim 34 29

Fics. 8-9. SEM of basidiospores (photos by Sérgio Santos).

8. Phaeocollybia nigripes. 9. P. megalospora.

176 ... Coimbra, Gibertoni & Wartchow

morphological character for identifying monophyletic groups below genus

level in Phaeocollybia.

Phaeocollybia megalospora var. megalospora, characterized by Singer (1987)

as also producing 2-sterigmate basidia and similar sized basidiospores, differs

in its yellowish to brownish stipe, pileipellis with a gelatinized ixocutis and

cutiform rather than pseudoparenchymatous hypodermium, ampullaceous

cheilocystidia, and more strongly verrucose basidiospores (Singer 1987). We

examined the holotype (Fics. 7, 9) of P megalospora, which we believe is the

same basidioma depicted in Singer (1987: Plate II, fig. 5), and noted that the

stipe was apparently broken and lacked the pseudorhiza (which typically bears

abundant tibiiform diverticula according to Norvell 1998), so that only one

very small (<7 x 2 um) tibiiform diverticulum was observed. The pileipellis

comprised gelatinized erect, non-incrusted, 1-3.5 um wide, yellowish hyphae

with the hypodermium comprising hyphae <5 um wide, radially oriented,

and with dark brown vacuolar pigments (Fig. 7). The basidiospores are very

similar in size 11-15 x (6-)6.5-8 um (L = 12.9 um; W = 7.2 um; Q = (1.43-)

1.50-2.00(-2.14); Q = 1.80) but more strongly verrucose (Fic. 9) than those

found in our species, supporting our conclusion that P megalospora var.

megalospora and P. nigripes are distinct species.

Another taxon with blackish tints in the basidioma is P singularis E. Horak &

Halling from Colombia (Horak & Halling 1991) and the Talamanca Mountains

in Costa Rica (Norvell pers. corr.), but it obviously differs in the blackish

lavender pileus, violaceous stipe, smaller (8-9.5 x 4.5-5 um) basidiospores, and

tibiiform cheilocystidia.

Phaeocollybia phaeogaleroides Norvell is another similar species with

relatively large basidiospores. It shares with P. nigripes the dark brown pileus

and stipe, clamp connections, and cylindrical cheilocystidia (Norvell 2002) but

differs in the hygrophanous pileus, shorter narrower basidiospores 9-12(-13)

x 5-6.5 um (L = 10 um; W = 5.8 um), 4-spored basidia, and longer (< 80 um)

subcapitate cheilocystidia (Norvell 2002).

Phaeocollybia longistipitata Halling & E. Horak, P odorata E. Horak,

P. pleurocystidiata Norvell & Redhead, P. singeri Guzman et al., and P. tentaculata

E. Horak are other slender stiped taxa that are segregated primarily by their

smaller basidiospores and lack of clamp connections (Horak 1977, Bandala et

al. 1989, Norvell & Redhead 2000, Halling & Horak 2008).

Phaeocollybia columbiana Singer from tropical rain forest of Colombia also

shares relatively large (10-12 x 6-7 um) basidiospores. However, it differs in

the spadiceous pileus with whitish centre, ochraceous brown stipe, ventricose

cheilocystidia, and absence of clamp connections (Singer 1970, Horak 1977).

Phaeocollybia amazonica Singer, from the Bolivian Amazon, differs from

P. nigripes macroscopically in pileus size (40-45 mm diam), ochraceous brown

Phaeocollybia nigripes sp. nov. (Brazil) ... 177

to rusty lamellae, and cinnamon stipe and microscopically in basidiospore

shape and larger (>30 x 6 um) cheilocystidia (Singer 1961, 1970).

Other lowland Amazonian Brazilian taxa — P. brasiliensis, P. elaeophylla,

P. flava, P. megalospora var. tetraspora — are easily segregated by their smaller

basidiospores (Singer 1987).

Here we also address the ecological status of Phaeocollybia nigripes. Earlier

studies implied Phaeocollybia as saprotrophic with the pseudorhizae arising

from buried wood (Smith 1957; Smith & Trappe 1972). Subsequently Redhead

& Malloch (1986), who studied the temperate P christinae (Fr.) R. Heim and

P. jennyae (P. Karst.) Romagn., concluded that Phaeocollybia is parasitic based

on the successful excavation of a pseudorhiza connected to a senescent spruce

rootlet. Norvell (1998), demonstrating that P. redheadii Norvell, P. kauffmanii

(A.H. Sm.) Singer, and several other taxa present ectomycorrhizal mantles

and Hartig-net in western hemlock and noble fir roots, emphasized that all

Pacific Northwest North American Phaeocollybia species are found in forests

with Tsuga, Picea, Abies, Pseudotsuga, Lithocarpus, Quercus, and/or Pinus, all

ectomycorrhizal-forming genera (Smith & Read 2008). Most recently, Norvell

& Exeter (2009) cite the presence of Hartig-net and high nitrogen isotope

readings presented by the studied species as the main characters supporting

the ectomycorrhizal nature of Phaeocollybia in the Pacific Northwest.

Neotropical phaeocollybias from Colombia and Costa Rica occur mostly

under the ectomycorrhizal tree genus Quercus (Singer 1987, Horak & Halling

1991, Halling & Horak 2008), while others from lowland Amazon forests

were found in “terra firme,’ a typical anectotrophic forest type according to

Singer & Aratijo (1979). In the case of PB nigripes, although no ectomycorrhizal

connection was detected, representatives of Caesalpiniaceae, Nyctaginaceae, and

Polygonaceae occurred in the collection area (Guedes 1998, Ferreira et al. 2007,

Souza et al. 2009); these families include genera shown to have ectomycorrhizal

associations in the tropical forests of Ecuador: e.g., Coccoloba (Polygonaceae)

and Guapira (Nyctaginaceae) (Tedersoo et al. 2010).

Artificial key for the Brazilian taxa of Phaeocollybia

1. Basidiospores small, 5-5.5 x 4.5-5 um; pileus yellow..................00. P. flava

1. Basidiospores longer, > 7 um long; pileus with brown tints...................00. 2

2. Basidiospores <10-15 um long; basidia mostly with two sterigmata .............. 3

2. Basidiospores with largest length not longer than 11.5 um; basidia with four

sterigmata (in P elaeophylla 1-4 sterigmata) ............ 0... cece eee ee eee -:

3. Stipe yellowish to brownish towards the base; cheilocystidia ampullaceous;

hypodermium cutiform made of hyphae 3-9 um wide

ET EE EN EE P. megalospora var. megalospora

178 ... Coimbra, Gibertoni & Wartchow

3°. Stipe dark brown with shiny metallic graphite-black apex; cheilocystidia narrowly

clavate to hyphoid (located in terminal position of chains of short-celled

hyphae); hypodermium pseudoparenchymatous made of elements of

DORD FPN SVS I. es Lele an beg avn, thy Bawa -ap g Monk hd Manny 4 Banka g Beawave gg avk's P. nigripes

4, Lamellae olive; basidiospores 8.5-10.3 x 4.5-6.5 um, cheilocystidia

heteromorphous 17-49 x 3.5-5.5 um, sometimes tibiiform-like .... P elaeophylla

4 Searivellae* Wak eTiG OHVecCOlO = (Liste sedi! les de, Ret ae, a EM ath aed eset pli 5

5. Pileus striate; basidiospores (7—)7.5-8.3 x 4.7-5.5 um; cheilocystidia ventricose

frequently capitate (tibiiform?) 25-32 x 6-7.5 um .............. P. brasiliensis

5. Pileus not striate; basidiospores 9.5-11.5 x 6-7 um, cheilocystidia clavate,

cylindrical or utriform of 40-50 x 3.8-6.5um .... P megalospora var. tetraspora

Acknowledgments

The authors would like to thank Dr. Lorelei L. Norvell for patiently reviewing all

previous manuscript drafts and teaching about this interesting genus. Dr. Clark L.

Ovrebo is also kindly acknowledged for reviewing our article. M.Sc. Juliano M. Baltazar

and M.Sc. Larissa Trierveiler-Pereira are thanked for help during the field trips, FACEPE

for providing a post-doctoral grant to FW (BFP 0100-2.03/09) and scholarship to VRMC

(BIC 0061-2.03/08) and partially financing this research (APQ 0444-2.03/08). Sergio

Santos, of the Department of Physics (Universidade Federal de Pernambuco, Brazil)

is acknowledged for the scanning electron micrographs of the basidiospores of both

species.

Literature cited

Bandala VM, Montoya L. 1994. Further investigations on Phaeocollybia with notes on infrageneric

classification. Mycotaxon 52: 397-422.

Bandala VM, Guzman G, Montoya L. 1989. Additions to the knowledge of Phaeocollybia (Agaricales,

Cortinariaceae) from Mexico with description of new species. Mycotaxon 35: 127-152.

Ferreira RLC, Marangon LC, Silva JAA, Rocha MS, Alves-Junior FT, Aparicio OS. 2007. Estrutura

fitossociolégica da mata ciliar do Agude do Meio, Reserva Ecolégica de Dois Irmaos, Recife-PE.

Magistra 19: 31-39.

Guedes MLS. 1998. A vegetacao fanerogamica da Reserva Ecoldégica Dois irmaos. In: Machado IC,

Lopes AV, Porto KC. (eds.). Reserva Ecolégica de Dois Irmaos: Estudos em um Remanescente

de Mata Atlantica em Area Urbana (Recife-Pernambuco-Brasil). Editora Universitaria da

UFPE, Recife.

Halling RE, Horak E. 2008. Phaeocollybia longistipitata sp. nov. from Costa Rica. North Am. Fungi

3: 177-185. http://dx.doi.org/10.2509/naf2008.003.00711

Heim R. 1931. Le genre Inocybe. Encycl. Mycol. 1: 1-429.

Horak E. 1977. Further additions towards a monograph of Phaeocollybia. Sydowia 29: 28-70.

Horak E, Halling RE. 1991. New records of Phaeocollybia from Colombia. Mycologia 83: 464-471.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10th ed. CAB International, Wallingford,

Matheny PB, Curtis JM, Hofstetter V, Aime MC, Moncalvo, J-M, Ge Z-W, Yang Z-L, Slot JC,

Ammirati JF, Baroni TJ, Bougher NL, Hughes KW, Lodge DJ, Kerrigan RW, Seidl MT, Aanen

DK, De Nitis M, Daniele GM, Desjardin DE, Kropp BR, Norvell LL, Parker A, Vellinga EC,

Vilgalys R, Hibbett DS. 2006. Major clades of Agaricales: a multilocus phylogenetic overview.

Mycologia 98: 982-995. http://dx.doi.org/10.3852/mycologia.98.6.982

Phaeocollybia nigripes sp. nov. (Brazil) ... 179

Norvell LL. 1998. Observations on development, morphology and biology in Phaeocollybia.

Mycol. Res. 102: 615-630. http://dx.doi.org/10.1017/S0953756297005431

Norvell LL. 2002. Phaeocollybia in western North America 3: New species P. phaeogaleroides and

P. rifflipes, with notes on the P festiva complex. Mycotaxon 81: 95-112.

Norvell LL, Exeter RL. 2009 ['2008’]. Phaeocollybia of Pacific Northwest North America. USDI

BLM/OR/WA/GI-08/100-1792, Salem, Oregon. 228 p.

Norvell LL, Redhead SA. 2000. Phaeocollybia in western North America 2: the vernal

P. pleurocystidiata sp. nov. and P. carmanahensis reconsidered. Mycologia 92: 984-911.

http://dx.doi.org/10.2307/3761593

Petersen G, Knudsen H, Seberg O. 2010. Alignment, clade robustness and fungal phylogenetics

- Crepidotaceae and sister families revisited. Cladistics 26: 62-71.

http://dx.doi.org/10.1111/j.1096.

Redhead SA, Malloch DW. 1986. The genus Phaeocollybia (Agaricales) in eastern Canada and its

biological status. Can. J. Bot. 64: 1249-1254. http://dx.doi.org/10.1139/b86-172

Ribeiro EMS, Ramos EMNE, Silva JSB. 2007. Impactos ambientais causados pelo uso publico em

areas naturais do Parque Estadual de Dois Irmaos, Recife-PE. Rev. Bras. Biociéncias 5 (suppl.

1): 72-74.

Singer R. 1961. Diagnoses fungorum novorum Agaricalium II. Sydowia 15: 45-83.

Singer R. 1970. Phaeocollybia (Cortinariaceae- Basidiomycetes). Flora Neotropica 4: 3-13.

Singer R. 1987. Phaeocollybia in the oak woods of Costa Rica, with notes on extralimital taxa.

Mycol. Helvetica 2: 247-366.

Singer R, Araujo IJS. 1979. Litter decomposition and ectomycorrhizal in Amazonian Forest. 1.

A comparison of litter decomposing and ectomycorrhizal Basidiomycetes in latosol-terra-firme

rain forest and white podzol campinarama. Acta Amazonica 9: 25-41.

Smith AH. 1957. A contribution toward a monograph of Phaeocollybia. Brittonia 9: 195-217.

http://dx.doi.org/10.2307/2804723

Smith AH, Trappe JM. 1972. The higher fungi of Oregon’s Cascade Head Experimental Forest and

vicinity. I. The genus Phaeocollybia (Agaricales) and notes and descriptions of other species in

the Agaricales. Mycologia 64: 1138-1153. http://dx.doi.org/10.2307/3758079

Smith SE, Read DS. 2008. Mycorrhizal symbiosis. 3th ed. Academic Press, New York.

Souza ACR, Almeida Jr. EB, Zickel CS. 2009. Riqueza de espécies de sub-bosque em um fragmento

florestal urbano, Pernambuco, Brasil. Biotemas 22: 57-66.

Tedersoo L, Sadam A, Zambrano M, Valencia R, Bahran M. 2010. Low diversity and high host

preference of ectomycorrhizal fungi in Western Amazonia, a neotropical biodiversity hotspot.

ISME Journal 4: 465-471. http://dx.doi.org/0.1038/ismej.2009.131

Tulloss RE, Ovrebo CL, Halling RE. 1992. Studies on Amanita (Amanitaceae) from Andean

Colombia. Mem. New York Bot. Gard. 66: 1-46.

Wartchow F. 2009. Volvariella cubensis: a rare neotropical agaric new to South America. Mycotaxon

107: 181-187. http://dx.doi.org/10.5248/107.181

Watling R. 1969. Colour identification chart. Her Majesty’s Stationery Office, Edinburgh.

MY COTAXON

http://dx.doi.org/10.5248/120.181

Volume 120, pp. 181-188 April-June 2012

Peziza michelii and its ectomycorrhizae with

Alnus nitida (Betulaceae) from Pakistan

T. ASHRAF’, M. HANIF? & A. N. KHALID?

Department of Botany, University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

CORRESPONDENCE TO *: *tayibamaan01@gmail.com,

*mhanif_r@hotmail.com, *drankhalid@gmail.com

ABSTRACT — Peziza michelii and its ectomycorrhizae with Alnus nitida are characterized

morphologically and molecularly by nrDNA ITS1-5.8S-ITS2 sequence analyses. Both the

fungus and its morphotype are new records for Pakistan.

Key worps — operculate asci, Pezizales, phylogeny

Introduction

Peziza Dill. ex Fr. (Pezizales, Pezizaceae) is a large, widely distributed

heterogeneous genus represented by 104 species (Kirk et al. 2008). Its trophic

status ranges from saprobic to mycorrhizal (Hansen et al. 2001).

Identification of pezizalean mycorrhizae using morphological or

conventional methods has remained difficult since they lack hyphal strands,

connecting apothecium, and diagnostic morphotypes. Generally pezizalean

ECM (ectomycorrhizae) possess a thin pseudoparenchymatous mantle,

infrequent emanating hyphae, and rhizomorphs, and clamp connections are

lacking (Agerer 1991, 2001). At the present time molecular tools are quite

helpful for ECM identification, and the ECM of many Peziza species have been

confirmed using such tools. Species identified as mycorrhizal with deciduous

trees include Peziza depressa Pers., P. michelii, P. ostracoderma Korf, P. succosa

Berk., and P. succosella (Le Gal & Romagn.) M.M. Moser ex Aviz.-Hersh. &

Nemlich (Tedersoo et al. 2006).

Betulaceae is represented in Pakistan by two genera, Alnus and Betula (Naisr

1975), for which there are no previous reports about ectomycorrhizal status

from Pakistan. Previously, P michelii has been reported as ectomycorrhizal

with Alnus sp., Picea abies (Tedersoo et al. 2009), Betula sp. (Tedersoo et al.

182 ... Ashraf, Hanif & Khalid

2008), Fagus orientalis (Bahram et al. 2012) Tsuga canadensis (McLenon-Porter

2008), and Tilia sp. (Lang et al. 2011). Mycorrhizal associations of Alnus with

P. michelii are confirmed molecularly in this study.

Ahmad et al. (1997) and Ashraf & Khalid (2012) have reported 89

pezizalean taxa (including nine Peziza species) from Pakistan, and ascocarp

surveys in Pakistan account for eight known to be ectomycorrhizal. Molecular

characterization of Pezizales and their ectomycorrhizae in Pakistan is in

progress. Our study, which documents association of P michelii with Alnus nitida

(Spach) Endl. in Asia for the first time, is the first attempt at morphological and

molecular characterization of ascomycete morphotypes from Pakistan.

Materials & methods

Ascomata were collected and photographed; necessary data was recorded in field

and specimens were dried with the help of a fan heater. Ascomata were rehydrated and

free hand sections were made in the laboratory. Microscopic (morpho-anatomical)

characters were noted at 10x and 40x and drawings were made with the help of camera

lucida.

Soil under A. nitida was sampled and washed to isolate, characterize, and identify

ECM. ‘The selected morphotypes were characterized morphologically following Agerer

(2001) and have been deposited in Herbarium of Botany Department (LAH), University

of the Punjab in Lahore.

DNA was extracted from dried ascomata and morphotypes using Extract N. Amp.

™ Plant kit (SIGMA). The nrDNA ITS1-5.8S-ITS2 region was amplified using fungal

specific primers pairs (ITS1F/ITS4) following manufacturer's protocol with denaturation

at 94°C for 4 min, followed by 35 cycles of 45 sec at 94°C, 45 sec at 54°C, and 1 min

30 sec at 72°C, and a final extension at 72°C for 2 min. After purification, the PCR

products underwent bidirectional sequencing by Macrogen (South Korea). GenBank

ITS sequences were BLAST-searched for sequence comparison and identification.

Selected sequences were aligned using Clustal W and corrected manually. All ambiguous

insertions and deletions were removed prior to further analyses. Sequences have been

accessioned in GenBank.

Results

Alnus nitida roots were found to be colonized by Peziza michelii. The BLAST

search of the ITS sequences from P michelii ascomata and morphotypes

associated with Alnus nitida obtained in the present study showed a 99%

identity and 98% query coverage match with P michelii from Denmark

(DQ200839.1). The identity percentage was calculated at 96.3, showing a

97% identity and minimum divergence of 1.5 and 0.8 compared with Peziza

michelii (DQ200838.1 and DQ200839.1) respectively (PLATE 1). The topology

and identification of P. michelii was confirmed by the neighbor-joining method

Peziza michelii +Alnus nitida ectomycorrhizae (Pakistan) ... 183

and maximum likelihood method (PLaTEs 2-3). Both analyses place P. michelii

with P. succosa and P. succosella as sister clade.

Peziza_michelii_ENA38845_Pakistan

Peziza_michelii_FR852088.1_Iran

Peziza_michelii_DQ200839.1_Denmark

Peziza_michelii_DQ200838.1_Denmark

Peziza_infossa_DQ974817.1_USA

Peziza_cf.__succosa_EU819417

Peziza_succosella_DQ200841.1_Denmark

Peziza_succosa_DQ200840.1_Denmark

Peziza_phyllogena_AY789329.1_USA

Peziza_ostracoderma_JN002180

Chromelosporium_carneum_FJ7911

Peziza_stuntzii_FJ268642.1_USA

Cazia_flexiascus_EU846309

Peziza_depressa_DQ200837

Mycoclelandia_arenacea_Q231745

Peziza_michelii_TA116_Pakistan

Divergence

Pate 1. Percent divergence and identity calculated by comparing sequence pairs reconstructed by

MegAlign (DNASTAR).

Cazia_flexiascus_EU846309

Mycoclelandia_arenacea_Q231745

Peziza_stuntzii_FJ268642.1_USA

Peziza_depressa_DQ200837

Peziza_ostracoderma_JN002180

Peziza_phyllogena_AY789329.1_USA

Chromelosporium_carneum_FJ7911

Peziza_infossa_DQ974817.1_USA

Peziza_cf.__succosa_EU819417

Peziza_succosella_DQ200841.1_Denmark

Peziza_succosa_DQ200840.1_Denmark

Peziza_michelii_ENA38845_Pakistan

Peziza_michelii_TA116_Pakistan

Peziza_michelii_DQ200839.1_Denmark

Peziza_michelii_DQ200838.1_Denmark

Peziza_michelii_FR852088.1_Iran

Sa ne er) Pee a |

295, 9 20- 15° 10, = JPG

Nucleotide Substitutions (x100)

29.6

PLATE 2. Phylogenetic tree based on ITS1-5.8S-ITS2 nucleotide sequences adjusted to 722 bases and

constructed by the neighbor-joining method using MegAlign. The scale indicates the percentage

of base difference (% divergence). Sequence data for phylogenetic analysis were taken from the

GenBank nucleotide sequence database for Peziza, Cazia, Chromelosporium, and Mycoclelandia.

184 ... Ashraf, Hanif & Khalid

99 - Peziza_michelii_ENA38845_Pakistan

50] L Peziza_michelii_TA116_Pakistan

41 || Peziza_michelii_FR852088.1_Iran

1001 Peziza_michelii_DQ200839.1_ Denmark

100 Peziza_michelii_DQ200838.1_Denmark

Peziza_succosella_DQ200841.1_ Denmark

100 Peziza_succosa_DQ200840.1_Denmark

53 Peziza_infossa_DQ974817.1_USA

85 Peziza_cf._succosa_EU819417.1_

67 Peziza_phyllogena_AY 789329.1_USA

47 Chromelosporium_cameum_FJ7911

Peziza_ostracoderma_JN002180.1

Mycoclelandia_arenacea_Q231745

Cazia_flexiascus_EU846309.1

40 Peziza_stuntzii_FJ268642.1_USA

25 Peziza_depressa_DQ200837.1

Cater o

PiaTE 3. Molecular Phylogenetic analysis was inferred by using the Maximum Likelihood method.

The tree with the highest log likelihood (-6466.0969) is shown. Bootstrap values are shown

against each branch. Initial tree(s) for the heuristic search were obtained automatically: maximum

parsimony method was used when common sites numbered <100 (or <1/4 total sites); otherwise

BIONJ method with MCL distance matrix was used. The tree is drawn to scale, with branch lengths

measured in number of substitutions per site. The analysis involved 16 sequences.

Taxonomy

Peziza michelii (Boud.) Dennis, Brit. Cup Fungi & Allies: 15 (1960) PLaTE 4

APOTHECIA cupulate, medium-sized, diam. up to 5 cm, deeply concave,

margins entire, but turning inwards, somewhat raised and torn, hymenium

reddish brown with purplish tinge, smooth, outer surface yellow to yellowish

brown, sessile to subsessile, solitary. HYMENIUM 290 um thick. Ascr cylindrical,

operculate, unitunicate, 8-spored, uniseriate, strongly amyloid at apex, but

diffusely blue along length, slightly narrowing near base, 260-278(-300) x

13-17 um. AscosporEs uniseriate, ellipsoid, biguttulate, ornamented, 15-17(-

19) x 9-10 um, ornamentation verrucose to warted: which appear elongated,

irregularly and widely spaced, 0.3-0.7um high. PARApHysES long, slender,

septate, with light brownish content, straight, 2-3 um wide, slightly thick up

to 5-6 um at tip. ExcrpuLum: Ectal excipulum a textura globulosa to angularis,

100-160 um thick, cells hyaline to slightly brown, thin-walled, diam. 10-22

Peziza michelii +Alnus nitida ectomycorrhizae (Pakistan) ... 185

PiatE 4. Peziza michelii: A. Apothecium. B. Apothecial section showing hymenium,

sub-hymenium, medullary, and ectal excipulum. C. Part of hymenium, showing asci

with ascospores and paraphyses. D. Ascospores. E. Excipulum. Scale bars: A = 0.5 cm;

B = 90 um; C = 30 um; D =6 um; E = 80 um

um; medullary excipulum a textura globulosa, 500 um thick, larger sub-globose

thin-walled cells, 22-40 um diam., interspersed with numerous interwoven

delicate hyphae <8 um diam.

MATERIAL EXAMINED: PAKISTAN: KHYBER PAKHTUNKHWA, Khanspur, Helipad,

Himalayan moist temperate forests, 2575 m (8205 ft) altitude, solitary, on ground, mossy

substrate, 21 Aug. 2010, T. Ashraf TA-116 (LAH 210810; GenBank JN836748).

Morphological description PLATE 5

ECTOMYCORRHIZAL SYSTEM simple, axis 4-5 x 1.0-1.3 mm. UNRAMIFIED

ENDS rounded to bent, bifurcate, 2-3 mm long and 0.8-1.2 mm in diam.,

186 ... Ashraf, Hanif & Khalid

PLATE 5. ECM of Peziza michelii: A-B. Habit. C. Pseudoparenchymatous outer mantle.

D. Pseudoparenchymatous inner mantle. E. Emanating hyphae. Scale bars: A=0.5 mm;

B= 0.5 mm; C-D = 2 um; E= 1.5 um.

younger tips creamy white, older tips brown to black. Texture of system felty to

velvety with matte luster, host tissue not visible under the sheath. RHIZOMORPH

absent. EMANATING HYPHAE rare, straight.

MANTLE pseudoparenchymatous in all layers, OUTER MANTLE

pseudoparenchymatous with irregular (ovoid to epidermoid) cells, hyphal cells

infrequent and inconspicuous, hyphae without clamps, light yellowish, no cell

Peziza michelii +Alnus nitida ectomycorrhizae (Pakistan) ... 187

PLATE 6. Cross section of Alnus nitida ectomycorrhizal root: m = mantle, ec = ectoderm, c = cortex,

en = endoderm, cc = central cylinder, hn = hartig net.

contents visible, cells 14-15 x 10-14um. INNER MANTLE pseudoparenchymatous,

cells colorless to light yellowish, cells roundish to irregular in shape, no matrix

material observed, cells 13-15 x 11-14 um.

MANTLE THICKNESS 36 um, < 5 layers thick, discernible, cells rectangular

to tangentially oval and/or tangentially elliptical. HARTIG NET paraepidermal,

followed by large cortical cells, endodermis and central cylinder (PLATE 6)

MATERIAL EXAMINED: PAKISTAN: KHYBER PAKHTUNKHWA, Khanspur, Helipad,

Himalayan moist temperate forests, 2575 m (8205ft) altitude, roots of Alnus nitida,

15 May 2010, M Hanif ENA38845 (LAH 150509; GenBank JN836749).

Discussion

The present work is the first report of Peziza michelii from Pakistan. This

species can be found frequently in Himalayan moist temperate forests of

Pakistan. Phylogenetically, P michelii clusters with the similar species from

USA, Iran, and Denmark.

Tedersoo et al. (2006) studied the morphotypes of P. michelii morphologically

and used molecular markers to identify the mycobiont. They found these

ectomycorrhizae to be solitary, rare, in small clusters, minute, conspicuous, and

yellow green to olive green. This finding differed from the present report and

morphotypes associated with Alnus nitida from Pakistan. Those morphotypes

we studied are simple, creamy white with brown to black older tips, and lacking

a rhizomorph. The anatomical features of the mantle surface also differ slightly

in both morphotypes, with a mantle surface varying from smooth to finely

granular in the morphotype reported by Tedersoo et al. (2006) versus the

A. nitida morphotype, which is felty to velvety with matte luster with almost

equal thickness (2-4 hyphal layers vs. 5 hyphal layers). Other mantle features in

both studies also varied. The morpho-anatomic features of both morphotypes

188 ... Ashraf, Hanif & Khalid

of Peziza michelii may vary because of interaction of mycobiont with different

photobionts.

Acknowledgements

The authors would like to thank Dr. Donald H Pfister (Harvard University Herbaria,

Cambridge USA) and Dr. Mohammad Bahram (University of Tartu. Estonia) for their

helpful comments and critical review of this manuscript.

Literature cited

Agerer R. 1991. Characterization of ectomycorrhiza. 25-73, in: JR Norris et al. (eds). Techniques

for the Study of Mycorrhiza. London, UK: Academic Press.

Agerer R. 2001. Exploration types of ectomycorrhizae. Mycorrhiza 11: 107-114.

http://dx.doi.org/10.1007/s005720100108

Ahmad §S, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Sultan Ahmad Mycological Society of

Pakistan, Lahore. 248 p.

Ashraf T, Khalid AN. 2012. New records of Pezizales from Pakistan. Mycotaxon 119: 301-306.

http://dx.doi.org/10.5248/119.301

Bahram M, Polme S, Koljalg U, Zarre S, Tedersoo L. 2012. Regional and local patterns of

ectomycorrhizal fungal diversity and community structure along an altitudinal gradient in the

Hyrcanian forests of northern Iran. New Phytol. 193(2): 465-473.

http://dx.doi.org/10.1111/j.1469-8137.2011.03927.x

Hansen K, Lzessge T, Pfister DH. 2001. Phylogenetics of the Pezizaceae, with an emphasis on Peziza.

Mycologia 93: 958-990. http://dx.doi.org/10.2307/3761760

Kirk PM., Cannon PF, David JC, Stalpers JA. 2008. Ainsworth and Bisby’s dictionary of the fungi,

10% ed. CABI: Wallingford, Oxon. 771 p.

Lang C, Seven J, Polle A. 2011. Host preferences and differential contributions of deciduous tree

species shape mycorrhizal species richness in a mixed central European forest. Mycorrhiza.

21:297-308. http://dx.doi.org/10.1007/s00572-010-0338-y

McLenon-Porter TM. 2008. Above and below ground fungal diversity in a hemlock-dominated

forest plot in southern Ontario and the phylogenetic placement of a new Ascomycota subphylum.

Ecology and Evolutionary Biology, University of Toronto.

Nasir YJ. 1975. Betulaceae. 1-5, in: E Nasir, SI Ali (eds). Flora of Pakistan. No. 95. Department of

Botany, Gordon College, Rawalpindi.

Tedersoo L, Hansen K, Perry BA, Kjoller R. 2006. Molecular and morphological diversity of

pezizalean ectomycorrhiza. New Phytol 170: 581-596.

http://dx.doi.org/10.1111/j.1469-8137.2006.01678.x

Tedersoo L, Jairus T, Horton BM, Abarenkov K, Suvi T, Saar I, Kéljalg U. 2008. Strong host

preference of ectomycorrhizal fungi in a Tasmanian wet sclerophyll forest as revealed by DNA

barcoding and taxon-specific primers. New Phytol 180: 479-490.

http://dx.doi.org/10.1111/j.1469-8137.2008.02561.x

Tedersoo L, Suvi T, Jairus T, Ostonen I, Pélme S. 2009. Revisiting ectomycorrhizal fungi of the

genus Alnus: differential host specificity, diversity and determinants of the fungal community.

New Phytol 182: 727-735. http://dx.doi.org/10.1111/j.1469-8137.2009.02792.x

MY COTAXON

http://dx.doi.org/10.5248/120.189

Volume 120, pp. 189-194 April-June 2012

Erysiphe javanica sp. nov.,

a new tropical powdery mildew from Indonesia

JAMJAN MEEBOON’, IMAN HIDAYAT? & SUSUMU TAKAMATSU’™*

"Department of Bioresources, Graduate School, Mie University, Tsu 514-8507, Japan

?Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences,

Cibinong 16911 West Java, Indonesia

* CORRESPONDENCE TO: takamatu@bio.mie-u.ac.jp

Axsstract — During the collection of tropical powdery mildews in Cibodas Botanical

Garden and Mount Tangkuban Perahu in Indonesia, we found interesting specimens on

Castanopsis javanica (Fagaceae). Phylogenetic analysis using a combination of 28S and ITS

rDNA sequences clearly showed that this fungus forms a distinct lineage among the Erysiphe

species found on hosts of the plant genus Castanopsis. Therefore, we consider this fungus a

new species of Erysiphe, described here as Erysiphe javanica. Differences between E. javanica

and closely related Erysiphe species are discussed.

Key worps — Asia, Brasiliomyces, Erysiphaceae, phytopathogenic fungi, tropic

Introduction

Species of Erysiphe R. Hedw. ex DC. on fagaceous plants have been reported as

a unique group of powdery mildews because they comprise not only fungi with

simple mycelioid appendages that are characteristic for Erysiphe sect. Erysiphe

(Braun & Takamatsu 2000), but also encompass taxa with different types of

chasmothecial appendages previously belonging to the genera Brasiliomyces

Viégas [rudimentary/mycelioid], Uncinula Lév. [uncinuloid], and Microsphaera

Lév. [dichotomously branched at the apex] (Braun & Takamatsu 2000,

Takamatsu et al. 2007, Divarangkoon et al. 2011). In addition, Divarangkoon

et al. (2011) reported that the lineage of fagaceous Erysiphe spp. consists of

not only taxa with multiple peridial cell layers but also species characterised

by a single peridial cell layer previously assigned to Brasiliomyces [i.e., E. trina

Harkn. (= Brasiliomyces trinus (Harkn.) R.Y. Zheng), E. asiatica Meeboon et al.,

and E. monoperidiata Meeboon et al. (Divarangkoon et al. 2011)]. These results

suggest that different appendage types and single-layered peridial cells evolved

on fagaceous hosts.

190 ... Meeboon, Hidayat & Takamatsu

Several authors (Zheng 1984, Braun 1987, To-anun et al. 2003) have discussed

the significance of the peridial cells in separating Brasiliomyces from Erysiphe.

Zheng (1984) noted that Brasiliomyces must be treated as an independent genus

due to having a single layer of peridial cells not differentiated into inner and

outer layers as described in the genus Cystotheca Berk. & M.A. Curtis. This

argument was supported by Braun (1987) based on the reexamination of the

type species, B. malvastri Viégas. Before 2011, eight Brasiliomyces species

had been described based on this morphological criterion (To-anun et al.

2003; Divarangkoon et al. 2011), of which there were recorded on fagaceous

host plants — B. trinus (= E. trina), B. cyclobalanopsidis K.C. Kuo et al.

(= E. cyclobalanopsidis (K.C. Kuo et al.) U. Braun), B. kumaonensis N. Ahmad et

al. (= E. kumaonensis (N. Ahmad et al.) U. Braun). Braun & Paul (2009) placed

these three species on fagaceous hosts in synonymy under Erysiphe, because

E. trina nested in the lineage of Erysiphe emend. Braun & Takamatsu (2000).

During a March 2011 visit to the Cibodas Botanical Garden and Mount

Tangkuban Perahu on Java Island (Indonesia), we found specimens

morphologically similar to E. asiatica and E. monoperidiata. Molecular analyses

showed that the rDNA sequences of the specimens form an independent lineage

separated from E. asiatica and E. monoperidiata sequences. We therefore

propose this fungus as a new species.

Materials & methods

Morphological examination

Specimens were collected in two locations of West Java province (Indonesia),

namely, Cibodas Botanical Garden, Bogor and Mount Tangkuban Perahu, Bandung.

All collections were conducted in March 2011. Details of host name, collection date,

place, and collector were recorded. Morphological examinations were carried out as

outlined in Divarangkoon et al. (2011). Mycelia and chasmothecia were stripped from

leaf surfaces with a clean needle, mounted on a microscope slide, and examined in

3% NaOH using a light microscope with phase contrast 10x, 20x, and 40x objectives.

Thirty chasmothecia, asci, and ascospores were measured per sample. Specimens were

deposited at the National Museum of Nature and Science (TNS), Japan; Mie University

Mycological Herbarium (MUMH), Japan; and Herbarium Bogoriense (BO), Indonesia.

Phylogenetic analysis

Whole-cell DNA was extracted from chasmothecia using the chelex method (Walsh

et al. 1991) as described in Hirata & Takamatsu (1996). The 5’-end of the 28S rDNA,

including the domains D1 and D2, and ITS region, including the 5.88 rDNA, were

amplified by polymerase chain reaction (PCR) and then sequenced using protocols

as described in Takamatsu et al. (2006). Representative sequences determined in this

study were deposited in DNA databases (DDBJ, EMBL, GenBank) under the accession

numbers of JQ220151-JQ220162. Sequences generated from the rDNA ITS region

and D1/D2 domains were aligned using MEGA 5 (Kumar et al. 2008) with Erysiphe

Erysiphe javanica sp. nov. (Indonesia) ... 191

and Brasiliomyces sequences retrieved from DNA databases (DDBJ, EMBL, GenBank).

The alignment was deposited in TreeBASE (http://www.treebase.org/) under the

accession number of $12121. Maximum parsimony (MP) analysis was done in PAUP*

4.0b10 (Swofford 2002) with the heuristic search option using the ‘tree-bisection-

reconstruction (TBR) algorithm. All sites were treated as unordered and unweighted,

with gaps treated as missing data. The strength of the internal branches of the resulting

tree was tested with bootstrap analysis using 1000 replications (Felsenstein 1985). Tree

scores, including tree length, CI, RI, and RC, were also calculated.

Taxonomy

Erysiphe javanica Meeboon & S. Takam., sp. nov. FIG. 1

MycoBank MB 563656

Similar to Erysiphe asiatica, but differing in 5-7-spored asci and occurrence on

Castanopsis javanica.

TYPE: on Castanopsis javanica (Blume) A. DC. (Fagaceae), Indonesia, West Java

province, Bogor, Cibodas Botanical Garden, 14 March 2011, J. Meeboon, I. Hidayat &

S. Takamatsu (Holotype: TNS-F-44236; isotype: MUMH 5153; ex-type rDNA sequences,

JQ220160 (28S), JQ220162 (ITS)).

Erymo.ocy: the species epithet refers to the island where the specimens were

collected.

Cotonies hypophyllous, persistent, forming irregular white patches on the

host surfaces. HypHAE hyaline, superficial, septate, branched, 3-6 um wide.

APPRESSORIA well-developed, coral-like, single or occasionally opposite in

pairs. CONIDIOPHORES and CONIDIA unknown. CHASMOTHECIA scattered to

Fic. 1. Erysiphe javanica sp. nov.: 1-3, chasmothecia; 4—6, asci with 5—7 spores. Bars = 50 um.

192 ... Meeboon, Hidayat & Takamatsu

gregarious, (41-)50-66(-69) um diam., globose, containing 2 asci; appendages

present, rarely absent, poorly developed, mycelioid, (8—)17-73(-95) x (2-)3-5

(-8), colourless, aseptate, thin-walled, smooth. PERtpIuM thin, one conspicuous

layer, yellowish to light brown, semitransparent. Asci globose to subglobose,

sessile or short-stalked, (30-)36-50(-76) x (26-)27-44(-53) um, 5-7-spored.

Ascosporss ellipsoid-ovoid, hyaline, (10-)11-24(-26) x (5-)8-14(-17) um.

ECOLOGY & DISTRIBUTION: on Castanopsis javanica, Indonesia.

ADDITIONAL COLLECTIONS EXAMINED: on Castanopsis javanica: INDONESIA. WEsT

JAVA PROVINCE, Bogor, Cibodas Botanical Garden, 14 March 2011, J. Meeboon, I.

Hidayat & S. Takamatsu (MUMH 5147, BO22660, BO22661, BO22662, BO22663);

Bandung, Mount Tangkuban Perahu, 12 March 2011, J. Meeboon, I. Hidayat &

S. Takamatsu (MUMH 5123, BO22655, BO22656, BO22657, BO22658, BO22659).

Phylogenetic analysis

In the 28S+ITS combination sequence data set, 1091 of 1277 total characters

used in the MP analysis were constant, 58 characters were variable and

parsimony-uninformative, and 128 characters were parsimony-informative.

A total of 240 equally parsimonious trees (TL = 212, CI = 0.934, RI = 0.924,

RC = 0.863) were generated by the MP analysis (Fic. 2). All Erysiphe species

found on Castanopsis or Lithocarpus form a monophyletic clade with 98%

bootstrap support. Erysiphe javanica, E. asiatica, and E. monoperidiata form

independent clades with 99%, 100% and 66% bootstrap supports, respectively.

69| Erysiphe monoperidiata MUMH4990

53 Erysiphe monoperidiata MUMH4991

Erysiphe monoperidiata MUMH4988

66l| Erysiphe monoperidiata MUMH4985

Erysiphe monoperidiata MUMH4986

Erysiphe monoperidiata MUMH4987

99| Erysiphe javanica MUMH5147

Erysiphe javanica MUMH5153

100 | Erysiphe asiatica MHMU4989

Erysiphe asiatica MUMH 4992

Erysiphe gracilis ex Quercus glauca AB022358

Erysiphe japonica ex Quercus AB022416

Erysiphe trina ex Quercus AB022351

100| Erysiphe necator ex Vitis DQ444327

Erysiphe necator ex Vitis DQ189089

~~~ 5 changes

Fic. 2. Phylogeny of Erysiphe javanica inferred from a combination of 28S + ITS rDNA sequences

using maximum parsimony (MP) method. Bootstrap support percentages (1000 replications;

>50%) are shown on the branches.

Erysiphe javanica sp. nov. (Indonesia) ... 193

TABLE 1. Morphological comparison of Erysiphe javanica, E. monoperidiata,

and E. asiatica.

CHARACTER E. javanica E. monoperidiata E. asiatica

COLONIES Hypophyllous Amphigenous, Hypophyllous

mainly epiphyllous

HYPHAE 3-6 4-6 4-6

(um diameter)

CHASMOTHECIA

(um diameter)

(41-)50-66.5(-69.5)

2 asci

(55.5-)58-82.5(-85)

2-4 asci

(51-)57-74(-78)

2 asci

APPENDAGE (8.5-)17.5-73.5(-95) x (15.5-)18-66(-75) x (31-)45-51(-66) x

(um) (2-)3.5-5.5(-8) (2.5-)3-6(-7.5) (4-)4.5-5(-5.5)

ASCI (30.5-)36—50.5(-76) x (34-)36-58(-61) x (45-)46-59(-62) x

(um) (26-)27.5-44(-53) (24-)28-49(-52) (38-)40-53(-57.5)

5—7-spored 4—6-spored 6—8-spored

ASCOSPORES (10.5-)11.5-24.1(-26) x (11-)12.5-25(-26) x (16-)18-25(-28) x

(um) (5-)8-14(-17) (6-)7.5-13(-14.5) (8.5-)9-15(-16.5)

Discussion

Among the seven Erysiphe species previously found on Castanopsis (Braun

1987; Divarangkoon et al. 2011), only E. asiatica and E. monoperidiata were

reported to have a chasmothecial peridium composed of a single cell layer

(Divarangkoon etal. 2011). Morphologically, there are only slight morphological

differences between E. javanica, E. asiatica, and E. monoperidiata (TABLE

1). Erysiphe javanica differs from E. monoperidiata in having hypophyllous

colonies, smaller chasmothecia (69 um diam. in E. monoperidiata), having only

2 asci (with 5-7 spores per ascus) per chasmothecium, and being found only

on C. javanica in Indonesia. In addition, the morphologically close E. asiatica

differs slightly in having 6-8 spored asci and being confined to C. diversifolia and

C. echinocarpa in Thailand. Although it is difficult to differentiate E. javanica

from E. asiatica and E. monoperidiata solely on morphology, our phylogenetic

analysis, using a combination of 28S and ITS rDNA sequences, clearly showed

that E. javanica forms a distinct lineage distinguishable from E. asiatica and

E. monoperidiata (Fic. 2).

Acknowledgments

This work was financially supported in part by a Grant-in-Aid for Scientific

Research (No. 23580061) from the Japan Society of the Promotion of Science to ST

and MONBUKAGAKUSHO: MEXT (Ministry of Education, Culture, Science, and

Technology) Scholarship of the Japanese Government awarded to JM. The authors

also thank the Research Center for Biology, Indonesian Institute of Sciences (LIPI) for

providing facilities during collection and examination of powdery mildew samples.

194 ... Meeboon, Hidayat & Takamatsu

Literature cited

Braun U. 1987. A monograph of the Erysiphales (powdery mildews). Beihefte zur Nova Hedwigia

89: 1-700.

Braun U, Paul YS. 2009. The Indian Erysiphaceae revised. Nova Hedwigia 89: 371-395.

Braun U, Takamatsu S. 2000. Phylogeny of Erysiphe, Microsphaera, Uncinula (Erysipheae) and

Cystotheca, Podosphaera, Sphaerotheca (Cystotheceae) inferred from rDNA ITS sequences—

some taxonomic consequences. Schlechtendalia 4: 1-33.

Divarangkoon R, Meeboon J, Monkhung S, To-anun C, Takamatsu S. 2011. Two new species of

Erysiphe (Erysiphales, Ascomycota) from Thailand. Mycosphere 2: 231-238.

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution

39: 783-791. http://dx.doi.org/10.2307/2408678

Hirata T, Takamatsu S. 1996. Nucleotide diversity of rDNA internal transcribed spacers extracted

from conidia and cleistothecia of several powdery mildew fungi. Mycoscience 37: 283-288.

http://dx.doi.org/10.1007/BF02461299

Kumar S, Nei M, Dudley J, Tamura K. 2008. MEGA: A biologist-centric software for

evolutionary analysis of DNA and protein sequences. Briefings in Bioinformatics 9: 299-306.

http://dx.doi.org/10.1093/bib/bbn017

Swofford DL. 2002. PAUP*: Phylogenetic Analysis Using Parsimony (*and Other Methods).

Sinauer, Sunderland, MA.

Takamatsu S, Matsuda S, Niinomi S, Havrylenko M. 2006. Molecular phylogeny supports a Northern

Hemisphere origin of Golovinomyces (Ascomycota: Erysiphales). Mycological Research 110:

1093-1101. http://dx.doi.org/10.1016/j.mycres.2006.07.005

Takamatsu S, Braun U, Limkaisang S$, Kom-un S, Sato Y, Cunnington JH. 2007. Phylogeny and

taxonomy of the oak powdery mildew Erysiphe alphitoides sensu lato. Mycological Research

111: 809-826. http://dx.doi.org/10.1016/j.mycres.2007.05.013

To-anun C, Limkaisang S, Fangfuk W, Sato Y, Braun U, Takamatsu S. 2003. A new species of

Brasiliomyces (Erysiphaceae) on Dalbergia cultrata var. cultrata from Thailand. Mycoscience 44:

447-451. http://dx.doi.org/10.1007/s10267-003-0140-1

Walsh SP, Metzger DA, Higuchi R. 1991. Chelex 100 as a medium for simple extraction of DNA for

PCR-based typing from forensic material. Biotechniques 10: 506-513.

Zheng RY. 1984. The genus Brasiliomyces (Erysiphaceae). Mycotaxon 19: 281-289.

MY COTAXON

http://dx.doi.org/10.5248/120.195

Volume 120, pp. 195-208 April-June 2012

Pisolithus: a new species from southeast Asia

and a new combination

C. PHosri?’, M.P. MARTIiN?, N. SUWANNASAL, P. SIHANONTH* & R. WATLING?

"Microbiology programme, Pibulsongkram Rajabhat University, Phitsanulok, 65000, Thailand

*Departamento de Micologia, Real Jardin Botanico, CSIC,

Plaza de Murillo 2, 28014, Madrid, Spain

*Department of Biology, Srinakharinwirot University, Bangkok, 10110, Thailand

‘Department of Microbiology, Faculty of Science, Chulalongkorn University,

Bangkok, 10330, Thailand

°Caledonian Mycological Enterprises,

Crelah, 26 Blinkbonny Avenue, Edinburgh, EH4 3HU, Scotland

*CORRESPONDENCE TO: cherd_phosri@yahoo.co.uk

ABSTRACT — Based on morphological and ITS nrDNA sequence analyses, a species

recognized in classical European literature as Lycoperdon capsulifer is transferred to Pisolithus

and P. orientalis is proposed as a new species. Pisolithus orientalis basidiomes, collected under

Pinus kesiya in Thailand, correspond to Pisolithus sp. 5 sensu Martin et al. Pisolithus sp. 4

sensu Martin et al. is shown to comprise two groups, one equivalent to P. arhizus as collected

from Pinus and Quercus forests from Italy and Spain and the other represented by the new

P. capsulifer from mixed pine-deciduous forests in England and France.

KEY worps — ectomycorrhizal fungi, gasteromycete, phylogenetic species, PT, taxonomy

Introduction

Pisolithus is one of the most widespread and cosmopolitan gasteromycete

genera ranging from temperate to tropical regions of the world. It has long

been considered as monotypic (Coker & Couch 1928, Cunningham 1942,

Pilat 1958) and is well-researched, primarily based on ectomycorrhizal

studies, as Pisolithus tinctorius (P. Micheli ex Pers.) Coker & Couch (Marx et

al. 1984, Burgess et al. 1995). Rauschert (1959) proposed that this species was

conspecific with Polysaccum arhizum Scop., which he recombined as Pisolithus

arhizus (Scop.) Rauschert. However, after molecular studies indicated that

P. tinctorius is a species complex (Anderson et al. 1998, Cairney et al. 1999,

Diez et al. 2001), Martin et al. (2002) proposed at least 11 phylogenetic species.

Several subsequent morphological and molecular studies have confirmed

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Pisolithus orientalis sp. nov. (Thailand) ... 197

the taxonomic rank of some old species [Pisolithus albus (Cooke & Massee)

Priest, P marmoratus (Berk.) E. Fisch., PB microcarpus (Cooke & Massee)

G. Cunn.] and established several new ones (Pisolithus abditus Kanch. et al.,

P. aurantioscabrosus Watling, P. indicus Natarajan & Senthil.). Nonetheless,

some clades recognised by Martin et al. (2002) remain without specific names.

Additional morphological studies and ITS nrDNA sequence analyses of

recently collected austro-asiatic, euro-asiatic, and paleotropical Pisolithus

specimens have been conducted to assign morphological species to the

Pisolithus sp. 5 phylogenetic lineage and to members of the AII clade of Martin

et al. (2002).

Materials & methods

Fungal isolates

The Pisolithus material used in this work included fresh, herbarium, or cultured

specimens sampled from a variety of geographical sites (TABLE 1). Basidiomes for

each isolate are retained in the herbarium at the Microbiology Programme, Faculty of

Science & Technology, Pibulsongkram Rajabhat University and BBH Thailand. Cultures

obtained from basidiomata were grown and maintained on Modified Melin Norkrans’

(MMN) medium (Marx & Kenny 1982) at 30°C with subculturing every 1-2 months.

Herbarium specimens were provided by BCN and E (Abbreviations for herbaria follow

Lanjouw & Stafleu 1964).

Molecular methods

Isolation, amplification, purification, and sequencing of DNA followed Phosri et al.

(2007). DNA was isolated using the E.Z.N.A. fungi DNA miniprep kit (Omega Biotech,

Doraville, Georgia, USA) as described by Martin & Garcia-Figueres (1999) or a DNeasy

Plant Mini Kit (Qiagen, Hilden, Germany). ITS regions were amplified with Ready-To-

Go™ PCR Beads (Amersham-Pharmacia Biotech, NJ, USA) (Martin & Winka 2000).

Primer pairs ITS1F/ITS4B and ITS1/ITS4 were used to amplify the ITS1+5.8S+ITS2

nrDNA gene cluster following White et al. (1990) and Gardes & Bruns (1993). Prior to

sequencing, products were cleaned using either E.Z.N.A. Clean kit (Omega Biotech) or

directly from the gel using QIAQuick PCR purification kit (QIAGEN, Chatsworth, CA).

Both strands were sequenced separately at Secugen S.L. (Madrid, Spain) or Macrogen

(South Korea). All samples were sequenced in both directions.

When needed, DNA was cloned using pGEM®-T Easy Vector System II cloning kit

(Promega Corporation, Madison, Wisconsin, USA) and purified with QIAPrep Spin

Mini prep (QIAGEN). Both strands were sequenced separately using vector specific

primers T7 and SP6 (Nag et al. 1988) at Secugen S.L. or Macrogen.

Sequence analysis

Navigator™ Sequence comparison software (Perkin Elmer Applied Biosystem) or

Sequencher™ version 4.2 (Gene Codes Corporations, Ann Arbor, Michigan, USA) was

used to assemble the consensus sequence from the two strands of the ITS nrDNA of

each isolate. Blastn searches with megablast option were used to compare the sequences

obtained against the sequences in the National Center of Biotechnology Information

198 ... Phosri & al.

(NCBI) nucleotide database. The new consensus sequences have been lodged in the

EMLB-EBI database with accession numbers indicated in TABLE 1.

Sequences obtained were compared with homologous sequences of Pisolithus spp.

retrieved from the EMBL Nucleotide Sequence Database with many published in Martin

et al. (2002), Kanchanaprayudh et al. (2003a,b), Moyersoen et al. (2003), and Reddy et

al. (2005) to represent all their described clades. Prior to 2009 sequences were aligned

using the SEQAPP software (Perkin Elmer Applied Biosystem) and thereafter using Se-

Al v2.0a11 Carbon (Rambaut 2002). The alignment was optimized visually. Alignment

gaps were indicated as “-” and ambiguous nucleotides were marked as “N”.

Phylogenetic analyses

The alignment was analysed using the programme PAUP 4.0b10 (Swofford 2003) and

MrBAYES v3.0 b4 (Huelsenbeck & Ronquist 2001) as described in Telleria et al. (2010).

Scleroderma citrinum Pers. (FM213344) and Suillus luteus (L.) Roussel (GU373495)

were included as outgroups. First, a maximum parsimony analysis (MP) was inferred

using the heuristic search option in PAUP. Gaps were treated as missing data. Branch

lengths equal to zero were collapsed to polytomies. Nonparametric bootstrap (BP)

support (Felsenstein 1985) for each clade was tested based on 10000 replicates, using

the fast-step option.

A second analysis was conducted using a Bayesian approach (Larget & Simon 1999,

Huelsenbeck & Ronquist 2001). It was performed assuming the HKY + G model as

suggested by hierarchical likelihood ratio test (hKRTs) and akaike information criterion

(AIC) in MrModeltest 2.3 (Posada & Crandall 1998, Nylander 2004).

A combination of both posterior probabilities and bootstrap proportion was used

to assess the level of confidence for a specific node (Lutzoni et al. 2004). The alignment

matrix and the 50% majority—rule consensus tree from the Bayesian analysis are available

in TreeBase (http://www.treebase.org/).

Recovered lineages and clades are named according to Martin et al. (2002). When a

specific name has been formally assigned to a clade, the name is included along with the

species number of Martin et al. (2002).

Results

Thirty-three new Pisolithus sequences have been generated, including those

obtained from the epitype (E00290168) of the new combination, P. capsulifer,

and the holotype (BBH28597) of the new species, P orientalis. These were

aligned with 67 additional sequences obtained from databases (GenBank and

UNITE; see Fics 1-2) and phylogenetically analyzed.

The ITS nrDNA dataset contains 97 sequences and 893 aligned positions, of

which 410 were constant, 177 parsimony uninformative, and 306 parsimony-

informative. Maximum parsimony analysis yielded 100 most parsimonious

trees (940 steps long, CI = 0.6681, HI = 0.3319, RI = 0.8968). One best tree

found by branch swapping is shown in Fic. 1, with bootstrap values on the

branches. The 50% majority-rule consensus tree from the Bayesian analyses is

in Fic. 2, with posterior probabilities indicated on the branches.

Pisolithus orientalis sp. nov. (Thailand) ... 199

Suillus luteus, GU373495

Scleroderma citrinum, ape 3344 F eaance ee fare Ralawsi

, Shorea macroptera, Malaysia A :

c AF415226, Shorea macroptera, Malaysia I P. aurantioscabrosus (P. species 11)

AY756113, Vateria indica, India, === P. indicus (P. species 13)

100 + AFO03915, Afzelia sp., Kenya ;

AF228653, Afzelia sp., Kenya | P. species 1

24PISOLI, Shorea sp., Thailand

A I AB099921, Shorea roxburgii, Thailand J ‘

100 |- AB099920, Shorea roxburgii, Thailand P. abditus (P. species 12)

31PISOLI, Shorea roxburgii, Thailand

AB099922, Dipterocarpus alatus, Thailand

AF003914, Eucalyptus camaldulensis, Kenya

AF374/19, Eucalyptus caiopnylia Australia»

, Eucalyptus calophylla, Australia f

99 IF AF004734, Eucalyptus sp., New South Wales P. marmoratus (P. species 2)

AF440866, Eucalyptus plantation, China

AF374665, auc nes patens, Australia

84 AF228641, Cistus ladanifer, Spain

ARHPIS3 (clones a-c), Cistus ladanifer, Spain .

400|| AF228642, Cistus ladanifer, Spain P. species 3

AF228643, Cistus ladanifer, Spain

AF228644, Cistus ladanifer, Spain ;

AF374624, Acacia sp., Australia —=—_—=_< P. species 10

AF347700, FUCAypUS, Brazil

AF004735, Eucalyptus sp., New South Wales

eee Busses she eee P. microcarpus

, Eucalyptus sp., Brazi :

95|L AF 140547, Eucalyptus sp.. Brazil (P. species 9)

64 AF374704, Eucalyptus dunni, Brazil

57 88 U62666, Eucalyptus citriodora, Brazil

AF374661, Eucalyptus camaldulensis, Australia | P ies'S

AF374708, Eucalyptus sp./Acacia sp., Australia 7SPECles

AF004732, Eucalyptus sp., New South Wales =< Pisolithus sp.

AF270786, Eucalyptus sp., Australia

AF374622, Acacia holosericea, Senegal

72\I— .13PISOLI, Eucalyptus camaldulensis, Thailand

14PISOLI, Eucalyptus camaldulensis, Thailand

86 15PISOLI, Eucalyptus camaldulensis, Thailand

AF228656, ueaipius sp., Spain

AF228655, Eucalyptus sp., Morocco

AF 228654, Eucalyptus sp., Morocco

B ABO099918, Thailand

99 AB099909, Eucalyptus camaldulensis, Thailand

AF416589, Eucalyptus tereticornis, India P. albus complex

AF440868, Eucalyptus sp., China (P. species 7)

AF374675, Eucalyptus sp., Queensland

AF004736, unknown potential host, New South Wales

AF004737, unknown potential host, New South Wales

17PISOLI, unknown potential host, Thailand

AJ629887 (12PISOLI), Eucalyptus camaldulensis, Thailand

20PISOLI, Eucalyptus sp., Hawaii, USA

35PISOLI, Melaleuca sp., Thailand

AB099911, Thailand

AF374638, Acacia mangium, Malaysia

21PISOLI, Eucalyptus Sp Australia ?

AF374646, Eucalyptus globulus, Australia === P. species 10

AF440867, Eucalyptus plantation, China === _- P. microcarpus

AF003916, Pinus caribaea, pale

AF228645, Quercus ilex/Q.coccifera, Spain

AF228646, Quercus ilex/C. ladanifer, Spain ; 5

72 posclls pins sp., sey P. tinctorius

, Pinus sp., i

AF 143234, unknown potential host, France (P. species 6)

19PISOLI, Pinus sp./Quercus sp., USA

AF 228647, Pinus caribaea, Kenya

AF374625, Pinus kesiya, Thailand

AF374679, Eucalyptus sp., China

AF374711, Pinus sp./Eucalyptus sp., China

AB099919, unknown potential host, Thailand

53 34PISOLI, Pinus kesiya, Thailand K .

pms enknown potent host Thailand P. orientalis

LI, Pinus kesiya, Thailan -

14PISOLI, Pinus kesiya, Thailand (P. species 5)

10PISOLI, Pinus kesiya, Thailand

33PISOLI, Pinus kesiya, Thailand

All 7PISOLI, Pinus kesiya, Thailand

8PISOLI, Pinus kesiya, Thailand

FM213365 (3PISOLI), Pinus sp., Spain

AF228648, Quercus ilex/Q. coccifera, Spain

92bisPISOLI (clone c), Will Wood, Sardinia, Italy

92bisPISOLI Go a, d), Will Wood, Sardinia, Italy

92bisPISOLI (clone f), Will Wood, Sardinia, Italy ;

AF228649, Q. ilex, Spain P. arhizus

AF228651, Q. ilex, Spain (P. species 4)

AF 228650, P. halepensis/Q. coccifera, Spain

54 AF 228652, P. halepensis, Spain

92bisPISOLI (clone b), Will Wood, Sardinia, Italy

6PISOLI, Pinus sp., Spain

AF374627, Pinus sp., South Africa ‘

AF374629, Pinus pumila/Betula ermanii, Japan

77 UDB001206, Pinus sp. forest, Sweden

51PISOLI, unknown potential host, England P. capsulifer

46PISOLI, unknown potential host, England F

954 S3PISOLI, mixed forest/Pinus sp., pranes (P. species 14)

49PISOLI, Pinus sylvestris/P. contorta, England

— 5changes

Fic. 1. Phylogenetic tree (one of 100 most parsimonious trees) obtained with a parsimony analysis

under heuristic search of ITS sequences of Pisolithus collections included in TABLE 1 and sequences

obtained from the GenBank and UNITE. Each branch is labeled with the DNA isolation code in

bold (new sequences) or with the accession number including the possible host and the country.

Numbers above branches represent bootstrap values. Clades and species number according to

previous authors.

200 ... Phosri & al.

In both analyses, Pisolithus sequences form a highly supported clade (BS =

84%, PP = 1.0) with at least 15 terminal assemblages. However, the relationships

between the clades are generally not well resolved and the main differences

between Fic. 1 and Fie. 2 are related to the position of clades from lineage AI

in Martin et al. (2002).

The basal clade (BS <50%, PP = 0.62) (lineage C, Martin et al. 2002) included

in our study contains Pisolithus aurantioscabrosus and P. indicus, which are

sister to the remaining Pisolithus taxa. These two taxa are restricted to native

dipterocarp forests in southeast Asia and India, respectively.

Lineage AI is separated into four clades, each with high support (BP = 99%

or PP = 1.0). Lineage AII also ramifies into at least four main groups, but these

possess weak support (BS < 50%, PP = 0.98). The four AII groups are Pisolithus

spp. 4, 5, and 6 of Martin et al. (2002), and one group here called Pisolithus sp.

14. Pisolithus sp. 6 (BS = 72%, PP = 0.99) consisted of eight Pisolithus sequences

from basidiomata found mainly in association with pine (Pinus caribaea

Morelet.) and oak (Quercus ilex L., Q. coccifera L.) in various regions ranging

from USA, Spain, Kenya, and France. We consider this clade to be P. tinctorius

s. St., sister to the Pisolithus sp. 5 clade formed by seven new isolates associated

with Pinus kesiya Royle ex Gordon in Thailand and five GenBank sequences.

The unifying features unique to these collections lead us to describe them below

as a new species, Pisolithus orientalis. The other two clades (Pisolithus spp. 4

and 14) in Lineage AII are sister to the clade formed by Pisolithus tinctorius

s. st. and P orientalis. Collections from the British Isles (46PISOLI, 49PISOLI

and 51pisoL1) and France (53pisoLi) clearly separate from Pisolithus sp. 4

(BS = 70%, PP = 1.0) and group together with sequences from Sweden

(UDB001206) and Japan (AF374629). Sequenced specimens from British Isles,

France, and Sweden have been analysed in parallel to basidiospore SEM studies.

The electron micrographs (Fic. 3) indicate that the spore ornamentation does

not resemble that of any other Pisolithus documented. The specific name

Pisolithus capsulifer has been assigned to Pisolithus sp. 14.

Discussion

In lineage AII, one clade (‘sp. 14’) comprises predominantly British material.

Pisolithus is very rare in the British Isles, where it is recorded from London,

Hampshire, Devon, and Norfolk. Most records are from Tertiary gravels and

sandy mineral soils and confined to the southernmost part, especially the

southeast of England. There is one record of Pisolithus from Ireland ‘on a bank

in a car park, under Castanea sativa and Pinus spp., well-drained peat over lying

sandy boulder-clay north slope of Knockmealdon Mountains, Kilballyboy Wd.,

South Tipperary (July 1984)’ (Ing 1995), but it is not recorded from Scotland.

The British sequences agreed with one each from France and Sweden. Here spore

morphology is also distinctive, but is it significant? Grand (1976) and Watling

Pisolithus orientalis sp. nov. (Thailand) ... 201

Suillus luteus, GU373495

Scleroderma citrinum, FM213344 pale: F

AY756113, Vateria indica, Indica =—=_ +P. indicus (P. species 13)

00 AF41 227, Shores macroptera, Malaysia } | P. aurantioscabrosus

, Shorea macroptera, Malaysia i

Al 4.00 - AF003915, Afzelia sp. Kenya % a (P. species 11)

AF228683, Afzelia sp. Kenya_ | P. species 1

A l AB099922, Dipterocarpus alatus, Thailand

24PISOLI, Shorea sp., Thailand 5 F

1.00 |} 31PISOLI, Shorea roxburgii, Thailand P. abditus (P. species 12)

AB099920, Shorea roxburgii, Thailand

AB099921 Shorea roxburgii, Thailand

AF228641, Cistus ladanifer, Spain. .

1.00 1.00 AF 228644, Cistus ladanifer, Spain ;

ARHPIS3, (clones a-c), Cistus ladanifer, Spain P. species 3

Al AF228643, Cistus ladanifer, Spain

AF228642, Cistus ladanifer, Spain.

0.88 4.00 AF374665, Eucalyptus patens, Australia

: AF003914, Eucalyptus camaldulensis, Kenya

‘AF374719, Eucalyptus caiophylla, Australia» P. marmoratus

AF440866, Eucalyptus plantation, China (P. species 12)

AF004734, Eucalyptus sp., New South Wales

AF004733, Eucalyptus sp., New South Wales ?

AF374646, Eucalyptus globulus, Australia P. species 10

0.96 AF440867 Eucalyptus plantation China P. microcarpu:

cacia sp., Australia 2. specie: ket

B AF374624, Acaci Austral P.

1.00 004732, Eucalyptus sp., New South Wales —_—_—=- Pisolithus sp.

8210.99 , Eucalyptus camaldulensis, Australia F

0.82 AF374661, Eucal /dulensis, Australia |

AF374708, Eucalyptus sp./Acacia sp., Australia P. species 8

0.99 AF374704, Eucalyptus dunni, Brasil

: 9.99] 4 oof + U62666, Eucalyptus citriodora, Brasil

AF 140547, Eucalyptus sp., Brasil. P. microcarpus

AF347700, Eucal lyptus sp., Brasil J

AF228657, Eucalyptus sp., Morocco (P. species 9)

1.00 AF 142991, Eucalyptus sp., Brasil

: AF004735, Eucalyptus sp., New South Wales

AB099911, Thailand é

20PISOLI, Eucalyptus sp.,. Hawaii, US

0.80/L 35PISOLI, Melaleuca sp., Thailand

AF374638, Acacia mangium, Australia_

17PISOLI, unknown potential host, Thailand __ :

AJ629887 (PISOLI12), Eucalyptus camaldulensis, Thailand

0.78 AF004737, unknown potential host, New South Wales

i AF004736, unknown potential host, New South Wales

21PISOLI, Eucalyptus sp., Australia

AF440868, Eucalyptus sp., China

AF374675, Eucalyptus 8p. Morocco _ “ P. albus complex

AB099909, Eucalyptus camaidulensis, Thailand (P. species 7)

0.99 AF416589, Eucalyptus tereticornis, India

AF270786, Eucalyptus sp., Australia

1.00 AF374622, Acacia holosericia, Senegal

AB099918, Thailand :

AF 228656, Eucalyptus sp., Spain

AF228655, Eucalyptus sp., Morocco

AF228654, Eucalyptus sp., Morocco _ ,

13PISOLI, Eucalyptus camaldulensis, Thailand

15PISOLI, Eucalyptus camaldulensis, Thailand

; ; 14PISOLI, Eucalyptus camaldulensis, Thailand

1.00 ARS TAGES. Pinus pumila/Betula ermanii, Japan

01206, Pinus sp. forest, Sweden -

0.91 4 Soe mown potential host, En fang eae P. capsulifer

; , Pinus sylvestris/P. contorta, Englan .

0.56 46PISOLI, unkncwn potential host, England (P. species 14)

L 53PISOLI, mixed forest/ Pinus sp., France

AF374627, Pinus SP South Africa

FM213365 (3PISOLI), Pinus sp., Spain

1.04 6PISOLI, Pinus sp, Spain ie i

92bisPISOLI (clone Wil Wood, Sardinia, Italia

92bisPISOLI (clone b), Will Wood, Sardinia, Italia P. arhizus

92bisPISOLI (clone c), Will Wood, Sardinia, Italia : 7

92bisPISOLI (clone a), Will Wood, Sardinia, Italia (P. species 4)

AF228648, Quercus ilex/Q. coccifera, Spain

All AF228650, Pinus halepensis/Q.coccifera, Spain

0.98 AF 228652, Pinus halepensis, Spain

, AF228649, Quercus ilex, Spain

AF228651, Quercus ilex, Spain

AF003916, Pinus caribaea, Kenya

19PISOLI, Pinus sp./Quercus sp., USA

0.99 }— AF 143234, unknown potential host, France H J

AF 143233, Pinus sp. USA P. tinctorius

AF 228647, Pinus caribaea, Kenya (P. species 6)

S5PISOLI, Pinus sp., Spain ; fi

1.00 AF228645, Quercus ilex/ Q. coccifera, Spain _

AF228646, Quercus ilex/Cistus ladanifer, Spain

AF374625, Pinus kesiya, Thailand

AF374679, Eucalyptus sp., China

0.69 9PISOLI, Pinus kesiya, Thailand

ABopoRt: panknown potential host, Thailand

inus kesiya, Thailan q .

TIPISOLI, Pinus kesiya, Thailand P. orientalis

10PISOLI, Pinus kesiya, Thailand A

TPISOLI, Pinus kesiva, thailand (P. species 5)

8PISOLI, Pinus kesiya, Thailand

AF374711, Pinus sp./Eucalyptus sp., China

34PISOLI, Pinus kesiya, Thailand

AB099919, unknown potential host, Thailand

0.1

Fic. 2. Phylogenetic tree obtained with a Bayesian Monte Carlo Markov (MCMC) analysis

assuming HKY + G model, as suggested by MrModeltest 2.3, of ITS nrDNA sequences of Pisolithus

collections included in TaBLE 1 and sequences obtained from the GenBank and UNITE. Each

branch is labeled with the DNA isolation code in bold (new sequences) or with the accession

number including the potential host and the country. Numbers above branches represent posterior

probabilities values. Clades and species numbers according to previous authors.

et al. (1999) note that basidiospore morphology should be considered carefully

as the mature spore ornamentation relies on spore development in which nurse

cells play a vital role and affect final surface features. Morphological differences

often have diagnostic importance for new morphological species, (e.g., Astraeus

odoratus; Phosri et al. 2004, 2007), despite instances of convergence.

202 ... Phosri & al.

Sowerby (1814), who provided an excellent illustration of a gasteroid fungus,

apparently coined the epithet “Lycoperdon capsuliferum based on the fact that

the fertile part of the fruiting body contained small spore capsules resembling

a fig fruit. Fries (1829: 54) recognized the significance of this collection and

how it differed from other species known to him mainly from the classical

literature. He formally proposed the superfluous new name Polysaccum

olivaceum in section Radiculosa for Sowerby’s species, differing from the other

members of the section in spore-mass colour and fruiting body shape. Because

of the morphological and habitat similarities shared by Sowerby’s fungus and

modern ‘sp. 14’ clade collection, we are confident in identifying this material as

Lycoperdon capsuliferum, which we transfer to Pisolithus.

Pisolithus capsulifer (Sowerby) Watling, Phosri & M.P. Martin, comb. nov. Fic. 3

MycoBank MB 519949 [PISOLITHUS SP. 14]

= Lycoperdon capsuliferum Sowerby, Col. Fig. Engl.

Fung., Suppl.(no. 31): pl. 425a/b. 1814.

= Polysaccum olivaceum Fr., Syst. Mycol. 3(1): 54. 1829 [nom. nov. superfl.].

Type: Coloured plate 425a/b in Sowerby, Col. Fig. Engl. Fung., Suppl.(no. 31), 1814

(lectotype designated here). ENGLAND, Berkshire, Sandhurst, 7 Sep 1993, leg. E.E.

Green, Wat. 25310 (epitype designated here, E00290168; GenBank FR748135)

BASIDIOMES turbinate to pyriform with subglobose to ellipsoid head 40-60

mm diam. or flattened at the apex > 90 mm broad, tapering into solid, firm,

irregular pseudostipe incorporating soil debris; pseudostipe 15-30(-90) mm

long often flattened and then 10-25 mm broad truncated downwards to 15 mm.

PERIDIUM wafer thin, > 1mm thick, dry, smooth, yellowish ochraceous with

olivaceous spots and faint netted appearance from outline of pseudoperidioles

below, then fragmenting to expose powdery spore-mass and disintegrating

pseudoperidioles. Lower part more compacted and with an olivaceous-

yellow hue, finally drying hard after weathering to become clinker-like.

PSEUDOPERIDIOLES pale then red-brown finally burnt umber, larger at apex

and compressed downwards, becoming pulverulent.

CLAMP-CONNECTIONS present. BASIDIOSPORES 6.8-8.2-10.6 um, globose,

spinose spines at first separate and 1.1-1.9-2.9 um long, before collapsing

to coalesce into groups and falling back onto spore surface to give a rivulose

effect.

ADDITIONAL SPECIMENS EXAMINED: FRANCE, 20 km. from Limoges, mixed

broadleaf-conifer forest, 1-Sep-1997, leg. E.E. Green, Wat. 25298 (E00159822).

UNITED KINGDOM, Hampsnirg, Ringwood, 12-Sep-1926, coll. E.J.H. Corner 81444

(E00075037); coll. E.J.H. Corner 81446 (E00075038); BERKSHIRE, Windsor Great Park,

Jul-1993, leg. EE. Green, Wat. 25311 (E00290167); Caesar’s Camp near Bracknell, Sep-

1970, leg. E.E. Green (E00159834); 9-Sep- 1973, leg. E.E. Green, Wat. 10118 (E00185017);

3-Oct-1981, leg. E.E. Green, Wat. 14216 (E00185018).

Pisolithus orientalis sp. nov. (Thailand) ... 203

LyS

Fic. 3. SEM of spore ornamentation of selected Pisolithus collections (bars = 1 pm). a-e:

P. capsulifer. From England—a-b: epitype E00290168 (Wat. 25310, 46pisoL1), c-d: E00185017

(Wat. 10118, 49PIsoL1); from France—e: E00159822 (Wat. 25298, 53pIsoLt). f: PB. orientalis.

Holotype BBH28597 (33PIsoLi).

ComMENTSs. The distribution of the pseudoperidioles in fresh material is clearly

expressed by Sowerby (1814) when he wrote, “at first strong and very rugged but

the congeries of seeds, if I may so-call them, were enveloped or compressed into

forms laying by each other, giving a reticulated appearance in some directions,

being mostly rather oblong, and of a dark brown colour...and looks like pollen

bursting from something analogous to anthers.”

204 ... Phosri & al.

Sete

/ & 426 C Ltda

‘ Sign Geos Ct E€ehlarcs

4) b

Me; éofecicleaeu Cafeticks ye recs Sov

So laps Hedddrr bet €

— te

Fic. 4. a: Sowerby’s illustration of Lycoperdon capsuliferum (arrow).

b: Pisolithus orientalis Holotype BBH28597 (33PIsoL!).

Pisolithus orientalis sp. nov. (Thailand) ... 205

Pisolithus orientalis Watling, Phosri & M.P. Martin sp. nov. Fia. 4

MycoBank MB519952 | PISOLITHUS SP. 5]

Differs from Pisolithus tinctorius by smooth pedicellate basidiomata with blackish brown

discs, smaller globose basidiospores ornamented by isolated groups of narrow cones

that adhere together to form secondary spines, and an association with Pinus kesiya.

Type: THAILAND. Chang Mai, Hot District, Bo Kaew pine plantation, under Pinus

kesiya, 11-Aug-2001 (holotype, BBH 28597; GenBank FR748148).

BASIDIOME: 35-55 mm high, pedicellate to broadly clavate, divided into

distinct head and cylindrical stipe, snuff-brown to cigar-brown with a black

shining surface around the head, surface smooth, not scaly. PERIDIUM: 24-45

mm broad, broadly ellipsoid to subglobose, rough, thin, brittle, snuff-brown,

breaking up at the apex to expose powdery contents and join up to stipe. STIPE:

13-24 x 10-15 mm woody, snuff-brown, smooth, cylindrical base of agglutinate

mycelium and soil, yellowish rhizomorphs when immature. GLEBA: snuff-

brown to ferruginous powdery mass formed by the breakdown of peridioles

seated in a small black pocket within the peridium. PERIDIOLEs: thin-walled,

ovoid-ellipsoid, smooth, clay-buff, 2 x 1 mm, at first clay-buff then more snuff-

brown or ferruginous as they mature, discrete, but later breaking down into

a snuff brown to ferruginous powdery mass, exuding sienna to fulvous fluid

when mounted in alkaline solution.

CONSTITUENT HYPHAE: intertwined, hyaline to ochraceous, thin-walled,

2.4-4.8 mm broad, septate without encrustation, clamp-connections present.

BASIDIA not seen. BASIDIOSPORES: globose, 5.8-9.0-10.7 um including spines,

heavy ornamentation consisting of isolated groups of connate, narrow cones

adhering together to form points, lacking connections.

ADDITIONALSPECIMENS EXAMINED: THAILAND, Chiangmai, 11-Aug-2001 (BBH28598;

GenBank FR748149), together with other specimens referred to P orientalis (sp. 5) in

TABLE 1.Other sequences obtained were directly from cultures.

The pedicellate, smooth basidiomes that become blackish brown at the disc

above the first maturing pseudoperidioles, the relatively small spores that

are covered with narrow cones that join together to form connate secondary

spines, and an association with Pinus kesiya distinguish Pisolithus orientalis

from other species in the genus. The distinctive spore ornamentation coupled

with the molecular data distinguishes this new taxon. The characters emphasis

how important it is to examine in detail the development of both basidiome

and basidiospores in Pisolithus.

Conclusion

Our study confirms that Pisolithus tinctorius sensu lato, as previously and

widely understood, covers several separate species. Molecular work linked

to scanning electron microscope studies of the basidiospores is proving

206 ... Phosri & al.

essential in unravelling speciation within Pisolithus. The marriage of data from

classical sources and our molecular and morphological studies confirms the

distinctiveness of the British, French, and Swedish collections and a new species

from SE Asia growing with Pinus kesiya.

Acknowledgements

We thank herbaria BCN, E for loan of specimens used and A.D. Parker for material

from his personal herbarium. Part of the work was supported by the Ministerio de

Educacién y Ciencica (CGL2006-12732-CO2-01/BOS). CP is indebted to the National

Research Council of Thailand (NRCT) and Royal Thai Government for financial support

and to the European Commission Human Potential Programme for supporting part of

this study at the Real Jardin Botanico de Madrid (BIODIBERIA). Authors also thank

peer reviewers for comments and suggestions.

Literature cited

Anderson IC, Chambers SM, Cairney JWG. 1998. Molecular determination of genetic variation

in Pisolithus isolates from a defined region in New South Wales, Australia. New Phytol. 138:

151-162. http://dx.doi.org/10.1046/j.1469-8137.1998.00894.x

Burgess T, Malajczuk N, Dell B. 1995. Variation in Pisolithus based on basidiome and basidiospore

morphology, culture characteristics and analysis of polypeptides using 1D SDS-PAGE. Mycol.

Res. 99: 1-13. http://dx.doi.org/10.1016/S0953-7562(09)80309-2

Cairney JWG, Chambers SM, Anderson IC. 1999. Pisolithus systematics-molecular methods

provide fresh insights. Mycologia 3: 31-35. http://dx.doi.org/10.1016/S0269-915X(99)80078-0

Coker WC, Couch JN. 1928. The Gasteromycetes of the eastern United States and Canada.

University of North Carolina Press, Chapel Hill. 201 p.

Cunningham GH. 1942. Gasteromycetes of Australia and New Zealand. Dunedin. 236 p.

Diez J, Anta B, Manjon JL, Honrubia M. 2001. Genetic variability of Pisolithus isolates associated

with native hosts and exotic Eucalyptus in the western Mediterranean region. New Phytol. 149:

577-587. http://dx.doi.org/10.1046/j.1469-8137.2001.00036.x

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evol. 39:

783-791. http://dx.doi.org/10.2307/2408678

Fries EM. 1829. Systema mycologicum 3(1): 1-259. Gryphiswaldae, Germany.

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidomycetes - applications

to the identification of mycorrhizae and rusts. Mol. Ecol. 1: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Grand LE. 1976. Distribution, plant associates and variation in basidiocarps of Pisolithus tinctorius

in the United States. Mycologia 68: 673-677. http://dx.doi.org/10.2307/3758989

Huelsenbeck JP, Ronquist F. 2001. MRBAYES: bayesian inference of phylogenetic trees. Bioinforma.

17: 754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

Ing B. 1995. Pisolithus in Ireland. Bull. Brit. Mycol. Soc. 19: 57-58.

http://dx.doi.org/10.1016/S0007-1528(85)80056-0

Kanchanaprayudh J, Zhou Z, Yomyart S. Sihanonth P, Hogetsu T, Watling R. 2003a. A new Species,

Pisolithus abditus, an ectomycorrhizal fungus associated with dipterocarps in Thailand.

Mycotaxon 88: 463-467.

Kanchanaprayudh J, Zhou Z, Yomyart S, Sihanonth P, Hogetsu T. 2003b. Molecular phylogeny

of ectomycorrhizal Pisolithus fungi associated with pine, Dipterocarp and Eucalyptus trees in

Thailand. Mycoscience 44: 287-294. http://dx.doi.org/10.1007/s10267-003-0110-7

Pisolithus orientalis sp. nov. (Thailand) ... 207

Lanjouw J, Stafleu FA. 1964. Index Herbariorum, part 1. The herbaria of the world, 5" edition,

Utrecht, Netherlands.

Larget B, Simon DL. 1999. Markov Chain Monte Carlo algorithms for the bayesian analysis of

phylogenetic trees. Mol Biol. & Evol. 16: 750-759.

Lutzoni FE, Kauff K Cox CJ, McLaughlin D, Celio G, Dentinger B, Padamsee M, Hibbett D, James

TY, Baloch E, Grube M, Reeb V, Hofstetter V, Schoch C, Arnold AE, Miadlikowska J, Spatafora

J, Johnson D, Hambleton S, Crockett M, Shoemaker R, Sung GH, Liicking R, Lumbsch T,

O’Donnell K, Binder M, Diederich P, Ertz D, Gueidan C, Hansen K, Harris RC, Hosaka K, Lim

YW, Matheny B, Nishida H, Pfister D, Rogers J, Rossman AM, Schmitt I, Sipman H, Stone J,

Sugiyama J, Yahr R, Vilgalys R. 2004. Assembling the fungal tree of life: progress, classification,

and evolution of subcellular traits. Amer. J. Bot. 91: 1446-1480.

http://dx.doi.org/10.3732/ajb.91.10.1446

Martin F, Diez J, Dell B, Delaruelle C. 2002. Phylogeography of the ectomycorrhizal Pisolithus

species as inferred from nuclear ribosomal DNA ITS sequences. New Phytol. 153: 345-357.

http://dx.doi.org/10.1046/j.0028-646X.2001.00313.x

Martin MP, Garcia-Figueres F. 1999. Colletotrichum acutatum and C. gloeosporioides cause

anthracnose on olives. Eur. J. Plant Pathol. 105: 733-741.

Martin MP, Winka K. 2000. Alternative methods of extracting and amplifying dna from lichens.

Lichenol. 32: 189-196. http://dx.doi.org/10.1006/lich.1999.0254

Marx DH, Kenny DS. 1982. Production of ectomycorrhizal fungus inoculum. 131-146, in: NC

Schenck (ed.). Methods and principles of mycorrhizal research. The American Phytopathological

Society, St Paul.

Marx DH, Cordell CE, Kenny DS, Mexal JG, Artman JD, Riffle JW, Molina RJ. 1984. Commercial

vegetative inoculum of Pisolithus tinctorius and inoculation techniques for development of

ectomycorrhizae on bare-rooted tree seedlings. For. Sci. monogr. 25: 1-101.

Moyersoen B, Beever, RE, Martin F. 2003. Genetic diversity of Pisolithus in New Zealand indicates

multiple long-distance dispersal from Australia. New Phytol. 160(3): 569-579.

http://dx.doi.org/10.1046/j.1469-8137.2003.00908.x

Nag DK, Huang HV, Berg DE. 1988. Bidirectional chain-termination nucleotide sequencing:

transposon Tn5seq1 as a mobile source of primer sites. Gene 64:135-145.

Nylander JAA. 2004. Mrmodeltest v2. Program distributed by author. Evolutionary biology centre,

Uppsala University.

Phosri C, Watling R, Martin MP, Whalley AJS. 2004. The genus Astraeus in Thailand. Mycotaxon

89: 453-463.

Phosri C, Martin MP, Sihanonth P, Whalley AJS, Watling R. 2007. Molecular study of the genus

Astraeus. Mycol. Res. 3: 275-286. http://dx.doi.org/10.1016/j.mycres.2007.01.004

Pilat A. 1958. Pisolithus. 575-582, in: Flora CSR B- 1: Gasteromycetes. Nakladatelstvi Ceskoslovenské

Akademie Véd. Prague. 862 p.

Posada D, Crandall KA. 1998. Modeltest: testing the model of DNA substitution. Bioinforma. 14:

817-818. http://dx.doi.org/10.1093/bioinformatics/14.9.817

Rambaut A. 2002. Se-al: sequences alignment editor v2.0al1. Edinburgh: Institute of evolutionary

biology, University of Edinburgh. http://tree.bio.ed.ac.uk/software/

Rauschert S. 1959. Beitrag zur Nomenklatur mitteleuropdischer Gasteromyceten. Zeits. Pilzkunde

25(2): 50-59.

Reddy MS, Singla S, Natarajan K, Senthilarasu G. 2005. Pisolithus indicus, a new species of

ectomycorrhizal fungus associated with Dipterocarps in India. Mycologia 97(4): 838-843.

http://dx.doi.org/10.3852/mycologia.97.4.838

Sowerby J. 1814. Coloured figures of English fungi or mushrooms. Supplement (no. 31). London, UK.

208 ... Phosri & al.

Swofford DL. 2003. PAUP” phylogenetic analysis using parsimony (*and other methods). version

4, Sinauer Associates, Sunderland, MA.

Telleria MT, Duefias M, Melo I, Hallenberg N, Martin MP. 2010. A re-evaluation of Hypochnicium

(Polyporales) based on morphological and molecular characters. Mycologia 102(6): 1426-1436.

http://dx.doi.org/10.3852/09-242

Watling R, Turnbull E, See LS. 1999. Pisolithus aurantioscabrosus Watl. (Pisolithaceae;

Basidiomycota)—an expanded view. Nova Hedwigia 69: 433-437.

White TJ, Bruns T, Taylor J. 1990. Amplification and direct sequencing of fungal rDNA genes for

phylogenetics. 315-322, in: MA Innes et al. (eds). PCR protocols. A guide to methods and

applications. San Diego, California: Academic Press, Inc.

MY COTAXON

http://dx.doi.org/10.5248/120.209

Volume 120, pp. 209-213 April-June 2012

Terriera simplex, a new species of Rhytismatales from China

X1A0-MING Gao! CHUN-TAO ZHENG! & YING-REN LIN”

' School of Life Science & ? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

*CORRESPONDENCE TO: *yingrenlin@yahoo.com

ABSTRACT —A new Terriera species found on fallen leaves of Trachelospermum jasminoides

(Apocynaceae) is described, illustrated, and designated as Terriera simplex. This taxon is

distinguished from its closest relatives by unbranched paraphyses and asci with truncate

or subtruncate apices. The type specimen is deposited in the Reference Collection of Forest

Fungi of Anhui Agricultural University, China (AAUF).

KEY worpDs —taxonomy, Ascomycota, morphology, vine plant

Introduction

The genus Terriera B. Erikss. was established by Eriksson (1970) for T. clado-

phila (Lév.) B. Erikss., belonging to Rhytismataceae (Kirk et al. 2008). Minter

(1996) provided a detailed description of the type species.

Over the years, however, some species which should belong to Terriera have

been mistakenly placed in Clithris (Fr.) Bonord., Dermascia Tehon, Hypoderma

De Not., and (particularly) Lophodermium Chevall. (Johnston 2001). Johnston

(2001), who divided Lophodermium on monocotyledonous plants into groups

A, B, C, and D, regarded Group B species as closer to the type species of Terriera

based on their special ascomatal structure. He therefore transferred 11 species

and 1 variety of Lophodermium Group B to Terriera as new combinations

and published three new species. The author also invalidly published a fourth

species, in that work, which he later validly published as T! samuelsii P.R.

Johnst. (Johnston 2003). In the paper by Ortiz-Garcia et al. (2003), Johnston

recombined Clithris minor Tehon as T: minor (Tehon) PR. Johnst. rDNA-ITS

sequence analyses by Ortiz-Garcia et al. (2003) indicated that Lophodermium

from pine hosts were distally related to Terriera and that two monophyletic

Terriera sequences formed a weakly supported sister group related to Lirula

macrospora (R. Hartig) Darker.

210 ... Gao, Zheng & Lin

Five Terriera species have previously been reported from China: T! brevis

(Berk.) P.R. Johnst., T. huangshanensis Z.Z. Yang et al., T. camelliae (Teng)

Y.R. Lin & Jiang L. Chen, T’ coacervata Y.R. Lin & Q. Zheng, and T: illiciicola

(S.J. Wang et al.) Q. Zheng & Y.R. Lin (Frohlich & Hyde 2000, Yang et al. 2011,

Chen et al. 2012, Zheng et al. 2012). In the present paper, we describe a new

species of Terriera on confederate jasmine, a vine from the Dabie Mountains in

Anhui Province, China.

Materials & methods

External characteristics of conidiomata and ascomata obtained from open ascomata

of the holotype specimen were observed under a dissecting microscope at 10-50x.

After rehydration in water for 10 min, 10-15 um thick sections of the conidiomata

and ascomata were cut using a freezing microtome. Microscopic examinations were

made in water, 5% KOH, Melzer’s reagent, or 0.1% (w/v) cotton blue in lactic acid. For

observing the outlines of ascomata and conidiomata in vertical section, sections were

mounted in lactic acid or cotton blue with pretreatment in water. Gelatinous sheaths

surrounding ascospores and paraphyses were observed in water or cotton blue in lactic

acid. The color of internal structures and ascospore contents were observed in water.

Measurements and drawings of asci, ascospores, and paraphyses (30 per specimen) were

made using material mounted in 5% KOH or Melzer’s reagent. Point and line integrated

illustrations of external shapes and internal structures of the conidiomata and ascomata

were prepared using a microscope drawing device.

Taxonomy

Terriera simplex Y.R. Lin, X.M. Gao & C.T. Zheng, sp. nov. FIGs 1-6

MycoBank MB 563610

Differs from T: cladophila in unbranched paraphyses and asci with truncate or

subtruncate apices and from T. minor by the presence of conidiomata and ascomata

with obtuse, rounded or slightly acute ends.

Type: on fallen leaves of Trachelospermum jasminoides (Lindl.) Lem. (Apocynaceae),

China, Anhui, Dabie Mountains, Tiantangzhai, alt. ca 700 m, 16 September 2005,

S.J. Wang and Y.R. Lin 2020 (Holotype AAUF 68128).

EryMo_oey: simplex, referring to the simple paraphyses, which are not branched near

the apex.

ZONE LINES absent.

Conipi1omarta on both sides of leaves, crowded or sometimes coalescent.

In surface view, conidiomata 130-200 um diam., rounded or subrounded,

black-brown in the centre, more or less concolorous with the substratum surface

elsewhere, slightly raising the leaf surface, discharging spores through a 3-5 um

diam. apical ostiole. In vertical section, conidiomata subepidermal, lenticular.

UPPER WALL very poorly developed, composed of tiny, thin-walled globose cells

1.6—2 um diam. BASAL WALL well developed, 12-17 um thick, consisting of 2-4

(-5) layers of angular cells 2—4.5 um diam. SUBCONIDIOGENOUS LAYER 4.5-8

Terriera simplex sp. nov. (China) ... 211

pa Ky .

os wie dgreleos

OG Fer a TOSS

SA 40 Sy OS

pice ke eaactd

Fics 1-6. Terriera simplex on Trachelospermum jasminoides. 1. A leaf bearing fruit bodies.

2. Conidiomata and ascomata observed under a dissecting microscope. 3. Ascoma in median

vertical section. 4. Portion of ascoma in median vertical section. 5. Paraphyses, asci and ascospores.

6. Conidioma in vertical section.

2D ae: Zheng & Lin

um thick, composed of textura angularis with nearly colorless, thin-walled

cells. CONIDIOGENOUS CELLS 6-11 x 2-3 um, subcylindrical, tapering to the

apex, proliferating sympodially. Conrp1a 2.5—4 x ca 1 um, cylindrical or nearly

elliptical, hyaline, unicellular.

Ascomarta in similar positions to conidiomata on the substratum, crowded

in irregular, light yellow to grayish-white, large bleached areas without

obvious edges, sometimes 2—6 ascomata coalescent. In surface view, ascomata

650-1000 x 350-480 um, elliptical to ovate, ends obtuse, rounded or slightly

acute, black, slightly shiny, with a clearly marked outline, moderately raising

the surface of the leaf, opening by a single longitudinal split which is sometimes

branched in the triangular ascomata. Immature ascomata seen as two parallel

black patches on the substratum surface rather than as a single more or less

elliptical one, with a pale central zone along future line of opening. Lips absent;

split extending 3/4—-4/5 the length of the ascoma. In median vertical section,

ascomata subepidermal, with epidermal cells becoming filled with fungal tissue

as ascomata develop, 120-135 um deep. COVERING STROMA 18-22 um thick

near the opening, black-brown, composed of textura angularis with thick-

walled cells 3-5.5 um diam. Along the edge of the ascoma opening, there is

a flattened, 8-12 um thick extension adjacent to the covering stroma, and

which comprises strongly carbonized tissue with no obvious cellular structure.

EXCIPULUM very poorly developed, arising from the inner layer of the basal

stroma, consisting of hyaline textura porrecta. BASAL STROMA 6-15 um thick,

dark-brown, composed of 2—3(—4) layers of 3-5 um diam., angular, thick-

walled cells. 20-32 um thick textura prismatica with somewhat thin-walled,

nearly colorless to grayish-brown cells exists between the covering stroma and

basal stroma. SUBHYMENIUM moderately developed, 7-12 um thick, consisting

of hyaline textura angularis and intricata. PARAPHYSES 95-110 x 1.2-1.5 um,

filiform, aseptate, not branched, occasionally gradually or abruptly swollen to

2.5-3 um near the apex where sometimes agglutinated, covered in a ca 1 um

thick gelatinous matrix. Asci ripening synchronously, 72-95(-105) x 4.8-5.2

uum, cylindrical, very short-stalked, thin-walled, apex truncate, subtruncate or

obtuse, without circumapical thickening, J-, 8-spored. AscospoREs arranged

in a fascicle, (45—)56-82 x 1-1.2 um, filiform, slightly tapered towards the

rounded base, hyaline, with a gelatinous sheath 0.8-1 um thick.

HOsT SPECIES & DISTRIBUTION: producing conidiomata and ascomata

on fallen leaves of Trachelospermum jasminoides; known only from the type

locality, Anhui, China.

COMMENTS—We place our new species in Terriera based on the presence of

a strongly carbonized extension adjacent to the ascoma opening and textura

prismatica in the corner between the covering and basal stroma and the absence

of lip cells.

Terriera simplex sp. nov. (China) ... 213

Terriera simplex is distinguished from the type species T: cladophila by its

sometimes coalescent and occasionally three-lobed ascomata, paraphyses with

unbranched upper parts, and asci with truncate or subtruncate apices. While

ascomata in T: cladophila are scattered and sometimes circular, the paraphyses

are frequently and irregularly branched and irregularly twisted or swollen at

the apex, and the tips of asci are rather rounded (Minter 1996). Terriera minor,

a widely distributed species, resembles T: simplex but differs in ascomata with

rounded ends that are not associated with conidiomata, paraphyses that branch

2-3 times in the upper 30-40 um, and sequentially maturing asci (Johnston

1988, 1989a,b).

Ascomata of T: simplex are mature on dead leaves. It is not known whether

this fungus occurs in living leaves as a mutualistic endobiont or as a parasite.

This needs to be investigated.

Acknowledgments

We thank the National Natural Science Foundation of China (No. 30870014) and

the Specialized Research Fund for the Doctoral Program of Higher Education of China

(No. 20070364002) for financial support. Thanks are also given to Dr D.W. Minter and

Dr M. Ye for pre-submission reviews leading to the improvement of our manuscript,

and to Dr S.J. Wang for the field investigations.

Literature cited

Chen JL, Lin YR, Hou CL, Wang SJ. 2012 [“2011”]. Species of Rhytismataceae on Camellia spp.

from the Chinese mainland. Mycotaxon 118: 219-230. http://dx.doi.org/10.5248/118.219

Eriksson B. 1970. On Ascomycetes on Diapensales and Ericales in Fennoscandia. Symb. Bot. Upsal.

19: 1-71.

Frohlich J, Hyde KD. 2000. Palm microfungi. Fungal Diversity Press. Hong Kong. 364 p.

Johnston PR. 1988. An undescribed pattern of ascocarp development in some non-coniferous

Lophodermium species. Mycotaxon 31: 383-394.

Johnston PR. 1989a. Lophodermium (Rhytismataceae) on Clusia. Sydowia 41: 170-179.

Johnston PR. 1989b. Rhytismataceae in New Zealand 2. The genus Lophodermium on indigenous

plants. New Zealand J. Bot. 27: 243-274.

Johnston PR. 2001. Monograph of the monocotyledon-inhabiting species of Lophodermium.

Mycol. Pap. 176: 1-239.

Johnston PR. 2003. Validation of Terriera samuelsii. Mycotaxon 87: 1-2.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi,

10" ed. CAB International. Wallingford. 771 p.

Minter DW. 1996. Terriera cladophila. IMI Descr. Fungi & Bact. no. 1296.

Ortiz-Garcia $, Gernandt DS, Stone JK, Johnston PR, Chapela IH, Salas-Lizana R, Alvarez-Buylla

ER. 2003. Phylogenetics of Lophodermium from pines. Mycologia 95: 846-859. http://dx.doi.

org/10.2307/3762013

Yang ZZ, Lin YR, Hou CL. 2011. A new species of Terriera (Rhytismatales, Ascomycota) from

China. Mycotaxon 117: 367-371. http://dx.doi.org/10.5248/117.367

Zheng Q, Lin YR, Yu SM, Chen L. 2012 [“2011”]. Species of Rhytismataceae on Lithocarpus spp.

from Mt Huangshan, China. Mycotaxon 118: 311-323. http://dx.doi.org/10.5248/118.311

MY COTAXON

http://dx.doi.org/10.5248/120.215

Volume 120, pp. 215-221 April-June 2012

The Leptogium juressianum complex in southeastern Brazil

Marcos J. KITAURA* & MARCELO P. MARCELLI?

‘UNESP, Instituto de Biociéncias, Depto de Botanica,

Distrito de Rubido Jr., Caixa Postal 510, Botucatu/SP, CEP 18618-970, Brazil

?Instituto de Botanica, Nucleo de Pesquisa em Micologia,

Caixa Postal 68041, Sdo Paulo/SP, CEP 04045-972, Brazil

*CORRESPONDENCE TO: junjimjk@gmail.com

AsBstTRACcT— Leptogium subjuressianum and L. subjuressianum var. caparoense are new

Brazilian taxa growing mostly in the southeastern mountains. They are differentiated from

European taxa by the ornamentation of the upper surface.

Key worps— lobules, margin, granular isidia

Introduction

Leptogium juressianum is the only species in Leptogium sect. Mallotium

producing isidia-like structures and with hairs confined mostly to the upper

surface (Jorgensen 1997, Tavares 1950). The other Leptogium species with

hairs chiefly on the upper surface — L. eriodermoides Arv. & P.M. Jorg. and

L. resupinans Nyl. Jorgensen 1997) — do not have ornamented thalli.

Such hairs, initially observed by Tavares (1950), are commonly considered

an important taxonomic characteristic and have a specific organization.

However, the organization is rarely described in detail and sometimes not even

mentioned (Swinscow & Krog 1988, Galloway & Jorgensen 1995).

Jorgensen & James (1983) described the hairs of L. juressianum as an

arachnoid tomentum while Jorgensen (1997) called them cobwebby hairs.

Tavares (1950) described the holotype from Portugal as producing isidiate or

lacinulate propagules, but later specimens with alternative propagules were

also identified as L. juressianum: cylindrical to complanate isidia and lacinulae

were noted for Ethiopian (Africa) collections (Swinscow & Krog 1988), western

European specimens produced granular to sublobulate isidia Jorgensen &

James 1983), and the upper surface of specimens from Chile were found to have

granular, sublobulate, or lacinulate isidia and isidiate-lacerate lobe margins

(Galloway & Jorgensen 1995).

216 ... Kitaura & Marcelli

Leptogium specimens collected during the first author's doctoral studies,

were initially identified as L. juressianum. Upon type revision and under our

descriptive protocol we discovered that the material represents new taxa, which

are described here, together with comments on L. juressianum.

Material & methods

This study is mainly based on material collected in the Serra da Mantiqueira, a major

mountain complex in southeastern Brazil, as well as on the isotype of L. juressianum

(UPS). We were unable to study the holotype because the Museu Nacional de Historia

Natural (LISU) in Lisboa, Portugal, did not respond to a loan request.

Comparative morphological and anatomical characteristics summarized in TABLE 1

are modeled after a table first assembled by Cunha (2007), who studied the Leptogium

species collected in forests (mainly mangroves) and urban littoral habitats of Sao Paulo

state. The character set has been modified to encompass the morphological variations

we observed in the Serra da Mantiqueira taxa.

The nature of the thalline surfaces and their ornamentation are considered important

taxonomic characters. The surfaces are described without and with magnification; the

system of folds and/or wrinkles and their size is determined by complex organization

of the hyphae (both cortical and internal), which remain constant in both dry and wet

specimens. Some folds are readily seen with the naked eye, but smaller folds (< 0.1 mm,

which appear smooth to the naked eye but can readily be seen under magnification =

10x) must be observed under the stereomicroscope.

Several species apparently bear similar propagules on the upper surface, but careful

observation has shown that their morphology and position are good species characters.

Thus descriptions of their form, dimensions, distribution, and ontogeny are described

in TABLE 1.

TABLE 1. Morphological comparison of Leptogium juressianum, L. subjuressianum,

and L. subjuressianum var. caparoense.

TAXON ORNAMENTATION ORNAMENT HAIR Dry THALLUS

DISTRIBUTION DISTRIBUTION COLOR*

L. juressianum Cylindrical Usually on Upper surface; Gray to bluish

when young to lobe margins parts of where glabrous

flattened when underside

fully developed

L. subjuressianum

var. subjuressianum

L. subjuressianum

var. caparoense

Granular isidia

and lobules

Granular isidia

* All whitish where hairy

Usually on

lobe margins

Abundant on

lamina; frequent

on lobe margins

Upper surface

and usually

on underside

margin

Frequent on

upper surface;

abundant on

underside

Brownish where

glabrous

Dark brown to

blackish where

glabrous

Leptogium subjuressianum sp. nov. (Brazil) ... 217

All specimens analyzed were dissected under a stereomicroscope with a razor blade

and the semi-permanent slides were mounted with solution of 50% glycerine.

Results & discussion

The name Leptogium juressianum has been attributed to all ornamented

(bearing isidia-like or lobule-like structures) specimens of Leptogium that

develop hairs mostly on the upper surface (Tavares 1950, Jorgensen &

James 1983, Jorgensen & Nash 2004). The detailed descriptions of thallus

and ornamentation produced according to our protocols have enabled us to

distinguish the Brazilian taxa. This was not possible using brief descriptions and

the characters commonly used to describe and differentiate Leptogium species.

Thallus color, hair distribution, and the nature of the vegetative propagules are

the principal diagnostic characters in this paper (TABLE 1).

Leptogium juressianum Tav., Port. Acta Biol., Ser. B, 3: 68, 1950. FIGs 1-2

Type: PORTUGAL, Minho [corresponding with the modern districts of Braga and

Viana do Castelo], Serra do Géres, between Pedra Bela and the Caldas, 700 m alt., on

Arbutus unedo, 16.V1.1947, leg. C. Tavares 2081 (holotype, LISU; isotype, UPS!).

THALLUS gray, Opaque, matt, gray to whitish when observed under the

stereomicroscope. Loses irregularly lacerate, to 3 mm wide, overlapping,

attached in points, convolute, upper surface smooth and velvety under the

naked eye, pubescent at 10x and higher magnifications (interlaced hairs that

are similar to sponge fibers); apices rounded, usually ascending to involute,

smooth or isidiate to lacinulate; lateral margin fimbriate (isidiate to lacinulate),

ascending, irregular; lower side bluish gray, smooth at different magnifications.

Istp1A absent. LACINULES smooth, cylindrical when young to flattened, ca.

0.08-0.30 x 0.03-0.08 mm, unbranched to irregularly branched but not coralloid,

erect, firm, concolorous with the thallus, usually marginal, dense, frequent to

abundant, grouped on the lamina. ATTACHED by hairs; rhizines absent; hapters

absent; hairs irregular, unbranched or branched, up to 125 um (ca. 9 cells) long,

dense on upper surface, frequent on lower side. APOTHECIA absent.

ANATOMY— THALLUS ca. 60 um thick, quadratic cells of the cortices ca.

7.5 x 5.0 um; columnar hyphae ca. 2.5 um thick, straight to inclined, 2(-3)

cells. CYANOBACTERIA blue, frequent to abundant, cell number not appraised,

spherical, 5 um diam.; gelatinous matrix poor, hyaline. Pycnip1a absent.

CoMMENTS— Leptogium juressianum is characterized by the isidiate to

lacinulate margins and hairs on the upper surface.

Originally described from Portugal, the species has been cited for western

Europe (Jorgensen & James 1983), southern Africa (Swinscow & Krog 1988),

and Chile, South America (Galloway & Jorgensen 1995), but always with

different type and/or position of propagules. Since the propagules are major

218 ... Kitaura & Marcelli

diagnostic characters, we believe the previously cited collections may represent

different taxa and that all that material needs re-evaluation

Leptogium subjuressianum Marcelli & Kitaura, sp. nov. Fics 3-4

MycoBAank 563829

Differs from Leptogium juressianum by granular isidia and presence of rounded lobules.

TyPE: Brazil, Rio Grande do Sul State, municipality of Tapes, on trunk of a tree, 5 m alt,

29.1.1994, leg. M.P. Marcelli 26459 (holotype, SP).

ErymMo oey: The specific epithet refers to the closely related species, L. juressianum, the

name first applied to the studied material.

THALLuS whitish or black (naked eye), opaque, matt, whitish brown when

observed under the stereomicroscope. LoBEs to 2 mm wide, irregularly

overlapping, attached in points, ascending; upper surface velvety when observed

with the naked eye, pubescent at 10x and higher magnifications; apices rounded,

ascending, involute to convolute, ornamented with small granular to lobulate

isidia; lateral margins lobulate, ascending or flat, undulate; lower side bluish

gray, smooth, covered by long hairs seen at 10x and higher magnification.

Isip1a granular, ca. 0.05 mm diam., unbranched, erect, firm, concolorous with

the thallus, rare on the lamina but dense at the margins. LoBuLEs with apices

rounded, ca. 0.5 mm diam., unbranched, erect, firm, concolorous with the

thallus, marginal, dense, abundant. ATTACHED by rhizines and hairs; hapters

absent; rhizines unbranched, beige, evenly distributed, frequent, formed from

agglutinated hairs; hairs interlaced, unbranched or branched, < 125 um (ca. 8

cylindrical cells) long, beige, present on both surfaces, abundant on the upper

surface and frequent on the lower surface. APOTHECIA absent.

ANATOMY— THALLUS ca. 55 um thick, upper quadratic cells ca. 10 x 10

um, lower quadratic cells ca. 5 x 5 um, columnar hyphae ca. 2.5 um thick,

straight and inclined, comprising 3-4 cells. CYANOBACTERIA blue, frequent,

cell number not appraised, cells spherical, ca. 5 um diam.; gelatin frequent,

colorless. PycNIDIA absent.

ADDITIONAL MATERIAL EXAMINED: BRAZIL. PARANA STATE, General Carneiro

Municipality, Fazenda Lageado Grande, s/d, S. Eliasaro 2892 (UPCB); SAo PAULO

STATE, Campos do Jordao Municipality, Parque Estadual de Campos do Jordao,

Araucaria/Podocarpus forest, on tree trunk inside the forest, 1400 m alt., 17.V1.1995, M.P.

Marcelli, A. Gugliotta & R. Maziero 28914 (SP); MINAS GERAIS STATE, Camanducaia

Municipality, on trunk, 21.X1.2008, M.J. Kitaura, M.P. Marcelli, A.E. Luchi & S.N. Inoue

1158 (SP), 1179 (SP); 22.X1.2008, M.J. Kitaura, M.P. Marcelli, A.E. Luchi & S.N. Inoue

1232 (SP), 1235 (H); Itamonte Municipality, BR-485, on trunk, 22.1.2009, M.J. Kitaura

& M.P. Marcelli 1468 (SP); Brejo da Lapa, on trunk, 23.1.2009, M.J. Kitaura & M.P.

Marcelli 1536 (SP); Espigtro SANTO STATE, Dores do Rio Preto Municipality, on trunk,

16.1X.2009, M.J. Kitaura & M.P. Marcelli 1792 (SP), 1804 (SP), 1825 (BOTU); on branch

fallen on the rock, 16.1X.2009, M.J. Kitaura & M.P. Marcelli 1911 (SP).

Leptogium subjuressianum sp. nov. (Brazil) ... 219

FiGuRES 1-6. Leptogium juressianum (isotype, UPS): 1. Thallus; 2. Propagules. L. subjuressianum

var. subjuressianum (holotype): 3. Thallus; 4. Propagules. L. subjuressianum var. caparoense

(holotype): 5. Thallus; 6. Propagules. Scale bars: 1, 3, 5 = 5 mm; 2, 4, 6 = 20 um.

220 ... Kitaura & Marcelli

ComMMENTS— Leptogium subjuressianum is characterized by the rounded

lobules on the thallus margin and spongioid hairs on the upper surface, which

usually appears whitish due to the presence of dense hairs. Both L. juressianum

and L. subjuressianum are covered by a dense layer of spongioid hairs, which

are interlaced and resemble fibers of sponges.

According to Tavares (1950), L. juressianum possesses only isidia and

lacinules on the margins. No rounded lobules were observed in any part of

the isotype. We did not find lacinules in any specimens of L. subjuressianum,

where only granular to lobulate isidia are present prior to the development of

lobules.

Leptogium subjuressianum is common in Brazil and is usually collected on

tree trunks. Specimens were collected from Tapes Municipality, Rio Grande

do Sul State (30°40'12"S 51°23'24"W) to Alto Caparaé Municipality in Minas

Gerais State (20°29'42"S 41°59'54"W).

Leptogium subjuressianum var. caparoense Kitaura & Marcelli, var. nov.

MycoBAnkK 563862 FIGs 5-6

Differs from Leptogium subjuressianum var. subjuressianum by the isidia distribution

and the presence of rhizomorphic hyphae on the lower surface.

Type: Brazil, Minas Gerais State, Municipality of Alto Caparad, Cachoeira Bonita,

on organic material on rock beside the waterfall, 17.1X.2009, leg. M.J. Kitaura & M.P.

Marcelli 1900 (holotype, SP; isotypes, H).

Erymo.oey: ‘The varietal epithet refers to the Caparad Range, a major mountain

complex in southeastern Brazil where the types were collected.

THALLUS whitish to blackish (naked eye), opaque, matt, whitish (hairy part) and

dark brown or black (glabrous part) when observed under the stereomicroscope.

Loses to 3 mm wide, irregular, overlapping, attached in points, ascending,

upper surface smooth or velvety when observed with the naked eye, pubescent,

smooth or ornamented at 10x and higher magnifications; apices rounded,

ascending or involute, usually smooth; lateral margins smooth to granular,

ascending, undulate; lower side usually brownish, with adhering substrata when

observed with the naked eye, smooth or less pubescent that upper surface at 10x

and higher magnifications. Isrp1a granular, ca. 0.1 mm diam., concolorous with

the thallus, firm, dense or abundant on the lamina and frequent at the margins.

LOBULES rounded, ca. 0.5 mm diam., unbranched, erect, firm, concolorous

with the thallus, marginal when the thallus is covered by hairs, frequent or

rare on the lamina. ATTACHED by rhizines and hairs; hapters absent; rhizines

unbranched, beige, evenly distributed, frequent, constituted from agglutinated

hairs; hairs interlaced, irregular, beige, dense on upper surface and frequent on

lower surface, cells cylindrical. APOTHECIA absent.

ANATOMY— THALLUS < 70 um thick, upper quadratic cells (wall slightly

blackish) ca. 10 x 10 um, lower quadratic cells ca. 7.5 x 7.5 um; columnar hyphae

Leptogium subjuressianum sp. nov. (Brazil) ... 221

ca. 2.5 um thick, straight to inclined, 4-celled. CYANOBACTERIA yellow to slight

blue, frequent, cells spherical, ca. 5 um diam.; gelatin frequent to scarce, yellow

next to upper cortex and colorless below. Pycnip1A absent.

ADDITIONAL MATERIAL EXAMINED: BRAZIL, Minas GERAIS STATE, Alto Caparad

Municipality, Cachoeira Bonita, saxicolous, 17.1X.2009, M.J. Kitaura & M.P. Marcelli

1902 (H), 1905 (SP); on the moss on the rock, 17.[X.2009, M.J. Kitaura & M.P. Marcelli

1904 (SP), 1932 (BOTU).

ComMMENTS— Leptogium subjuressianum var. caparoense is characterized by

granular isidia on both the lamina and margins (older parts) and the blackish

or dark brown colored thallus (when not covered by spongioid hairs).

The lobules common on the margins of L. subjuressianum var. subjuressianum

were not observed in L. subjuressianum var. caparoense (TABLE 1).

Furthermore, the isidia in L. subjuressianum var. caparoense are mainly

laminal but primarily marginal in L. subjuressianum var. subjuressianum. Also,

L. subjuressianum var. caparoense has rhizomorphic hyphae covering the entire

lower surface whereas in var. subjuressianum they are present mainly at the

margins of the lobes.

All specimens of L. subjuressianum var. caparoense were collected in an

especially humid place on wet rocks beside a waterfall inside Caparaé National

Park. We attribute to this the abundance of rhizomorphic hyphae and spreading

of the propagules and hairs and prefer to retain the taxon at the varietal level.

Acknowledgments

The authors wish to thank Dr. J.A. Elix and Dr. R. Liicking for the critical revision of

the manuscript and valuable suggestions. M. Kitaura is grateful to FAPESP (2008/51072-

3) for a PhD grant and M.P. Marcelli to CNPq for a research grant.

Literature cited

Cunha IPR. 2007. Fungos liquenizados do género Leptogium (Ascomycetes) no litoral sul do Estado

de Sao Paulo. UNESP, Instituto de Biociéncias (Master Thesis), Botucatu. 101 p.

Galloway DJ, Jorgensen PM. 1995. The lichen genus Leptogium (Collemataceae) in southern

Chile, South America. 227-247, in: FJA Daniels et al. (eds). Flechten Follmann. Contribution

to Lichenology in Honour of Gerhard Follmann. Geobotanical and Phytotaxonomical Study

Group, Botanical Institute, University of Cologne, Germany.

Jorgensen PM. 1997. Further notes on hairy Leptogium species. Symbolae Botanicae Upsalienses

32(1): 113-130.

Jorgensen PM, James PW. 1983. Studies on some Leptogium species of western Europe. Lichenologist

132109125,

Jorgensen PM, Nash III TH, 2004. Leptogium. 330-350, in: TH Nash III et al. (eds). Lichen Flora

of the Great Sonoran Desert Region. Vol. 2, Lichens Unlimited, Arizona State University,

Arizona.

Swinscow TDV, Krog H. 1988. Macrolichens of East Africa. British Museum (Natural History).

London.

Tavares CN. 1950. Liquenes da Serra do Géres. Portugaliae Acta Biologica, Ser. B, 3: 1-189.

MY COTAXON

http://dx.doi.org/10.5248/120.223

Volume 120, pp. 223-230 April-June 2012

Coriolopsis psila comb. nov. (Agaricomycetes) and

two new Coriolopsis records for Brazil

GEORGEA S. NOGUEIRA-MELO’, PRISCILA S. DE MEDEIROS’;

ALLYNE C. GoMES-SILVA’, LEIF RYVARDEN?, HELEN M.P. SOTAO?

& TATIANA B. GIBERTONI’

* Universidade Federal de Pernambuco, Departamento de Micologia

Av. Nelson Chaves s/n, CEP 50760-420, Recife, PE, Brazil

*Museu Paraense Emilio Goeldi, Coordenagao de Botanica,

Caixa Postal 399, CEP 66040170, Belém, PA, Brazil

>University of Oslo, Department of Botany, P. O. Box 1045, Blindern, N-0316, Oslo, Norway

*CORRESPONDENCE TO: georgeacomea@hotmail.com

AsBstTRACT — The new combination Coriolopsis psila is proposed and C. brunneoleuca and

C. hostmannii are reported as new to Brazil. Descriptions of these collections and a key to the

eight accepted Coriolopsis species reported from Brazil are provided.

Key worps — Polyporaceae, diversity, C. byrsina, C. caperata, C. floccosa

Introduction

Coriolopsis Murrill (Murrill 1905) comprises about 17 valid species (Kirk et al.

2008). The genus is cosmopolitan, although several species are restricted to the

temperate or tropical zones (Ryvarden & Johansen 1980, Ryvarden & Gilbertson

1993, Nunez & Ryvarden 2001, Dai 2011, Dai et al. 2011). It is characterized by

the annual (rarely perennial) pileate or sessile (rarely resupinate) basidiomata

with great color variation. The hyphal system is trimitic with clamp connections

on generative hyphae, cystidia are absent, and basidiospores are cylindrical,

smooth, thin-walled, non-amyloid, oblong—ellipsoid, and hyaline to pale brown

(Ryvarden & Johansen 1980, Ryvarden 1991, Ryvarden & Gilbertson 1993).

Only seven Coriolopsis species have previously been reported from Brazil:

C. aspera (Jungh.) Teng, C. badia (Berk.) Murrill, C. burchellii (Berk. ex Cooke)

Ryvarden, C. byrsina, C. caperata, C. floccosa, and C. gallica (Fr.) Ryvarden

(Ryvarden 1988, Gugliotta & Abrahao 2011).

To increase the knowledge of polypore diversity in Brazil, we record two

additional species, propose a new combination, and provide identification keys

to all known Brazilian species.

224 ... Nogueira-Melo & al.

Material & methods

In the Brazilian Amazonia, field trips were undertaken in Para State from July 2006

to February 2008 in the Floresta Nacional de Caxiuana (1°42'3"S 51°31'45"W) and in

Rondonia State from 2007 to 2008 in the Estagao Ecoldgica de Cunia (8°04'S 63°31’ W)

and Parque Natural Municipal de Porto Velho (8°45'S 63°54’W). In Pernambuco

State, Brazilian mangroves were surveyed from March 2009 to March 2010 in Maria

Farinha (7°51'24.8"S 34°50'32.7"W), Itamaraca (7°46'52.6"S 34°52'53.3"W), Maracaipe

(8°32'22.8"S 35°00'29.1” W), and Rio Formoso (8°41'20.8"S 35°06'06.6" W).

Both recent collections and specimens deposited in INPA, O, RB, and SP were

studied. Specimens were analyzed macro- (shape, color, hymenial surface) and

micromorphologically (hyphal system, presence/absence and measurements of sterile

structures and basidiospores). Slides were prepared with 5% KOH (stained with 1%

aqueous phloxine) or Melzer’s reagent (Ryvarden 1991). Color designation followed

Watling (1969). The material was incorporated to HFSL, MG, O, and URM.

Results & discussion

After identification of new collections and revision of herbaria, eight

Coriolopsis species are reported for Brazil. The presence of C. gallica is not

confirmed, as previous reports actually represent Hexagonia hydnoides (Sw.)

M. Fidalgo (Tavares 1939, URM 608/IPA 281) or Trametes sp. (Bononi et al.

1984, SP 156751). Coriolopsis badia is a dubious species because it is considered

a synonym of either Phellinus badius (Cooke) G. Cunn. (http://www.cbs.knaw.

nl, http://www.mycobank.org) or C. aspera (Ryvarden & Johansen, 1980). The

only record of C. badia in Brazil (Gibertoni et al. 2004, URM 77849) represents

a Trametes sp.

Coriolopsis brunneoleuca and C. hostmannii are new occurrences for the

country. Coriolopsis psila, here transferred from Fomes, is probably a new

occurrence for the Brazilian Amazonia. Coriolopsis byrsina represents a new

record for the states of Amazonas, Rondénia, and Roraima, C. caperata for

Amazonas, and C. floccosa for Acre, Amazonas, and Mato Grosso. These new

occurrences were studied also from collections deposited in INPA, RB and

SP, underscoring the importance of herbarium revisions and accessibility of

herbaria records to the knowledge of fungal diversity.

Coriolopsis brunneoleuca (Berk.) Ryvarden, Norw. Jl Bot. 19: 230 (1972).

BASIDIOMATA annual, effused-reflexed, gregarious, coriaceous, < 5.5 cm,

1.5 cm wide and to 1 mm thick. ABHYMENIAL SURFACE tomentose, dull, cigar

brown (16) to fuscous black (36) with concentric zones of dark color. MARGIN

entire, acute, concolorous with the abhymenial surface. Context homogeneous,

fibrous, < 100 um thick, milky coffee (28) to cinnamon (10). HYMENIAL SURFACE

with pores angular, 2-3 per mm, dissepiments 50-100 um thick, snuff brown

(17) to clay buff (32). HyPHAL sysTEM trimitic; generative hyphae hyaline,

branched, clamped, thin-walled, 1.5-2.5 um diam.; skeletal hyphae yellowish to

Coriolopsis in Brazil... 225

pale brown, thick-walled, 4-7.5 um diam., dextrinoid; binding hyphae golden-

yellow, almost solid, 3-5 um diam. Cystip1a absent. Basidia not observed.

Basipiosporss cylindrical, hyaline, thin-walled, smooth, inamyloid, 7-11 x

3-4 um.

EcoLoGy & DIsTRIBUTION: On deciduous wood. According to Ryvarden & Johansen

(1980), the species is pantropical. It is new to Brazil.

SPECIMENS EXAMINED: BRAZIL. PaRA, Floresta Nacional de Caxiuana, VIII.2007, leg.

PS Medeiros et al. PS122, PS226, PS342 (MG195152, MG195155, MG195182).

REMARKS: A cigar brown to fuscous black pileus surface, rather large shallow

pores, and broad dextrinoid skeletal hyphae characterize the species.

Coriolopsis byrsina (Mont.) Ryvarden, Norw. Jl Bot. 19: 230 (1972).

DEscrIPTION: This species is characterized by small pores, soft rusty-brown

basidiomata, and ellipsoid to sub-cylindrical basidiospores, 9-14 x 4.5-5 um.

For further details see Ryvarden & Johansen (1980).

EcoLoGy & DISTRIBUTION: On deciduous wood. In tropical Africa, America, and Asia

(Ryvarden & Johansen 1980; Dai 2011). In Brazil, previously known from the states of

Acre, Mato Grosso, Rio Grande do Sul, and Sao Paulo (Gugliotta & Abrahao 2011);

newly reported from Amazonas, Ronddénia, and Roraima.

SPECIMENS EXAMINED: BRAZIL. Acre: Rio Branco, 24.1X.1980, leg. B. Lowy et al.

241 (INPA 183806); AMAZONAS: Presidente Figueiredo, 18.1.1983, leg. M.A Jesus 123

(INPA 183806); Maro Grosso: Aripuana, 23.IV.1978, leg. M.A. Sousa 427 (INPA

75606); RONDONIA: Jaru, Reserva Bioldgica de Jaru, 13.V.1987, leg. M. Capelari et al.

(SP 211995); location not determined, 4.VII.1968, leg. K.P. Dumont et al. 143 (INPA

65179); 3.111984, leg. GJ. Samuels 55 (INPA 129095); RorAIMA: locality unknown,

1.XII.1977, leg. I.J. Aratijo et al. 757 (INPA 78476); SAo PAuLo: Itaicy, 06.VII.1957, leg.

not determined (URM 7880, as Polystictus byrsinus).

Coriolopsis caperata (Berk.) Murrill, N. Amer. Fl. 9(2): 77 (1908).

DESCRIPTION: Coriolopsis caperata is characterized by effused-reflexed to

pileate darkly colored basidiomata and a zonate abhymenial surface in different

shades of brown. For further details see Ryvarden & Johansen (1980).

ECOLoGy & DISTRIBUTION: On deciduous wood. Tropical Africa and America (Ryvarden

& Johansen 1980). In Brazil, it is reported for the states of Acre, Alagoas, Amapa,

Bahia, Espirito Santo, Mato Grosso, Minas Gerais, Para, Paraiba, Parana, Pernambuco,

Rio Grande do Norte, Rond6énia, Roraima, Rio de Janeiro, Rio Grande do Sul, Santa

Catarina, Sao Paulo, Sergipe (Baltazar & Gibertoni 2009, Gugliotta & Abrahao 2011),

and now for Amazonas State.

SPECIMENS EXAMINED: BRAZIL: Acre: Sena Madureira, 27.1X.1968, leg. G. T. Prance

et al. 7595 (INPA 24394); AMAZONAS: Barcelos, II.1984, leg. G. J. Samuels 72 (INPA

129121); Borba, 6.V.1985, leg. K.F. Rodrigues et al. 490 (INPA 129022, as Coriolus

versicolor); Manaus, 1.V.1977, leg. M. A. Souza 149 (INPA 74657, as Coriolopsis polyzona);

6.X.1985, leg. K.E. Rodrigues et al. 800 (INPA 137086); 5.X.1927, leg. P. Occhioni (SP

25530); 23.XII.1983, leg. G. Guzman et al. (SP 193577); Manicoré, IV.1985, leg. K.F.

Rodrigues et al. 91 (INPA 128914, as Coriolopsis sp); Nova Aripuana, 29.IV.1985, leg.

226 ... Nogueira-Melo & al.

K.F. Rodrigues et al. 410, 389 (INPA 129009, as C. polyzona, INPA 129005, as Polyporus

sp.); Tapuruquara, 22.1.1978, leg. I. J. Araujo et al. 973 (INPA 78745, as Hexagonia

sp.); Tefé, 11. VII.1973, leg. E. Lleras et al. 16611 (INPA 39800); Cachoeira de Taruma,

5.X.1927, leg. P. Occhioni (RB 217241); Serra de Araga, 29.11.1984, leg. G. J. Samuels

(RB 24785); Manicoré, Estrada do Estanho, 20.IV.1985, leg. K.F Rodrigues (RB 238166);

Novo Aripuana, Vila do Apuy, 29.IV.1985, leg. K.F. Rodrigues et al. (RB 238334); Distrito

Agropecuario da Suframa, 03.X.1985, leg. N. Rodrigues et al. (RB 237012, RB 240587);

PaRA: Belém, 13.IX.1968, leg. B. Santiago (SP 106707); 3.VI.1980, leg. V. L. Bononi (SP

177467); Melgaco, VIII.2007, leg. T. B. Gibertoni (URM 79656 on Xylopia sp; URM

79653 on Pouteria sp.; URM 79650 on Nectandra sp.; URM 79649 on Licania sp.; URM

79646 on Parkia sp.; URM 79647 on Sclerolobium sp.; URM 79648 on Eschweilera sp.;

URM 79652 on Dinizia excelsa); Redengao, 20.VIII.1984, leg. N. Rodrigues (RB 224462);

Oriximina, 28.VI.1980, leg. V.L. R. Bononi et al. 638, 241 (INPA 103609, as Coriolus sp.,

INPA 103335, as Trametes pinsita); Itaituba, [X.1977, leg. M.A. Sousa et al. 25 (INPA

74631, as Hexagonia caperata); PERNAMBUCO: Escada, 26.X.1954, leg. not determined

(URM 1010, as Polystictus occidentalis); Recife, 24.V1I.1957, leg. not determined (URM

12468, as P. caperatus); Tapera, 1932, leg. not determined (URM 762, as P. caperatus);

RONDONIA: Caracarai, 3.VII.1968, leg. G.T. Prance et al. 5497 (INPA 22257); Campo

Novo, X.1979, leg. R.H. Petersen 155 (INPA 110517, as Polyporus sp.); Jaru, 2.X.1986,

leg. M. Capelari & R. Maziero (SP 211273); Vilhena, X.1979, leg. R. H. Petersen 218

(INPA 110706, as Polyporus sp.); Porto Velho, Estacao Ecoldgica de Cunia, I1.2007, leg.

A. C. Gomes-Silva 33 (URM 78898); II.2008, leg. A. C. Gomes-Silva 222 (URM 78906);

11.11.2010, leg. A.C. Gomes-Silva et al. 815 (URM 82837); Parque Natural Municipal

de Porto Velho, VII.2007, leg. A. C. Gomes-Silva 59 (URM 78900); VII.2008, leg. A.C.

Gomes-Silva 603 (URM 79552); 20.V1.2009, leg. A.C. Gomes-Silva et al. 738 (URM

82833); 29.1.2010, leg. A.C. Gomes-Silva et al. 941 (URM 82831); 3.11.2011, leg. A.C.

Gomes-Silva et al. 1089 (URM 82903); Fazenda Mucuim, VII.2007, leg. A.C. Gomes-

Silva 91 (URM 78901); RoraimMa: Alto Alegre, 21.V1.1986, leg. K.E Rodrigues et al.

1083 (INPA 143425); Caracarai, 16.X1.1977, leg. I .J. Araujo et al. 438 (INPA 76933, as

Hexagona sp.).

Coriolopsis floccosa (Jungh.) Ryvarden, Norw. Jl Bot. 19(3-4): 230 (1972).

DESCRIPTION: The umber brown tomentose to hirsute or even velutine

pileus surface and fairly large pores are diagnostic for this species. For further

details see Ryvarden & Johansen (1980).

EcoLoGy & DISTRIBUTION: On deciduous wood. Pantropical (Ryvarden & Johansen

1980). In Brazil, reported for the states of Alagoas, Bahia, Para, Paraiba, Parana,

Pernambuco, Rio Grande do Sul, Rondénia, Roraima, Santa Catarina and Sao Paulo

(Gugliotta & Abrahao 2011). Newly recorded for Acre, Amazonas, and Mato Grosso.

SPECIMENS EXAMINED: BRAZIL: Acre: location not determined, 3.X.1980, leg. B.

Lowy et al. 1069 (INPA 100886, as Polyporus sp.); AMAZONAS: Manaus, 10.X.1989,

leg. R. Carvalho 1332 (INPA 192503, as Coriolopsis sp.); 21.V1.1985, leg. M. A. Jesus

755 (INPA 185984); 10.[X.1977, leg. M. A. Sousa & I. J. Aratjo 219 (INPA 74661, as

Coriolus maximus); Itacoatiara, 21.XII.1966, leg. G. T. Prance et al. 3778 (INPA 19403,

as Coriolopsis sp.); 10.1X.1980, leg. B. Lowy et al. 129 (INPA 100040, as Polyporus sp.);

Bauia: Conde, 08.11.1955, leg. A.C. Batista (URM 1242, as Polystictus pinsitus); Amaro,

23.V.1957, leg. not determined (URM 7450, as Polystictus membranaceus); MATo

Coriolopsis in Brazil ... 227

Grosso: Aripuana, 23.IV.1978, leg. M.A. Sousa et al. 431 (INPA 75610); PARA: Belém,

17.1V.1979, leg. I.J. Araujo 1239 (INPA 102851); Itaituba, 3.X.1977, leg. M.A. Sousa et

al. 140 (INPA 74538, as Coriolopsis sp.); PERNAMBUCO: Recife, 23.VII.1955, leg. not

determined (URM 12498, as P. membranaceus); RIO GRANDE DO SUL: Sao Leopoldo,

1932, leg. J. Rick (URM 9023, as Polystictus rigens); RORAIMA: Alto Alegre, 16.V1.1986,

leg. E.S.S. Silva et al. 493 (INPA 154958); Boa Vista, 19.VII.1989, leg. M.A. Jesus 880

(INPA 186185, as Coriolopsis sp.); 1.X1I.1977, leg. L.L.J. Aguiar et al. 750 (INPA 78469,

as Coriolopsis sp.); 31.1.1984, leg. G.J. Samuels 39 (INPA 129089); RONDONIA: Porto

Velho, Estacao Ecolégica de Cunia, VII.2007, leg. A.C. Gomes-Silva 204 (URM 78909);

Parque Natural Municipal de Porto Velho, VII.2007, leg. A.C. Gomes-Silva 186 (URM

79477, as Coriolopsis rigida); Fazenda Mucuim, VII.2007, leg. A.C. Gomes-Silva 258

(URM 79478, as C. rigida).

Coriolopsis hostmannii (Berk.) Ryvarden, Syn. Fung. 23: 39 (2007).

BASIDIOMATA perennial, sessile, hard and brittle when dry, dimidiate to

flabelliform, solitary, projecting 2-4 cm, 3-5 cm wide and 0.5-1.5 mm thick.

ABHYMENIAL SURFACE glabrous, dull to subshiny, zonate with brown and purple

zones, hazel (27) to drab (33). MARGIN acute, concolorous with abhymenial

surface. CONTEXT homogeneous, < 0.5 mm thick, fulvous (12) to cigar brown

(16). Tubes concolorous with context, < 0.5 mm long. HYMENIAL SURFACE with

pores round, 6-7 per mm, dissepiments 50-100 um thick, cigar brown (16).

HYPHAL SYSTEM trimitic, generative hyphae hyaline, clamped, thin-walled,

1.5-2.5 um diam.; skeletal hyphae yellowish to pale brown, thick-walled, 4-7.5

um diam.; binding hyphae golden-yellow, almost solid, 3-5 um diam. CysTIDIA

absent. BAsIp1A not observed. Bastp1osPorEs cylindrical, hyaline, thin-walled,

inamyloid, 8-10 x 2.5-4.0 um.

EcoLoGy & DISTRIBUTION: On deciduous wood. Neotropical, from Southern United

States and south to Venezuela (Ryvarden & Johansen 1980). It is new to Brazil.

SPECIMENS EXAMINED: BRAZIL. PERNAMBUCO: Paulista, Manguezal de Maria Farinha,

on dead Rhizophora mangle, 20.VII.2009, leg. G.S. Nogueira-Melo et al. NM003 (URM

82147); on dead Avicennia schaueriana, 31.1.2010, leg. G.S. Nogueira-Melo et al. NM002

(URM 82146); Itamaraca, Manguezal do Canal de Santa Cruz, on live A. schaueriana,

25.V1.2009, leg. G. S. Nogueira-Melo et al. NM002, NM025 (URM 82134, URM 82138);

on dead A. schaueriana, 10.III.2009, leg. G.S. Nogueira-Melo et al. NM001, NM004

(URM 82136, 82145); 26.VI.2009, leg. G.S. Nogueira-Melo et al. NM003, NMO07,

NMO011 (URM 82144, URM 82143, 82137); 26.VII.2009, leg. G.S. Nogueira-Melo et

al. NM002 (URM 82139); 22. VIII.2009, leg. G.S. Nogueira-Melo et al. NM006, NM012

(URM 82141, URM 82140); 17.1.2010, leg. G.S. Nogueira-Melo et al. NM002 (URM

82135); 19.1I1.2010, leg. G.S. Nogueira-Melo et al. NM004 (URM 82142).

REMARKS: Coriolopsis hostmannii may be confused with C. aspera, which has

similar hyphae and basidiospores. However, C. hostmannii has smaller pores

(3-4 pores per mm in C. aspera) and smooth pilei. The zonate abhymenial

surface may cause the species to be mistaken for Funalia polyzona (Pers.)

Niemela, although Funalia usually has a distinct variably zoned tomentum.

228 ... Nogueira-Melo & al.

Coriolopsis psila (Lloyd) Ryvarden, comb nov.

MycosBank MB561160

= Fomes psila Lloyd, Syn. Fomes: 233 (1915). [BPI!].

= Phylloporia psila (Lloyd) Ryvarden, Norw. Jl Bot. 19: 235 (1972).

BASIDIOMATA annual to perennial, sessile, solitary, sessile, pileus dimidiate

to flabelliform, < 6 cm in diam. and 2 cm thick with the tomentum, the latter

as a dense mat of intertwined hairs in reddish brown colours, < 1 cm thick.

CONTEXT cigar brown (16) to umber (18), < 4 mm thick in lower denser part,

no black zone towards the cottony dense hair surface mat, tubes single layer

or stratified as in the type, individual layer 3-4 mm thick. HYMENIAL SURFACE

rusty-brown; dissepiments entire, relatively thick; pores round, 6-7 per mm,

almost invisible to the naked eye. HYPHAL SYSTEM trimitic, generative hyphae

often collapsed and difficult to observe, hyaline, thin-walled, but always with a

distinct lumen often with secondary adventitious septa, < 8 um in diam., binding

hyphae irregular in outline, frequently branched or with a few long tapering

branches, thick-walled, yellow to light brown, < 6 um in diam. BASIDIOSPORES

cylindrical, 9-12 x 3-4.5 um, hyaline, thin-walled and non-amyloid.

EcoLocy & DISTRIBUTION: On deciduous wood. Known from Brazil and Mexico. The

type locality in Brazil is not detailed but J. Rick, who collected the material, used to work

in Rio Grande do Sul State, South Brazil. If so, it is probably a new occurrence for the

Brazilian Amazonia and Northeast Brazil.

SPECIMENS EXAMINED: BRAZIL. AMAzoNAS: Manaus, 20.IV.1985, leg. M.A. Jesus

709 (INPA 185947, as Perenniporia sp.); 2.V.1990, leg. M.A. Jesus 1384 ([NPA 192652,

as Microporus sp.); Manicoré, 20.IV.1985, leg. K.E Rodrigues 233 (INPA 128955, as

Polyporus sp.); BAHIA: location not determined, date not determined, leg. C. Torrend

(O 10486); Gongogi, V.1910, leg. C. Torrend, s.n. (URM 8893, as Trametes ocellata Berk.

& M.A. Curtis); RONDONIA: location not determined, 21.[X.1980, leg. B. Lowy et al. 230

(INPA 185947, as Perenniporia sp.); 20.V.1987, leg. M. Capelari et al. (SP 212021).

EXTRALIMITAL SPECIMEN EXAMINED: MEXICO. Oaxaca: Tehuantepec, 13.V1.1976,

leg. A.L. Welden 3606 (O 10488).

REMARKS: Coriolopsis psila is a conspicuous species due the dense thick mat

of entangled reddish brown hairs covering the pileus. Superficially it reminds

one of H. hydnoides, which, however, has black and stiffer hairs on the pileus.

Basidiospores of the two species are similarly cylindrical, but those of H.

hydnoides are longer (12-14 um). The generic concepts among polypores

with a trimitic hyphal system producing white rot (Coriolopsis, Hexagonia,

Pycnoporus, Trametes) are unclear and are based in part on basidioma color, a

character of dubious value at the generic level. Further DNA sequencing will

reveal the true phylogeny within these genera. For the time being we place F.

psila in Coriolopsis due to its brown context and a spore length that is closer to

other Coriolopsis species than in Hexagonia.

The combination in Phylloporia was based on the presence of a black line

between the hairy cover of the pileus and context, a prominent character for

Coriolopsis in Brazil ... 229

Phylloporia species. However Phylloporia was later characterized by small

ellipsoid basidiospores and a parasitic life strategy (Wagner & Ryvarden 2002),

while C. psila is saprophytic and has larger basidiospores. Seemingly a biological

convergent character, the hairy pilear covering occurs in many polypore genera

and is apparently an adaption to resist drying.

Key to the species of Coriolopsis recorded in Brazil

la/Basidiomata mostlysessile:. ir... sa. <s awe haa le 2 hale a Oka + Mao os uae Oe ee OS 2

1b Basidiomata-mostly-efhused=retlexed iia dys ssi-s dis steadringcion dine drs Pedy s ea dared 5

Das Weices~ led spel alii titer Bet. weet art ea aes Game! aed eet baRMags hoRteks (ecient Lacie ball 3

2D. ORES O= 7? DE MIE: ef DFImGd AsRGd As RG EM st ME MOR Alder UR Alden carer Pedces 4

3a. Abhymenial surface finely tomentose at the margin, pores angular,

1-2 per mm, basidiospores not known ................ ee eee eeee C. burchellii

3b. Abhymenial surface with scrupose tuft of agglutinated hairs, pores round,

3-4 per mm, basidiospores cylindrical 9-16 x 3-5 um .............. C. aspera

4a, Abhymenial surface glabrous, basidiospores 8-10 x 2.5-4 um ....... C. hostmannii

4b. Abhymenial surface with a dense mat of intertwined hairs,

basidiospores 9-12 GAs 3 se,-une, ste ines ie tuna in en nue tee C. psila

5a. Basidiospores oblong ellipsoid to subcylindrical, 10-14 x 4.5-6 um,

hyaline. tosvery; palesellowiShiss.tccerm vtscrem, Fst, wala rites tarn ities eral on dg C. byrsina

5b. Basidiospores ellipsoid to cylindrical, fusiform, hyaline, 7-11(-14) x 2-4 um..... 6

6a. Context distinctly duplex, tobacco-brown and shiny................... C. floccosa

6b. Context mostly homogeneous, dark brown to bay and dull..................... 7

7a. Context distinctly darker than the tubes, hymenial surface bay to deep brown,

pores round to angular, (2-)4-5 permm..................00. C. brunneoleuca

7b. Context dark date-brown concolorous with tubes, hymenial surface ochraceous,

to cinnamon to deep chocolate brown, pores round, 3-5 permm ....C. caperata

Acknowledgments

We would like to thank Dr. YC Dai and Dr. P. Buchanan for critically reviewing this

manuscript; Ana Cristina R. Souza, curator of the HFSL, for support during the field

trips of ACGS; Carlos Franciscon, curator of INPA and Adriana Gugliotta, curator of SP,

for the loan of exsiccates; Anibal Alves de Carvalho Junior for support during the visit

to RB; the staff of the ECFPn and of the MPEG for support during the field trips of PSM

and TBG; and the Programa de Pesquisa em Biodiversidade (PPBio) da Amazonia for

support during the field trips of PSM. Further, we acknowledge the Conselho Nacional

de Desenvolvimento Cientifico (CNPq) for the master scholarship of ACGS, GSNM and

PSM; the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (Capes) for

the doctorate scholarship of ACGS; the Instituto Internacional de Educagao do Brasil

230 ... Nogueira-Melo & al.

(IEB) and the Gordon and Betty Moore Foundation for the Scholarship of Studies on

Amazonia Conservation (BECA) to ACGS, PSM and TBG; the Dottorato di Ricerca in

Ecologia Sperimentale e Geobotanica (Universita degli Studi di Pavia, Italy), the Pés-

Graduacao em Biologia de Fungos (UFPE, Brazil), the Instituto Nacional de Ciéncia e

Tecnologia - Herbario Virtual de Plantas e Fungos (CNPQ 573883/2008-4) and FACEPE

(APQ 0433-2.03/08) for partially financing this study.

Literature cited

Baltazar JM, Gibertoni TB. 2009. A checklist of the aphyllophoroid fungi (Basidiomycota) recorded

from the Brazilian Atlantic Forest. Mycotaxon 109: 439-422.

http://dx.doi.org/10.5248/109.439

Bononi VLR. 1984. Basidiomicetos do Cerrado da Reserva Biolégica de Moji-Iuacu, SP.Rickia 11:

1-25.

Dai YC. 2012. Polypore diversity in China with an annotated checklist of Chinese polypores.

Mycoscience 53: 49-80. http://dx.doi.org/10.1007/s10267-011-0134-3

Dai YC, Cui BK, Yuan HS, He SH, Wei YL, Qin WM, Zhou LW, Li HJ. 2011. Wood-inhabiting fungi

in southern China 4. Polypores from Hainan Province. Ann. Bot. Fennici 48: 219-231

Gibertoni TB, Ryvarden L, Cavalcanti, MAQ. 2004. Poroid fungi (Basidiomycota) of the Atlantic

Rain Forest in Northeast Brazil. Syn. Fung.18: 33-43.

Gilbertson RL, Ryvarden L. 1986. North American polypores 1. Fungiflora, Oslo.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the Fungi. 10 ed. CABI

Publishing, Surrey. 771 p.

Nufiez M, Ryvarden L. 2001. East Asian polypores 2. Polyporaceae s. lato. Synopsis Fungorum 14:

170-522,

Ryvarden L. 1991. Genera of Polypores - Nomenclature and taxonomy. Synopsis Fungorum 5:

1-363.

Ryvarden L, Gilbertson RL. 1993. European Polypores, part 2. Synopsis Fungorum 7: 394-743.

Ryvarden L, Johansen I. 1980. A preliminary polypore flora of East Africa. Oslo, Fungiflora, 630 p.

Tavares I. 1939. Catalogo dos fungos de Pernambuco. Bol. Secr. Agric. (Pernambuco) 4(1): 1-33.

Wagner T & Ryvarden L. 2002: Phylogeny and taxonomy of the genus Phylloporia (Hymenochaetales).

Mycol Progress 1:105-116. http://dx.doi.org/10.1007/s11557-006-0009-8

Wang B, Cui BK, Li HJ, Du P, Jia BS. 2011. Wood-rotting fungi in eastern China 5. Polypore

diversity in Jiangxi Province. Ann. Bot. Fennici 48: 237-246.

MY COTAXON

http://dx.doi.org/10.5248/120.231

Volume 120, pp. 231-237 April-June 2012

A new species of Postia (Basidiomycota) from Northeast China

Bao-Kai Cur & Hat-Jiao Li

Institute of Microbiology, P.O. Box 61, Beijing Forestry University,

Beijing 100083, China

*CORRESPONDENCE TO: baokaicui@yahoo.com.cn

ABSTRACT — A new polypore, Postia subplacenta sp. nov. from Northeast China, is described

and illustrated. It is characterized by resupinate basidiocarps, angular pores (3-5 per mm)

with a cream buff to pale cinnamon-buff pore surface, and cylindrical basidiospores (4.2-6.0

x 1.9-2.4 um) that are negative in Melzer’s reagent and Cotton Blue. A key to accepted species

of Chinese Postia is supplied.

Key worps — lignicolous, poroid, fungi, Fomitopsidaceae, taxonomy

Introduction

Postia Fr. (Polyporales, Basidiomycota) is an important genus of brown-rot

fungi that are widely distributed in the northern hemisphere and mostly grow

on gymnosperm wood. Some species of the genus are economically important,

such as Postia guttulata (Peck) Jiilich and P lactea (Fr.) P. Karst., which have

been used as medicine in China (Dai et al. 2009a).

Some mycologists consider Postia a synonym of Oligoporus Bref. (Gilbertson

& Ryvarden 1987, Nufiez & Ryvarden 2001, Ryvarden & Gilbertson 1994),

while others regard it as an independent genus (Dai & Hattori 2007, Larsen

& Lombard 1986, Niemela et al. 2004, Renvall 1992, Wei & Dai 2006). The

genus is characterized by an annual growth habit, a monomitic hyphal system

with clamp connections, and thin-walled, allantoid to cylindrical or ellipsoid

basidiospores. Twenty-eight species have previously been known from China

(Cui et al. 2008, Dai 2009, Dai et al., 2003, 2004, 2007a,b, 2009b, Dai & Penttila

2006, Li et al. 2007, 2008, Wang et al. 2009, 2011, Wei & Dai 2006, Wei & Qin

2010, Yuan & Dai 2008, Yuan et al. 2010).

During a study on the taxonomy and diversity of wood-rotting fungi in

Northeast China, two Postia specimens were found that represented one

previously undescribed species, which we propose as P. subplacenta. In addition,

we provide an identification key to the Postia species thus far reported from

China.

2352-,.. Cui & Li

Materials & methods

The studied specimens were deposited in the herbarium of Beijing Forestry University

(BJFC). The microscopic procedure follows Dai (2010) and Cui et al. (2011). In

presenting the variation in the size of the spores, we exclude 5% of measurements (given

in parentheses) from each end of the range. The following abbreviations are used: IKI =

Melzer’s reagent, IKI- = negative in Melzer’s reagent, KOH = 5% potassium hydroxide,

CB = Cotton Blue, CB- = acyanophilous, L = mean spore length (arithmetic average of

all spores), W = mean spore width (arithmetic average of all spores), Q = variation in

the L/W ratios between the specimens studied, n = number of spores measured from

given number of specimens. Sections were studied at magnifications up to x1000 using

a Nikon Eclipse E 80i microscope and phase contrast illumination. Drawings were made

with the aid of a drawing tube. Special colour terms follow Petersen (1996).

Taxonomy

Postia subplacenta B.K. Cui, sp. nov. FIG. 1

MycoBank MB 563784

Differs from Postia placenta in the cream-buff to pale cinnamon-buff pore surface, wider

subicular hyphae, smaller basidiospores, and absence of gloeopleurous hyphae.

Type. — China. Jilin Province, Antu County, Changbaishan Nature Reserve, on fallen

trunk of Pinus, 8.VII.2011, Cui 10001 (holotype, BJFC).

EryMoLocy — subplacenta (Lat.) denotes a similarity with Postia placenta.

FRuITBODY — Basidiocarps annual, resupinate, corky when fresh, corky to

fragile when dry, up to 5 cm long, 3 cm wide, and 2.5 mm thick at the center.

Pore surface white to cream-buff when fresh, cream-buff to pale cinnamon-

buff when dry; pores angular, 3-5 per mm; dissepiments thin, entire to lacerate.

Margin indistinct, narrow to almost lacking. Subiculum cream to cream-buff,

corky when dry, <0.3 mm thick. Tubes paler than pore surface, pale cream-buff,

fragile when dry, <2.2 mm long.

HYyPHAL STRUCTURE — Hyphal system monomitic; generative hyphae

bearing clamp connections, IKI-, CB-; tissues unchanged in KOH.

SUBICULUM — Generative hyphae hyaline, thin- to mostly slightly thick-

walled, occasionally branched, interwoven, 3-7 um in diam.

TuBes — Generative hyphae hyaline, slightly thick- to distinctly thick-

walled, with a wide to narrow lumen, occasionally branched, interwoven, 2-5

tum in diam. Cystidia and cystidioles absent. Basidia clavate with four sterigmata

and a basal clamp connection, 12-18 x 4-5 um; basidioles in shape similar to

basidia, but slightly smaller.

Spores — Basidiospores cylindrical, hyaline, thin-walled, smooth, IKI-,

CB-, (4.0-)4.2-6.0(-6.9) x 1.9-2.4(-2.5) um, L = 5.07 um, W = 2.11 um,

Q = 2.37-2.45 (n = 60/2).

TYPE OF ROT — Brown rot.

ADDITIONAL SPECIMEN EXAMINED — CHINA. HEILONGJIANG PROVINCE, YICHUN,

Fenglin Nature Reserve, on stump of Pinus, 1. VIII.2011 Cui 9818 (BJFC).

Postia subplacenta sp. nov. (China) ... 233

10 ym

Fic. 1. Postia subplacenta (holotype), microscopic structures.

a: Basidiospores. b: Basidia and basidioles. c: Hyphae from trama. d: Hyphae from subiculum.

REMARKS — Postia subplacenta resembles P. placenta (Fr.) M.J. Larsen & Lombard

in having resupinate basidiocarps, similar pores, and cylindrical basidiospores;

however, P. placenta has a salmon-pink pore surface, gloeopleurous hyphae,

narrow subicular hyphae, and larger basidiospores (5.5-7 x 2-2.5 um;

Gilbertson & Ryvarden 1987, Ryvarden & Gilbertson 1994).

Postia subplacenta is similar to P. rancida (Bres.) M.J. Larsen & Lombard,

which differs in having rancid taste and larger basidiospores (6-8 x 2-3 um;

Gilbertson & Ryvarden 1987).

Postia obliqua Y.L. Wei & W.M. Qin, recently described from Southwest

China (Wei & Qin 2010), may be confused with P subplacenta in its resupinate

234 ... Cui & Li

basidiocarps and similar pores and basidiospores (4.8-6.3 x 2.0-2.5 um)

but differs in its very large basidiocarps with oblique tubes that usually form

imbricate pseudopilei and the presence of gloeopleurous-like hyphae.

Macroscopically, Postia subplacenta looks somewhat like Oligoporus rennyi

(Berk. & Broome) Donk, which, however, has chlamydospores and produces

basidiospores that are oblong-ellipsoid and thin- to slightly thick-walled

(Ryvarden & Gilbertson 1994).

Key to species of Postia in China

. Basidiocarps effused-reflexed, pileate or stipitate ............ 0. . eee eee eee 2

J BasidiGcarps Tesupinates, ows v6 Gea eh oes GS eG ae BGR a eG Ree SG Sx ea eBe Lage ta 23

, Basidiocarps stipitate or substipitate::.. «<i. + saihece a dinar a diner theta heeees aes 3

. Basidiocarps effused-reflexed or pileate ..... cece eee eee eens 4

SUPOTES PL SHPST MIMD 5.25.5}. 4. cgtiabea chelated ties loa atte P. subundosa Y.L. Wei & Y.C. Dai

Pores: 3=5:per mii. |... 24). 4g hide gale ea dine P. ceriflua (Berk. & M.A. Curtis) Jiilich

. Basidiocarps with distinct grey to bluish tints ............ 0... cece eee eee eee 2

. Basidiocarps white, cream, yellowish or brown ............ 0.00. c cece eee eee eee 7

AD IIEATY STO SO CLIN an chew Rho Rae ha le, ere Mend tN P. alni Niemela & Vampola

Omevinnospenny, Wo Sa, ha, Moh, Rab, eoie » okie emis Wuohicy ations wabanhery a tieate. att 6

. Basidiospores; <1.8-wm wide 2.0... 2.00 oes ow Pel ae Jace oe P. caesia (Schrad.) P. Karst.

. Basidiospores >1.8 um wide ................20005 P. luteocaesia (A. David) Julich

. Basidiocarps becoming brown when bruised or when dry ..................0.. 8

. Basidiocarps unchanged when bruised or dry ............ 0.0 cece eee eee eee ee 10

Molo acyStidia PRESENt «oc p ions eek bance thee ees P. leucomallella (Murrill) Jiilich

. GOSOCYSUIC IAD SEN tse fap. cwsse don conse tor asosce dan. seraplat Sure gl atsse, pebin:dnsraal ates deh deu ed eaeh Tucows 9

. Basidiosporesr<ls6 pmwide 062.60 eon Soha eect hee the P lateritia Renvall

Basidigspores:2 9-7 Unik Wide! oo aoa skate meee meee eee 2 P. fragilis (Fr.) Jalich

. Basidiocarps chalky when dry ................... P. calcarea Y.L. Wei & Y.C. Dai

. Basidiocarps fragile, corky or woody hard when dry ...............0....0005 11

HCY STIAIT PECSONE YS cn elit. darehs teats naa ale teats tmkhs nants toma hs baer ban aie 12

MEV StH IA <A DG CIE ia dlrs dhe cies fy Fovgion dy tenes dus poten Fy pate as pckerg Pyle chew byt peckow snyas php sdyht gE 14

Gv sticlia thin=wealled ; fu, siete. totes mekbey Soke 4 P. amurensis Y.C. Dai & Penttila

PE yotid ia thick swale: ae. o actus spe tarace teanache deavache tawnche tearaghadearaces ae Okee 13

. Cystidia amyloid in Melzer’s reagent ..... P. pileata (Parmasto) Y.C. Dai & Renvall

. Cystidia inamyloid in Melzer’s reagent ................ P. balsamea (Peck) Jiilich

POLES DIAOeT Os POE TINS (ies ie eae wg a ad gd dg P. undosa (Peck) Jiilich

Rovesssinaller; > 3. periinis 4.07 Pit.) Rist o8! Wn! Wah PN Ae WW tam D Bel! oe 15

15.

16.

17.

18.

eee

19.

20.

—_—_ —_

22;

23.

24.

24,

25:

26.

26;

27,

28.

Ze:

205

2S,

Postia subplacenta sp. nov. (China) ... 235

Gloeocystidiapresent:..6 2. ces 2. lek Rsk, Re. FENG. SEE on SER EBS i RS cn ES 16

COCOGVSEIAT AA DSER so 35 cect crease ore i eee tere Ee en een eed Nera, ae 17

Hyphal pegs abundant .................... P. gloeocystidiata Y.L. Wei & Y.C. Dai

Hyphal-pegs.absetit. (e520 Gin 2.506.250 28 since a P. qinensis Y.C. Dai & Y.L. Wei

Piléal surface more. or less: prvkewhencfréshicy, soacts seats seacts soadts seats o oats: 18

Pileal surface-never putk when: fteshi ag 40 ack 9 esp cto Het acdeg Sh poe Shp toe Fy pee gasp al 19

Basidiospores sds57pim- wide 1.8. i. weds wees 8 P. persicina Niemela & Y.C. Dai

Basidiospores: 15: (it Wide" w.s00,"t-caccts tearacts saracte dive P. cana H.S. Yuan & Y.C. Dai

Pilealtstikface-7Onate<w 28 pot.2o Pito8 Bite whet tat 08s P. zebra Y.L. Wei & Y.C. Dai

Pileall SUPEACE AZ ON ALC op tartes Biemer thy Besma tin Meaney Wemed ee Beaman Mendel NeLuadd nahtonade wiktada beth 20

Basidiocarps mild, upper surface greyish brown........ P. tephroleuca (Fr.) Jitlich

Basidiocarps bitter, upper surface cream or yellow-brownish ................. 21

b, BASTCAOSPOTES! 32 SNL WIE” s0s-s0- notary pots wild Ges atlas Sed cwie ota ve Gte sae a BS P. guttulata

FAIS AST IOS PONG SES ce MUTA OVI cs ic PR ol Ud Baad Sad ta a LOGI ad alr ca dete heen 22,

Basidiocarps woody hard when dry .................06. P. stiptica (Pers.) Julich

Basidiocarps-trasileswhetirdry Sasss0-e5tass aoe ie Siluueesta we oed ale Ba P. lactea

Basidiocarps becoming reddish to rusty brown when touched......... P lateritia

Basidiocarpsunchanged when touched 2.64 .-..csge i tage v atetage Be ge Hee lage Bley 24

Basidiospores mostly >2 um wide, cystidia absent ................... 0.0 25

Basidiospores mostly <2 um wide, cystidia present .................. 00000 28

Basidiocarps salmon pink; basidiospores 2-3 um wide ............... P. placenta

Basidiocarps white or cream; basidiospores 2-2.5 um wide ...............4-. 26

Basidiocarps suitti-ranicid-srielll, ch. sey tho isem adler whe My ele ake ghtwsbie P. rancida

Basidiocarps wati@ut, PAN CTS Stivell sys, 1dr Heer Diese ace anda dae Seas tae ee aero oe 27

Basidiocarps very large, gloeopleurous hyphae present ................ P. obliqua

Basidiocarps small, gloeopleurous hyphae absent ................. P. subplacenta

Cystidia thick-walled spa 52s cic Papa ese et oR gt PR neh lo clo gh ta io P. pileata

Gysticia- thin: walledee i. 0 ten FOUN ie FE Aes wR nue eee A MRE WSS ee ne, AEe ha, 29

Pores 5-6 per mm; basidiospores mostly <1 um wide ..... P. simanii (Pilat) Jiilich

Pores 3-4 per mm; basidiospores mostly >1 um wide

pie Reet eee Oe ON MEN ote CONE ole toh? P. hibernica (Berk. & Broome) Jiilich

Acknowledgements

Special thanks are due to Mr. Chang-Lin Zhao (China) for help in field collection.

We express our gratitude to Drs. Michal TomSovsky (Czech Republic) and Yu-Lian

Wei (China) who reviewed the manuscript. The research was financed by the National

Natural Science Foundation of China (Project No. 31170018) and Program for New

Century Excellent Talents in University (NCET-11-0585).

236 ... Cui & Li

Literature cited

Cui BK, Yuan HS, Dai YC. 2008. Wood-rotting fungi in eastern China 1. Polypores from Wuyi

Mountains, Fujian Province. Sydowia 60: 25-40.

Cui BK, Li HJ, Dai YC. 2011. Wood-rotting fungi in eastern China 6. Two new species of

Antrodia (Basidiomycota) from Mt. Huangshan, Anhui Province. Mycotaxon 116: 13-20.

http://dx.doi.org/10.5248/116.13

Dai YC. 2009. A checklist of polypores in China. Mycosystema 28: 315-327.

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 131-343.

http://dx.doi.org/10.1007/s13225-010-0066-9

Dai YC, Hattori T. 2007. Postia japonica (Basidiomycota), a new polypore from Japan. Mycotaxon

102: 113-118.

Dai YC, Penttila R. 2006. Polypore diversity of Fenglin Nature Reserve, northeastern China.

Annales Botanici Fennici 43: 81-96.

Dai YC, Harkénen M, Niemela T. 2003. Wood-inhabiting fungi in southern China 1. Polypores

from Hunan Province. Annales Botanici Fennici 40: 381-393.

Dai YC, Wei YL, Wang Z. 2004. Wood-inhabiting fungi in southern China 2. Polypores from

Sichuan Province. Annales Botanici Fennici 41: 319-329.

Dai YC, Cui BK, Huang MY. 2007a. Polypores from eastern Inner Mongolia, northeastern China.

Nova Hedwigia 84: 513-520. http://dx.doi.org/10.1127/0029-5035/2007/0084-0513

Dai YC, Cui BK, Yuan HS. 2007b. Notes on polypores from Gansu and Qinghai Province, northwest

China. Cryptogamie Mycologie 28: 177-187.

Dai YC, Yang ZL, Cui BK, Yu CJ, Zhou LW. 2009a. Species diversity and utilization of medicinal

mushrooms and fungi in China (Review). International Journal of Medicinal Mushrooms 11:

287-302. http://dx.doi.org/10.1615/IntJ MedMushr.v11.i3.80

Dai YC, Yuan HS, Wang HC, Yang F, Wei YL. 2009b. Polypores (Basidiomycota) from Qin Mts. in

Shaanxi Province, central China. Annales Botanici Fennici 46: 54-61.

Gilbertson RL, Ryvarden L. 1987. North American polypores 2. Fungiflora, Oslo. 434-885 pp.

Larsen MJ, Lombard FE. 1986. New combinations in the genus Postia Fr. (Polyporaceae). Mycotaxon.

26: 27-1)-273;

LiJ, Xiong HX, Zhou XS, Dai YC. 2007. Polypores (Basidiomycetes) from Henan Province in central

China. Sydowia 59: 125-137.

Li J, Xiong HX, Dai YC. 2008. Polypores from Shennongjia Nature Reserve in Hubei Province,

Central China. Cryptogamie Mycologie 29: 267-277.

Niemela T, Dai YC, Kinnunen J, Schigel DS. 2004. New and in North Europe rare polypore species

(Basidiomycota) with annual, monomitic basidiocarps. Karstenia 44: 67-77.

Nujiez M, Ryvarden L. 2001. East Asian polypores 2. Polyporaceae s. lato. Synopsis Fungorum 14:

165-522.

Petersen JH. 1996. Farvekort. The Danish Mycological Society’s colour-chart. Foreningen til

Svampekundskabens Fremme, Greve. 6 p.

Renvall P. 1992. Basidiomycetes at the timberline in Lapland 4. Postia lateritia n. sp. and its rust-

coloured relatives. Karstenia 32: 43-60.

Ryvarden L, Gilbertson RL. 1994. European polypores 2. Synopsis Fungorum 7: 394-743.

Wang B, Dai YC, Cui BK, Du P, Li HJ. 2009. Wood-rotting fungi in eastern China 4. Polypores from

Dagang Mountains, Jiangxi Province. Cryptogamie Mycologie 30: 233-241.

Wang B, Cui BK, Li HJ, Du P, Jia BS. 2011. Wood-rotting fungi in eastern China 5. Polypore

diversity in Jiangxi Province. Annales Botanici Fennici 48: 237-246.

Postia subplacenta sp. nov. (China) ... 237

Wei YL, Dai YC. 2006. Three new species of Postia (Aphyllophorales, Basidiomycota) from China.

Fungal Diversity 23: 391-402.

Wei YL, Qin WM. 2010. Two new species of Postia from China. Sydowia 62: 165-170.

Yuan HS, Dai YC. 2008. Polypores from northern and central Yunnan Province, southwestern

China. Sydowia 60: 147-159.

Yuan HS, Dai YC, Wei YL. 2010. Postia cana sp. nov. (Basidiomycota, Polyporales) from Shanxi

Province, northern China. Nordic Journal of Botany 28: 629-631.

http://dx.doi.org/10.1111/j.1756-1051.2010.00849.x

MY COTAXON

http://dx.doi.org/10.5248/120.239

Volume 120, pp. 239-246 April-June 2012

Three new Caeoma species on Rosa spp. from Pakistan

N.S. AFSHAN”™, A.N. KHALID? & A.R. NIAZI’

™Centre for Undergraduate Studies & *Department of Botany, University of the Punjab,

Quaid-e-Azam Campus, Lahore, 54590, Pakistan

*CORRESPONDENCE TO: pakrust@gmail.com

ABSTRACT — Three representatives of the anamorphic genus Caeoma —C. ahmadii on Rosa

microphylla; C. khanspurense and C. rosicola on Rosa webbiana— are described as new rust

species from Pakistan. This first report of Caeoma raises the number of known anamorphic

rust genera from the country to five.

Key worps — Khanspur, Mansehra, Phragmidium

Introduction

The genus Caeoma Link is traditionally used for species having sori that

lack obvious bounding structures and that produce catenulate spores with

intercalary cells. This contrasts with the genus Aecidium Pers., which has a cup-

shaped sorus with a well-developed peridium. Similar sori are found in the

aecial state of Melampsora Castagne and the uredinia of Chrysomyxa Unger,

Coleosporium Lév.,and other genera (Cummins & Hiratsuka 2003). The aecia of

Phragmidium are (usually) Caeoma-type with catenulate spores or (less often)

Uredo-type (Petrova & Denchev 2004) with verrucose or echinulate aeciospores.

Cummins & Hiratsuka (2003) refer to the anamorphic genus Lecythea Léveillé

the Phragmidium species with aecia corresponding to Caeoma UI of Sato &

Sato (1985). Because Hennen et al. (2005) regard Lecythea as confusing and

not in use, we consider Caeoma the appropriate anamorph for accommodating

species with Phragmidium aecia.

So far, 22 genera and 417 species of rust fungi have been described or reported

from Pakistan (Afshan et al. 2008a,b,c,d, 2009a,b, 2010a,b, 2011a,b; Sultan et al.

2008; Afshan & Khalid 2009; Khalid & Afshan 2009; Iqbal et al. 2008, 2009),

including four genera of anamorphic rust fungi, Aecidium, Monosporidium

Barclay, Peridermium (Link) J.C. Schmidt & Kunze, and Uredo Pers. Although

no representatives of Caeoma have been reported previously from Pakistan,

three new Caeoma species occurring on Rosa are described here.

240 ... Afshan, Khalid & Niazi

Materials & methods

Freehand sections of infected tissue and spores were mounted in lactophenol and

gently heated to boiling point. The preparations were observed under a NIKON YS

100 microscope and photographed with a digipro-Labomed and a JSM5910 scanning

electron microscope. Drawings of spores and paraphyses were made using a Camera

Lucida (Ernst Leitz Wetzlar, Germany). An ocular micrometer was used for spore

dimensions, with at least 25 spores measured for each spore stage. The rusted specimens

have been deposited in the Botany Department herbarium, University of the Punjab,

Lahore (LAH).

Taxonomy

Figs. A-C: Caeoma ahmadii (holotype), SEM micrographs. A: Aecidium containing

aeciospores and paraphyses. B: Aeciospores. C: Detail of an aeciospore showing verrucose

wall ornamentation (paraphyses visible in background).

Caeoma spp, nov. (Pakistan) ... 241

Caeoma ahmadii Afshan, Khalid & Niazi, sp. nov. Fies. A-E

MycoBank MB 564295

Differs from Phragmidium tuberculatum in larger aeciospores that are hyaline with

orange-yellow contents.

TyPE: PAKISTAN, KHYBER-PAKHTUNKHAWAH (KPK), Ayubia National Park, at 2135 m

a.s.l., on Rosa microphylla Desf., stage I, 18 June, 2008, N.S. Afshan & A.N. Khalid. NSA

#151A (Holotype, LAH NSA1114).

ErymMoLoecy: Named after the world renowned mycologist, Sultan Ahmad.

SPERMOGONIA, UREDINIA and TELIA not found. Agecta hypophyllous, on leaves,

bright yellow to orangish yellow, rounded or oblong, scattered, 0.1-0.3 x 0.09-

0.10 mm. PARAPHYSES numerous, erect to suberect, clavate, hyaline with light

yellow granules, 50-70 um long and 8-12 um wide. AEcIOosPoREs globose to

subglobose or ellipsoid to broadly ellipsoid, (23—)26-31 x 28-36 um; wall 1-1.5

um thick, hyaline with orange-yellow contents, densely verrucose, with 5-8

scattered pores.

y eines.

. vesting,

wep Tonner nero

2 etm cone set nae « aeneeeee ott

Seater

Fite nna,

Sze eg,

a neces senmanp meres tet r

ite apie tslapatdien-en

ae:

monemesemngahontene mt

Sanne

SSIS

Figs. D-E: Caeoma ahmadii (holotype), lucida drawings.

D: Aeciospores showing wall ornamentation. Scale bar= 9 um.

E: Apices of paraphyses. Scale bar = 12 um.

ComMENTs: Aeciospores of C. ahmadii are larger than those of Phragmidium

tuberculatum J.B. Mull. (18-24 x 20-30 um), Ph. mucronatum (Pers.) Schtdl.

(17-21 x 20-28 um), Ph. montivagum Arthur (16-19 x 21-26 um), Ph. rosae-

arkansanae Dietel (16-23 x 23-27 um), Ph. speciosum (Fr.) Burrill (16-24 x

242 ... Afshan, Khalid & Niazi

24-35 um), and Ph. rosae-pimpinellifoliae Dietel (15-20 x 18-27 um). They also

differ in color and wall ornamentation from those of Ph. tuberculatum.

Fics. F-G: Caeoma khanspurense (holotype), lucida drawings.

F: Apices of clavate paraphyses. Scale bar = 20 um. G: Mature aeciospores. Scale bar = 10 um.

Caeoma khanspurense Khalid, Afshan & Niazi, sp. nov. Fics. F-G

MycoBank MB 564317

Differs from Phragmidium mucronatum and Ph. tuberculatum in larger aeciospores with

verrucose wall ornamentation.

Type: PAKISTAN, KHYBER-PAKHTUNKHAWAH (KPK), Helipad, Khanspur, at 2135 m

a.s.l., on Rosa webbiana Wall. ex Royle, stage I, 23 May 2006, N.S. Afshan & A.N. Khalid.

NSA #03 (Holotype, LAH NSA1116).

ErymMo.ocy: Named after the type locality, Khanspur.

SPERMOGONIA, UREDINIA and TELIA not found. Agecta hypophyllous or

caulicolous, on the veins, stems and petioles, scattered, rounded or oblong,

0.3-0.9 x 0.09-0.10 mm, orange yellow when fresh, surrounded by clavate

paraphyses. AEcrosporEs globose, subglobose or ellipsoid to obovoid, pale

yellow to orange-yellow, verrucose, occasionally with a plateau-shaped base,

23-32 x 23-41 um, borne on one or two basal cells. PARAPHYSES clavate,

hyaline, 70-105 um long, 8-12 um wide, with uniformly thin walls.

ComMENTs: Aeciospores of C. khanspurense are larger than those of

Phragmidium mucronatum (17-21 x 20-28 um) and Ph. tuberculatum (18-24

Caeoma spp, nov. (Pakistan) ... 243

x 20-30 um); their verrucose ornamentation further distinguishes them from

the aeciospores of Ph. mucronatum.

The broader aeciospores of C. khanspurense differ from those of C. rosicola

(reported on the same host), characterized by verrucose to echinulate, 19-28

um broad aeciospores.

Aeciospores with a verrucose wall ornamentation with a plateau-shaped

base distinguish C. khanspurense from Phragmidium rosae-pimpinellifoliae with

smaller aeciospores (15-20 x 18-27 um).

Fic. H: Caeoma rosicola, lucida drawing.

Aeciospores. Scale bar = 10 um.

Caeoma rosicola Afshan, Niazi & Khalid, sp. nov. Fics. H-J

MycoBank MB 564318

Differs from other Caeoma species in larger aeciospores and lack of paraphyses.

Type: PAKISTAN, KHYBER-PAKHTUNKHAWAH (KPK), Helipad, Khanspur, at 2135 m

a.s.l., on Rosa webbiana Wall. ex Royle, stage I, 23 May 2006, N.S. Afshan & A.N. Khalid.

NSA #02 (Holotype, LAH NSA1117).

ErymMo.Locy: Named after the host genus, Rosa.

SPERMOGONIA, UREDINIA and TELIA not found. AEcta hypophyllous or

petiolicolous, mostly on the veins, petioles, branches and fruits, causing

malformations, yellowish orange to bright orange, scattered. PARAPHYSES

not observed. AECIOSPORES globose to subglobose or ellipsoid to ovoid; wall

hyaline with orange yellow contents, echinulate to verrucose, 19-28 x 21-41

um, germ pores 1-4, mostly equatorial.

ComMENTs: Larger aeciospores and the lack of paraphyses differentiate C.

rosicola from other rust species reported on Rosaceae: Phragmidium rosae-

pimpinellifoliae (15-20 x 18-27 um), Ph. rosae-arkansanae (16-23 x 23-27 um),

244 ... Afshan, Khalid & Niazi

Figs. I-J: Caeoma rosicola (holotype), SEM micrographs. I: Aeciospores. J: Detail of

aeciospore showing echinulate wall ornamentation.

Caeoma spp, nov. (Pakistan) ... 245

Ph. mucronatum (17-21 x 20-28 um), and Ph. montivagum (16-19 x 21-26

um). Phragmidium montivagum is further distinguished by aeciospores with 8

scattered germ pores.

Acknowledgements

We are highly obliged to Higher Education Commission (HEC) of Pakistan for

providing financial support. We sincerely thank Dr. Marcin Piatek (W. Szafer Institute of

Botany, Polish Academy of Sciences, Poland) and Dr. Omar Paino Perdomo (Dominican

Society of Mycology Santo Domingo, Dominican Republic) for their valuable suggestions

to improve the manuscript and acting as presubmission reviewers.

Literature cited

Afshan NS, Khalid AN. 2009. New records of Puccinia and Pucciniastrum from Pakistan. Mycotaxon

108: 137-146. http://dx.doi.org/10.5248/108.137

Afshan NS, Berndt R, Khalid AN, Niazi AR. 2008a. New graminicolous rust fungi from Pakistan.

Mycotaxon 104: 123-130.

Afshan NS, Khalid AN, Javed H. 2008b. Further additions to the rust flora of Pakistan. Pakistan

Journal of Botany 40(3): 1285-1289.

Afshan NS, Khalid AN, Niazi AR. 2008c. New records of graminicolous rust fungi from Pakistan.

Pakistan Journal of Botany 40(3): 1279-1283.

Afshan NS, Khalid AN, Niazi AR. 2008d. New records and distribution of rust fungi from Pakistan.

Mycotaxon 105: 257-267.

Afshan NS, Iqbal SH, Khalid AN, Niazi AR. 2009a. A new anamorphic rust fungus with a new

record of Uredinales from Azad Kashmir, Pakistan. Mycotaxon 112: 451-456.

http://dx.doi.org/10.5248/112.451

Afshan NS, Khalid AN, Iqbal SH, Niazi AR, Sultan A. 2009b. Puccinia subepidermalis sp. nov. and

new records of rust fungi from Pakistan. Mycotaxon 110: 173-182.

http://dx.doi.org/10.5248/110.173

Afshan NS, Khalid AN, Niazi AR. 2010a. Three new species of rust fungi from Pakistan. Mycological

Progress http://dx.doi.org/10.1007/s11557-010-0655-8

Afshan NS, Khalid AN, Iqbal SH, Niazi AR, Sultan A. 2010b. Puccinia anaphalidis-virgatae, a new

species and a new variety of rust fungi from Fairy Meadows, Northern Pakistan, Mycotaxon

112: 483-490. http://dx.doi.org/10.5248/112.483

Afshan NS, Khalid AN, Iqbal SH, Niazi AR. 201 1a. Puccinia species new to Azad Jammu & Kashmir,

Pakistan, Mycotaxon: 116: 175-182.

Afshan NS, Khalid AN, Niazi AR, Iqbal SH. 2011b. New records of Uredinales from Fairy Meadows,

Pakistan. Mycotaxon: 115: 203-213. http://dx.doi.org/10.5248/115.203

Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi. Third ed. The American

Phytopathological Society. APS Press, St. Paul, MN.

Hennen JF, Figueir edo MB, Carvalho AA, Hennen PG. 2005. Catalogue of the species of plant rust

fungi (Uredinales) of Brazil. FAPESP, CNPq, FAPERJ, NSF, USDA.

Iqbal SH, Khalid AN, Afshan NS, Niazi AR. 2008. Rust Fungi on Saccharum species from Pakistan.

Mycotaxon 106: 219-226.

Iqbal SH, Afshan NS, Khalid AN, Niazi AR, Sultan A. 2009. Additions to the rust fungi of Fairy

Meadows, Northern Areas of Pakistan. Mycotaxon 109: 1-7. http://dx.doi.org/10.5248/109.1

246 ... Afshan, Khalid & Niazi

Khalid AN, Afshan NS. 2009. Additions to the graminicolous rust fungi of Pakistan. Mycotaxon

108: 175-183. http://dx.doi.org/10.5248/108.175

Petrova RD, Denchev CM. 2004. A taxonomic study of Phragmidiaceae (Uredinales) in Bulgaria.

Mycologia Balcanica 1: 95-115.

Sato T, Sato S. 1985. Morphology of the aecia of the rust fungi. Trans. Br. Mycol. Soc. 85: 223-238.

Sultan MA, Haq I, Khalid AN, Mukhtar H. 2008. Two new anamorphic rust fungi from northern

areas of Pakistan. Mycotaxon 105: 23-27.

MYCOTAXON

http://dx.doi.org/10.5248/120.247

Volume 120, pp. 247-251 April-June 2012

Nomenclatural status and morphological notes on

Tubifera applanata sp. nov. (Myxomycetes)

D.V. LEONTYEV’™ & K.A. FEFELOV’

‘Department of Biotechnology, Kharkiv Zooveterinary Academy,

Akademichna str. 1 Mala Danylivka, Kharkiv 62341 Ukraine

*The Institute of Plant and Animal Ecology,

Vosmogo Marta str. 202, Yekaterinburg 620144 Russia

* CORRESPONDENCE TO: protista@mail.ru

Asstract — Tubifera applanata sp. nov. is proposed to validate “Tubulifera applanata” nom.

inval. The species diagnosis and some notes on its morphology are provided. At 0.40-0.65 mm

diam., individual sporothecae are somewhat larger than in T. ferruginosa and T. microsperma

and smaller than those in T. casparyi. Circular ornamentations on the inner peridial surface

in T: applanata are larger than previously noted, reaching a size up to 2.9 um.

Key worps — morphometry, SEM, species delimitation

Introduction

In 2001, C. Lado stated that the invalid generic name “Tubulifera” O.F. Mull.

1775 had been validated as Tubulifera O.F. Mill. ex Jacq. 1779, a name with

priority over Tubifera J.F. Gmel. 1792 (Lado 2001). On this understanding,

Leontyev & Fefelov (2009) proposed a new species, Tubulifera applanata.

Subsequently, Lado (2011) indicated that, contrary to previous opinion,

Jacquin’s publication failed to validate the genus “Tubulifera.” This means that

all species names published in “Tubulifera,” including “Tubulifera applanata,”

are also invalid (ICBN [Vienna Code] Article 43.1). We therefore propose a

validation of “Tubulifera applanata” as Tubifera applanata sp. nov. and provide

additional morphological data.

Materials & methods

A total of 13 specimens of Tubifera applanata were examined (CWU MR 001, 038a,

038b, 039, 054, 058, 074, 077, 121, 122, 123, 125, 126). Additionally, 8 specimens of

T. ferruginosa (Batsch) J.E. Gmel. (CWU MR 006, 023, 012, 091, 095, 103, 113, 114), 4 of

T. microsperma (Berk. & M.A. Curtis) G.W. Martin (CWU MR 008, 041, 049, 153), and

3 of T: casparyi (Rostaf.) T. Macbr. (CWU MR 049, 181, 183) were used for comparison.

All the specimens were collected during 2003-2010 in the East Forest-Steppe region

248 ... Leontyev & Fefelov

of Ukraine (Gomolsha Forests National Nature Park, Kharkiv region). Material was

deposited in the herbarium of the V.N. Karasin National University of Kharkiv (CWU),

the subsection of Reticulariaceae (MR).

The specimens were examined using a stereoscopic microscope MBS-9 and scanning

electron microscope (SEM) Jeol JSM-6060 (30 kV, gold evaporation).

The diameters of intact closed sporothecae were measured from above by a

micrometer. After drawing an imaginary line corresponding to the largest fruit body

diameter on the surface of the pseudoaethalium, we measured 30 sporothecae along this

line from the edge of the pseudoaethalium. The diameters of annular ornamentations on

the inner peridial surface were measured using the SEM scale.

Means, standard deviations, and standard errors were calculated using Stat Soft

Statistica 8.0 and Microsoft Excel 2003 software to describe the morphological variety.

Results & discussion

Tubifera applanata Leontyev & Fefelov, sp. nov. FIG. 1c-D

MycoBank MB 561713

“Tubulifera applanata” Leontyev & Fefelov, Bol. Soc. Mycol. Madrid 33: 119. 2009

Differs from Tubifera ferruginosa by its large pseudoaethalia, flat sporothecal tips,

annular embossments on the internal peridial surface, and salmon color of young

fructifications.

Type: Ukraine, Kharkiv region, Gomolsha Forests National Nature Park, Zadinetske

forestry, edge of pine forest near ‘Bile Ozero’ Camp (49°36'50.69"N 36°21'06.15”E), on

the bark of fallen trunk of Pinus sylvestris L., 14.07.2003, leg. D.V. Leontyev (Holotype,

CWU MR 039).

EryMmo oey: applanatus (Lat.) = flat; referring to the flattened sporothecal tips.

PSEUDOAETHALIUM large, (12—)23-37(-73) mm long, (7-)16.5-27.5(-38) mm

wide, 4-6 mm in height, oval or irregular as seen from above, flat pulvinate, rust-

brown. SPOROTHECAE Straight, clearly prismatic from mutual pressure. Tips

OF SPOROTHECAE 0.4—0.65 mm diam., flat or slightly convex, 5-6 angular or

almost circular, equal in height, forming a smooth surface of pseudoaethalium.

PERIDIUM opaque, light brown, lateral walls of the sporothecae often plicate.

INTERNAL SURFACE OF PERIDIUM covered with annular embossments

(0.4-)1.5-2.9 um diam., quite distant from each other. COLUMELLA absent

or not reaching the top of the sporothecae, thick, with color of spore mass.

PSEUDOCAPILLITIUM absent or scanty, filiform, appearing as occasionally

branched strings. HyPOTHALLUS spongy, poorly developed or absent. SPORES

in mass rust-brown, brownish by transmitted light, 4.9-6.3(-7.5) um diam.,

banded reticulate. PLAsmopIUM salmon or rusty-cream. In coniferous forests,

on bark, litter and slightly decomposed wood.

Notes on the morphology

Tubifera applanata is characterized by the prismatic shape of the individual

sporothecae, their flat hexagonal tips, ring-like ornamentations on the inner

peridial surface, and the salmon color of young fructifications. Although we

Tubifera applanata sp. nov. ... 249

first noted that quantitative parameters such as pseudoaethalium size and

spore diameter distinguished T: applanata from related species (Leontyev &

Fefelov 2009), we did not realize that individual sporothecal diameters might

also serve as a diagnostic criterion. Moreover, there were no data referring to

the diameter of sporothecae in other Tubifera species (Nannenga-Bremekamp

1991, Neubert et al. 1993, Ing 1999).

The study of sporothecae size in Tubifera applanata, T: ferruginosa, T. micro-

sperma, and T. casparyi has shown that all species have significant differences

(p<0.01) from one another. Measurements show that individual sporothecae

are somewhat larger in T: applanata than in T. ferruginosa and T. microsperma

and smaller than those in T’ casparyi (TABLE 1).

TABLE 1. Sporothecal diameters in four Tubifera species.

T. applanata T. ferruginosa T. microsperma T. casparyi

Range of specimen means (1m) | 464-633. =| = 322-474 330-398 512-692

Mean (um)

Standard deviation [sd] (tm)

Rounded variation, for use in species

descriptions (mm)

04-065 | 03-05 | 03-045 | 0.5-0.7

Although the difference in sporothecae size between T. applanata and

T. ferruginosa is quite small (0.1-0.2 mm), it can be seen with the naked eye.

We propose to use this criterion as an additional character in descriptions of

Tubifera species.

Peridium ultrastructure

Nelson et al. (1982) introduced a new diagnostic criterion into Tubifera

taxonomy — the type of inner peridial ornamentation, noting that in T’ micro-

sperma the inner surface of the peridium is ornamented by so-called ‘rimmed

craters. In Ukrainian specimens of T: microsperma, the ornamentation agrees

with this description (Fic. 1A-B).

In 2009 the SEM-study of T: applanata showed ‘peridial rings, similar to

those of T! microsperma but much lower and more distant from each other

(Leontyev & Fefelov 2009). Both ‘peridial rings’ and ‘rimmed craters’ appear to

be remnants of channels used by the plasmodium for active water evacuation

(R. McHugh, pers. comm.).

We previously indicated the outer ‘peridial ring’ diameter in T’ applanata as

0.4-0.9 um (Leontyev & Fefelov 2009). Improved SEM techniques demonstrate

that in the type specimen, these structures reach 1.6-2.9 um diam. (FIG. 1C-D),

significantly larger than similar structures in T. microsperma. In T: applanata,

rings < 1 um diam. are usually less prominent than the larger ones.

250 ... Leontyev & Fefelov

ZE9R8 Wit

Figure 1. A-B: Tubifera microsperma [CWU MR 135]—ornamentation of the inner peridium

surface illustrating ‘rimmed craters’ C-D: Tubifera applanata [CWU MR 039, holotype]—

ornamentation of the inner peridium surface illustrating ‘peridial rings’ and intricate filaments.

Tubifera applanata sp. nov. ... 251

This study has shown that the peridial surface of T. applanata is ornamented

by thin intricate filaments, appearing to be composed of dried slime (Fic. 1D).

Such filaments are especially abundant in the bottom of the ‘peridial rings’

Acknowledgments

We express our sincere gratitude to David W. Mitchell (East Sussex, England),

Carlos Lado (Royal Botanic Garden, Madrid, Spain), and Shaun Pennycook (Landcare

Research, Auckland, New Zealand) for valuable comments and corrections; and to

Yana V. Belousova (Kharkiv State Zooveterinary Academy) for the assistance in the

measurements of sporothecae.

Literature cited

Ing B. 1999. The Myxomycetes of Britain and Ireland. An identification handbook. Richmond.

Lado C. 2001. Nomenmyx: a nomenclatural taxabase of Myxomycetes. Cuad. Trab. Fl. Micol. Iber.

1621-221.

Lado C. 2011. The nomenclatural status of the genus Tubifera (Myxomycetes). Taxon 60: 221-222.

Leontyev DV, Fefelov KA. 2009. Tubulifera applanata. The new species from Eastern Europe and

Northern Asia. Bol. Soc. Mycol. Madrid 33: 115-127.

Nannenga-Bremekamp NE. 1991. A guide to temperate Myxomycetes. Biopress Ltd., Bristol.

Nelson RK, Scheetz RW, Alexopoulos CJ. 1982. Taxonomic studies in the myxomycetes. V.

Significance of peridial and spore ornamentations in the genus Tubifera, with a revised key to

the species. Mycologia 74: 541-548. http://dx.doi.org/10.2307/3792740

Neubert H, Nowotny W, Baumann K. 1993. Die Myxomyceten Deutschlands und des angrenzenden

Alpenraumes unter besonderer Beriicksichtigung Osterreichs. Bd. 1. Gomaringen, Baumann

Verlag.

MY COTAXON

http://dx.doi.org/10.5248/120.253

Volume 120, pp. 253-267 April-June 2012

Paralepistopsis gen. nov. and Paralepista

(Basidiomycota, Agaricales)

ALFREDO VIZZINI* & ENRICO ERCOLE

Dipartimento di Scienze della Vita e Biologia dei Sistemi - Universita degli Studi di Torino,

Viale Mattioli 25, I-10125, Torino, Italy

*CORRESPONDENCE TO: alfredo. vizzini@unito. it

ABSTRACT — Paralepistopsis, a new genus in Agaricales, is proposed for the rare toxic species,

Clitocybe amoenolens from North Africa (Morocco) and southern and southwestern Europe

and C. acromelalga from Asia (Japan and South Korea). Paralepistopsis is distinguished from

its allied clitocyboid genera by a Lepista flaccida-like habit, a pileipellis with diverticulate

hyphae, small non-lacrymoid basidiospores with a smooth slightly cyanophilous and

inamyloid wall, and the presence of toxic acromelic acids. Combined ITS-LSU sequence

analyses place Paralepistopsis close to Cleistocybe and Catathelasma within the tricholomatoid

clade. Our phylogenetic analysis further supports Lepista subg. Paralepista (= Lepista sect.

Gilva) as an independent clitocyboid evolutionary line. We recognize the genus Paralepista,

for which we propose twelve new combinations.

KEY worps — Agaricomycetes, erythromelalgia/acromelalgic syndrome, Clitocybe sect.

Gilvaoideae, /catathelasma clade

Introduction

The genus Clitocybe (Fr.) Staude traditionally encompassed saprobic agarics

that produce fleshy basidiomata with often adnate-decurrent lamellae, convex

to funnel-shaped pilei, usually a whitish to pinkish yellow spore print, and

smooth non-amyloid basidiospores (Kihner 1980, Singer 1986, Bas 1990,

Raithelhuber 1995, 2004).

Recent molecular studies that included a significant number of Clitocybe

species (Moncalvo et. al. 2002, Redhead et al. 2002, Matheny et al. 2006, Vizzini

et al. 2010a,b, 2011) have shown that taxa in this traditional genus do not form

a monophyletic group but rather a heterogeneous artificial set of disparate and

(in many cases) phylogenetically unrelated taxa (the so-called clitocyboid fungi

or Clitocybe s.l.).

Clitocybe amoenolens is a rare and rather localized species known thus far

only from Morocco, southern France, northern and central Spain, and central

254 ... Vizzini & Ercole

Italy (Malencon & Bertault 1975, Bon 1987, Poumarat & Neville 1993, Contu

et al. 1999, Moreau et al. 2001, Martinez et al. 2010). It was responsible, first

in France (Fourré 1997, Charignon & Garcin 1998, Moreau et al. 2001, Saviuc

et al. 2001, 2002) and then in Italy (Leonardi et al. 2002, Marinetti & Recchia

2005), for induced erythromelalgia (= acromelalgia syndrome sensu Saviuc et

al. 2001), a poisoning syndrome caused by the ingestion of C. acromelalga in

Japan (Nakamura et al. 1987). This syndrome is characterized by varying degrees

of tingling sensations, followed by intense burning pain in the extremities

but predominantly in the feet (Saviuc & Danel 2006). Clitocybe amoenolens

was confused with edible mushrooms in the Lepista flaccida complex (e.g.,

L. flaccida (Sowerby) Pat., L. lentiginosa (Fr.) Bresinsky, L. gilva (Pers.) Pat.) and

with Infundibulicybe gibba (Pers.) Harmaja (Fourré 1997, Moreau et al. 2001).

Clitocybe amoenolens shows features intermediate between Clitocybe s.s.

(smooth spores) and Lepista subg. Paralepista (cream spore-print, spotted

pileus, lamellae separable from context, and cyanophilic spores released in

tetrads), making its generic position uncertain.

Using recent French and Italian collections of C. amoenolens, we investigated

its phylogenetic position within the clitocyboid fungi through morphological

and molecular analyses and expanded its known geographic distribution.

Materials & methods

Morphology

Macromorphological features were described from fresh specimens. Microscopical

preparations from dried material were rehydrated in 3% KOH and stained in Congo red,

Cresyl Blue, Cotton Blue and Melzer’s reagent.

Basidiospore measurements are based on means of 120 spores from prints (four

collections), stained in Melzer’s reagent. The basidia width was measured at the

widest part and the length from the apex (sterigmata excluded) to basal septum. The

following abbreviations are used in text: L = number of entire lamellae; | = number of

lamellulae between each pair of entire lamellae; Q = the quotient of length and width

of the spores in side view; Qm = average quotient. Colour terms in capital letters (e.g.

Pale Ochraceous-Buff) are those of Ridgway (1912). Herbarium acronyms follow

Thiers (2011). Author citations follow Index Fungorum (http://www.indexfungorum.

org/authorsoffungalnames.htm). All examined collections are housed at TO. The new

genus and new combinations are deposited in MycoBank (http://www.mycobank.org/

DefaultPage.aspx ).

DNA extraction, PCR amplification, and DNA sequencing

Genomic DNA was isolated from 1 mg of herbarium material (TaBLE 1) using the

DNeasy Plant Mini Kit (Qiagen, Milan Italy) according to the manufacturer’s instructions.

Universal primers ITS1F/ITS4 were used for the ITS region amplification (White et al.

1990, Gardes & Bruns 1993) and primers LROR/LR7 for the LSU rDNA amplification

(Vilgalys & Hester 1990, Vilgalys lab unpubl., http://www.botany.duke.edu/fungi/

Paralepistopsis gen, nov. and Paralepista ... 255

TABLE 1. Collections newly sequenced in this study.

GENBANK ACC. NUMBERS SOURCE, COUNTRY.

> >

pREClE® ITS LSU DATE, COLLECTOR

Paralepistopsis amoenolens 1 JQ585653 JQ585654 if Reuerics Pe st

Paralepistopsis amoenolens 2 JQ585655 — 2 Pena ree a

Paralepista flaccida 1 JQ585656 JQ585657 a Ron te ake

Paralepista flaccida 2 JQ585658 JQ585659 ae en

Paralepista gilva JQ585660 JQ585661 eee eaten

09/11/2011, A. Vizzini

mycolab). Amplification reactions were performed in a PE9700 thermal cycler (Perkin-

Elmer, Applied Biosystems) following Vizzini et al. (2010b). PCR products were purified

with the AMPure XP kit (Beckman) and sequenced by DINAMYCODE srl (Turin, Italy)

and MACROGEN Inc. (Seoul, Republic of Korea). Sequences were assembled and edited

with the phred/phrap/consed software suite. The sequences were submitted to GenBank

and their accession numbers are reported in TABLE | and Ficure 1.

Sequence alignment and phylogenetic analysis

The sequences obtained in this study were checked and assembled using Geneious

v5.3 (Drummond et al. 2010) and compared to those available in GenBank (http://www.

ncbi.nlm.nih.gov/Genbank/) using the blastn algorithm. Based on the blastn results,

sequences were selected according to the outcomes of recent phylogenetic studies on

Agaricales (Matheny et al. 2006, Binder et al. 2010, Vizzini et al. 2011). A combined

ITS and LSU sequence analysis was carried out using sequences from the same strain

or specimen. Xeromphalina campanella (GU320006 and GU320009) was used as

outgroup. Alignments were generated using MAFFT (Katoh et al. 2002) with default

conditions for gap openings and gap extension penalties. The sequence alignment, its

manual adjustment, and the best-fit models estimation follow Vizzini et al. (2010b).

The GTR+G and GTR+G substitution models were used in the ITS and LSU analyses,

respectively. A partitioned matrix was used in all the analyses. Molecular-phylogenetic

analyses were performed using the Bayesian Inference (BI) and Maximum Likelihood

(ML) approaches. BI using Monte Carlo Markov Chains (MCMC) was carried out

with MrBayes 3.1.2 (Huelsenbeck and Ronquist 2001). Four incrementally heated

simultaneous MCMC were run over 5.000.000 generations, under model assumption.

Trees were sampled every 500 generations resulting in an overall sampling of 10.001

trees. The “burn-in” value was evaluated using Tracer 1.5 (Rambaut & Drummond 2007).

The first 20% of trees was discarded as “burn-in”. For the remaining trees, a majority rule

consensus tree showing all compatible partitions was computed to obtain estimates for

Bayesian Posterior Probabilities (BPP). ML estimation was performed through RAxML

v.7.0.4 (Stamatakis 2006) with 1.000 bootstrap replicates (Felsenstein 1985) using the

256 ... Vizzini & Ercole

LL ema Callistosporium graminicolor 0Q484065, AY745702

Callistosporium xanthophyllum AF325667, AF261406

— Callistosporium luteoolivaceum AF 325666, AF261405

0.9/- Pseudoomphalina pachyphylla Hm191750, #

oe Entoloma abortivum F 3940730, AF223169

4/100 Entoloma sinuatum Gu289652, AY207199

Less —— = —__ Entoloma cetratum £U784213, AY700180

Entoloma prunuloides 0206983, AY691891

199 citopilus giovanellae £F 413030, EF413027

Clitopilus prunulus 0202272, AY700181

4/100 ->——- Rhodocybe fallax AF357018, AF223165

Rhodocybe mundula Da494694, AY700182

400 ->—— Hypsizygus tessulatus Da917653, DQ917664

—— Ossicaulis lignatilis 09825426, AF261397

Tricholomella constricta 0Q825429, AF223188

Lyophyllum palustris AF357043, AF223199

a7 Lyophyllum sykosporum AF357051, AF357073

i Lyophyllum boudieri 00825427, 00825430

atoo Clitocybe rhizophora JF907812, #

Clitocybe vermicularis JF907813, #

— Clitocybe vermicularis JF907817, #

400

4/100

Clitocybe subditopoda 0a202269, AY691889

litocybe candicans DQ202268, AY645055

1100 r Clitocybe puberula FM877682, FM877681

Clitocybe puberula FM877683, FM877680

Clitocybe ditopa JF907805, #

Clitocybe phaeophthalma Fms77684, FM877679

-/95¢ Clitocybe odora #, AY206714

0.92/- Clitocybe phyllophila #, AY207157

Clitocybe ruderalis #, AY207159

Collybia tuberosa AF274376, AF261385

0.84). Clitocybe fragrans JF907811, AY207153

er Lepista nuda AF357062, AY207223

a Lepista personata AF241522, # 7 7

epista saeva #, AY207224

rel Lepista Sordida FJ770391, AY207225 Clitocybe/L epista

{C Clitoc ‘be metachroa JF907806, AY207155

ar Clitocybe vibecina JF907821, AY207160

09507 -— Lepista irina 0Q221109, DQ234538

0.76/- (= Se ees Lepista Ovispora #, GQ289207

Clitocybe dealbata AF357061, AF223175

70 Clitocybe nebularis AF357063, AF223217

L_usoil Gitocybe nebularis DQ486691, DQ457658

Clitocybe nebularis AY521248, AY586685

1n00+ Clitocybe amoenolens 1 JQ585653, JQ585654

sr i Paralepistopsis

223§ "coy teu amoenotens 2 sastets pistopsis | /catathelasma clade

Cleistocybe vernalis EF416917, EF416916

Catathelasma imperiale Gag81498, AF261402

Catathelasma ventricosum 0a486686, DQ089012

Tricholoma focale AF319437, TFU76460

Tricholoma imbricatum AF319426, TIU76458

Lise Tricholoma pardinum AF319427, TPU76462

Tricholoma venenatum AF319433, TVU76463

Neohygrophorus angelesianus 00494678, DQ470814

—f__,_,, Clitocybe alkaliviolascens JF907807, #

100, Infundibulicybe gibba Da490635, DQ457682

Clitocybe geotropa #, AY207154

4 Omphalina pyxidata JN944402, JN944403

Pseudoclitocybe cyathiformis JF926522, JF926523

0.6107

L___ioof '— Pseudoclitocybe expallens JF926524, JF926525

ove M

197

1/99

193

lusumecia bettlakensis JF926520, JF926521

4/98 pepista faccida #, eNeelerd

epista flaccida 2 40585658, JQ585659 H

-— Lepista Haccids 1 JOBSSESH: JQ585657 | P. are al lepista

7 1/100" ——————- Lepista gilva JQ585660, JQ585661

4/400 Cc Arthenia auriscalpium AAU66428 (ITS and LSU fragments)

0.99/61 ; Arrhenia lobata ALU66429 (ITS and LSU fragments)

400 -——— My cena galericulata Da404392, AY647216

Mycena plumbea 00494677, DQ470813

Xeromphalina campanella Gu320006, GU320009

0.04 expected change per sites

FicurE 1. Tricholomatoid clade. Bayesian phylogram obtained from the combined ITS-LSU

sequence alignment. Support values for clades that are supported in either the Bayesian (Posterior

Probabilities values - BPP) or Maximum likelihood (ML Bootstrap percentage - MLB) analyses

are indicated. BPP > 0.70 and MLB > 50% are given above branches. Numbers (1, 2) refer to the

Paralepistopsis and Paralepista collections reported in TABLE 1.

GTRGAMMA algorithm for both ITS and LSU to perform a tree inference and search

for a good topology. Support values from bootstrapping runs (MLB) were mapped on

the globally best tree using the “-fa” option of RAxML and “-x 12345” as a random seed

to invoke the novel rapid bootstrapping algorithm. Only BPP values over 0.70 and MLB

over 50% are reported in the resulting tree (Fic. 1). Pairwise % identity values of ITS

sequences were calculated using MEGA 5.0 (Tamura et al. 2011).

Results

Molecular results

The combined dataset comprises a total of 71 taxa (including 66 from

GenBank) and is 2412 base pairs long. The ITS and LSU datasets are 796 and

1616 base pairs long, respectively. Topologies of the combined ITS and LSU

Bayesian and Maximum Likelihood trees are congruent (Fic. 1).

In both analyses the two Clitocybe amoenolens collections (ITS pairwise

identity value = 99.9%) clearly cluster with the C. acromelalga collection

(BPP 1 and MLB 100%) in the /catathelasma clade, a monophyletic group

Paralepistopsis gen, nov. and Paralepista ... 257

in the tricholomatoid clade (Matheny et al. 2006, Ammirati et al. 2007). The

ITS pairwise identity value between the C. acromelalga and C. amoenolens

sequences is 94.4%; accepting an intraspecific variability lower than 3% (Nilsson

et al. 2008), C. amoenolens and C. acromelalga should be considered distinct

species. Cleistocybe vernalis Ammirati et al. cluster sister to the C. acromelalga/

C. amoenolens pair (BPP 0.82); Catathelasma Lovejoy is basal to this group with

low BPP and MLB values.

Clitocybe amoenolens and C. acromelalga have no phylogenetic relationship

with other Clitocybe species. Accordingly, we propose to establish for them a

new genus, Paralepistopsis.

Lepista flaccida and L. gilva are not phylogenetically related to the other

Lepista species.

Taxonomy

Paralepistopsis Vizzini, gen. nov.

MycoBank MB 564340

A Paralepista differt sporis levis, haud subglobosis, atque praesentia acidi acromelalgici

qua de causa venenatae species sunt. et in stuctura molecularis (ITS-spatiis internis

transcriptis et LSU DNA).

TYPE SPECIES — Clitocybe amoenolens Malencon

EryMOLoGy — named in reference to its resemblance to Paralepista species.

Basidiomata agaricoid (with distinct pileus, lamellae and stipe), resembling

those of Paralepista gilva, veils absent, spore-print whitish to cream,

basidiospores thin-walled, smooth, inamyloid and slightly cyanophilous, pileal

surface a cutis of repent to interwoven, cylindrical hyphae, clamp-connections

present, no sarcodimitic texture in any part of the basidioma. On the ground,

never on wood.

Paralepistopsis amoenolens (Malencon) Vizzini, comb. nov. Fic. 2

MycoBank MB 564341

= Clitocybe amoenolens Malencon, in Malencon & Bertault, Flore des champignons

superieurs du Maroc 2 - Trav. Inst. Sci. Chérifien, Sér. Bot. Biol. Vég. 33: 141 (1975).

SELECTED DESCRIPTIONS — Malencon & Bertault (1975: 138-141); Moreau et al. (2001:

99-100, 101-103).

SELECTED ICONOGRAPHY — Malencon & Bertault (1975: pl. 8); Poumarat & Neville

(1993: 48); Martinez et al. (2010: fig. 2, p. 104).

PitEus (2-)3.5-7(-8) cm diam., fleshy, sub-elastic, hemispherical to convex

at first, becoming plano-convex and applanate, finally plano-concave, at times

broadly umbonate; margin narrowly inrolled and decurved at first, remaining

inrolled for a long time, then expanding to become somewhat wavy, shortly

sulcate-striate or corrugated; not hygrophanous, occasionally appearing

258 ... Vizzini & Ercole

hygrophanous when water soaked; surface slightly viscid when moist, at first

entirely whitely pruinose, then pubescent-pruinose only at margin, typically

diffracted-scaly near the disc, sometimes corrugated, wrinkled and areolate,

later subglabrous often with more or less concentrically arranged watery, drop-

like spots, especially near margin (as many Lepista species); at first cream-beige

coloured (Capucine Buff, Pale Ochraceous-Salmon, Pale Ochraceous-Buff),

then pinkish beige to rusty orange (Ochraceous-Buff, Zinc Orange) towards

the centre. LAMELLAE crowded to close (L = 40-48(-50), 1 = (0—)1-2(-3)),

thick, interspersed with lamellulae, (2-)3-4(-5) mm broad, decurrent, easily

separable from the pileus context, at times intervenose or forked towards the

stipe, at first whitish then yellowish ochre (Ivory Yellow, Chamois) to pinkish

beige (Pale Ochraceous-Salmon, Pale Ochraceous-Buff); edges even, entire,

concolorous. STIPE (2.5—)3-3.5(-5) cm long, 0.7-1.3 cm thick, short, central

or subexcentric, equal or with a somewhat enlarged base, straight or recurved,

concolorous with the pileus or slightly paler, minutely white pruinose at apex,

glabrous elsewhere, stuffed with white medulla, becoming hollow; the base

often with copious whitish tomentum with adhering Pinaceae needles and

woody debris. CONTEXT 4-11(-14) mm thick at disc, elastic, white in the

pileus, whitish cream (Capucine Buff) in the stipe cortex, unchanging; taste

mild, fungoid, subfarinaceous, slightly bitter-farinaceous after long mastication;

odour strong, aromatic, floral, reminiscent of Inocybe corydalina, Tricholoma

caligatum, Lepista irina, or Entoloma ameides. SPORE PRINT whitish to pale

cream (Light Buff, Pale Pinkish Buff).

BASIDIOSPORES (3.8—)4.0-5.4(-5.6) x (2.3-)3.2-4.0(-4.3) um (n = 120),

on average 4.8 x 3.4 um, Q = 1.3-1.7, Qm = 1.42, broadly ellipsoid, hyaline,

thin-walled, smooth, slightly cyanophilous, inamyloid, non-dextrinoid, usually

with only a single oil drop and a distinct truncated apiculus up to 0.7 um

long, mostly adhering together in tetrads in dried specimens. Basrip1A (25-)

30-37(-38) x 5-6(-7) um, cylindro-clavate, usually four-spored, occasionally

two-spored, sterigmata up to 5 um long. HYMENOPHORAL TRAMA regular in

young stages, but subirregular in mature basidiomata, consisting of hyaline,

elongated, cylindrical hyphae 4-6(-8) um broad. PLEUROcystTiD1A absent.

LAMELLA EDGES fertile, with rare, scattered cells, not well differentiated from

basidia, 15-45(-60) x 2.5-5(-7) um, cylindric to subfusiform or sublageniform,

often curved and flexuous, sometimes forked at apex, hyaline, thin-walled.

PILEIPELLIS duplex: upper layer (suprapellis) a soon disappearing, slightly

gelatinous thin cutis (10-30 um thick), of cylindrical hyphae, 1-3 um broad;

lower layer (subpellis) (150-350 um thick) composed of densely arranged

parallel to slightly interwoven hyphae (4-6 um in diam.), terminal elements

scattered, erect and repent cylindrical to fusiform-lageniform, sometimes

with short lateral outgrowths. PILEITRAMA consisting of cylindrical or slightly

Paralepistopsis gen, nov. and Paralepista ... 259

FiGuRE 2. Paralepistopsis amoenolens. a. Basidiomata. b. Diffracted-scaly pileus with minute

squamules. c. Areolate-corrugated pileus. d. Sulcate-striate pileus margin (red arrow), lamellae,

and stipe. e. Basidiomata. [a—b from TO AV2007; c-d from TO AV2006 ; e from TO AV2004.

Bars = 1 cm]

260 ... Vizzini & Ercole

inflated, smooth hyphae, (4-)5-9(-12) um broad, irregularly arranged, and

with aeriferous lacunae. STIPITIPELLIS a cutis consisting of hyphae 3-4 um

broad; towards the stipe apex with numerous thin-walled, flexuous, cylindrical

to subfusiform caulocystidioid elements, 20-35 x 3-6 um. STIPITITRAMA

formed by hyphae 6-7 um broad. THROMBOPLEROUS HYPHAE (= oleiferous

hyphae sensu Clémencon 2004) present especially in subpellis, pileus and

hymenophoral trama. PIGMENT (yellowish) usually parietal and intracellular

(cytoplasmatic) in the pileipellis; sometimes minutely incrusting or forming

extracellular masses and granules. CLAMP CONNECTIONS present at nearly all

septa.

HABITAT & DISTRIBUTION: scattered, gregarious, occasionally subcaespitose

on pinaceous needle-beds and debris, often together with Lepista flaccida, on

calcareous soil. Autumn. Very rare; known only from Morocco, France, Spain,

and Italy.

MATERIAL STUDIED: FRANCE, SAvorgE, HAUTE-MAURIENNE VALLEY, Lanslebourg-

Mont-Cenis, 02 Sept 2011, litter of Picea abies, 1400 m asl, leg. G. Moretto (TO AV2004);

14 Oct 2011, leg. G. Moretto (TO AV2005). - ITALY, PrEDMONT, HiGH SusaA VALLEY,

Salbertrand, Parco Naturale del Gran Bosco di Salbertrand, 20 Oct 2011, litter of Picea

abies, 1500 m asl, leg. A. Vizzini (TO AV2006); Sauze d’Oulx, Parco Naturale del Gran

Bosco di Salbertrand, 12 Nov 2011, litter of Larix decidua, 1550 m asl, leg. S. Anselmino

(TO AV2007).

Clitocybe amoenolens was originally described from Morocco, in the Middle

Atlas, growing among Cedrus atlantica litter in a high-altitude cedar forest

(1600-1700 ma.s.l.) mixed with Ilex aquifolium and Quercus ilex, on calcareous

soil (Malencon & Bertault 1975). It has recently been found in southern France

(Bon 1987, Poumarat & Neville 1993) and in the Maurienne Valley (Charignon

& Garcin 1998, Fourré 1997) in coniferous forests (Pinus sylvestris, Larix

decidua, Picea abies) and always on calcareous soil.

Clitocybe amoenolens has been responsible for several erythromelalgic-type

poisonings in the Maurienne Valley (Savoie, France) (Moreau et al. 2001, Saviuc

& Danel 2006). It also has been found in the Abruzzi region (Centre Italy) under

Pinus nigra (Contu et al. 1999), P. nigra, and Cedrus spp. (Leonardi et al. 2002)

and P. nigra and Larix decidua (Leonardi & Maggi 2007), and poisoning cases

referable to C. amoenolens have been recognised in this region (Leonardi et al.

2002, Marinetti & Recchia 2005). Finally, the species has been reported from

Spain (Martinez et al. 2010), where it was collected in the autonomous regions

of La Rioja (Picea abies) and Castilla-La Mancha (Pinus pinaster and Cupressus

arizonica or P. nigra and Quercus petraea), close to the north and centre of

Spain, respectively.

Our collections are the first record from northern Italy. According to our

observations and bibliographic data, this species seems strictly restricted to

Pinaceae in higher altitude thermophilic forests on calcareous soils.

Paralepistopsis gen, nov. and Paralepista ... 261

Paralepistopsis acromelalga (Ichimura) Vizzini, comb. nov.

MycoBank MB564342

= Clitocybe acromelalga Ichimura, Bot. Gaz. (Tokyo) 65: 110 (1918).

SELECTED DESCRIPTIONS — Ichimura (1918: 110); Moreau et al. (2001: 109-111).

SELECTED ICONOGRAPHY — Guez (2010: (http://www.mycodb.fr/fiche.php?genre=Clit

ocybe&espece=acromelalga ).

HABITAT & DISTRIBUTION: Clitocybe acromelalga, described from Japan

(Ichimura 1918, Imazeki & Hongo 1957, Imazeki et al. 1988; Romagnesi 1989,

Guez 1990), also occurs in South Korea (Lee & Hong 1985). It was reported

as growing on both angiosperm (Phyllostachys bambusoides, Acer palmatum,

Zelkova serrata) and gymnosperm (Cryptomeria japonica) litter.

Phylogeny and specific delimitation

In our combined ITS-LSU phylogenetic tree (Fic. 1) Clitocybe amoenolens

and its sister C. acromelalga are not closely related to C. nebularis (Batsch)

P. Kumm., the type of the genus Clitocybe (Redhead et al. 2002), nor to other

Clitocybe species or allied taxa. As these two species represent a new phyletic

line of clitocyboid fungi, it seems most appropriate to transfer them to the new

genus Paralepistopsis.

Based on its habit, coloured lamellae and small spores, Bon (1997) and

Moreau et al. (2001) placed C. amoenolens traditionally in subg. Clitocybe

sect. Gilvaoideae Harmaja, where it occupies an isolated position. Contu et

al. (1999), focusing on hymenial features (basidia longer than 30 um), placed

it in subgen. Hygroclitocybe Bon sect. Clavipedes Harmaja, a subgenus shown

in recent molecular analyses (Redhead et al. 2002, Vizzini et al. 2011) to be

artificial and heterogeneous.

The traditionally defined Lepista (Fr.) W.G. Sm. —clitocyboid fungi with

a pinkish yellow spore print, usually separable lamellae, and inamyloid

cyanophilous ornamented [verruculose to spiny] basidiospores (Singer 1986,

Bon 1997, Consiglio & Contu 2003)— is a polyphyletic genus (Fic. 1). The

species of Lepista subg. Paralepista (= Lepista sect. Gilva), which combine very

crowded decurrent lamellae with subglobose to largely ellipsoidal spores, are

not closely related either to Lepista s.s. or to other taxa in the tricholomatoid

clade. Consequently we accept this lineage as a distinct genus and propose

Paralepista for Lepista flaccida, L. gilva and allies. Following Bigelow (1985),

Bon (1991), and Raithelhuber (2004), we list below all the taxa accepted in

Paralepista:

Paralepista Raithelh., Gattung Clitocybe 1: 17 (1981).

TYPE SPECIES — Agaricus inversus Scop.

= Lepista subg. Paralepista (Raitelh.) Bon, Doc. Mycol. 26(102): 18 (1996).

= Clitocybe sect. Eulepistae Singer, Ann. Mycol. 41: 40 (1943).

262 ... Vizzini & Ercole

= Lepista sect. Gilva Harmaja, Karstenia 18: 53 (1978).

“Lepista sect. Eulepista” Konrad & Maubl., Icon. Select. Fung. 6(10): 350 (1936), nom. inval.

“Lepista sect. Inversae” Singer & Clémencon, Nova Hedwigia

23: 310 (1973 [“1972”]), nom. inval.

Paralepista abdita (Dérfelt) Vizzini, comb. nov.

MycoBank MB 564343

= Lepista abdita Dorfelt, Boletus 1(2): 37 (1997).

Paralepista ameliae (Arcang.) Vizzini, comb. nov.

MycoBAnk MB 564344

= Clitocybe spinulosa var. ameliae Arcang., Nuovo Giorn. Bot. Ital. 21: 434 (1889).

Paralepista biformis (Peck) Vizzini, comb. nov.

MycoBANnkK MB 564345

= Clitocybe biformis Peck, Bull. N.Y. St. Mus. 150: 25 (1911).

Paralepista concentrica Raithelh., Metrodiana 23: 122 (1996).

Paralepista femoralis (H.E. Bigelow) Vizzini, comb. nov.

MycoBank MB 564346

= Clitocybe femoralis H.E. Bigelow, Sydowia 36: 14 (1983).

Paralepista flaccida (Sowerby) Vizzini, comb. nov.

MycoBank MB 564347

= Agaricus flaccidus Sowerby, Col. Fig. Engl. Fungi 2: pl. 185 (1799).

Paralepista flaccida var. fibrillosa (Malencon) Vizzini, comb. nov.

MycoBank MB 564348

= Clitocybe flaccida var. fibrillosa Malencon, in Malencon & Bertault,

Flore des champignons superieurs du Maroc 2 - Trav. Inst.

Sci. Chérifien, Sér. Bot. Biol. Vég. 33: 157 (1975).

Paralepista gilva (Pers.) Vizzini, comb. nov.

MycoBANnkK MB 564349

= Agaricus gilvus Pers., Syn. Meth. Fung.: 448 (1801).

“Paralepista gilva” Raithelh., Metrodiana 23: 117 (1996), nom. inval.

Paralepista inversa (Scop.) Raithelh., Gattung Clitocybe 1: 17 (1981).

= Agaricus inversus Scop., Fl. Carniol., Ed. 2, 2: 445 (1772).

Paralepista lentiginosa (Fr.) Vizzini, comb. nov.

MycoBank MB 564350

= Agaricus lentiginosus Fr., Epicr. Syst. Mycol.: 69 (1838).

Paralepista maculosa (Sacc.) Vizzini, comb. nov.

MycoBank MB 564352

= Agaricus maculosus Peck, Bull. Buffalo Soc. Nat. Sci. 1:

45 (1873), nom. illegit., non Pers. (1801).

= Clitocybe maculosa Sacc., Syll. Fung. 5: 183 (1887).

Paralepistopsis gen, nov. and Paralepista ... 263

Paralepista pseudoparilis (Enderle & Contu) Vizzini, comb. nov.

MycoBank MB 564353

= Lepista pseudoparilis Enderle & Contu, Beitr. Kenn. Pilze Mittel. 13: 12 (2000).

Paralepista repanda (Raithelh.) Raithelh., Metrodiana 23: 121 (1996).

= Lepista repanda Raithelh., Metrodiana 14: 21 (1986 [“1985”]).

Paralepista shafferi (H.E. Bigelow) Vizzini, comb. nov.

MycoBank MB 564354

= Clitocybe shafferi H.E. Bigelow, Beih. Nova Hedwigia 81: 339 (1985).

Paralepista splendens (Pers.) Vizzini, comb. nov.

MycoBank MB 564355

= Agaricus splendens Pers., Syn. Meth. Fung.: 452 (1801).

Discussion

Paralepistopsis species are characterized by a habit (decurrent and crowded

lamellae) and colours (ochre-orange tinges) reminiscent of Paralepista or

Infundibulicybe Harmaja, a whitish to cream spore print, smooth cyanophilic

spores often arranged in tetrads in dried specimens and rarely exceeding

5(-6) um in length. Paralepista differs in having strongly ornamented spores

(Raithelhuber 1995, 2004); Infundibulicybe is distinguished by smooth

lacrymoid spores with confluent bases and cyanophobic spore walls (Harmaja

2003).

Paralepistopsis clusters with Cleistocybe Ammirati et al. and Catathelasma

in the /catathelasma clade. Because of the low resolution and lack of BPP and

MLB support within the tree, a more precise, accurate position for the new

genus could not be suggested. The presence of decurrent lamellae, confluent

pileus and stipe, pale reddish brown colouration, and growth on soil are

characters shared by Paralepistopsis, Cleistocybe, and Catathelasma. Cleistocybe

and Catathelasma are distinguished from Paralepistopsis mainly by a partial

veil, divergent to interwoven hymenophoral trama, and larger cyanophobic

spores (Ammirati et al. 2007, Vizzini 2009); in addition Catathelasma spores

are amyloid (Singer 1986).

Paralepistopsis amoenolens is delimited by a unique combination of macro-

/micromorphological and chemical features, such as i) a strong aromatic, floral

odour reminiscent of Tricholoma caligatum, Inocybe corydalina, Lepista irina,

and Entoloma ameides caused by volatile metabolites identified by Fons et al.

(2006) as methyl-(E)-cinnamate (also a key odorant of T. caligatum), methyl-

benzoate, (E)-nerolidol, and methylanthranilate; ii) lamellae easily separating

from the pileus context; iii) a cream coloured spore print; iv) smooth cyanophilic

spores often arranged in tetrads; v) basidia reaching 35-40 um; vi) pileipellis

hyphae with short diverticula; vii) abundant thromboplerous hyphae; viii) and

the presence of the toxic metabolite, acromelic acid A, a powerful neurotoxic

264 ... Vizzini & Ercole

amino acid responsible for erythromelalgic poisoning and _ structurally

homologous with kainic acid (a strong agonist of non-N-methyl-D-aspartate

glutamate receptor subtypes) and domoic acid (Bessard et al. 2004).

Paralepistopsis acromelalga differs from P. amoenolens morphologically in a

darker pileus and stipe, a pileus that soon becomes depressed, more crowded

lamellae, a different odour, thromboplerous hyphae occurring only rarely,

smaller spores (Ichimura 1918, Romagnesi 1989, Guez 1990, Miyauchi 1998,

Moreau et al. 2001), and a more complex metabolite pattern (presence of

acromelic acids A—-E with 19 other toxins; Konno et al. 1983, 1988; Fushiya et

al. 1990, 1992; Saviuc & Danel 2006). Additionally, our analyses show only a

91% pairwise ITS sequence identity between P. acromelalga and P. amoenolens.

Singer (1986) transferred P acromelalga to the heterogeneous genus Neoclitocybe

Singer based on the presence of rare diverticulate hyphae in the pileipellis.

Based on their small spores and the Paralepista-like habit, C. gilvaoides

Kauffman and C. gracilis (H.E. Bigelow & A.H. Sm.) Harmaja (sect. Gilvaoideae)

from the coniferous forests of North America and Scandinavia (Bigelow 1985,

Harmaja 1969) may also belong to Paralepistopsis, but more recently collected

specimens are needed to perform molecular and biochemical analyses.

Acknowledgements

We would like to thank Vladimir Antonin (Moravian Museum, Brno, Czech

Republic), Joe Ammirati (University of Washington, Seattle, USA), and Shaun Pennycook

(Auckland, New Zealand) for their pre-submission reviews. Our most sincere thanks

are due to Marco Contu (Olbia, Italy) for helpful suggestions and comments.

Literature cited

Ammirati JE, Parker AD, Matheny PB. 2007. Cleistocybe, a new genus of Agaricales. Mycoscience

48: 282-289. http://dx.doi.org/10.1007/s10267-007-0365-5

Bas C. 1990. Tricholomataceae R. Heim ex Pouz. 65-70, in: C Bas et al. (eds). Flora Agaricina

Neerlandica 2. A.A. Balkema, Rotterdam.

Bessard J, Saviuc P, Chane-Yene Y, Monnet S, Bessard G. 2004. Mass spectrometric determination

of acromelic acid A from a new poisonous mushroom: Clitocybe amoenolens. J. Chromatogr.

A. 1055: 99-107. http://dx.doi.org/10.1016/j.chroma.2004.08.133

Bigelow HE. 1985. North American species of Clitocybe. Part Il. Beih. Nova Hedwigia 81:

281-471.

Binder M, Larsson K.-H, Matheny PB, Hibbett DS. 2010. Amylocorticiales ord. nov. and Jaapiales

ord. nov.: early diverging clades of Agaricomycetidae were dominated by corticioid forms.

Mycologia 102: 865-880. http://dx.doi.org/10.3852/09-288

Bon M. 1987. Quelques espéces intéressantes étudiées au stage FMDS de Saint Germain Monts

dOr. Bull. Fed. Mycol. Dauphiné-Savoie 105: 28-30.

Bon M. 1997. Les clitocybes, omphales et ressemblants. Doc. Mycol., mémoires hors-série 4:

1-174.

Charignon Y, Garcin R. 1998. Un nouveau champignon toxique en France. Bull. Féd. Mycol.

Dauphiné-Savoie 149: 11-14.

Paralepistopsis gen, nov. and Paralepista ... 265

Clémencon H. 2004. Cytology and plectology of the Hymenomycetes. Bibl. Mycol. 199: 1-488.

Consiglio G, Contu M. 2003. Il genere Lepista in Italia. Riv. Micol. 46: 131-176.

Contu M, Signorello P, Anastase A. 1999. Clitocybe amoenolens Mal. in Abruzzo con osservazioni

sulla sua posizione sistematica. Micol. Veget. Medit. 48: 16-18.

Drummond AJ, Ashton B, Buxton $, Cheung M, Cooper A, Duran C, Field M, Heled J, Kearse M,

Markowitz S, Moir R, Stones-Havas S, Sturrock S, Thierer T, Wilson A. 2010. Geneious v5.3.

Available from http://www.geneious.com/.

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution

39: 783-791. http://dx.doi.org/10.2307/2408678

Fons F, Rapior S, Fruchier A, Saviuc P, Bessiere J. 2006. Volatile composition of Clitocybe amoenolens,

Tricholoma caligatum and Hebeloma radicosum. Cryptogam. Mycol. 27: 45-55.

Fourré G. 1997. Intoxications: un sosie du Lepista inversa a produit en Savoie un terrifiant

syndrome...japonais. Bulletin de la Société Mycologique du Massif d’Argenson 16: 6-11.

Fushiya S, Sato S, Kazasawa T, Kusano G, Nozoe S. 1990. Acromelic acid C. A new toxic constituent

of Clitocybe acromelalga: a efficient isolation of acromelic acids. Tetrahedron Lett. 31:

3901-3904. http://dx.doi.org/10.1016/S0040-4039(00)97501-4

Fushiya S, Sato S, Kera Y, Nozoe S. 1992. Isolation of acromelic acids D and E from Clitocybe

acromelalga. Heterocycles 34: 1277-1280. http://dx.doi.org/10.3987/COM-92-6065

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes - application

to the identification of mycorrhizae and rusts. Mol. Ecol. 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Guez D. 1990. Apergu sur la flore mycologique du Japon. Bull. Fed. Mycol. Dauphiné-Savoie 116:

12-16;

Harmaja H. 1969. The genus Clitocybe (Agaricales) in Fennoscandia. Karstenia 10: 5-168.

Harmaja H. 2003. Notes on Clitocybe s. lato (Agaricales). Ann. Bot. Fenn. 40: 213-218.

Huelsenbeck JP, Ronquist FE. 2001. MrBayes: Bayesian inference of phylogeny. Bioinformatics 17:

754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

Ichimura T. 1918. A new poisonous mushroom. Bot. Gaz. (Tokyo) 65: 109-111.

http://dx.doi.org/10.1086/332195

Imazeki R, Hongo T. 1957. Coloured illustrations of mushrooms of Japan, vol. I. Hoikusha Publ.,

Osaka.

Imazeki R, Otani Y, Hongo T. 1988. Fungi of Japan. Yama-Kei publ., Tokyo.

Katoh K, Misawa K, Kuma K, Miyata T. 2002. MAFFT: a novel method for rapid multiple sequence

alignment based on fast Fourier transform. Nucl. Acids Res. 30: 3059-3066.

http://dx.doi.org/10.1093/nar/gkf436

Konno K, Shirahama H, Matsumoto T. 1983. Isolation and structure of acromelic acid A and B.

New kainoids of Clitocybe acromelalga. Tetrahedron Lett. 24: 939-942.

http://dx.doi.org/10.1016/S0040-4039(00)81571-3

Konno K, Hashimoto K, Ohfune Y, Shirahama H, Matsumoto T. 1988. Acromelic acids A and B.

Potent neuroexcitatory amino acids isolated from Clitocybe acromelalga. J. Am. Chem. Soc.

110: 4807-4815. http://dx.doi.org/10.1021/ja00222a044

Kithner R. 1980. Les Hyménomycetes agaricoides. Bull. Soc. Linn. Lyon 49, num. spéc.:1-1027.

Lee JY, Hong SW. 1985. Illustrated flora and fauna of Korea. No. 28. Mushrooms. Ministry of

Culture and Education, Korea.

Leonardi M, Maggi S. 2007. Nuova segnalazione per Clitocybe amoenolens in Abruzzo. Micol.

Veget. Medit. 22: 80.

Leonardi M, Ciulli G, Pacioni G, Recchia G. 2002. Una intossicazione collettiva da Clitocybe

amoenolens riconducibile alla sindrome acromelalgica. Micol. Veget. Medit. 17: 133-142.

266 ... Vizzini & Ercole

Malencon G, Bertault R. 1975. Flore des champignons supérieurs du Maroc, tome 2. Trav. Inst. Sci.

Chérifien, Sér. Bot. Biol. Vég. 33: 1-540.

Marinetti V, Recchia G. 2005. Nuovi casi di sindrome acromelalgica in Abruzzo. Boll. Gr. micol. G.

Bres. (n.s.) 48: 39-43.

Martinez F, Martinez R, Meléndez A, Pérez Del Amo CM. 2010. Clitocybe amoenolens. Primera cita

para Espajia. Bol. Soc. Micol. Madrid 34: 103-112.

Matheny PB, Curtis JC, Hofstetter V, Aime MC, Moncalvo JM, Ge ZW, Yang ZL, Slot JC, Ammirati

JE, Baroni TJ, Bougher NL, Hughes KW, Lodge DJ, Kerrigan RW, Seidl MT, Aanen DK, DeNitis

M, Daniele GM, Desjarden DE, Kropp BR, Norvell LL, Parker A, Vellinga EC, Vilgalys R,

Hibbett DS. 2006. Major clades of Agaricales: a multi-locus phylogenetic overview. Mycologia

98: 982-995. http://dx.doi.org/10.3852/mycologia.98.6.982

Miyauchi S. 1998. Comparison Clitocybe acromelalga with Clitocybe sp. collected in France.

Rapport of the Nagaoka University of Technology, sect. Bio-Ingeneering Kamitomioka 1603-1,

Nagaoka, Japan.

Moncalvo J-M., Vilgalys R, Redhead SA, Johnson JE, James TY, Aime MC, Hofstetter V,

Verduin SJW, Larsson E, Baroni TJ, Thorn RG, Jacobsson S, Clémencon H, Miller OK Jr.

2002. One hundred and seventeen clades of euagarics. Mol. Phylogenet. Evol. 23: 357-400.

http://dx.doi.org/10.1016/S1055-7903(02)00027-1

Moreau PA, Courtecuisse R, Guez D, Garcin R, Neville P, Saviuc P, Seigle-Murandi F. 2001. Etude

taxinomique d’une espéce toxique: Clitocybe amoenolens Malengon. Cryptogam. Mycol. 22:

95-117. http://dx.doi.org/10.1016/S0181-1584(01)80003-8

Nakamura K, Shoyama F, Toyama J, Tateishi K. 1987. Empoisonnement par le Dokou-sassa-ko

(Clitocybe acromelalga). Jpn. J. Toxicol. 1: 35-39.

Nilsson RH, Kristiansson E, Ryberg M, Hallenberg N, Larsson K-H. 2008. Intraspecific ITS

variability in the Kingdom Fungi as expressed in the International Sequence Databases and its

implications for molecular species identification. Evol. Bioinf. 4: 193-201.

Poumarat S, Neville P. 1993. Espéce de la zone du Quercus ilex au Maroc, montagnarde en France,

Clitocybe amoenolens Malencon. Bull. Fédér. Assoc. Mycol. Médit. 4: 16-19.

Redhead SA, Lutzoni F, Moncalvo JM, Vilgalys R. 2002. Phylogeny of agarics: partial systematics

solutions for core omphalinoid genera in the Agaricales (Euagarics). Mycotaxon 83: 19-57.

Romagnesi H. 1989. Curiosité mycologique: un champignon tortionnaire japonais: Clitocybe

acromelalga Ichimura. Bull. Soc. Mycol. France 105: 131-132.

Raithelhuber J. 1995. Trichterlinge mitteleuropas. Metrodiana 22(2): 52-94.

Raithelhuber J. 2004. Mitteleuropdische Trichterlinge. Gattungen Clitocybe, Pseudolyophyllum und

Paralepista. JH. Raithelhuber Aufenseiterverlag, Stuttgart.

Rambaut A, Drummond AJ. 2007. Tracer v1.4. [Available from http://beast.bio.ed.ac.uk/Tracer].

Ridgway R. 1912. Color standards and color nomenclature. Washington, D.C., published privately

(by the author). 43 p. + 53 color pls.

Saviuc PF, Danel VC, Moreau PA, Guez DR, Claustre AM, Carpentier PH, Mallaret MP,

Ducluzeau R. 2001. Erythromelalgia and mushroom poisoning. Clin. Toxicol. 39: 403-407.

http://dx.doi.org/10.1081/CLT-100105162

Saviuc PF, Danel VC, Moreau PA, Claustre AM, Ducluzeau R, Carpentier PH. 2002. Erythermalgie

soudaine: cherchez le champignon! Rev. Méd. Interne 23: 394-399.

http://dx.doi.org/10.1016/S0248-8663(02)00576-3

Saviuc PF, Danel VC. 2006. New syndromes in mushroom poisoning. Toxicol. Rev. 25: 199-209.

http://dx.doi.org/10.2165/00139709-200625030-00004

Singer R. 1986. The Agaricales in modern taxonomy, 4th ed. Koeltz Scientific Books, Koenigstein.

Paralepistopsis gen, nov. and Paralepista ... 267

Stamatakis A. 2006. RAxML-VI-HPC: Maximum Likelihood-based phylogenetic analyses with

thousands of taxa and mixed models. Bioinformatics 22: 2688-2690.

http://dx.doi.org/10.1093/bioinformatics/btl446

Tamura K, Peterson D, Peterson N, Stecher G, NeiM, Kumar S. 2011. MEGA5: molecular evolutionary

genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony

methods. Mol. Biol. Evol. 28: 2731-2739. http://dx.doi.org/10.1093/molbev/msr121

Thiers B. 2011. (continuously updated). Index Herbariorum: a global directory of public herbaria

and associated staff. New York Botanical Garden's Virtual Herbarium.

http://sweetgum.nybg.org/ih/

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several Cryptococcus species. J. Bacteriol. 172: 4238-4246.

Vizzini A. 2009. Due nuove combinazioni nel genere Cleistocybe (Basidiomycota, Agaricomycetes),

con validazione di Hygrophorus pleurotoides. Micol. Veget. Medit. 24: 95-98.

Vizzini A, MusumeciE, Murat C. 2010a. Trichocybe, anew genus for Clitocybe puberula (Agaricomycetes,

Agaricales). 2010. Fung. Diver. 42: 97-105. http://dx.doi.org/10.1007/s13225-010-0030-8

Vizzini A, Contu, M., Musumeci, E. and Ercole E. 2010b. A new taxon in the Infundibulicybe gibba

complex (Basidiomycota, Agaricales, Tricholomataceae) from Sardinia (Italy). Mycologia 103:

203-208. http://dx.doi.org/10.3852/10-137

Vizzini A, Contu M., Ercole E. 2011. Musumecia gen. nov. in the Tricholomatoid clade

(Basidiomycota, Agaricales) related to Pseudoclitocybe. Nord. J. Bot., Article first published

online: 20 OCT 2011. http://dx.doi.org/10.1111/j.1756-1051.2011.01169.x

White TJ, Bruns TD, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR Protocols. Academic

Press, London.

MYCOTAXON

http://dx.doi.org/10.5248/120.269

Volume 120, pp. 269-276 April-June 2012

Three new species of Septobasidium (Septobasidiaceae)

from Hainan Province in China

SUZHEN CHEN?” & LIN Guo'*

'State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing

100101, China

?Ocean University of China, Qingdao 266003, China

* CORRESPONDENCE TO: guol@im.ac.cn

AsstRact —Three new Septobasidium species are described: S. capparis on Capparis

membranifolia associated with Andaspis sp., S. reevesiae on Reevesia longipetiolata associated

with Lepidosaphes sp., and S. dacrydii on Dacrydium pierrei associated with Pinnaspis sp.

They were collected from Hainan Province, China.

Key worps —Pucciniomycetes, Septobasidiales, taxonomy

To date, nine new species and three new Chinese records of Septobasidium

have been discovered from Hainan Province (Chen & Guo 2011b,c,d, 2012, Lu

& Guo 2009a, 2010b,c). An additional three new species of Septobasidium are

reported as follows:

Septobasidium capparis S.Z. Chen & L. Guo, sp. nov. Fics 1-6

FUNGAL NAME FN570003

Differs from Septobasidium euryae-groffii by its thicker hymenium and larger basidia.

Type: China, Hainan Province, Changjiang, Baomeiling, alt. 250 m, on Capparis

membranifolia Kurz (Capparaceae), associated with Andaspis sp. (Diaspididae),

12.IV.2011, L. Guo 11599 (HMAS 263233, holotype).

Erymo.ocy: The epithet refers to the substrate plant genus, Capparis.

Basidiomata on branches, resupinate, 10-24 cm long, 1.5-4 cm wide, pale

cinnamon-brown or brown; margin determinate, surface smooth, cracked at

maturity. In section 1500-2000 um thick, composed of 3-8 layers. Subiculum

brown, 40-70 um thick. Pillars brown, 80-100 um high, 70-130 um wide,

branched outwards to form a hyphal layer, 500-550 um high. From hymenial

layer the fungal hyphae renew growth successively to form hyphal layers and

270 ... Chen & Guo

Fic. 1. Septobasidium capparis (HMAS 263233, holotype). Basidia and basidiospores.

hymenia. Hymenium brown, 100-170 um thick. Basidia arising directly from

the hyphae without a probasidial cell, cylindrical, straight or slightly curved,

4-celled, 45-56 x 8-12 um, hyaline or brown. Sterigmata conical, 6-8 x 3-5

um, hyaline or brown. Basidiospores reniform, 16-20 x 4.5-8 um, hyaline.

Haustoria consisting of irregularly coiled hyphae.

ComMENTSs: Septobasidium capparis is similar to S. euryae-groffii C.X. Lu &

L. Guo, which differs by its thinner hymenium (70-110 um) and smaller basidia

(20-45 x 5-8 um; Lu & Guo 2010a).

Septobasidium spp. nov. (China) ... 271

Fics. 2-6 Septobasidium capparis (HMAS 263233, holotype). 2. Basidioma on branch.

3-4. Basidia (arrows). 5. Haustoria. 6. Section of basidioma.

272 ... Chen & Guo

Fic. 7. Septobasidium reevesiae (HMAS 263427, holotype). Basidia and basidiospores.

Septobasidium reevesiae S.Z. Chen & L. Guo, sp. nov. Figs 7-12

FUNGAL NAME FN570004

Differs from Septobasidium henningsii by its upright pillars and thicker stratified

hymenial layer.

Type: China, Hainan Province, Bawangling Natural Reserve, alt. 1030 m, on Reevesia

longipetiolata Merr. & Chun (Sterculiaceae), associated with Lepidosaphes sp. (Diaspididae),

25.X1.2010, Y.E. Zhu & FE. He 518 (HMAS 263427, holotype).

Erymo.oey: The epithet refers to the substrate plant genus, Reevesia.

Basidiomata on branches, resupinate, 7.5-8.5 cm long, 3-6 cm wide, cinnamon-

brown; margin white, determinate, surface smooth, at first frequently forming

orbiculate or discoideous patches, confluent in the old stage. The dense pillars

are barely visible, especially near the margin. In section 1650-2200 um thick.

Subiculum brown, 30-50 um thick. Pillars fasciculate, brown, 700-1170 um

high, 40-100 um wide. Hymenial layer hyaline or brown, 730-1000 um thick,

often forming 2-4 strata, with a brown, 2-7 um thick horizontal layer between

the hymenia, and closely packed parallel upright hyphae. Basidia arising directly

from the hyphae without a probasidial cell, cylindrical, straight or slightly

curved, 4-celled, 37-55 x 8-13(-18) um, hyaline or brown; wall 1-2(-3) um

thick. Sterigmata conical, 9-11(-23) x 3-4.5 um. Basidiospores reniform,

15-16x5-6 um. Haustoria consisting of irregularly coiled hyphae.

ComMENTs: Septobasidium reevesiae is similar to S. henningsii Pat., which differs

by its thinner hymenial layer (60-400 um high) and characteristic slanting,

entangled, anastomosing pillars (Couch 1938).

Septobasidium spp. nov. (China) ... 273

mn IT mt

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10 ym

Fics. 8-12. Septobasidium reevesiae (HMAS 263427, holotype). 8. Basidiomata on branches.

9. Stratified hymenial layer. 10. Basidia (arrow). 11. Haustoria. 12. Section of basidioma.

274 ... Chen & Guo

Fic. 13. Septobasidium dacrydii (HMAS 263232, holotype). Basidia and basidiospores.

Septobasidium dacrydii S.Z. Chen & L. Guo, sp. nov. Figs 13-18

FUNGAL NAME FN570005

Differs from Septobasidium apiculatum by its thicker section with pillars.

Type: China, Hainan Province, Bawangling Natural Reserve, alt. 1050 m, on Dacrydium

pierrei Hickel (Podocarpaceae), associated with Pinnaspis sp. (Diaspididae), 24.X1.2010,

Y.F. Zhu & FE. He 529 (HMAS 263232, holotype).

Erymo oey: The epithet refers to the substrate plant genus, Dacrydium.

Basidiomata on branches, resupinate, 35 cm long, 1-6 cm wide, cinnamon-

brown or dark brown; margin determinate, surface smooth, peeled off in the

old stage, dark brown pillars and hyphal layers emerging. In section 1450-2000

um thick. Subiculum brown, 45-70 um thick. From the subiculum forming

pillars or hyphal layers. Pillars brown, higher or shorter, 850-1200 um high,

35-110 um wide, or 100-200 um high, 40-80 um wide. Hyphal layer, 700-800

uum high. Pillars and hyphal layers successively formed from the hyphal layer.

Hymenium hyaline, 85-140 um thick. Basidia arising directly from the hyphae

without a probasidial cell, cylindrical or clavate, straight or slightly curved,

3-celled, 30-40 x 9-10 um, hyaline. Sterigmata conical, 10-15 x 4-5 um,

hyaline. Basidiospores cylindrical or fusiform, 21-26x5-6.5 um, hyaline.

Haustoria consisting of irregularly coiled hyphae.

ComMENTs: Septobasidium dacrydii is similar to S. apiculatum Couch ex L.D.

Gomez & Henk, which differs by its thinner section (250-550 um thick) and

absence of pillars (Couch 1938).

Including the three new species reported in this paper, 46 Septobasidium species

have now been reported in China (Sawada 1933, Couch 1938, Teng 1963, Tai

Septobasidium spp. nov. (China) ... 275

14 15

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aad booed ha

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UATE

Fics. 14-18. Septobasidium dacrydii (HMAS 263232, holotype). 14. Basidioma on branch.

15. Basidia (arrow). 16. A basidiospore. 17. Haustoria. 18. Section of basidioma.

276 ... Chen & Guo

1979, Kirschner & Chen 2007, Lu & Guo 2009a,b,c, 2010a,b,c, 2011, Lu et al.

2010, Chen & Guo 201 1a,b,c,d, 2012).

Acknowledgements

The authors would like to express their deep thanks to Drs. Eric H.C. McKenzie

(Auckland, New Zealand) and Shuanghui He (Beijing Forestry University) for serving

as pre-submission reviewers, to Dr. Shaun Pennycook (Auckland, New Zealand)

for nomenclatural review, to Prof. Zhenyu Li (Institute of Botany, Chinese Academy

of Sciences) and Mr. Qing Chen (Bawangling Natural Reserve, Hainan Province)

for identifying the host plants, to Prof. Sanan Wu (Beijing Forestry University) for

identifying the scale insects, and to Mrs. Xiangfei Zhu for inking in line drawings. This

study was supported by the foundation of Ministry of Science and Technology of the

People’s Republic of China (No. 2006FY110500-5).

Literature cited

Chen SZ, Guo L. 2011la. Septobasidium sichuanense sp. nov. (Septobasidiaceae) from China.

Mycotaxon 115: 481-484. http://dx.doi.org/10.5248/115.481

Chen SZ, Guo L. 2011b. Septobasidium atalantiae sp. nov. (Septobasidiaceae) and S. henningsii new

to China. Mycotaxon 117: 291-296. http://dx.doi.org/10.5248/117.291

Chen SZ, Guo L. 201 1c. Septobasidium saurauiae sp. nov. (Septobasidiaceae) and S. pseudopedicellatum

new to China. Mycotaxon 118: 283-288. http://dx.doi.org/10.5248/118.283

Chen SZ, Guo L. 2011d. Septobasidium glycosmidis and S. albiziae spp. nov. (Septobasidiaceae)

from Hainan Province. Mycosystema 30: 861-864.

Chen SZ, Guo L. 2012. Three new species and three new Chinese records of Septobasidium

(Septobasidiaceae). Mycosystema 31 (in press).

Couch JN. 1938. The genus Septobasidium. Univ. of North Carolina Press, Chapel Hill. 480 p.

Kirschner R, Chen CJ. 2007. New reports of two hypophyllous Septobasidium species from Taiwan.

Fung. Sci. 22(1,2): 39-46.

Lu CX, Guo L. 2009a. Septobasidium maesae sp. nov. (Septobasidiaceae) from China. Mycotaxon

109: 103-106. http://dx.doi.org/10.5248/109.103

Lu CX, Guo L. 2009b. Two new species of Septobasidium (Septobasidiaceae) from China. Mycotaxon

109: 477-482. http://dx.doi.org/10.5248/109.477

Lu CX, Guo L. 2009c. Septobasidium annulatum sp. nov. (Septobasidiaceae) and Septobasidium

kameii new to China. Mycotaxon 110: 239-245. http://dx.doi.org/10.5248/110.239

Lu CX, Guo L. 2010a. Three new species of Septobasidium (Septobasidiaceae) from Gaoligong

Mountains in China. Mycotaxon 112: 143-151. http://dx.doi.org/10.5248/112.143

Lu CX, Guo L. 2010b. Two new species of Septobasidium (Septobasidiaceae) and S. pallidum new to

China. Mycotaxon 113: 87-93. http://dx.doi.org/10.5248/113.87

Lu CX, Guo L. 2010c. Two new species of Septobasidium (Septobasidiaceae) from Hainan province

in China. Mycotaxon 114: 217-223. http://dx.doi.org/10.5248/114.217

Lu CX, Guo L. 2011. Two new species of Septobasidium (Septobasidiaceae) from Gaoligong

Mountains in China. Mycotaxon 116: 395-400. http://dx.doi.org/10.5248/116.395

Lu CX, Guo L, Wei JG, Li JB. 2010. Two new species of Septobasidium (Septobasidiaceae) from

southern China. Mycotaxon 111: 269-274. http://dx.doi.org/10.5248/111.269

Sawada K. 1933. Descriptive catalogue of the Formosan fungi. Part VI. Rep. Dept. Agric. Govt. Res.

Inst. Formosa 61: 1-99.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing. 1527 p.

Teng SC. 1963. Fungi of China. Science Press, Beijing. 808 p.

MY COTAXON

http://dx.doi.org/10.5248/120.277

Volume 120, pp. 277-286 April-June 2012

Characterization of the causal agent of poplar anthracnose

occurring in the Beijing region

ZHENG Lr, YING MEI LIANG? & CHENG MING TIAN”

'Key Laboratory for Silviculture and Conservation of Ministry of Education,

Beijing Forestry University & ?Museum of Beijing Forestry University,

Beijing 100083, P. R. China

* CORRESPONDENCE TO: chengmt@bjfu.edu.cn

ABSTRACT — Twenty fungal isolates derived from infected poplar leaves collected in four

Beijing regional districts during 2009 and 2010 were examined for morphological and cultural

characteristics. Multi-gene phylogenetic sequence analyses of the rDNA ITS (ITS1-5.8S-ITS2)

region and the B-tubulin2, partial actin (ACT), glyceraldehyde-3-phosphate dehydrogenase

(GPDH), and glutamine synthetase (GS) genes were performed for these isolates. The

morphological and cultural evaluations and DNA sequence analyses demonstrated that 16

isolates represented Colletotrichum gloeosporioides while the remaining four isolates, all from

Shi Jingshan district, represent a new species, Colletotrichum populi, which is described and

illustrated.

KEY worps — disease, multi-locus phylogeny, taxonomy

Introduction

The genus Colletotrichum causes anthracnose disease on a wide range of

plants worldwide. The main Colletotrichum species causing anthracnose of

forest trees include C. gloeosporioides, C. acutatum, and C. crassipes, which

may cause infections of leaves and fruits of plants such as poplar, China fir,

and paulownia, often leading to premature defoliation (Xu et al. 2004). Of

these, C. gloeosporioides is the most common, affecting a wide range of tree

hosts. The taxonomy of C. gloeosporioides is extremely complex because of its

heterogeneous nature and instability in cultural characteristics. For example,

more than 600 C. gloeosporioides synonyms have been identified (von Arx

1957). Different experimental conditions and measurement errors may have

a marked influence over the recorded sizes of conidia. Identification results

obtained through traditional morphological examination are thought to be

unreliable (Cai et al. 2009) when compared to molecular methods, and several

new species, such as C. asianum and C. cordylinicola (Prihastuti et al. 2009,

278 ... Li, Tian & Liang

TABLE 1. Isolates of Colletotrichum used in study.

ISOLATE No. Host ORIGIN SAMPLING DATE

1C5-7 Populus xbeijingensis Chang ping 2010.08

C1-5-1 Populus xbeijingensis Chang ping 2010.08

2C5-8 Populus xbeijingensis Chang ping 2010.08

25-9 Populus xbeijingensis Chang ping 2010.08

2C5-14 Populus xbeijingensis Chang ping 2010.08

3C5-3 Populus xbeijingensis Chang ping 2010.08

4C5-2 Populus xbeijingensis Chang ping 2010.08

C2-5 Populus xbeijingensis Chang ping 2010.07

Y3-4 Populus cathayana Yan qing 2010.08

Y3-6 Populus cathayana Yan qing 2010.08

Yp1-7 Populus cathayana Yan qing 2010.08

Bh7-2 Populus xbeijingensis Yan qing 2010.09

Bh9-1 Populus cathayana Yan qing 2010.09

Bh12-2 Populus xbeijingensis Yan qing 2010.09

Dy10-2 Populus nigra Yan qing 2010.09

HMBFU191 Populus nigra var. italica Shi Jingshan 2009.10

HMBFU163 Populus nigra var. italica Shi Jingshan 2009.10

HMBFU173 Populus nigra var. italica Shi Jingshan 2009.10

HMBFU141 Populus nigra var. italica Shi Jingshan 2009.10

M1-6 Populus nigra Mi yun 2010.08

Phoulivong et al, 2010a), have recently been separated from C. gloeosporioides

based on the results from molecular analysis.

Colletotrichum gloeosporioides (teleomorph Glomerella cingulata) has been

reported as the main pathogen of poplar anthracnose in China and North

America (He et al. 1990, Newcombe 2000). Colletotrichum graminicola, which

usually infects monocotyledonous plants such as cereals or grasses, may also

infect poplar in India (Dipak et al. 1999). Most studies that are based on

morphology may be contradicted by molecular data (Phoulivong et al. 2010b).

The present study was carried out to determine the Colletotrichum species

associated with poplar anthracnose in Beijing area through morphological

examination and molecular methods.

Materials & methods

Isolation of Colletotrichum species

Twenty Colletotrichum isolates were obtained from lesions on infected leaves of

different poplar species collected in four districts in Beijing: Chang ping, Yan qing,

Mi yun, and Shi Jingshan (TaBLeE 1). These isolates are deposited at the Mycological

Herbarium of Beijing Forestry University (BJFC).

Three 5x5 mm samples cut from the interface of healthy and diseased tissue

were surface sterilized by dipping in 70% ethanol for 1 minute, then in 1% sodium

hypochlorite for 3-4 minutes and washing in three changes of sterile water. The leaf

segments were then placed onto the surface of potato dextrose agar (PDA) and incubated

Colletotrichum populi sp. nov. (China) ... 279

at room temperature (28°C). The growing edges of fungal hyphae developing from the

leaf segments were transferred onto a new PDA plate after 1-2 days. Each isolate was

purified through single spore isolation and was maintained on a new PDA plate at 28°C

for further study.

Morphological studies

Mycelial discs (5 mm diam) derived from the edge of a 7-day old culture were

transferred onto PDA and incubated at 28°C in the dark. Each colony radius was

measured daily after the 2nd day for 4 days to determine the growth rate by averaging

the daily growth (cm per day) of the colonies. Colony characters were also recorded

and described. After 7 days a spore suspension (5x10° conidia/ml) was made from each

culture and the size and shape of 30 conidia were measured by light microscopy at a

magnification of 40x. A drop of the spore suspension was placed on a slide, which was

placed in a Petri dish containing water-soaked filter paper to keep moist. After 24 hours

(at 28°C) conidial germination was observed, and the size and shape of 20 appressoria

were measured.

Statistical data (morphology and growth rate) were analyzed using SPSS software

version 11.5.0 (SPSS Inc., USA).

DNA extraction and PCR amplification

Mycelium was scraped from the surface of 7-day old cultures of each isolate and dried

using sterilized filter paper. Genomic DNA was extracted using the modified CTAB

method. Extracted DNA was electrophoresed on 1% agarose gel to check genomic DNA

quality.

Using polymerase chain reaction (PCR), DNA was amplified from the complete

rDNA-ITS (ITS) region and the $-tubulin (TUB2), partial actin (ACT), glutamine

synthetase (GS), and glyceraldehyde-3-phosphate dehydrogenase (GPDH) genes of

each Colletotrichum isolate. The primer pairs for the amplified genes are ITS1+ITS4

(White et al. 1990), Bt2a+Bt2b (Glass et al. 1995), ACTF+ACTR (Carbone & Kohn

1999), GSF1+GSR1 (Guerber et al. 2003), GDF1+GDRI1 (Peres et al. 2008) respectively.

Amplification was performed in 25 wL volume with 1 uL (20 ng) DNA template, 2 uL

dNTP (2.5 mmol/L), 2.5 uL 10xPCR buffer, 0.5 uL of each primer (10 uLmol/L), 0.15 uL

Taq DNA polymerase and 18.35 uL ddH,O. The PCR profile consisted of denaturation at

95°C for 3 min, followed by 34 cycles at 95°C for 1 min, 55°C for 30 s, 72°C for 1 min and

a final cycle at 72°C for 10 min. PCR products were electrophoresed on 1% agarose gel

and DNA sequencing was carried out by the Shanghai Invitrogen Biological Technology

Co. Errors were adjusted manually with BioEdit software version 7.0.5.3 (© Tom Hall).

All isolate and DNA sequence accession (DDBJ, EMBL, GenBank) numbers are listed

in TABLE 2.

Phylogenetic analysis

The sequences of Colletotrichum isolates were aligned using Clustal X (1.81).

Alignments were manually adjusted to allow maximum alignment and maximum

sequence similarity, and gaps were treated as missing data.

For parsimony analysis, PAUP version 4.0b10 was used and heuristic search was

performed with 1000 repeats of random addition sequences in Stepwise-Addition

Option and TBR swapping algorithm in Branch Swapping Option. Confidence limits

280 ... Li, Tian & Liang

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Colletotrichum populi sp. nov. (China) ... 281

for the branches based on parsimony criteria were estimated by bootstrap analysis of

1000 replicates. Phylogenetic trees were constructed from distance matrix values by

the maximum parsimony (MP) method. Describing trees was based on tree length,

consistency index (CI), retention index (RI), rescaled consistency index (RC) and

homoplasy index (HI). Constructed trees were viewed by TreeView software. The

GenBank accession numbers used as references in this study are listed in TABLE 2.

Results

Morphological and cultural characterization

The isolates could be divided into two groups (TABLE 3) based on differences

in cultural characteristics and appressorial morphology. Group 1 consisted of

16 isolates with colonies varying from white or grey to dark brown and turning

dark grey or olive-green with time; aerial mycelium white or grey (PLATE

1a-B). These cultures produced abundant orange conidial masses around the

inoculation point. The conidia were cylindrical, obtuse at both ends or slightly

tapered at one end, possessing 2-3 vacuoles and measuring 9.8-22.0 x 2.9-6.9

um (PLATE lc). Appressoria were generally round or ovate and 4.9-15.9 x

4.4-9.8 um (PLATE ID).

TABLE 3. Morphological and cultural characteristics of 20 Colletotrichum isolates. *

C. gloeosporioides C. populi

(Group 1) (Group 2)

ISOLATES TESTED 16 4

CONIDIA

SHAPE Cylindrical Cylindrical

LENGTH (um) 15.8 + 1.9 (9.8-22.0) 15.1 + 2.0 (9.8-17.7)

WIDTH (um) 4.9 + 0.4 (2.9-6.9) 5.2 + 0.9 (3.0-7.3)

APPRESSORIA

SHAPE Circular or ovate Irregular, edge lobed

LENGTH (um) 9.44 2.1 (4.9-15.9) 9.4 + 1.6 (7.3-12.2)

WIDTH (um) 5.9 + 1.0 (4.4-9.8) 6.2 + 1.2 (4.9-9.3)

COLONY Light, dark grey or brown; Aerial mycelium cottony,

aerial mycelium grey white,

with orange conidial masses with indistinct conidial masses

DaILy GROWTH (cm/d) 0.72 0.08 (0.56-0.88) 0.73 0.05 (0.62—0.80)

* Measurements are presented as mean + sd (range).

Group 2 consisted of four isolates collected from Shi Jingshan district. The

colonies were white to light grey in color and covered with white cottony aerial

mycelia, with indistinct conidia masses on the surface of colonies. Conidia

were similar to those of Group 1 in shape (cylindrical) and size (9.8-17.7 x

3.0-7.3 um), but had 1-2 vacuoles. Appressoria were irregular in shape with

lobed edges and measured 7.3-12.2 x 4.9-9.3 um.

There were no significant differences in growth rate between the two groups,

both showing an average daily growth of ca. 0.72 cm/d.

282 ... Li, Tian & Liang

Multi-locus phylogeny

The sequence information obtained from five genes and the phylogenetic

tree information based on each gene sequence are shown in TABLE 4. Of the

five genes, GS and GPDH had the highest rates of parsimony informative

characters (PIC), GS comprising 439 (40.1%) PIC out of a total 1095 characters

and GPDH 114 (38.6%) PIC out of 295 characters.

The phylogenetic tree was constructed on the basis of combined data of the

five genes (PLATE 2). The best tree contained 2593 characters with NPIC=681,

CI=0.865, RI=0.853, HI=0.135, RC=0.738 and Length=1712, showing that the

two morphologically distinct groups were placed into two separate branches.

Group 1 clustered with C. gloeosporioides with a bootstrap value of 100%.

Group 2 was distinctive from C. fructicola (bootstrap value = 79%) and other

members in ‘gloeosporioides complex’ (bootstrap values > 90%), suggesting the

probability of a new species.

TABLE 4. Gene regions amplified.

DNA SEQUENCE LENGTHS PHYLOGENETIC TREE

bee (BP) NPIC/CHARACTERS (%)

ITS 534-537 8.0

TUB2 440-441 18.4

ACT 241 20.1

GPDH 221-230 38.6

GS 981-988 40.1

Taxonomy

Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. PLATE 1

Lesions on leaves of poplar as black spots distributed randomly or in

circles with a dark brown border, orange conidial masses sometimes observed.

Conidia cylindrical, 9.8-22.0 x 2.9-6.9 um, obtuse at ends, hyaline, smooth.

Appressoria ovate or circular, 4.9-15.9 x 4.4-9.8 um. Colonies on PDA with

white, grey, dark grey, or olive grey mycelium and orange sporulation. Reverse

brown or olive-green with dark spots.

PiaTE 1. Colletotrichum gloeosporioides isolates (Group 1): cultural and morphological characters.

A: Colony of isolate 2C5-14; B: Colony of isolate BH7-2; C: Conidia of isolate M1-6; D: Clavate or

circular appressoria of isolate C1-5-1. Bars = 10 um.

Colletotrichum populi sp. nov. (China) ... 283

Group |

2C5-14

BH12-2

Colletotrichum populi HMBFU163

Colletotrichum ee HMBFU191 Group 2

Colletotrichum populi HMBFU173

Colletotrichum populi HMBFU141

Colletotrichum fructicola BPD-I12

Colletotrichum siamense BML-115

Colletotrichum jasmine-sambac LLTA-01

Colletotrichum asianum BPD-14

Colletotrichum kahawae IMI319418

Colletotrichum cordylinicola LC0856

olletotrichum horii TSG002

Colletotrichum acutatum CBS294.67

Colletotrichum simmondsii BRIP28519

(New Species)

Colletotrichum boninense C05016

_10

PLaTE 2. Phylogenetic tree based on the combination of sequences of ITS, TUB2, ACT, GPDH,

and GS genes (MP.)

Colletotrichum populi C.M. Tian & Zheng Li, sp. nov. PLATE 3

MYCOBANK 564436

Lesions with black spots in circle and a light brown border, sometimes sunken. Differs

from Colletotrichum fructicola by grey-white PDA colonies (reverse grey white or pale

yellow) and from C. gloeosporioides by longer and irregularly lobed appressoria formed

from conidia.

Type: China. Beijing, Shi Jingshan District, from leaves of Populus nigra var. italica, 25th

October 2009, coll. C.M. Tian, HMBFU191 (Holotype BJFC, lyophilized PDA culture).

ETyMo_ocy: populi, referring to the host species.

On leaves of poplar producing circular black spots with a dark brown border.

Colonies on PDA at first white, becoming grey-white, reverse grey white or pale

yellow, in 5 days at 28°C, daily growth 0.62-0.80 cm/day (mean = 0.73 + 0.05,

n=5). Aerial mycelium cottony, white with few conidial masses, setae absent in

culture. Conidia 9.8-17.7 x 3.0-7.3 um (mean = 15.1 + 2.0 x 5.2 + 0.9, n = 30),

conidia smooth-walled, hyaline, and cylindrical with obtuse ends. Appressoria

7.3-12.2 x 4.9-9.3 um (mean = 9.4 + 1.6 x 6.2 + 1.2, n = 20), formed from

conidia, brown to dark brown, irregularly lobed in shape.

ADDITIONAL COLLECTIONS EXAMINED: CHINA. BEING, Shi Jingshan District, from

leaves of Populus nigra var. italica, 25th October 2009, coll. C.M. Tian, HMBFU161,

HMBFU141, HMBFU173 (BJEFC, lyophilized PDA cultures).

284 ... Li, Tian & Liang

aay

rn |

ih =

PLaTE 3. Colletotrichum populi isolates (Group 2): cultural and morphological characters of

holotype (isolate 1-9-1). A: Colony; B: Conidia; C & D: irregular appressoria. Bars = 10 um.

ComMENTs: Colletotrichum populi can be separated from the other species

by both morphological and molecular characters (TABLE 5). The similar

C. asianum grows more slowly and produces narrower conidia (8.7-20.3

x 3-4.7 um). The new species shares similar colony characteristics with

C. boninense, C. gloeosporioides, and C. kahawae, all of which produce shorter

and more rarely lobed appressoria. The genetically most closely related C. fructi-

cola is distinguished by colonies that are grayish green in reverse with white

edges. Colonies of C. hymenocallidis, which has similarly sized conidia and

appressoria, appear denser in the central zone and produce fusiform conidia.

C. jasmine-sambac differs by its colony with fimbriate margin.

Discussion

Both Colletotrichum gloeosporioides and C. populi have been found to cause

anthracnose on poplars in Beijing region but can be distinguished by differing

colony and appressoria morphologies as well as by molecular data derived from

sequencing five genes. Appressorial size and shape have been used to distinguish

some Colletotrichum species. For example, C. gloeosporioides was distinguished

from C. acutatum by the shape of appressoria (Du et al. 2005). Crouch & Beirn

(2009) pointed out that the size and shape of appressoria might be effective

with the combination of host species during their work on anthracnose of

cereals and grasses

Although rDNA-ITS gene is not an ideal marker for intraspecific relation-

ships, it is very useful in many cases for reconstruction of interspecific

relationships (Cai et al. 2009). Other gene sequences have been examined in

the phylogenetic analysis of Colletotrichum. GS and calmodulin (CAL) were

found to be useful when studying anthracnose on coffee berries (Prihastuti et

al. 2009). Compared with six gene regions, GPDH, CAL and ACT were useful

for inferring the relationship of C. gloeosporioides sensu lato (Cai et al. 2009).

Whilst there is no defined set of genes to be used for taxonomic studies of

Colletotrichum, molecular data from a wider range of genes would give better

insights into this group of fungi.

TABLE 5. Colletotrichum populi and closely related species.

Colletotrichum populi sp. nov. (China) ... 285

SPECIES COLONY ConrDIA (tm) eee conc REFERENCE

Gandini Be Pte Cyl. with obtuse ends, eae ish Prihastuti

HieMlascen 8.7-20.3 x 3-4.7 gas elin ad et al. 2009

White aerial

mycelium Irregularly Jouji et al

C. boninense y { Cyl, 11.5-17 x 4-7 shaped, J :

reverse cream to 2003

6-17 x 4-15

orange

: Ovoid, occ.

: White; in age grey Ca lawn ene ObuISe te clavate, oft Prihastuti

C. fructicola sl. rounded,

at the center 85-16 x 3.5-5 complex, et al. 2009

6-9 x 5.5-7

a Coes wine ee Cyl. base truncate, Glare Sutton

C. gloeosporioides dark grey, aerial obovate,

4 12-17 x 3.5-6 1992

mycelium 6-20 x 4-12

cs White; later pale Fusiform, straight, ends Ovate, occ. Yang et al.

C. hymenocallidis a ih obtuse clavate,

grey with circles 14-20 x 5-6.5 711% 5-75 2009

White aerial Cyl, straight with Shape variable, :

ow ; . ; : Wikee et al.

C. jasmine-sambac —_ mycelium, with obtuse ends, hyaline, ovoid, clavate, sl. 2010

fimbriate margin 13-15 x 3.5-4 irregular, brown

F : Cyl. straight with Irregular, edge F

a erie cae pbnuse-cnds, eee nes

¥ 9.8-22.0 x 2.9-7.3 7.3-12.2 x 4.9-9.3

Grey erences Straight, cyl., guttulate, Circular to sl. Piihaetuti

C. kahawae barker alse: are apex obtuse, irregular, et al. 2009

PON ee TGA TRS BS 4.5-10 x 7.5

Acknowledgments

We wish to thank Professor M.H. Pei and Dr. C.J. You for text amendments. We are

also grateful to Professor Q. Lu (Chinese Academy of Forestry Sciences, Beijing) and Dr

E.H.C. McKenzie (Landcare Research, Auckland, New Zealand) for their constructive

reviews and valuable suggestions. This research was funded by Forestry Commonweal

Industry Scientific Research Plan (no. 201004003).

Literature cited

von Arx JA. 1957. Die Arten der Gattung Colletotrichum Cda. Phytopathologische Zeitschrift 29:

413-468.

Cai L, Hyde KD, Taylor PWJ, Weir B, Waller J, Abang MM, Zhang JZ, Yang YL, Phoulivong S, Liu

ZY, Prihastuti H, Shivas RG, McKenzie EHC, Johnston PR. 2009. A polyphasic approach for

studying Colletotrichum. Fungal Diversity 39: 183-204.

Carbone I, Kohn LM. 1999. A method for designing primer sets for speciation studies in filamentous

ascomycetes. Mycologia 91(3): 553-556. http://dx.doi.org/10.2307/3761358

Crouch JA, Beirn LA. 2009. Anthracnose of cereals and grasses. Fungal Diversity 39: 19-44.

Dipak S, Chandra JP, Singh AB. 1999. Response of fungicides and antibiotics against anthracnose of

poplar caused by Colletotrichum graminicola. Indian Forester 125: 566-572.

286 ... Li, Tian & Liang

Du MZ, Schardl CL, Vaillancourt LJ. 2005. Using mating-type gene sequences for improved

phylogenetic resolution of Colletotrichum species complex. Mycologia 97: 641-658.

http://dx.doi.org/10.3852/mycologia.97.3.641

Glass NL, Donaldson GC. 1995. Development of primer sets designed for use with PCR to amplify

conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61:

1323-1330:

Guerber JC, Liu B, Correll JC, Johnston PR. 2003. Characterization of diversity in Colletotrichum

acutatum sensu lato by sequence analysis of two gene introns, mtDNA and intron RFLPs and

mating compatibility. Mycologia 95(5): 872-895. http://dx.doi.org/10.2307/3762016

He W, Yang W, Shen RX. 1991. The preliminary study on anthracnose of Populus beijingensis. Forest

Pest and Disease 4: 7-9.

Hyde KD, Cai L, Cannon PF, Crouch JA, Crous PW, Damm U, Goodwin PH, Chen H, Johnston

PR, Jones EBG, Liu ZY, McKenzie EHC, Moriwaki J, Noireung P, Pennycook SR, Pfenning

LH, Prihastuti H, Sato T, Shivas RG, Tan YP, Taylor PWJ, Weir BS, Yang YL, Zhang JZ. 2009.

Colletotrichum-names in current use. Fungal Diversity 39: 147-182.

Jouji M, Toyozo S, Takao T. 2003. Morphological and molecular characterization of Colletotrichum

boninense sp. nov. from Japan. Mycoscience 44: 47-53.

http://dx.doi.org/10.1007/s10267-002-0079-7

Newcombe G. 2000. Inheritance of resistance to Glomerella cingulata in Populus. Canadian Journal

of Forest Research 30: 639-644.

Peres NA, MacKenzie SJ, Peever TL, Timmer LW. 2008. Postbloom fruit drop of citrus and key

lime anthracnose are caused by distinct phylogenetic lineages of Colletotrichum acutatum.

Phytopathology 98(3): 345-352. http://dx.doi.org/10.1094/PHY TO-98-3-0345

Phoulivong S, Cai L, Chen H, McKenzie EHC, Abdelsalam K, Chukeatirote E, Hyde KD. 2010a.

Colletotrichum gloeosporioides is not a common pathogen on tropical fruits. Fungal Diversity

44: 33-43. http://dx.doi.org/10.1007/s13225-010-0046-0

Phoulivong S, Cai L, Parinn N, Chen H, Abd-Elsalam K, Chukeatirote E, Hyde KD. 2010b. A new

species of Colletotrichum from Cordyline fruticosa and Eugenia javanica causing anthracnose

disease. Mycotaxon 114: 247-257. http://dx.doi.org/10.5248/114.247

Prihastuti H, Cai L, Chen H, McKenzie EHC, Hyde KD. 2009. Characterization of Colletotrichum

species associated with coffee berries in Chiang Mai, Thailand. Fungal Diversity 39: 89-109.

Sutton BC. 1992. The genus Glomerella and its anamorph Colletotrichum. 1-26, in: JA Bailey et al.

(eds). Colletotrichum - Biology, Pathology and Control, CAB International.

White TJ, Bruns T, Lee S, Taylor JW. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR Protocols: a guide to

methods and application. San Diego, Academic Press.

Wikee S, Cai L, Pairin N, McKenzie EHC, Su YY, Chukeatirote E, Thi HN, Bahkali AH, Moslem

MA, Abdelsalam K, Hyde KD. 2011. Colletotrichum species from jasmine (Jasminum sambac).

Fungal Diversity 46: 171-182. http://dx.doi.org/10.1007/s13225-010-0049-x

Xu HM, Chen JY, Xiao DL. 2004. Progress in study on anthracnose from forest tree. Hubei Forestry

Science and Technology 130: 40-42.

Yang YL, Liu ZY, Cai L, Hyde KD, Yu ZN, McKenzie EHC. 2009. Colletotrichum anthracnose of

Amaryllidaceae. Fungal Diversity 39: 123-146.

MY COTAXON

http://dx.doi.org/10.5248/120.287

Volume 120, pp. 287-290 April-June 2012

Sympodioplanus yunnanensis, a new aquatic species

from submerged decaying leaves

GUANG-ZHU YANG, Kat-PING Lu, YUE YANG, LI-Bo Ma,

MIN QIAO, KE-QIN ZHANG & ZE-FEN Yu*

‘Laboratory for Conservation and Utilization of Bio-resources,

Key Laboratory for Microbial Resources of the Ministry of Education,

Yunnan University, Kunming, Yunnan, 650091, PR. R. China

*CORRESPONDENCE TO: zfyuqm@hotmail.com

ABSTRACT — Sympodioplanus yunnanensis sp. nov. from submerged leaves collected from

Gengma county, Lincang city, was found when researching aquatic hyphomycetes in Yunnan.

The new species is well defined as a Sympodioplanus by its sympodial proliferation with

multiple and crowded conidiogenous loci in the upper conidiophores and elongated conidia.

It differs in conidial shape and septation from the type species, S. capensis.

KEY worpDs — anamorphic fungi, taxonomy

Introduction

Fungal diversity in southern China is high, and many anamorphic fungi

collected in Yunnan Province have been published (Ma & Zhang 2007a,b,c,

Shang & Zhang 2007, Wang & Zhang 2007, Ma et al. 2008, 2010, 2011, Zhang

et al. 2009a,b,c, 2011a,b).

Sinclair et al. (1997) erected Sympodioplanus R.C. Sinclair & Boshoff for

anamorphic fungi that exhibit sympodial proliferation with multiple and

crowded conidiogenous loci in the upper part of the brown conidiophore, and

elongated conidia with septa. Up to now, the monotypic genus comprised only

S. capensis R.C. Sinclair & Boshoff isolated from dead decorticated wood. We

add here a second species, S. yunnanensis, isolated from submerged decaying

leaves.

Materials & methods

In 2010, a culture was isolated from leaves of a dicotyledonous plant submerged in

a river in Yunnan Province, China. A 2 x 5 cm rotten leaf was spread on the surface

of CMA (20 g cornmeal, 18 g agar, 40 mg streptomycin, 30 mg ampicillin, 1000 mL

distilled water) and incubated for ten days; single conidia were isolated using a sterilized

288 ... Yang & al.

PLATE 1. Sympodioplanus yunnanensis (holotype: YMF1.03797). A-F. Conidia. G. Conidiophores

bearing conidia. H-I. Conidiophores showing moderate protuberance in the apex region.

Scale bars: A-F = 10 um, G =25 um, H-I = 10um.

Sympodioplanus yunnanensis sp. nov. (China) ... 289

toothpick viewed under a CX31 microscope and cultivated on CMA in Petri plates.

Morphological observations were made from CMA after incubation at 25°C for one week;

pure cultures and a permanent slide were deposited in the herbarium of Laboratory for

Conservation and Utilization of Bio-resources, Yunnan University, Kunming, Yunnan,

PR. China (YMF).

Taxonomy

Sympodioplanus yunnanensis G.Z. Yang & Z.F. Yu, sp. nov. PLATE 1

MycoBank MB 563548

Differs from Sympodioplanus capensis by its larger conidia with more numerous septa.

Type: PR China, Yunnan Province, Lincang city, Meihua mountain, 23°38'N 99°22’E,

elev. 1925 m, in a river on submerged leaves of an unidentified dicotyledonous plant,

Dec 2010, G.Z. Yang (Holotype: YMF 1.03797; ex-type culture YMF 1.03797).

ETYMOLOGY: yunnanensis refers to the province in which the species was found.

Colonies effuse, brown, sparse, hairy, often inconspicuous, attaining 30 mm

diam after 10 days on CMA at 25°C. Mycelium mostly immersed in the

substrate, composed of branched, septate, hyaline to brown, smooth, 1.0-3.0

uum wide hyphae. Conidiophores macronematous, mononematous, solitary

or in groups, erect, simple, straight, septate, brown, paler towards the apex,

smooth, < 70-205 um long, 4.5-6.7 um wide. Conidiogenous cells holoblastic,

polyblastic, integrated and terminal, sympodial, thin-walled, bearing multiple,

closely approximated protuberances at the upper cells, including the apex. Scars

are flat in relation to the adjacent conidiogenous cell wall and not thickened,

0.5-1.0 um wide. Conidia solitary, smooth, mostly spindle with truncate base,

occasionally constricted at septa, at the apex mostly slightly attenuate, 38-56 x

6.0-10.8 um, thin-walled, mostly 7-, sometimes 6-, 5-, or 4-septate.

ComMENTs: Sympodioplanus yunnanensis was easily determined as a

Sympodioplanus by its similarity to the type species (S. capensis) in exhibiting

sympodial proliferation with closely placed multiple conidiogenous loci in the

upper conidiophore that are flat and equally thick as the conidiophore wall,

forming solitary terminal and lateral conidia with cell walls similar to those

of the upper conidiophore, and in producing elongated conidia with eusepta.

However, the conidia of S. capensis are smaller (13-16(-19) x 2.5-4 um) and

have fewer septa (3(-5)-septate).

Acknowledgements

This work was jointly financed by National Natural Science Foundation Program

of PR China (31160008, 31060008), Grants from the Young Academic and Technical

Leader Raising Foundation of Yunnan Province (2010CI020). We are very grateful to

Drs. X.G. Zhang and H.-O. Baral for critically reviewing the manuscript and providing

helpful suggestions to improve this paper.

290 ... Yang & al.

Literature cited

Ma J, Zhang XG. 2007a. Three new species of Corynespora from China. Mycotaxon 99: 353-358.

Ma J, Zhang XG. 2007b. Taxonomic studies of Sporidesmium from China. Mycotaxon 99:

367-371.

Ma J, Zhang XG. 2007c. Two new species of Sporidesmium from Yunnan, China. Mycotaxon 101:

73-76.

Ma J, Zhang K, Zhang XG. 2008. Two new species of the genus Minimelanolocus in China.

Mycotaxon 104: 147-151.

Ma IG, Ma J, Zhang YD, Zhang XG. 2010. A new species of Spadicoides from Yunnan, China.

Mycotaxon 113: 255-258. http://dx.doi.org/10.5248/113.255

Ma LG, Ma J, Zhang YD, Zhang XG. 2011. Taxonomic studies of Endophragmiella from southern

China. Mycotaxon 117: 279-285. http://dx.doi.org/10.5248/117.279

Shang ZQ, Zhang XG. 2007. Two new Corynespora species from Jiangsu, China. Mycotaxon 100:

155-158.

Sinclair RC, Boshoff S, Eicher A. 1997. Sympodioplanus, a new anamorph genus from South Africa.

Mycotaxon 64: 365-374.

Wang XM, Zhang XG. 2007. A new species of Corynespora from Yunnan, China. Mycotaxon 101:

79-81.

Zhang K, Fu HB, Zhang XG. 2009a. Taxonomic studies of Minimelanolocus from Yunnan, China.

Mycotaxon 109: 95-101. http://dx.doi.org/10.5248/109.95

Zhang K, Ma LG, Zhang XG. 2009b. New species and records of Shrungabeeja from southern

China. Mycologia 101: 573-578. http://dx.doi.org/10.3852/09-006

Zhang K, Ma J, Wang Y, Zhang XG. 2009c. Three new species of Piricaudiopsis from southern

China. Mycologia 101: 417-422. http://dx.doi.org/10.3852/08-147

Zhang YD, Ma J, Ma LG, Zhang XG. 2011a. Two new species of Taeniolina from southern China.

Mycological Progress 11: 71-74. http://dx.doi.org/10.1007/s11557-010-0729-7

Zhang YD, Ma J, Wang Y, Ma LG, Castafieda-Ruiz RF, Zhang XG. 2011b. New species and

record of Pseudoacrodictys from southern China. Mycological Progress 10: 261-265.

http://dx.doi.org/10.1007/s11557-010-0696-z

MY COTAXON

http://dx.doi.org/10.5248/120.291

Volume 120, pp. 291-294 April-June 2012

Phaeotrichoconis crotalariae,

endophytic on Vitis labrusca in Brazil

THAIS EMANUELLE FEIJO DE LIMA”, JOSE LUIZ BEZERRA

& MARIA AUXILIADORA DE QUEIROZ CAVALCANTI*

Departamento de Micologia, Universidade Federal de Pernambuco,

Rua Nelson Chaves, s/n, Cidade Universitaria, Recife, 50670-901, Brazil

* CORRESPONDENCE TO: thaisfeijo@gmail.com, xiliamac@gmail.com

ABSTRACT — Phaeotrichoconis crotalariae was isolated during studies with endophytic fungi

on healthy leaves of Vitis labrusca in Pernambuco, Brazil. This is a new host for P. crotalariae,

and the second report of this fungus from Brazil.

KEY worps — Dematiaceae, Vitaceae, taxonomy

Introduction

The genus Phaeotrichoconis was proposed by Subramanian (1956) after

Salam & Rao (1954) reassessed Trichoconis crotalariae originally described on

leaves of Crotalaria verrucosa L. The new genus was described as an anamorphic

dematiaceous fungus with phragmosporic acrogenous brown elongated

fusiform conidia with a long appendix.

Phaeotrichoconis crotalariae has a pantropical distribution in Africa, Asia,

Australia, North America, and South America. It is reported to cause foliar flecks

on Acacia spp. and has also been recorded on Alternanthera, Chrysalidocarpus,

Cocos, Crotalaria, Cyperus, Elaeis, Marsilea, Oryza, Rotala, and Xanthium

(Chase 1982, Ellis 1971, Miller 1997, Mishra et al. 1972, Mohanan & Sharma

1988, Old et al. 1996, Ramakrishnan et al. 1972). In Brazil, it has been reported

from Ceara State as a leaf endophyte of Copernicia prunifera (Freire & Bezerra

2001).

Phaeotrichoconis crotalariae was found in healthy leaves of Vitis labrusca

in the Siriji Valley (municipalities of Sao Vicente Férrer and Macaparana),

Pernambuco, Brazil. This paper presents a description of the fungus in culture.

292 ... Lima, Bezerra & Cavalcanti

Pate 1. Phaeotrichoconis crotalariae.

A. sclerotia; B. conidiophore; C-D. young conidia; E-H. mature conidia.

Phaeotrichoconis crotalariae endophytic on Vitis ... 293

Materials & methods

During the February 2010 dry season, healthy mature leaves of Vitis labrusca were

collected from forest areas of the municipalities of Sao Vicente Férrer and Macaparana,

Pernambuco.

In the laboratory, each leaf was washed gently in running water with soap. Leaf discs

were cut with a sterile metallic cork punch (6 mm diam.), decontaminated with 70%

alcohol for 30 sec and hypochlorite sodium solution (NaOCl) at 2% for 2.5 min, and twice

washed with sterilized distilled water in order to remove hypochlorite excess (Petrini

1996; modified technique). Six surface sterilized discs were transferred in triplicate to

each Petri dish containing Potato Dextrose Agar (PDA) + chloramphenicol (50 mg.L")

to prevent bacterial growth. The plates were incubated at room temperature (28° + 2°C)

and observed daily during 15 days for colony development. For asepsis control, 50 uL of

water used to remove hypochlorite was plated in PDA to confirm surface disinfection

(Pereira et al. 1993). Species identification was based on macro- and micro-structural

characteristics of the colony according to Subramanian (1956) and Ellis (1971). One

isolated P. crotalariae, lyophilized and conserved in mineral oil, is maintained in the

Culture Collection Depot at Universidade Federal de Pernambuco (URM).

Results

Five isolates of P. crotalariae were obtained from the dry season collection

(February 2010).

Phaeotrichoconis crotalariae (M.A. Salam & P.N. Rao) Subram. Proc.

Indian Acad. Sci., B 44: 2 (1956) PLATE 1

Colony on malt extract agar, incubated for eight days at room temperature

(28 + 2°C), showed well-developed, flocculate aerial mycelium, at first

light orange, turning gray upon reaching maturity, reddish brown reverse.

Conidiophores macronematous, mononematous, unbranched, straight or

flexuous, brown, smooth, 19.2-141.6 x 3.6-4,8 um. Polytretic conidiogenous

cells, terminal, integrated, merged, sympodial, cylindrical, with dark scars.

Conidia enteroblastic, acrogenous, solitary, dry, fusiform to obclavate,

elongated, rostrate, usually with 5-6 transverse septa, thick walled, smooth,

with a dark brown scar at the base, 45-79.2 x 12-17.5 um. Appendix long,

hyaline to pale brown, narrow (2.4 um diam.), 35-220.8 um long, 0-1 thin

septum. Brown-dark sclerotia present.

SPECIMEN EXAMINED: BRAZIL. PERNAMBUCO: SAO VICENTE FERRER, in healthy mature

leaves of Vitis labrusca L. cv. Isabel, 11 Feb 2010, Lima (URM 6360 [culture conserved

in mineral oil and lyophilized]).

Notes: This is the second record of Phaeotrichoconis crotalariae from Brazil,

and the first time it has been described on artificial culture medium. The fungus

has not previously been recorded from Vitis.

Acknowledgments

The authors thank Francisco C.O. Freire (Embrapa Agroindustria Tropical,

Brazil),and Edna D.M. Newman Luz (Ceplac/Cepec/Sefit, Brazil) for critical review.

294 ... Lima, Bezerra & Cavalcanti

Literature cited

Chase AR. 1982. Dematiaceous leaf spots of Chrysalidocarpus lutescens and other palms in Florida.

Plant Dis. 66: 697-699. http://dx.doi.org/10.1094/PD-66-697

Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, England.

608p.

Freire FCO, Bezerra JL. 2001. Foliar endophytic fungi of Ceara State (Brazil): a preliminary study.

Summa Phytopathol. 27(3): 304-308.

Miller JW. 1997, Plant pathology. Tri-ology Tech Report 36: 11-12.

Mishra B, Prakash O, Misra AP. 1972. Incidence of Phaeotrichoconis crotalariae on Cyperus iria

Linn. from India. Science and Culture 38: 371-372.

Mohanan C, Sharma JK. 1988. Diseases of exotic acacias in India. J Trop Forestry 4: 357-361.

Old KM, Harwood CE, Robson KJ, Haines MW, Solomon DJ. 1996. Foliar pathogens of tropical

acacias in Australia. 11-19, in: KSS Nair et al. (eds). Impact of Diseases and Pests in Tropical

Forests. Proceedings of IUFRO Symposium, Pecchia, Kerala, India.

Pereira JO, Azevedo JL, Petrini O. 1993. Endophytic fungi of Stylosanthes. Mycologia 85: 362-364.

http://dx.doi.org/10.2307/3760696

Petrini O. 1996. Ecological and physiological aspects of host-specificity in endophytic fungi.

87-100, in: SC Redlin, LM Carris (eds). Endophytic fungi in grasses and woody plants. St Paul:

American Phytopathological Society Press.

Ramakrishnan CK, Menon MR, Devi LR. 1972. Alternanthera sessilis R.Br. A new host of

Phaeotrichoconis crotalariae (Salam and Rao) Subramanian comb.nov. Curr. Sci. 41: 751-752.

Salam MA, Rao PN. 1954. A new species of Trichoconis on L. from Hyderabad-Dn. J Indian Bot

Soc 33: 189-191.

Subramanian CV. 1956. Phaeotrichoconis, a new genus of the Dematiaceae. Proc. Indian Acad. Sci.,

B 44:1-2.

MY COTAXON

http://dx.doi.org/10.5248/120.295

Volume 120, pp. 295-300 April-June 2012

Studies on Wrightoporia from China 1.

A new species from Hunan Province, South China

JIA-JIA CHEN* & Hal-You Yu

Institute of Microbiology, P.O. Box 61, Beijing Forestry University, Beijing 100083, China

*CORRESPONDENCE TO: freejoly@yahoo.com.cn

AxBstRAcTt — Wrightoporia nigrolimitata sp. nov. is described from Hunan Province,

southern China. It is characterized by an annual growth habit, small pileate basidiocarps, a

brown to black line between the tubes and context, a monomitic hyphal system with clamp

connections, nondextrinoid hyphae, and finely asperulate amyloid basidiospores. A key to

accepted Chinese Wrightoporia species is supplied.

Key worps — Bondarzewiaceae, polypore, taxonomy, wood-inhabiting fungi

Introduction

Pouzar (1966) established Wrightoporia Pouzar for W. lenta (Overh. &

J. Lowe) Pouzar, the type species. This genus is characterized by resupinate to

pileate basidiocarps, annual to perennial growth habit, monomitic to trimitic

hyphal system, and amyloid asperulate basidiospores (Pouzar 1966, Ryvarden

1982). Of the 39 species described in or transferred to the genus worldwide

(Buchanan & Ryvarden 2000; David & Rajchenberg 1987; Hattori 2003, 2008;

Johansen & Ryvarden 1979; Lindblad & Ryvarden 1999; Loguercio-Leite et al.

1998; Nunez & Ryvarden 2001; Rajchenberg & David 1990; Ryvarden 1975,

1987, 2000), 14 species have been found in China (Cui & Dai 2006; Dai 2012;

Dai & Cui 2006; Dai et al. 2011).

Many new polypores have been recently described due to research on the

diversity of wood-inhabiting fungi in southern China (Cui & Dai 2008, 2011;

Cui et al. 2009, 2010, 201 1abc; Dai et al. 2010, 2011; Du & Cui 2009; Jia & Cui

2011; Li & Cui 2010). During our study of wood-rotting fungi from Mangshan

Forest Park, Hunan Province, we found one Wrightoporia species with a

monomitic hyphal structure that cannot be identified to any known species.

We describe it as a new species in this paper and provide an identification key

to the 15 Wrightoporia species recorded in China.

296 ... Chen & Yu

Materials & methods

Sections were studied microscopically according to Dai (2010) at magnifications

< 1000x using a Nikon Eclipse E 80i microscope with phase contrast illumination.

Drawings were made with the aid of a drawing tube. To present spore size variation,

the 5% of measurements excluded from each end of the range are given in parentheses.

Basidiospore spine lengths are not included in the measurements. Abbreviations include

IKI = Melzer’s reagent, IKI- = negative in Melzer’s reagent, KOH = 5% potassium

hydroxide, CB = Cotton Blue, CB- = acyanophilous, L = mean spore length (arithmetic

average of all spores), W = mean spore width (arithmetic average of all spores),

Q = variation in the L/W ratios between the specimens studied, and n = number of

spores measured from given number of specimens. Special colour terms follow Petersen

(1996). The studied specimens were deposited in herbaria as cited below.

Taxonomy

Wrightoporia nigrolimitata Jia J. Chen, sp. nov. FIG. 1

MycoBank MB 564036

Differs from other Wrightoporia species by its small pileate basidiocarps, presence of

a black layer between tubes and context, a monomitic hyphal system, and generative

hyphae that are nondextrinoid and dissolve in KOH.

Type — China. Hunan Province, Yizhang County, Mangshan Forest Park, on angiosperm

stump, 25.VI.2007, Li 1697 (holotype, BJFC; isotype, IFP).

EryMoLocy — nigrolimitata (Lat.): referring to the black line separating the tubes and

context.

FruiTBopy — Basidiocarps annual, pileate, hard corky upon drying, without

odor or taste; pileus semi-circular to irregularly formed, projecting up to 1 cm

long, 2 cm wide, 4 mm thick at base. Pileal surface straw-yellow to honey-

yellow at base, buff-yellow towards the margin, azonate, slightly sulcate or not;

margin obtuse. Pore surface cream-buff to pinkish buff when dry; pores round

to angular, 4-7 per mm; dissepiments thin to slightly thick, entire. Context buff

to cinnamon-buff, hard corky, up to 1 mm thick. Tubes concolourous with pore

surface, fibrous-tough, up to 3 mm long; a brown to black line present between

tubes and context.

HyPHAL STRUCTURE — Hyphal system monomitic; generative hyphae

bearing clamp connections, IKI- or weakly dextrinoid in the black layer, CB-,

hyphae in trama and context dissolved in KOH; hyphae in the black layer

become reddish to dark brown in KOH.

CONTEXT — Generative hyphae hyaline, thin- to slightly thick-walled,

frequently branched, regularly arranged, 1.5-8 um in diam.

TuBEs — Generative hyphae hyaline, thin- to slightly thick-walled,

frequently branched, subparallel to parallel along the tubes, 2-7 um in diam;

cystidia absent, fusoid cystidioles present, hyaline, thin-walled, 13-19 x 3.5-5

um; basidia clavate, bearing four sterigmata and a basal clamp connection,

Wrightoporia nigrolimitata sp. nov. (China) ... 297

|» rx. E> ©

Oo ©

Kt 5

ee ee

pA OE.

<P,

<)>

Est

ro)

“VOUDY

re Go

10 pm

Fic. 1. Microscopic structure of Wrightoporia nigrolimitata (drawn from the holotype).

a: Basidiospores. b: Basidia and basidioles. c: Cystidioles.

d: Hyphae from trama. e: Hyphae from context.

13-25 x 4-6 um; basidioles in shape similar to basidia, but slightly smaller.

Spores — Basidiospores ellipsoid, hyaline, thin-walled, mostly bearing one

or two small guttules, finely asperulate, amyloid, CB-, 3-4 x 2-2.7(-2.9) um,

L = 3.43 um, W = 2.42 um, Q = 1.42 (n = 30/1).

REMARKS — Although most Wrightoporia species possess a dimitic hyphal

system, only three others have a monomitic hyphal system: W. flava (Ryvarden)

298 ... Chen & Yu

A. David & Rajchenb., W. novae-zelandiae Rajchenb. & A. David, and

W. porilacerata Log.-Leite et al. (David & Rajchenberg 1987, Loguercio-Leite et

al. 1998, Rajchenberg & David 1990, Ryvarden 1975).

Wrightoporia porilacerata also possesses pileate basidiocarps and similar

sized (3-3.5 x 2-2.5 um) basidiospores but is distinguished from W. nigrolimitata

by distinctly larger basidiocarps (< 13.5 x 8.7 x 2.6 cm), larger pores (1-3 per

mm), no black line between tubes and context, and interwoven tramal hyphae

(Loguercio-Leite et al. 1998).

Like W. nigrolimitata, W. flava has pileate basidiocarps, nondextrinoid

generative hyphae, and thin-walled asperulate amyloid basidiospores. However,

W. flava is distinguished by possessing gloeoplerous hyphae, interwoven tramal

hyphae, and lack of a black layer (David & Rajchenberg 1987, Ryvarden 1975).

Except for a monomitic hyphal system, W. novae-zelandiae is unique in

Wrightoporia by combining a gossypine brittle resupinate basidiocarp with

gloeoplerous hyphae (Rajchenberg & David 1990).

OTHER SPECIMENS EXAMINED — Wrightoporia flava. GABON. OGoovE-IvINDo

PROVINCE, Makokou, Ipassa, XII.1974, LY 1733 (LY). TANZANIA. TANGA PROVINCE,

Usambara Mts. Amani, Tanga distr., 18.11.1973, Ryvarden 10594 (O). MALAWI.

SOUTHERN PROVINCE, Makwawa, 16.XI]I.1981, B.M. 342 (O).

— W. novae-zelandiae. NEW ZEALAND. AUCKLAND, Waitakere Ranges, Kitekite

Track, on Leptospermum sp., 19.IV.1989, P.K. Buchanan 89/114 (paratype, PDD 55206).

— W. porilacerata. BRAZIL. PARANA StTaTE, Parangua, Guaraguacu, fazenda

Sambaqui, on decayed dicotyledon trunk, 22.V1.1993, A.A.R. De Meijer 2805 (O).

Key to species of Wrightoporia in China

PRE phial Syston tn GO CRMC pa, scieg eis cheg ph feken hh Seba ti foe db Med hes W. nigrolimitata

Litiyphalsystemndimiitic- OrgQcunittie eo. ee a hon Me kee hat Nee bie dhe Rar een banged 2

2 Generative hyphae-withoutclainp:conmections 2%: 1108-1 ii.0%2 bites ites YE 3

2, Generative hyphae with clamp connections. 0.85 vo kes hones abe eee Gaee as 5

3. Basidiocarps pileate; basidiospores >7 um long... . W. radicata G.Y. Zheng & Z.S. Bi

3. Basidiocarps resupinate to effused-reflexed; basidiospores <7 um long............ 4

4. Basidiocarps without rhizomorphs .......... W. casuarinicola Y.C. Dai & B.K. Cui

4; Basidiocarps with rhizOmorpns. 44.4 ja.. 5.6.5 esas esse beets eS W. rubella Y.C. Dai

5: Basidiospores: > 5:9: (unr longa. 4. oe ia ee enone nee eg sleet S W. lenta

DoDASIMOSPOLES, So 94 [ FIA TONS creek onthe cael h gaia dn cued geal) g seca erates erates eee 6

Go SKe letal iyo Ae Geer UNGIG La pct iene ty aes ca, ean Oe ie cal Bea aly a Penna tama he natn 7

Gs Skeletal hyplaedexeinG ide eee en en eee ee ee ee Ok ete d tie oe dee ec lel 8

7. Cystidia and gloeocystidia absent....................006. W. aurantipora T. Hatt.

Pakystidiatand, gloeoeystidia presents. sact:"sceuctss Sewn Bs Seewe Baes W. austrosinensis Y.C. Dai

SiGloeoplerous-hyp lide pEeSent «shee vtcrmew, Wieew Mehr Piedra Web den eRe tern neterdern ef ion ber te 9

SGlOsGplerausihyplige ADSeMt ss. yee res re sania aight armen a orien dated nw the CaN ease 10

Wrightoporia nigrolimitata sp. nov. (China) ... 299

9. Basidiocarps membranous; tramal skeletal hyphae 1-2 um in

PAINS Ws. Os Sete No a Go RN Nas bes Nb ebay A bo W. avellanea (Bres.) Pouzar

9. Basidiocarps tough; tramal skeletal hyphae 2-2.5 um in diam .. W. borealis Y.C. Dai

10. Basidiospores >3.2 um wide ................ W. unguliformis Y.C. Dai & B.K. Cui

LOU BAsiGsOsPOles. <32 NP WIeL «5 se tlenk wetlina tytn satire seth Nasa ten aside oe nmabireet waine 11

Ph Skeletalthyphae ioteneruste dr 5 fi oy odie ng Lid aeek dinner d ald peek ai penen 42 behets tupetet 12

IL Skeletal Ny phaerenceusted sts ft Me Adri. tRGot UeMte! LeRG wtamlor LEM URN id 13

12. Basidiocarps annual; tramal skeletal hyphae 2-4 um in

UAT ra Stas 5 iia lc ee es tae pete gies Rg ws sete al W. africana I. Johans. & Ryvarden

12. Basidiocarps perennial; tramal skeletal hyphae 4-8 um in

CHATS, corde ott scien coe Seed Vas yo ane Ya W. tropicalis (Cooke) Ryvarden

13. Skeletal hyphae in the trama inflated, up to 6-9 um . W. luteola B.K Cui & Y.C. Dai

13. Skeletal hyphae in the trama not inflated, up to5um............ 0c. eee eee 14

14. Generative hyphae with both clamp connections and simple septa; cystidioles

ADSOMES we leet iapten h.So tan talaga the atta tee W. gillesii A. David & Rajchenb.

14. Generative hyphae only with clamp connections; fusoid cystidioles present

thy Wheat, Btbetln Hekate, Hevaty ween WEN eee 8 W. japonica Nufiez & Ryvarden

Acknowledgements

We express our gratitude to Drs. Hai-Sheng Yuan (IFP, China) and Xiao-Yong

Liu (HMAS, China) who reviewed the manuscript. The research was financed by the

National Natural Science Foundation of China (Project No. 31170018) and Program for

New Century Excellent Talents in University.

Literature cited

Buchanan PK, Ryvarden L. 2000. New Zealand polypore fungi: six new species and a redetermination.

New Zealand Journal of Botany 38: 251-263.

http://dx.doi.org/10.1080/0028825x.2000.9512682

Cui BK, Dai YC. 2006. Wrightoporia (Basidiomycota, Aphyllophorales) in China. Nova Hedwigia 83:

159-166. http://dx.doi.org/10.1127/0029-5035/2006/0083-0159

Cui BK, Dai YC. 2008. Skeletocutis luteolus sp. nov. from southern and eastern China. Mycotaxon

104: 97-101.

Cui BK, Dai YC. 2011. A new species of Pyrofomes (Basidiomycota, Polyporaceae) from China.

Nova Hedwigia 93: 437-441. http://dx.doi.org/10.1127/0029-5035/2011/0093-0437

Cui BK, Dai YC, Bao HY. 2009. Wood-inhabiting fungi in southern China 3. A new

species of Phellinus (Hymenochaetales) from tropical China. Mycotaxon 110: 125-130.

http://dx.doi.org/10.5248/110.125

Cui BK, Dai YC, Yuan HS. 2010. Two new species of Phylloporia (Basidiomycota, Hymenochaetaceae)

from China. Mycotaxon 113: 171-178. http://dx.doi.org/10.5248/113.171

Cui BK, Du P, Dai YC. 2011a. Three new species of Inonotus (Basidiomycota, Hymenochaetaceae)

from China. Mycological Progress 10: 107-114. http://dx.doi.org/10.1007/s11557-010-0681-6

Cui BK, Tang LP, Dai YC. 2011b. Morphological and molecular evidences for a new species of

Lignosus (Polyporales, Basidiomycota) from tropical China. Mycological Progress 10: 267-271.

http://dx.doi.org/10.1007/s11557-010-0697-y

Cui BK, Zhao CL, Dai YC. 2011c. Melanoderma microcarpum gen. et sp. nov. (Basidiomycota) from

China. Mycotaxon 116: 295-302. http://dx.doi.org/10.5248/116.295

300 ... Chen & Yu

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 31-343.

http://dx.doi.org/10.1007/s13225-010-0066-9

Dai YC. 2012. Polypore diversity in China with an annotated checklist of Chinese polypores.

Mycoscience 53: 49-80. http://dx.doi.org/10.1007/s10267-011-0134-3

Dai YC, Cui BK. 2006. Two new species of Wrightoporia (Basidiomycota, Aphyllorales) from

southern China. Mycotaxon 96: 199-206.

Dai YC, Cui BK, Liu XY. 2010. Bondarzewia podocarpi, a new and remarkable polypore from

tropical China. Mycologia 102: 881-886. http://dx.doi.org/10.3852/09-050

Dai YC, Cui BK, Yuan HS, He SH, Wei YL, Qin WM, Zhou LW, Li HJ. 2011. Wood-inhabiting

fungi in southern China. 4. Polypores from Hainan Province, Annales Botanici Fennici 48:

209-231.

David A, Rajchenberg M. 1987. A reevaluation of Wrightoporia and Amylonotus (Aphyllophorales,

Polyporaceae). Canadian Journal of Botany 65: 202-209. http://dx.doi.org/10.1139/b87-027

Du P, Cui BK. 2009. Two new species of Megasporoporia (Polyporales, Basidiomycota) from tropical

China. Mycotaxon 110: 131-138. http://dx.doi.org/10.5248/110.131

Hattori T. 2003. Type studies of the polypores described by E.J.H Corner from Asia and West

Pacific Areas. VI. Species described in Tyromyces (3), Cristelloporia, Grifola, Hapalopilus,

Heterobasidion, Ischnoderma, Loweporus, and Stecchericium. Mycoscience 44: 453-463.

http://dx.doi.org/10.1007/s10267-003-0139-7

Hattori T. 2008. Wrightoporia (Basidiomycota, Hericiales) species and their allies collected in Japan.

Mycoscience 49: 56-65. http://dx.doi.org/10.1007/s10267-007-0389-x

Jia BS, Cui BK. 2011. Notes on Ceriporia (Basidiomycota, Polyporales) in China. Mycotaxon 116:

457-468. http://dx.doi.org/10.5248/116.457

Johansen I, Ryvarden L. 1979. Studies in the Aphyllophorales of Africa. 7. Some new genera and

species in the Polyporaceae. Transactions of the British Mycological Society 72: 189-199.

http://dx.doi.org/10.1016/S0007-1536(79)80031-5

Li HJ, Cui BK. 2010. A new Trametes species from Southwest China. Mycotaxon 113: 263-267.

http://dx.doi.org/10.5248/113.263

Lindblad I, Ryvarden L. 1999. Studies in neotropical polypores. 3. New and interesting Basidiomycetes

(Poriales) from Costa Rica. Mycotaxon 71: 335-359.

Loguercio-Leite C, Gerber AL, Ryvarden L. 1998. Wrightoporia porilacerata, a new species of pore

fungi from Southern Brazil. Mycotaxon 67: 251-255.

Nujiez M, Ryvarden L. 2001. East Asian polypores, vol 2. Synopsis Fungorum 14: 170-522.

Petersen JH. 1996. The Danish Mycological Society’s colour-chart. Foreningen til

Svampekundskabens Fremme, Greve.

Pouzar Z. 1966. Studies in the taxonomy of the polypores 1. Ceska Mykologie 20: 171-177.

Rajchenberg M, David A. 1990. A new species of Wrightoporia Pouz. (Polyporaceae, Basidiomycetes).

New Zealand Journal of Botany 28: 185-186.

http://dx.doi.org/10.1080/0028825X.1990.10412356

Ryvarden L. 1975. Studies in the Aphyllophorales of Africa. 2. Some new species from East Africa.

Nordic Journal of Botany 22: 25-34.

Ryvarden L. 1982. Synopsis of the genus Wrightoporia. Nordic Journal of Botany 2: 145-149.

http://dx.doi.org/10.1111/j.1756-1051.1982.tb01174.x

Ryvarden L. 1987. New and noteworthy polypores from tropical America. Mycotaxon 28:

525-541.

Ryvarden L. 2000. Studies in neotropical polypores 7. Wrightoporia (Hericiaceae, Basidiomycetes)

in tropical America. Karstenia 40: 153-158.

MYCOTAXON

http://dx.doi.org/10.5248/120.301

Volume 120, pp. 301-307 April-June 2012

A new species of Bipolaris from Iran

ABDOLLAH AHMADPOUR', ZEINAB HEIDARIAN', MARYAM DONYADOOST-

CHELAN’, MOHAMMAD JAVAN-NIKKHAH’ & TAKAO TSUKIBOSHI’

"Department of Plant Protection, Faculty of Agricultural Sciences & Engineering,

University College of Agriculture and Natural Resources,

University of Tehran, Karaj, Iran

? National Institute of Livestock and Grassland Science,

Senbonmatsu 768, Nasushiobara, Tochigi 329-2793, Japan

* CORRESPONDENCE TO: ahmadpour.abdollah@gmail.com

ABSTRACT— Bipolaris salkadehensis sp. nov., isolated from the leaves of Sparganium

erectum (Thyphaceae) and Cladium mariscus (Cyperaceae) collected in Iran, is described

and illustrated. The new species is characterized by straight to slightly curved subcylindrical

to fusoid conidia with end cells demarcated by thick dark septa and with monopolar and

bipolar germination. Morphological data and ITS+5.8SrDNA sequence analyses support

B. salkadehensis as a distinct species of Bipolaris.

Key worps — morphology, taxonomy, fungus, Cochliobolus, phylogeny

Introduction

Bipolaris species are known to infect many kinds of grasses causing spots or

flecks on leaves or stems and sometimes cause diseases on herbaceous plants

such as Cactaceae and Musaceae (Ellis 1971, Sivanesan 1987). Since Sivanesan

(1987) monographed the graminicolous species of the genus, several new

Bipolaris species have been described (Chiang et al. 1989, Peng & Lu 1989,

Sisterna 1989, Alcorn 1990, Sivanesan 1992, Alcorn 1996, Chen et al. 2000, Zhu

et al. 2000, Deng & Zhang 2002, Jiang & Zhang 2008, Zhang & Li 2009). Recent

molecular phylogenetic analyses have shown that Bipolaris and Curvularia are

closely related to each other and comprise a different clade from Exserohilum

(Berbee et al. 1999, Olivier et al. 2000). The rDNA-ITS sequences of (particularly

the internally transcribed spacers ITS1 and ITS2) are useful tools for resolving

taxonomic relationships within Bipolaris.

In this study, we describe a new species of Bipolaris, which we compare both

morphologically and phylogenetically with other species.

302 ... Anmadpour & al

Materials & methods

Collection and isolation of the fungus

Plant material was obtained from the Salkadeh village, Khoy city, West Azerbaijan

province, in northwestern Iran in the summer of 2010. Leaf samples of Sparganium

erectum L. (Thyphaceae) and Cladium mariscus (L.) Pohl (Cyperaceae) with brown oval

to elliptical lesions with approximately 2-3 mm in length were collected and stored dry

in a refrigerator at 5°C until use. A single spore produced on the surface-sterilized leaf

under Nuv light on 12 h diurnal cycle at room temperatures 23 + 2°C (Sivanesan 1987)

was transferred to tap water agar with autoclaved wheat straw (TWA+wheat straw) plates.

Five isolates were obtained: Bi-1-3 from S. erectum, and Bi-4—5 from C. mariscus. For

morphological inspection each isolate was transferred to Petri dishes containing PDA

using a thin glass needle and incubated at 23 + 2°C under darkness; species descriptions

are based on 10-14 day old cultures. Measurement and microphotographs were taken

from slide mounts in lactophenol and lactophenol cotton blue using an Olympus light

microscope (model BH2). The fungal isolates were compared with the descriptions

of Ellis (1971, 1976), Sivanesan (1987) and reported species after 1987. Dried culture

vouchers have been deposited in the herbarium in the Department of Plant Protection,

Faculty of Agricultural Sciences & Engineering, University College of Agriculture and

Natural Resources, University of Tehran, Karaj, Iran (TUPP).

Molecular phylogenetic analyses

Whole genomic DNA was extracted from the mycelium of each isolate grown on

V8 juice agar by homogenization in a standard sodium dodecyl sulfate (SDS) detergent

lysis buffer followed by a phenol : chloroform extraction and precipitation in ethanol

with sodium acetate (Sambrook et al. 1989). The ITS1+5.8S+ITS2 rDNA region was

amplified using the polymerase chain reaction (PCR) conditions and the ITS1 and

ITS4 primer pair (White et al. 1990). Purified PCR products were sequenced by ABI

PRISM 3100 automated sequencers (Applied Biosystems, Foster City, CA, USA). For

phylogenic comparison, the GenBank sequences of 26 Bipolaris species, 5 Curvularia

species, 2 Exserohilum species, 2 Drechslera species, and Alternaria alternata (Fr.) Keissl.

(as outgroup) were included (Berbee et al. 1999). The DNA sequences were aligned using

Clustal X version 1.8 (Thompson et al. 1997). Further visual alignments were done in

Sequence Alignment (Se-Al) Editor version 2.0 (Rambaut 2000). The data were analysed

phylogenetically using distance methods. The distance matrix was calculated using

Kimura's two parameter method (Kimura 1980) and analyzed with the neighbor-joining

(NJ) method (Saitou & Nei 1987) using the program PAUP* 4.0 beta 10 (Swofford 2002).

Bootstrap values were generated with 1000 replicate heuristic searches to estimate

support for clade stability of the consensus tree using the same program. Sequences

have been deposited in the DNA Data Bank of Japan (DDBJ).

Results

New species

Bipolaris salkadehensis Ahmadpour & Heidarian, sp. nov. Fic. 1

MycoBaAnk MB 564565

Differs from Bipolaris cynodontis in conidia with both monopolar and _ bipolar

germination and conidial end cells that are demarcated by thick dark septa.

Bipolaris salkadehensis sp. nov. (Iran) ... 303

Fic. 1. Bipolaris salkadehensis. A, B. Conidiophores. C-G. Conidia. H-J. Germinating conidia.

Scale bars = 10 um.

Type: Iran. West Azerbaijan, Khoy City, Salkadeh village, on infected leaves of Sparganium

erectum, 20 Sep 2010, coll. A. Ahmadpour Bi-1 (Holotype, TUPP1366 [dried culture];

DDBJ sequence, AB675490).

ETyMOLoGy: referring to the type locality.

Colonies in “[WA+wheat straw’ agar brown, velvety, floccose. Hyphae brown,

septate, branched, smooth, 5-7 um wide. Stromata not formed. Conidiophores

single or fasciculate in small groups, simple, pale brown to brown, cicatrized

with scars often inflated and smooth, multiseptate, 225-590 um long, swollen

to 7.5-15 um diam at base, then narrowing to 5-8.5 um diam (middle) and

5-7.5 um diam (apex). Conidia pale brown to dark brown, subcylindrical to

fusoid, occasionally obclavate to clavate, straight to slightly curved, smooth,

end cells rounded, very pale and demarcated by thick, dark septa, (4—)6-8

(-10) distoseptate, (32—)53-80(-93) x 12-15 um, hilum <2 um diam, sometimes

slightly protruding. Germination of conidia is monopolar and bipolar. The

primary septum in developing conidia is submedian, the second delimiting the

basal cell, and the third distal.

304 ... Anmadpour & al

Cultural characteristics: Colony velvety, floccose, olivaceous-brown on

PDA.

ADDITIONAL MATERIAL EXAMINED: IRAN. WEST AZERBAIJAN, Khoy City, Salkadeh

village, on infected leaves of Cladium mariscus, 20 Sep 2010, coll. A. Ahmadpour Bi-4

(TUPP1362; DDBJ sequence, AB675491) and Bi-5, on infected leaves of Sparganium

erectum, Bi-2-3.

ComMENTs: Its conidial morphology clearly establishes our specimen as

belonging to Bipolaris. It is morphologically similar to B. cynodontis, B. setariae

(Sivanesan 1987), B. sesuvii (Zhang & Li 2009), and B. fusca (Jiang & Zhang

2008). Bipolaris cynodontis is distinguished from the new species by its smaller

conidia, absence of thick dark septa demarcating the end cells demarcated by

thick dark septa, and with only bipolar germination (Sivanesan 1987). The

conidia of B. setariae are somewhat similar — fusoid to navicular, pale to mid

golden brown, 5-10 distoseptate, (45-)50-70(-100) x 10-15 um (Sivanesan

1987) — but with end cells demarcated by unthickened hyaline septa and paler

than those of B. salkadehensis. The conidia of B. sesuvii are olivaceous-brown

to brown, basal cell concolorous or slightly pale, subcylindrical to broadly

fusoid and rounded at ends, straight, smooth, 5-9 distoseptate, 52-77 x

13-16 um (Zhang & Li 2009) and are narrower and have end cells demarcated

by unthickened hyaline septa. The conidia of B. fusca, which are shorter and

wider than those of B. salkadehensis, are straight or occasionally slightly

curved, smooth, cylindrical or ellipsoidal, (3—)5-7(-11) distoseptate, 31-67 x

11-20 um, yellowish brown to dark brown, with end cells often very pale and

demarcated by thick dark septa (Jiang & Zhang 2008).

Phylogenetic analysis & discussion

PCR products of ITS+5.8S rDNA sequences were 506bp for isolate Bi-1

and 505bp for isolates Bi-2-5. Since the Bi-2-5 sequences matched completely,

the Bi-1 sequence was registered as DDBJ AB675490 and the Bi-4 as DDBJ

AB675491. According to the neighbor-joining tree derived from ITS+5.8S

rDNA sequences made in this study, the B. salkadehensis isolates formed

a monophyletic group with a 95% bootstrap value close to B. cynodontis

(AF163093) (Fic. 2). However, the 98.2% (497/506 bp) sequence similarity

between B. salkadehensis and B. cynodontis indicates that those two species

clearly differ. Moreover, despite similar conidial morphologies, B. salkadehensis

formed a different group from B. setariae and B. sesuvii (Fic. 2).

Berbee et al. (1999) used the ITS+5.8S rDNA sequences and a portion of the

glyceraldehyde-3-phosphate dehydrogenase sequences to evaluate Cochliobolus

(anamorphs Bipolaris) and proposed that Bipolaris be divided further, separating

species with large, canoe-shaped, gently curving conidia (Cochliobolus Group

1) from those with short, either straight or curved conidia lacking a gentle

curve along the whole spore length (Cochliobolus Group 2), which intermix

Bipolaris salkadehensis sp. nov. (Iran) ... 305

Gipolaris zeicola AF158110

Bipolaris victoriae AF 158109

Bipolaris urochloae AF071334

Gipolaris heveae AB179835

Bipolaris peregianensis AF 158111

Bipolans oryzae X18122

95, Bipolaris salkadehensis Bi-1 AB675490

Bipolaris salkadehensis Bi-4 AB675491

Gipolaris cynodontis AF163093

Gipolaris stenospila AB119837

Bipolaris luttrelhi AFO71350 Cochliobolus

Bipolaris melinidis AF071350 Group 1

Bipolaris zeae AF081452

Bipolaris sorokiniana AF071329

Gipolans sorghicola AF071332

Bipolans setariae EF452444

Bipolaris setariae GU290228

Bipolaris bicolor AF120260

Bipolaris maydis AF158108

Bipolaris sacchari AF071318

82 Bipolanis australiensis AF081450

88 Bipolans hawaiensis AFO71324

Gipolaris perotidis AF071320

Cunulana gladioli AF071337

Bipolaris ravenelii AF071321

Cumulania lunata AF071339

2 Cunularia gudauskasii AF071337 es px ti i : ¢ ae

Cunlana afinis AF071335

400 98| Sipolaris indica AF081449

85 Bipolans sesuvi EF 175940

Sipolanis portulacae AY004778

98 Bipolaris kusanoi AF071352

78 Bipolaris nodulosa GU073110

Cumularia clavata AF071336

400 Exserohilum minor AF071341

Exserohilum rostratum AF071342

This study

100

99 Drechslera erythrospila AY004782

Drechslera tritici-repentis AF071348

Alternana alternata AF071346

+1 0.02 substitutions/site

Fic. 2. A neighbor-joining tree inferred from the ITS+5.8S rDNA sequences from 35 taxa. Bootstrap

values for 1000 replicates are shown on the branches, and rDNA-ITS DDBJ accession numbers

stand after the species names. The branch length is proportional to the number of base changes as

indicated by the scale bar. Alternaria alternata (AF071346) is an outgroup.

306 ... Anmadpour & al

with Curvularia species. All B. salkadehensis isolates used in this study clustered

in the Cochliobolus Group 1 subclade, which contains Bipolaris species large,

canoe-shaped, gently curving conidia. Although the conidial morphology of

B. salkadehensis is similar to B. cynodontis, B. setariae (Cochliobolus Group 1),

and B. sesuvii (Cochliobolus Group 2), the phylogenetic analysis did not show

a close relationship. The combined molecular-morphological analysis supports

B. salkadehensis as an unreported new species.

Although the teleomorph was not observed in cultures or on the natural

host, the B. salkadehensis teleomorph relationships can be compared by using

differences or similarity in conserved DNA sequences (e.g. ITS, Gpp, BRN1

and mating type genes) with other known Bipolaris species to provide a more

reliable classification system at the generic and species levels (Berbee et al.

1999, Shimizu et al. 1998, Turgeon 1998). The MAT phylogenetic trees, along

with the ITS and Gpp trees, can be used as a database from which to look back

into evolutionary history in order to understand how different reproductive

lifestyles arose (Turgeon 1998).

Both morphological and phylogenetic evidence supports B. salkadehensis

as a previously unreported species separate from other known Bipolaris.

This is the first report of Bipolaris on the hosts S. erectum (Thyphaceae) and

C. mariscus (Cyperaceae).

Acknowledgments

The authors wish to special thanks to Dr. Meng Zhang (College of Plant Protection,

Henan Agriculture University, P. R. China) for kind guidance. We are greatly indebted

to Dr. Guang-yu Sun (College of Plant Protection, Northwest A&F University, P. R.

China) and Mr. Takuya Sakoda (Yokohama Plant Protection Station, Japan) for their

critical reviewing of the manuscript. We also appreciate the corrections by Dr. Shaun

Pennycook, Nomenclatural Editor, and suggestions by Dr. Lorelei L. Norvell, Editor-in-

Chief. This work was financially supported by University of Tehran.

Literature cited

Alcorn JL. 1990. Additions to Bipolaris, Cochliobolus and Curvularia. Mycotaxon 39: 361-392.

Alcorn JL. 1996. Cochliobolus heliconiae sp. nov. (Ascomycota). Australian Systematic Botany 9:

813-817. http://dx.doi.org/10.1071/SB9960813

Berbee ML, Pirseyedi M, Hubbard S. 1999. Cochliobolus phylogenetics and the origin of known,

highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase

gene sequences. Mycologia 91: 964-977. http://dx.doi.org/10.2307/3761627

Chen WQ, Swart WJ, Nieuwoudt TD. 2000. A new species of Bipolaris from South Africa.

Mycotaxon 76: 149-152.

Chiang MY, Leonard K, Van Dyke C. 1989. Bipolaris halepense: a new species from Sorghum

halepense (johnsongrass). Mycologia 81: 532-538. http://dx.doi.org/10.2307/3760128

Deng H, Zhang TY. 2002. Taxonomic studies of Bipolaris from China I. Mycosystema 21: 327-333.

http://dx.doi.org/CNKI:SUN:JWXT.0.2002-03-005

Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew.

Bipolaris salkadehensis sp. nov. (Iran) ... 307

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew.

Kimura M. 1980. A simple method for estimating evolutionary rate of base substitutions through

comparative studies of nucleotide sequences. Journal of Molecular Evolution 16: 111-120.

http://dx.doi.org/10.1007/BF01731581

Jiang YL, Zhang TY. 2008. New species of Bipolaris, Scolecobasidium and Torula from soil.

Mycotaxon 104: 135-140.

Olivier C, Berbee ML, Shoemaker RA, Loria R. 2000. Molecular phylogenetic support from

ribosomal DNA sequences for origin of Helminthosporium from Leptosphaeria-like

loculoascomycete ancestors. Mycologia 92: 736-746. http://dx.doi.org/10.2307/3761430

Peng JH, Lu JY. 1989. Studies on Bipolaris, Drechslera and Exserohilum. Journal of Nanjing

Agricultural University 12: 46-53. http://dx.doi.org/cnki:ISSN:1000-2030.0.1989-04-009

Rambaut A. 2000. Se-Al: sequence alignment editor. Department of Zoology, University of Oxford,

Oxford, UK.

Saitou N, Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic

trees. Molecular Biology Evolution 4: 406-425.

Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular cloning: a laboratory manual, 2"’ Ed. Cold

Spring Harbor Laboratory, Cold Spring Harbor, NY.

Shimizu K, Tanaka C, Peng YL, Tsuda M. 1998. Phylogeny of Bipolaris inferred from nucleotide

sequences of Brn1, a reductase gene involved in melanin biosynthesis. Journal of General

Applied Microbiology 44:251-258. http://dx.doi.org/10.2323/jgam.44.251

Sisterna MN. 1989. Two new species of Bipolaris. Plant Pathology 38: 98-100. http://dx.doi.

org/10.1111/j.1365-3059.1989.tb01433.x

Sivanesan A. 1987. Graminicolous species of Bipolaris, Curvularia, Drechslera, Exserohilum and

their teleomorphs. Mycological Papers. No. 158.

Sivanesan A. 1992. New Bipolaris, Curvularia and Exserohilum species. Mycological Research 96:

485-489. http://dx.doi.org/10.1016/S0953-7562(09)81095-2

Swofford DL. 2002. PAUP*. Phylogenetic analysis using parsimony (* and other methods). Version

4, Sinauer, Sunderland, MA.

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The Clustal X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 25: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Turgeon BG. 1998. Application of mating type gene technology to problems in fungal biology. Annual

Review of Phytopathology 36: 115-137. http://dx.doi.org/10.1146/annurev.phyto.36.1.115

White TJ, Bruns T, Lee SB, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: M Gelfand et al. (eds). PCR protocols: a guide to

methods and applications. Academic, San Diego, California.

Zhang JZ, Li MJ. 2009. A new species of Bipolaris from the halophyte Sesuvium portulacastrum in

Guangdong Province, China. Mycotaxon 109: 289-300. http://dx.doi.org/10.5248/109.289

Zhu MQ, Sun GY, Zhang TY. 2000. Studies on the genus Bipolaris of Guangxi province. Acta

Agriculturae Boreali-occidentalis Sinica 9: 32-34.

http://dx.doi.org/CN KI:SUN:XBNX.0.2000-03-008

MY COTAXON

http://dx.doi.org/10.5248/120.309

Volume 120, pp. 309-316 April-June 2012

Notes on Xylophallus xylogenus (Phallaceae, Agaricomycetes)

based on Brazilian specimens

LARISSA TRIERVEILER-PEREIRA* & ROSA MARA BORGES DA SILVEIRA

Depto. de Botanica, Universidade Federal do Rio Grande do Sul

Av. Bento Goncalves 9500, Porto Alegre-RS, 91501-970, Brazil

*CORRESPONDENCE TO: It_pereira@yahoo.com.br

ABSTRACT — From morphological analysis of Phallus pygmaeus specimens collected in

Brazil and reexamination of the holotype, we conclude that this species is a synonym of the

previously described Xylophallus xylogenus and agree with the recently proposed synonymy.

We describe new specimens of X. xylogenus from Northeastern Brazil and provide color

photos and a key for the species of the small xylophilous members of Phallus, including

Xylophallus.

KEY worDs — gasteromycetes, herbarium revision, Mutinus, stinkhorns, tropical fungi

Introduction

Phallus pygmaeus was described from the tropical rainforest in Northeastern

Brazil, growing on decaying wood (Baseia et al. 2003). The species is

characterized by small basidiomata (< 15 mm high), a smooth receptacle with

minute perforate apex, and lignicolous habitat. The remarkably small size of

this phalloid is not exclusive to P pygmaeus, as other small species are known

from the tropics, e.g., Phallus tenuis (E. Fisch.) Kuntze, P minusculus Kreisel

& Calonge, and P. drewesii Desjardin & B.A. Perry (Calonge & Kreisel 2002,

Calonge 2005, Desjardin & Perry 2009).

Xylophallus xylogenus is a taxon with a problematic generic position.

Originally described as Phallus xylogenus from Cayenne (French Guiana),

its author (Montagne 1855) placed the species in sect. Mutinus, while some

years later Schechtendal (1861) classified it in Phallus sect. Xylophallus Schltdl.

Fischer (1898-99), who first proposed the combination Mutinus xylogenus,

later erected the genus Xylophallus (Schltdl.) E. Fisch. for the species (Fischer

1933a). Saenz et al. (1972) also contributed to the morphology and ontogeny of

the species based on collections from Costa Rica. Since its original description,

X. xylogenus has been ambiguously illustrated by different authors (Fic. 1).

310 ... Trierveiler-Pereira & Silveira

Fic. 1. Xylophallus xylogenus, redrawn from original publications:

A-B. Montagne (1855), as Phallus xylogenus. C. Fischer (1933).

D. Saenz et al. (1972). E. Baseia et al. (2003), as Phallus pygmaeus.

Cheype (2010), who recently described and provided colour photos of

X. xylogenus specimens from the type locality, proposed synonymizing

P. pygmaeus with X. xylogenus.

During field expeditions in the State of Pernambuco, Northeastern Brazil,

specimens initially determined as P pygmaeus were collected in abundance

Xylophallus xylogenus in Brazil... 311

from rotten logs. Our analysis of this material and reexamination of the

P. pygmaeus holotype leads us to confirm Cheype’s (2010) synonymy.

Materials & methods

Field expeditions were carried out in 2008 and 2009 in two reserves in the State

of Pernambuco, Northeast Region of Brazil: Reserva Privada do Patriménio Natural

(RPPN) Carnijé (Moreno; 8°10'00"S 35°05'15"W) and Parque Dois Irmaos (Recife;

8°07'30"S 34°52'30"W). Both reserves, at altitudes at 20-180 m, are relict fragments

of the Brazilian Atlantic rainforest. The sampled basidiomata were transported to the

laboratory in plastic boxes (Lodge et al. 2004). Both fresh and dried basidiomata were

examined.

Macroscopic measurements and colors are based on fresh material. Colors are coded

according to Kornerup & Wanscher (1978). Observations of microscopic characters

were made under a light microscope on glass slides mounts (in 5% KOH) prepared from

dried specimens. Voucher specimens are deposited in URM (Thiers 2011).

Taxonomy

Xylophallus xylogenus (Mont.) E. Fisch., Nat. Pflanzenfam., 2 Aufl., 7a: 96. 1933.

FIGS 1-3

= Phallus (Mutinus) xylogenus Mont., Ann. Sci. Nat., Bot., Sér. 4, 3: 137. 1855.

= Mutinus xylogenus (Mont). E. Fisch., Nat. Pflanzenfam. 1(1**): 290. 1899.

= Phallus pygmaeus Baseia, Mycotaxon 85: 78. 2003.

BASIDIOMATA gregarious; when immature globose, subglobose to ovoid,

sometimes tapering towards the base, glabrous, smooth or groovy, 2.5-3.5 mm

high x 2.5-4 mm broad, yellowish brown (5D5, 5E5) at the apex, becoming

lighter near to the base; at maturity, 6-14 mm _ high, including the volva.

Votva yellowish brown, with irregular dehiscence, basal hyphal strand simple

or multiple, whitish, penetrating the woody substrate or forming a network

above it. PsEUDOSTIPE cylindrical, hollow inside, 5-7 mm high x 1.5-2.5

mm broad, translucent white, indusium absent; externally ornamented by a

reticulated structure composed of rectangular alveoles with prominent edges.

RECEPTACLE conic-campanulate, surface smooth, adnate to the pseudostipe,

with round or umbilicate apex, non or minutely perforate at maturity, 2.5-4

mm high x 2.5-4 mm broad, concolor with pseudostipe; receptacle margin

smooth to denticulate. GLEBA gelatinous, grayish green (1D5) to olive (1F7),

very foetid.

BASIDIOSPORES ellipsoid, smooth, thin-walled, hyaline or with greenish

tints, 3-4 x 1.5 um. VoLtva composed of isodiametric to irregularly rectangular

pseudoparenchymatous hyphae, hyaline, yellowish to brownish, slightly

thick-walled, 7-15 x 6-13 um. PSEUDOSTIPE AND RECEPTACLE composed by

isodiametric pseudoparenchymatous hyphae, hyaline, thin-walled or slightly

thick-walled, 12-45 x 12-32 um.

312 ... Trierveiler-Pereira & Silveira

Y Ya tat

Fic. 2. Xylophallus xylogenus in situ.

A. Brazilian basidiomata (scale = 12 mm). B. Costa Rican basidiomata (photo by Clark L. Ovrebo).

Xylophallus xylogenus in Brazil ... 313

ECOLOGY & DISTRIBUTION: on rotten wood in French Guiana (Montagne

1855, Cheype 2010), French Antilles (Guadeloupe; Cheype 2010), Suriname

(Fischer 1933b), Costa Rica (Saenz et al. 1972, Saenz & Nassar 1982, Calonge et

al. 2005), Brazil (as P pygmaeus; Baseia et al. 2003, Baseia et al. 2006, Leite et al.

2007), Peru and Ecuador (Gémez & Gazis 2006).

SPECIMENS EXAMINED: BRAZIL. PERNAMBUCO, GUuRJAU, 28.VI.2002, leg. Baseia &

Gibertoni (URM 77078, holotype of Phallus pygmaeus); MORENO, RPPN Carnijé,

17.V1.2008, leg. Trierveiler-Pereira 10, 11 (URM 80261, 80262); 08.VII.2008, Trierveiler-

Pereira 83, 84 (URM 80264, 80265); 14.VIII.2008, Trierveiler-Pereira 186, 187 (URM

80269, 80270); 23.1X.2008, Trierveiler-Pereira 148, 149 (URM 80267, 80268); 16.X.2008,

Trierveiler-Pereira (URM 80271); 23.1.2009, Baltazar & Coimbra 206 (URM 80272);

12.III.2009, Trierveiler-Pereira 207, 208 (URM 80273, 80274); 21.V.2009, Trierveiler-

Pereira 226, 227 (URM 80275, 80276); ReciFE, Parque Dois Irmaos, 07.VII.2008,

Trierveiler-Pereira 67 (URM 80263); 12.VIII.2008, Trierveiler-Pereira 142 (URM

80266).

REMARKS: Desjardin & Perry (2009) noted that P xylogenus was a long forgotten

name. Various authors have illustrated it differently (Fic. 1). Montagne’s (1855)

illustration (Fic. 1B), which is representative, depicts its xylophilous gregarious

habit, while Montagne’s detailed figure of a single basidiome (Fic. 1A) is quite

imaginary, showing the receptacle margin distant from the pseudostipe and the

pseudostipe with a shallow reticulum. The illustration in Fischer (1933a), based

on original material, is more realistic and the pseudostipe with deep alveoles is

well represented (Fra. 1C).

Lloyd’s (1907) photo of the original material shows that the upper basidiome

of the type was not well preserved, leading him to describe the species as having

a “capitate, globose mass of gleba”. Dennis (1970) repeated this information,

probably based on Lloyd's description. Lloyd’s belief that Xylophallus was a

synonym of Mutinus has been wrongly propagated through the literature

during the 20th century.

Saenz et al. (1972) accompany their good detailed description of X. xylogenus

from Costa Rica with an illustration of the receptacle with an umbilicate apex

(Fic. 1D) and perforated apex (approximately 100 um in diam). Baseia et al.

(2003) distinctly illustrated the receptacle pore (Fic. 1E). Among the examined

materials, we found specimens with rounded to umbilicate apices, minutely

perforated or not. We believe that the perforated apex probably represents the

mature receptacle.

Calonge et al. (2005), basing their observations on Dring (1973), cited X.

xylogenus as cosmopolitan. However, Dring noted distribution only for the

entire genus Mutinus, not for this species in particular. According to Gomez

& Gazis (2006) and elsewhere, although X. xylogenus may be common in

neotropical forests, the species rarely collected due to its minute size and

ephemeral nature.

314 ... Trierveiler-Pereira & Silveira

Fic. 3. Xylophallus xylogenus.

A. Single basidiome (scale = 2.5 mm). B-D. Microscopical structures (scale = 10 um):

B. Pseudoparenchymatic hyphae from the pseudostipe.

C. Pseudoparenchymatic hyphae from the volva. D. Basidiospores.

We agree with Saenz et al. (1972) in separating X. xylogenus from Mutinus,

since the gleba develops on a receptacle externally modified from the

pseudostipe. Although the xylophilous habit of X. xylogenus is observed in

other Phallus species, we prefer to retain the species in Xylophallus until further

molecular analyses are carried out.

Xylophallus xylogenus in Brazil ... 315

Key to small xylophilous phalloid species

la. Receptacle smooth, minutely perforated; pseudostipe with rectangular alveoles

PIN COLEO PICS) oF voter Muh Ae tere roe man uirh, Muel nied oth Xylophallus xylogenus

1b. Receptacle reticulate, clearly perforated with a conspicuous pore; pseudostipe

mintitely reticulate-Orspongy~ 264 2.500 2.n Goth Gn Pee teh 8S cle c8S ae tee ao 2

2a. Pseudostipe yellow, basidiomata 70-100 mm high (China, Japan, Indonesia,

Srp Wan kes, Aro pial FATE CA) 4 at actg dst oxteg ty isgeten dressed even Hest geen dyitgctt Phallus tenuis

2b. Pseudostipe white, basidiomata smaller, < 45 mm high.....................0.. 3

3a. Pseudostipe spongy, 25-33 mm high; basidiospores 2-3 um long (Tanzania,

CASIET MATIC Ae A 8) F589 Ate RO RL Ee aa MOE oe P. minusculus

3b. Pseudostipe reticulate-lacunose, 20-45 mm high; basidiospores 3-3.5(-3.8) um

lotisstSao Pome Western AaPiCa)S 5 ct. Yecacecn iva Ma Hevea Nova che Bona dba yesua’s P. drewesii

Acknowledgments

We thank RPPN Carnijé and Parque Dois Irmaos (Pernambuco, Brazil) for permission

to collect; Juliano M. Baltazar and Victor R.M. Coimbra for helping during field work;

PPGBF-UFPE and Dra. Leonor Costa Maia, curator of Herbarium URM; CNPq and

CAPES (Brazil) for providing fellowship to the first author. We express sincere gratitude

to Clark L. Ovrebo (U.S.A.) for his photo and Gabriel Grilli (Argentina) for his assistance

in scanning photo slide. We are grateful to Marc Stadler (Germany), Jean-Louis Cheype

(France) and Juliano M. Baltazar (Brazil) for suggestions to improve the manuscript;

Jacques Fournier (France) for sending useful literature. Dr. Eduardo R. Nouhra (IMBIV,

Cordoba, Argentina), Maria Luciana H. Caffot (IMBIV, Cordoba, Argentina) and Taiga

Kasuya (University of Tsukuba, Ibaraki, Japan) are acknowledged for critically reading

the manuscript.

Literature cited

Baseia IG, Gibertoni TB, Maia LC. 2003. Phallus pygmaeus, a new minute species from a Brazilian

tropical rainforest. Mycotaxon 85: 77-79.

Baseia IG, Calonge FD, Maia LC. 2006. Notes on the Phallales in the Neotropics. Bol. Soc. Micol.

Madrid 30: 87-93.

Calonge FD. 2005. A tentative key to identify the species of Phallus. Bol. Soc. Micol. Madrid 29:

Calonge FD, Kreisel H. 2002. Phallus minusculus sp. nova from tropical Africa. Feddes Repert.

113(7-8): 600-602.

Calonge FD, Mata M, Carranza J. 2005. Contribuicién al catalogo de los Gasteromycetes

(Basidiomycotina, Fungi) de Costa Rica. Anales del Jardin Botanico de Madrid 62(1): 23-45.

Cheype J-L. 2010. Phallaceae et Clathrus récoletés en Guyane francaise. Bulletin Mycologique et

Botanique Dauphiné-Savoie 197: 51-66.

Dennis RWG. 1970. Fungus flora of Venezuela and adjacent countries. Kew Bulletin Additional

Series 3. Vaduz, J. Cramer.

Desjardin DE, Perry BA. 2009. A new species of Phallus from Sao Tomé, Africa. Mycologia 101(4):

545-547. http://dx.doi.org/10.3852/08-166

316... Trierveiler-Pereira & Silveira

Dring DM. 1973. Gasteromycetes, in Ainsworth GC, Sparrow FK, Sussman AS (eds.) The Fungi

IVB. New York, Academic Press, pp. 451-478.

Fischer E. 1898-99. Phallineae. 276-296, in: A Engler, K Prantl (eds). Die natirlichen

Pflanzenfamilien, Teil 1(1**).

Fischer E. 1933a. Phallineae. 76-108, in: A Engler, K Prantl (eds). Die natirlichen Pflanzenfamilien,

2 Aufl., Vol. 7a.

Fischer E. 1933b. Gastromyceteae Stahelianae. Ann. Mycol. 31(3): 113-125.

Goémez LD, Gazis R. 2006. Dos Gasteromycetes (Basidiomycotina, Fungi) del Peru. Brenesia 65: 71.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3rd ed. London, Eyre Methuen.

Leite AG, Silva BDB, Araujo RS, Baseia IG. 2007. Espécies raras de Phallales (Agaricomycetidae,

Basidiomycetes) no Nordeste do Brasil. Acta Botanica Brasilica 21(1): 119-124.

http://dx.doi.org/10.1590/S0102-33062007000100011

Lloyd CG. 1907. Mycological Notes 26. Concerning the Phalloids. Mycological Writings 2:

325-337.

Lodge DJ, Ammirati JA, O’Dell TE, Mueller GM, Huhndorf SM, Wang CJ, Stokland JN, Schmit

JP, Ryvarden L, Leacock PR, Mata M, Umania L, Wu Q, Czederpiltz DL. 2004. Terrestrial and

lignicolous macrofungi. 127-172, in: GM Mueller et al. (eds). Biodiversity of fungi. Inventory

and monitoring methods. Boston, Elsevier Academic Press.

Montagne C. 1855. Cryptogamia Guayanensis. Ann. Sci. Nat., Bot., Sér. 4, 3: 91-144.

Saenz JA, Nassar M. 1982. Hongos de Costa Rica: Familias Phallaceae y Clathraceae. Revista Biol.

Trop. 30(1): 41-52.

Saenz JA, Nassar M, Morales MI. 1972. Contribution to the study of Xylophallus xylogenus.

Mycologia 64(3): 510-520. http://dx.doi.org/10.2307/3757868

Schlechtendal DFL. 1861. Eine neue Phalloidee, nebst Bemerkungen uber die ganze Familie

derselben. Linnaea 31: 101-194.

Thiers B. 2011 [continuously updated]. Index Herbariorum: a global directory of public herbaria

and associated staff. New York Botanical Garden's Virtual Herbarium.

http://sweetgum.nybg.org/ih/ [accessed September 2011]

MYCOTAXON

http://dx.doi.org/10.5248/120.317

Volume 120, pp. 317-329 April-June 2012

Plectania seaveri (Ascomycota, Pezizales),

a new discomycete from Bermuda

MATTEO CARBONE ?*, CARLO AGNELLO ” & SCOTT LAGRECA °

’ Via Don Luigi Sturzo 173, I-16148, Genova, Italy

? Via Antonio Gramsci 11, I-72023, Mesagne (BR), Italy

° Plant Pathology Herbarium, Dept. of Plant Pathology & Plant Microbe Biology,

Cornell University, Ithaca, NY 14853, USA

* CORRESPONDENCE TO: Matteocarb@hotmail.com

ABSTRACT— Plectania seaveri sp. nov. is described after revision of Bermudian material

identified by Seaver under the name Bulgaria melastoma. The only known host for this new

species is the endemic Bermudian tree Juniperus bermudiana (Bermuda cedar), a species

whose numbers have greatly reduced over the past 70 years due to the introduction of invasive

scale insects. As this new fungus has not been collected since 1940, it may now be extinct.

Color pictures of fresh and dried specimens, microscopic morphology, and a black and

white drawing of hymenial elements are included. Related, excluded, and doubtful species of

Plectania are also discussed.

KEY wWoRDS— ascomycetes, Donadinia, taxonomy

Introduction

When the senior authors (MC, CA) began their revision of the genus

Plectania Fuckel, they realized that the Bermudian collections reported by

Seaver (1928, 1942) and Seaver & Waterston (1946) under the name Bulgaria

melastoma [i.e., Plectania melastoma (Sowerby) Fuckel] required further study.

After publication of Carbone et al. (2012) and Agnello & Carbone (2012),

the need to re-examine these collections was even clearer. These early studies

strongly indicated that Seaver’s Bermudian collections might represent a new

taxon, and so, joined by the third author (SLG), we examined all Bermudian

material of this taxon in the Plant Pathology Herbarium of Cornell University

(CUR):

Our revision rapidly confirmed that Bermudian material of Bulgaria

melastoma in CUP does not belong to B. melastoma but in fact represents a

strikingly different, undescribed species. We here name this species in honour

318 ... Carbone, Agnello & LaGreca

PiaTE 1. FJ. Seaver in his office at NY Botanical Garden (photographed 1930-35). Photo by

H.M. Fitzpatrick, from the Cornell University Plant Pathology Herbarium photograph collection.

of the original researcher for most of this material, the American mycologist

Fred Jay Seaver (PLATE 1).

Materials & methods

Microscopic characters are based on dry specimens. Two optical microscopes were

used: Olympus CX41 trinocular and Optika B353 trinocular with plan-achromatic

objectives 4x, 10x, 40x, 60x, and 100x in oil immersion. Microscopic pictures were made

using a Nikon 4500 camera and a Nikon Coolpix. Primary mounting media were Melzer’s

reagent, cotton blue, and Congo red. Water mounts were used for all measurements

and observations of pigments. At least 30 spores were measured from each examined

apothecium. SEM observations were made using a Hitachi S-4000 scanning electron

microscope (SEM) at the University of Florida Core EM facility (Gainesville, Florida,

USA). A piece of the hymenium from the holotype was mounted on aluminum stubs

with adhesive graphite tabs and sputter coated with a gold-palladium alloy for 60 s.

Taxonomy

Plectania seaveri M. Carbone, Agnello & LaGreca, sp. nov. PL. 2-6

MycoBank MB 563704

“Bulgaria melastoma” sensu Seaver, N. Amer. Cup-fung.,

Operc.: 198, 1928, Bermudian records.

Plectania seaveri sp. nov. (Bermuda) ... 319

Differs from Plectania melastoma in the absence of macroscopic orange-red warts, more

stipitate apothecia, and ellipsoid and differently ornamented spores; from P. nannfeldtii

and P helvelloides in a very different spore ornamentation; and from P lusitanica in

larger spores and more pronounced ornamentation.

Type: Bermuda, Walsingham, ca. 150 fruitbodies on bark of Juniperus bermudiana,

20 January 1922, leg. H.H. Whetzel, Bermuda Fungi no. 188 [Holotype, CUP-Whetz.

B.F.-0188, as Bulgaria melastoma].

Erymo.oey: In honor of the American mycologist, Fred Jay Seaver.

MACROCHARACTERS (based on dried specimens) — APOTHECIA gregarious to

subcespitose, mostly long stipitate, nearly globose and closed at first, expanding

as they mature; base attached to substrate with very numerous dark black,

threadlike filaments (subiculum); cup <18 mm across and <6 mm high; margin

mostly entire: HyMENIUM blackish to black, apparently smooth; EXTERNAL

SURFACE blackish to black, with a dark brownish to purplish-amaranth shade,

rough, gently wrinkled; stipe <16 mm high and <2 mm thick, solid, black; the

length varying depending on the point of growth from the substrate; FLESH

leathery, darker near external surfaces.

MICROCHARACTERS — ASCI 350-400 x 14-15 um, cylindrical, operculate,

with a slightly excentric operculum, inamyloid, eight-spored, with walls <1 um

thick and a tapered, flexuous base with a generative hypha that is connected

more or less distantly from the basal septum; PARAPHYSES not, or only slightly,

exceeding the length of the asci, 1.5-2(-2.5) um wide, cylindrical, septate,

sometimes anastomosing, branched, pale brownish, multiguttulate by very small

oil drops; the apex usually more or less diverticulate and lobed; an extracellular,

brownish, amorphous pigment present in the upper part (observed in water

mounts), uniting them in bundles; seraE (hymenial hairs) cylindrical, as long

as the paraphyses, (3-)3.5-4.5 um wide, with a simple to slightly subcapitate

apex and single septum at the very base, pale brownish; sporgs ellipsoid, 22-27

x 10-12 um, most frequently 23-25 x 10-11 um, very few spores observed

<31 um long, Q = (1.8-)2-2.7(-3), subhyaline to light yellowish, filled with

a granular content and sometimes with few slightly bigger oil drops, walls <1

um thick; spore sculpturing of small but well-defined round warts <0.8 um

broad and 0.6 um high, visible also at low magnifications in optical microscopy;

confirmed by the SEM, showing round isolated warts; very young spores are

round, smooth and thick walled; suBHYMENIUM composed of a dense textura

intricata of cylindrical, frequently septate, hyphae, with thickened, more or

less dark brown walls; at low magnifications, it appears uniformly brownish to

brown and not so sharply differentiated from the upper layer of the medullary

excipulum; MEDULLARY EXCIPULUM of textura intricata with cylindrical,

septate hyphae, 5-11 um wide, with walls on average 0.5-0.8 um thick; two

main layers can be recognized: (i) an upper one with more brownish, denser,

less gelified hyphae with a parallel orientation to the hymenial surface, (ii) a

320 ... Carbone, Agnello & LaGreca

PLaTE 2. Plectania seaveri. Upper row: CUP-033234: fresh specimens. Photo by FJ. Seaver:

Bottom row: dried specimens of holotype collection. Photos by M. Carbone. Bar = 10 mm valid

for bottom row only.

middle-lower one with lighter to yellowish, more loosely interwoven hyphae

immersed ina subgelatinous matrix; ECTAL EXCIPULUM of textura subglobulosa-

angularis to textura angularis made up of elements <20 um wide and/or high,

very dark brown due to the colored walls and the presence of an incrusting

Plectania seaveri sp. nov. (Bermuda) ... 321

brown pigment. In the very outer part the pigmentation becoming more

amber-like and crystalline; EXTERNAL HAIRS mainly of two types, although

intermediate forms occurring; (i) cylindrical, septate, hyaline, thin walled,

heavily encrusted hyphoid hairs, <2.5 um wide; (ii) cylindrical, infrequently

septate, <6 um wide, very long, mainly straight, smooth to very finely encrusted

(only in the basal part) by the same kind of crystalline pigment, and brown

due to an epimembranaceous pigmentation, with walls thickened <0.8 um; in

non-squashed mounts, they are mainly lying on the external surface, and it is

not easy to determine their entire length; susicuLUM made up of cylindrical,

sometimes very slightly flexuous and/or notched, 6-9 um wide, <3 mm (or

more) long, septate hyphae, brown due to an epimembranaceous pigmentation,

with walls thickened <0.8 um, mainly smooth but in places a very light greenish

extracellular pigment is present.

ECOLOGY & DISTRIBUTION — gregarious to subcespitose, on bark of

Bermuda cedar (Juniperus bermudiana L.). January and February. Known only

from the Bermuda Archipelago.

ADDITIONAL COLLECTIONS EXAMINED: Mature collections: BERMUDA, Agricultural

Station, eleven fruitbodies on bark of Juniperus bermudiana on ground, 4 February

1926, leg. HLH. Whetzel, Ogilvie & FJ. Seaver (CUP-033235); six fruitbodies on J.

bermudiana, 15 January 1926, leg. H.H. Whetzel, Ogilvie & FJ. Seaver (CUP-034615).

Burts Island, four fruitbodies on fallen tiny twigs of J. bermudiana, 9 February 1926, leg.

H.H. Whetzel, Ogilvie & FJ. Seaver (CUP-033236).

Immature collections: BERMUDA, Agricultural Station, on bark of Juniperus

bermudiana on ground, 12 January 1926, leg. H.H. Whetzel, Ogilvie & FJ. Seaver (CUP-

033231); on bark of living J. bermudiana, 28 January 1926, leg. H.H. Whetzel, Ogilvie &

EJ. Seaver (CUP-033234). Harrington house, on fallen bark of J. bermudiana on ground,

23 January 1926, leg. H.H. Whetzel, Ogilvie & FJ. Seaver (CUP-033233).

Discussion

PRELIMINARY NOTES — The first hint of the existence of these Bermudian

collections can be found in Seaver’s note (1928) under Bulgaria melastoma:

“What appears to be this species is very abundant in Bermuda on the bark

of Bermuda cedar, Juniperus bermudiana, but never on anything else. Since

the host is an endemic species, the fungus may also be found to differ and

be itself endemic. The writer has not yet had the opportunity to clear up this

point but reference will be made to it in later publications” In Seaver (1942),

again under Bulgaria melastoma: “Range extended to Bermuda (FJ. Seaver &

H.H. Whetzel)”. And lastly, in Seaver & Waterston (1946): > However one of us

(Seaver 1928: 198, 1942: 320) has shown that the Bermuda species is referable

to Bulgaria melastoma (Sow.) Seaver. Figure 1 (upper) shows this species on

rotten bark of Juniperus bermudiana L., from roots of living trees exposed at

soil level, Walsingham, Bermuda, Jan. 20, 1922, H.H. Whetzel Bermuda Fungi

No. 188. This plant is characterized by apothecia which are frequently stipitate,

322 ... Carbone, Agnello & LaGreca

PLATE 3. Plectania seaveri (holotype). a—b: spores in lactic Cotton blue. c-e: spores in Congo

red. f-h: spores in water mount. Bars: a, b, d, e, h = 10 um; c = 2,5 um; f, g = 5 um. Photos by

M. Carbone.

with almost smooth, hyaline, ellipsoidal spores, 20-25 x 9-10 uw”. In Waterston

(1947) we find only a list of the collections studied here, under Bulgaria

melastoma, and nothing is added from a taxonomic point of view.

Korf (1957) seems to have been the first mycologist to realize that these

collections might represent a new taxon: “...the species of “Bulgaria” described

by Seaver and Waterston (1946: 182) from Bermuda, which appears to be

undescribed...” While revising the cited material we found that Denison (Korf’s

student) also noted Seaver’s misinterpretation, for in collection CUP-033235,

there is a handwritten annotation noting, “This is not Bulgaria (Plectania)

melastoma. Note, for example, the delicately sculptured spores, the larger

apothecia and longer stipe. W C Denison 2 Nov 1961”.

Manyyears before, Boedijn (1932) suggested that Seaver’s Bermudian Bulgaria

melastoma (Seaver 1928) might be conspecific with Sarcosoma thwaitesii (Berk.

& Broome) Petch [= Galiella thwaitesii (Berk. & Broome) Nannf.], mainly

Plectania seaveri sp. nov. (Bermuda) ... 323

because Petch (1910) recorded S. thwaitesii from Ceylon, growing on Juniperus

bermudiana. Although Seaver & Waterston (1946) definitely misinterpreted

Bulgaria melastoma, they demonstrated as well that Boedijn (1932) was wrong.

Galiella thwaitesii is undoubtedly a very distinct and different taxon. For further

details about Galiella thwaitesii see Le Gal (1953).

SPECIES DELIMITATION — Plectania melastoma, in its original sense, is definitely

a very different species from the material studied here. As demonstrated by

Agnello & Carbone (2012) the macroscopic, vivid orange-red, external warts

are always present, and still visible to the naked eye even after 150 years; in

addition, the apothecia are at most substipitate and never definitely stipitate

as in P. seaveri. Microscopically, there are more numerous and immediately

appreciable differences. For example, in P. seaveri the spore sculpturing is

delicate but, at the same time, very distinct and pronounced. It is easily visible

in water mounts (1000x, oil immersion). In contrast, in P melastoma, the spores

are far less sculptured and have very different warts that are barely visible,

even in 1000x Cotton blue mounts. The spore shapes are also quite different

— mostly subfusoid in P melastoma, ellipsoid in P. seaveri. Lastly, P melastoma

produces only one kind of external hairs and a differently coloured crystalline

pigmentation that is mostly a vivid orange-red-amber, not pale brownish to

dark amber as in P seaveri.

Unlike Plectania melastoma, P. seaveri surely belongs to Plectania sect.

Donadinia (Bellem. & Mel.-Howell) M. Carbone & Agnello. In this section,

Carbone et al. (2012) recognize only three stipitate species with more or less

defined ornamented ellipsoid spores: P. helvelloides (Donadini et al.) Donadini,

P. nannfeldtii Korf, and P. lusitanica (Torrend & Boud.) M. Carbone et al. As

pointed out by Carbone et al. (2012), while all these species have extremely

similar gross macroscopic morphologies, excipular anatomies, and external

hairs, they can clearly be distinguished based on the combination of spore

size and spore sculpturing. In fact, if on one hand we observe that spore

measurements show a continuum from a minimum of 16 um (in Plectania

lusitanica) to a maximum of 38 um (in Plectania nannfeldtii), on the other hand

we are presently unaware of any populations that show a simultaneous overlap

of spore size range and sculpturing. SEM and optical observations also support

this morphological distinction.

The well-known American “snowbank mushroom, Plectania nannfeldtii, can

be immediately differentiated by its different ecology, geographical distribution,

longer spores, and different episporium (e.g. Seaver 1928; Miller 1967; Smith et

al. 1981; Arora 1986; Tylutki 1993; Li & Kimbrough 1995). Plectania lusitanica

surely shares a similar ornamentation but differs in its smaller spores and a

much less pronounced spore sculpturing that is barely visible in optical

microscopy (data taken from many spore prints). Lastly, P helvelloides spores

324 ... Carbone, Agnello & LaGreca

PiateE 4. Plectania seaveri (holotype). a—-b: spores. c: pole of the spore. d: spore sculpturing.

Bars a—b = 5 um; c-d = 1 um. SEM courtesy of Mark Whitten and the Electron Microscopy Core

Facility at the University of Florida (Gainesville, USA).

(22-28 x 9-13 um; Donadini et al. 1973, Carbone et al. 2012), which might be

regarded as the closest to P. seaveri in size, have a different ornamentation type

— relatively easy to see in cotton blue (1000x), but smaller, differently shaped,

and not easily detectable in outline.

The spore sculpturing seen in Plectania seaveri is extremely similar to that

found in Galiella Nannf. & Korf, which, however, produces very different

fleshy large gelatinous apothecia; for a quick survey, see Boedijn (1932, as

Sarcosoma), Le Gal (1953, as Sarcosoma), Korf (1957), Cao et al. (1992), and

Zhuang & Wang (1998). The same can be said for the genus Neournula Paden

& Tylutki, previously placed in Sarcosomataceae Kobayasi but now transferred

into a different family, Chorioactidaceae Pfister. Neournula does indeed possess

similarly warted spores but is distinct from Plectania in many other features

(see Pfister et al. 2008).

Plectania seaveri sp. nov. (Bermuda) ... 325

p eh

see

Pate 5. Plectania seaveri (holotype). a: section of the apothecium; b: medullary excipulum in water

mount; c: bundles of paraphyses; d: seta (hymenial hair); e: ascus tip; f: medullary excipulum in Congo

red (magnified); g—h: hyphoid excipular hairs; i: excipular hairs; j: ectal excipulum; k: subiculum;

1: encrusted cells of the margin; m: crystals of excipular pigment. Bars: a = 100 um; b = 20 um;

c, e; f, j = 10 um; d, g, h, i, k, 1 = 5 um. Photos by M. Carbone.

EXTRALIMITAL AND DOUBTFUL SPECIES — Mention must be made here of two

other Plectania species described from the Americas. Plectania coelopus (Mont.)

Sacc. was described from Chile and, according to the iconotype and original

description, it surely belongs to sect. Donadinia. We agree with Le Gal (1958)

and Cabello (1988) that P. coelopus must be regarded as a doubtful species until

326 ... Carbone, Agnello & LaGreca

PLaTE 6. Plectania seaveri (holotype). a: section of the cup; b: spores; c: ascus; d: paraphyses;

e: seta (hymenial hair); f: excipular hairs. Bars: a = 100 um; b = 10 um; c-f = 10 um. Drawing by

C. Agnello.

its real identity is investigated and/or it is typified with topotypical (or other

Chilean) material.

Another doubtful American taxon is Plectania rimosa Peck, described from

California (Peck 1903). Seaver (1942) treated it as a “doubtful species’, and due

to the (apparent) lack of original material, he stated that this species might

represent Paxina corium (O. Weberb.) Seaver [= Helvella corium (O. Weberb.)

Massee]. As the protologue does not fit Helvella corium, we disagree and regard

Plectania rimosa as a nomen dubium.

Plectania seaveri sp. nov. (Bermuda) ... 327

During our literature searches, we have found other unclear species

seemingly in need of revision (e.g., Neournula helvelloides sensu Zhuang &

Wang (1998) and Plectania nannfeldtii in the sense of the Chinese authors Teng

(1963), Tai (1979), and Bi et al. (1990) and the Japanese author Otani (1973).

Future studies will address the misapplications of these names.

Eco.toecy — All known Plectania seaveri specimens were collected from rotting

bark of Juniperus bermudiana, an endemic Bermudian tree that historically

dominated the upland forests of the Bermuda Archipelago (Thomas 2004). Most

collections of this fungus were made in the 1920s, with the last known specimen

collected in 1940 (Waterston, 1947). A search of 125 fungus specimens from the

last major Bermudian mycological expedition collected by Richard Korf and

colleagues during the week of January 17-23, 1980 (unpublished; specimens

deposited at CUP) revealed no additional P seaveri specimens.

Between 1946 and 1953, introduced scale insects decimated the Bermuda

cedar, killing an estimated 95% (Phillips 1984), thus deforesting the islands and

altering the ecology of many other Bermudian species in major ways (Thomas

2004). With no known Plectania seaveri specimens collected since 1940, it

seems probable that this fungus is now extinct. While it may still occur on

some of the dead, non-decorticated trunks of Bermuda cedar that now dot the

Bermudian landscape (and perhaps on the few living Bermuda cedars which

still persist), it seems likely that the fate of P seaveri may eventually follow that

of other endemic Bermudian organisms dependent on Bermuda cedar, such as

the Bermuda cicada (Tibicen bermudiana) and the ochre-banded looper moth

(Semiothisa ochrifascia) (Sterrer 1998).

As far as we know, all species of Plectania sect. Donadinia, including

P. seaveri, associate as saprophytes with coniferous trees. Future collections of

representatives of this section would help us understand whether this coniferous

association is taxonomically relevant.

FINAL CONSIDERATIONS — This study demonstrates that Bulgaria melastoma as

interpreted by Seaver (1928) needs an in-depth revision. Paden & Tylutki (1969)

observed that “in Seaver, P milleri will key out to P melastoma (as Bulgaria

melastoma). We can speculate that, at least in the case of Plectania seaveri,

Seaver might have misinterpreted the encrusting pigment of the external

surface, indicating that he was not aware of the true diagnostic characters of

P. melastoma. In any case, we are convinced that Seaver misapplied the name

P. melastoma to other specimens, such as one from Puerto Rico (CUP-PR-

001281, examined as part of the current study), and probably also the Jamaican

collection cited by Seaver & Waterston (1946). The latter specimen, supposedly

deposited in NY, could not be located during a recent visit to that herbarium

(SLG pers. obs., April 2, 2012). Our future studies will attempt to answer these

lingering questions.

328 ... Carbone, Agnello & LaGreca

Acknowledgments

We are grateful to Mark Whitten and the Electron Microscopy Core Facility at the

University of Florida (Gainesville, USA) for generously performing the SEM work for

this study. Dick Korf and Nicolas VanVooren both reviewed a draft of this manuscript;

we thank them for their helpful comments. Barbara Thiers and Stephen Sinon are

thanked for their attempts to track down Seaver’s original plates of Bulgaria melastoma

in the Seaver Collection housed at the Mertz Library, New York Botanical Garden. We

are thank Ellen Bloch (NY) for providing key information about certain specimens

cited in this paper, Lucy Klebieko and Mike Bevans (NY) for their helpful assistance

in scanning the herbarium photograph for Fic. 1, and the Curator of the Herbarium of

Natural History of Venice (MCVE) for arranging the loan of the examined collections.

Literature cited

Agnello C, Carbone M. 2012 ['2011’]. Appunti di studio su Plectania melastoma. Rivista di

Micologia 54(4): 315-337.

Arora D. 1986. Mushrooms demystified. Ten speed Press, Berkeley. 960 p.

Bi ZS, Zheng GY, Li TH, Wang Y-Z. 1990. Macrofungus flora of the mountainous district of North

Guangdong. Guangdong Science & Technology Press, Guangzhou, China. 734 p.

Boedijn KB. 1932. The genus Sarcosoma in Netherlands India. Bulletin du Jardin botanique de

Buitenzorg, 3" series, 12(2): 273-279.

Cabello MN. 1988. Estudio sistematico del suborden Sarcoscyphineae (Pezizales, Ascomycotina)

empleando técnicas numéricas. Boletin de la Sociedad Argentina de Botanica 25(3-4):

395-413.

Cao J-Z, Fan L, Liu B. 1992. Notes on the genus Galiella in China. Mycologia 84(2): 261-263.

Carbone M, Agnello C, Baglivo A, Peri¢ B, De Giorgi D. 2012 ['2011’]. Studio comparato delle

specie lungamente stipitate del genere Plectania. Mycologia Montenegrina 14: 7-38.

Donadini JC, Berthet P, Astier J. 1973. Un Urnula nouveau: Urnula helvelloides (Discomycetes

operculés). Bulletin mensuel de la Société linnéenne de Lyon 42(3): 37-40.

Korf RP. 1957. Two bulgarioid genera: Galiella and Plectania. Mycologia 49: 107-111.

http://dx.doi.org/10.2307/3755734.

Le Gal M. 1953. Les Discomycétes de Madagascar. Paris. 465 p.

Le Gal M. 1958. Discomycétes du Maroc. I. Un Urnula nouveau, Urnula megalocrater Malencon

et Le Gal sp. nov. Etude de lespéce suivie dune révision des caractéres des genres Urnula et

Sarcosoma. Bulletin de la Société mycologique de France 74: 155-177.

Li LT, Kimbrough JW. 1995. Spore wall ontogeny in Pseudoplectania nigrella and Plectania

nannfeldtii (Ascomycotina, Pezizales). Canadian Journal of Botany 73(11): 1761-1767.

http://dx.doi.org/10.1139/b95-188.

Miller OK Jr. 1967. Notes on western fungi. I. Mycologia 59: 504-512.

http://dx.doi.org/10.2307/3756769.

Otani Y. 1973. On Pseudoplectania and Plectania collected in Japan. Reports of the Tottori

Mycological Institute 10: 411-419.

Paden JW, Tylutki EE. 1969. Idaho discomycetes I. Mycologia 61: 683-693.

http://dx.doi.org/10.2307/3757460.

Peck CH. 1903. New species of fungi. Bulletin of the Torrey Botanical Club 30( 2): 95-101.

http://dx.doi.org/10.2307/2478879.

Petch T. 1910. Revisions of Ceylon fungi (Part II). Annals of the Royal Botanic Gardens (Peradeniya)

4: 372-444,

Plectania seaveri sp. nov. (Bermuda) ... 329

Pfister DH, Slater C, Hansen K. 2008. Chorioactidaceae: a new family in the Pezizales (Ascomycota)

with four genera. Mycological Research 112: 513-527.

http://dx.doi.org/10.1016/j.mycres.2007.11.016.

Phillips B. 1984. Bermuda cedar: survival or extinction. Bermuda's Heritage: 150-151.

Seaver FJ. 1928. North American cup-fungi (Operculates). New York. 284 p.

Seaver FJ. 1942. North American cup-fungi (Operculates). Supplemented edition. New York.

377 p.

Seaver FJ, Waterston JM. 1946. Contributions to the mycoflora of Bermuda IV. Mycologia 38(2):

180-201. http://dx.doi.org/10.2307/3755060.

Smith AH, Smith HV, Weber NS. 1981. How to know the non-gilled mushrooms. 2™ Edition,

Pictured Key Nature. 336 p.

Sterrer W. 1998. How many species are there in Bermuda? Bulletin of Marine Science 62:

809-840.

Tai FL. 1979. Sylloge Fungorum Sinicorum. China, Beijing Science Press. 1527 p.

Teng SC. 1963. Chung-kuo Ti Chen-chun. (Fungi of China). Peiping, Science Press. 586 p.

Thomas MLH. 2004. The natural history of Bermuda. Bermuda Zoological Society, Bermuda.

255 p.

Tylutki EE. 1993. Mushrooms of Idaho and the Pacific Northwest Vol. I, Discomycetes. University

of Idaho Press. 133 p.

Waterston JM. 1947. The fungi of Bermuda. Bermuda Dept. of Agriculture. 305 p.

Zhuang W-Y, Wang Z. 1998. Sarcosomataceous discomycetes in China. Mycotaxon 67: 355-364.

MY COTAXON

http://dx.doi.org/10.5248/120.331

Volume 120, pp. 331-342 April-June 2012

New species of Entoloma (Basidiomycetes, Agaricales)

from Kerala State, India

C.K. PRADEEP’ , K.B. VRINDA’*, SHIBU P. VARGHESE! & T.J. BARON?’

"Tropical Botanic Garden & Research Institute,

Palode, Trivandrum, Kerala, 695562, India

*Department of Biological Sciences, State University of New York,

College at Cortland, New York, USA.13045

*CORRESPONDENCE TO: drckpradeep@hotmail.com

ABSTRACT — Three new species of Entoloma (E. suaveolens, E. crassum, and E. aurantio-

quadratum) are described, illustrated, and discussed based on collections made from Kerala

State, India.

KEY WORDS — agaric diversity, under-explored, Western Ghats

Introduction

Entoloma is one of the largest genera of Agaricales, with more than 1500

species known worldwide (Noordeloos 1981a). However, vast areas are still

under-explored, especially in Africa, South America, India, and South East

Asia (Noordeloos & Morozova 2010). Recent studies of the genus in Kerala

State, India, by Manimohan et al. (1995, 2002, 2006) documented 39 species, of

which 26 are new to science, indicating that Entoloma is especially rich in the

state. While documenting the agaric diversity of Western Ghats of Kerala, the

authors collected several specimens of Entoloma. Our detailed study of these

collections has resulted in three new species, which are described, illustrated,

and discussed here.

Materials & methods

Gross morphological descriptions are based exclusively on fresh materials

collected from Kerala State, India. Color coding follows Kornerup & Wanscher (1978).

Microscopic characters were studied from hand cut sections of dried basidiomata

revived in a 3% KOH aqueous solution, stained with 1% Congo red, and examined

under a Leica DME 1000 compound microscope. The mean quotient (Q) of spore length

divided by spore width was calculated from measurements of 30 basidiospores; the hilar

332 ... Pradeep & al.

appendix was included in the measurement of the spore length. Line drawings were

made with assistance of an attached drawing tube. All specimens cited were collected

by the authors unless otherwise indicated. Holotypes are deposited at the Herbarium of

the Royal Botanic Gardens, Kew (K), and all isotypes and additional materials examined

are deposited at the Mycological Herbarium of Tropical Botanic Garden and Research

Institute, Trivandrum (TBGT).

Taxonomy

Entoloma suaveolens C.K. Pradeep & K.B. Vrinda, sp. nov. PLATE 1A-B; FIG. 1

MycoBank MB 564454

Differs from other blue Entoloma species in combining an appressed squamulose pileus

and stipe with the lack of hymenial cystidia and the presence of a strong fragrant odor.

Type -— India, Kerala State, Trivandrum District, Palode, TBGRI campus, 25 Sep 1996,

Pradeep 3621 (Holotype, K(M) 172393).

ETyMOLocy -suaveolens refers to the fragrant odor of the basidiomes

PiLEus 20-53 mm diam., convex with a broad obtuse umbo; surface ink blue,

dark blue to greyish blue (20F4/21F4/21F8/22F4/21D5) at disc with greyish

violet to dark blue (18E5/19E4/19E5/19E6) elsewhere, appressed squamulose

throughout, often cracked to expose the white underlying context in dry

season; squamules often washed off during heavy rain towards margin to

appear subsquamulose, dry; margin straight, entire. LAMELLAE adnate, white,

cream to greyish orange (443/4B4/5A2/5B3/6A2/6B4), < 11 mm wide, close to

crowded, with lamellulae of different lengths; edge concolourous, entire. STIPE

35-114 x 3-10 mm, central, cylindric, curved, fistulose to narrowly hollow,

brittle, tapering up from a subclavate base; surface mostly concolorous with

pileus or greyish violet (19D5), greyish blue (20D5), appressed squamulose

throughout; base white with white mycelial mat and mycelial cords. CONTEXT

white, up to 3 mm, soft. OporR strong, distinct, pleasant, fragrant, similar to

holy ash. Spore PRINT pink.

BasIp1osPpores 10-12 x 5-7.5(-8) um, avL = 11, avW = 6.77, Q = 1.3-1.7,

avQ = 1.65, heterodiametric ovate, 5-7 facets in profile. BAstp1A 32-42.5 x 8.5-14

um, clavate, 4-spored; sterigmata up to 4.5 um long. LAMELLA EDGE fertile.

CHEILOCYSTIDIA and pleurocystidia absent. HYMENOPHORAL TRAMA regular;

hyphae 2.5-16(-28) um wide, thin-walled, hyaline. PILEAL TRAMA interwoven;

hyphae similar to hymenophoral trama. PILEIPELLIs a trichoderm with clavate

to cylindro-clavate elements, 62.5-147 x 10-14 um, thin-walled with bluish

(in water) contents which dissolve in 3% KOH. STIPITIPELLIS a cutis of loosely

arranged septate thin-walled hyphae, 2.4-4.8 um wide, with pale greyish

plasmatic contents. CAULOCYSTIDIA in tufts present on the upper part of the

stipe, 41-67 x 5-9 um, cylindro-clavate, thin-walled with pale brown vacuolar

contents. CLAMP CONNECTIONS and oleiferous hyphae present in all parts.

Entoloma spp. nov. (India) ... 333

1S)

Fic. 1: Entoloma suaveolens. A, basidia; B, basidiospores;

C, trichodermial elements; D, caulocystidia. Scale bars = 10 um.

334 ... Pradeep & al.

HABIT & HABITAT — Solitary, scattered on soil among decayed litter in

evergreen forest, June-December.

ADDITIONAL SPECIMENS EXAMINED — INDIA, KERALA STATE, TRIVANDRUM DISTRICT,

Palode, TBGRI campus: 16 Sep 1997, TBGT 3887; 15 Oct 2001, TBGT 6770; 8 Jun 2005,

TBGT 8907; 4 Nov 2005, TBGT 9425; 17 Jul 2007, TBGT 10427; 25 Jul 2008, TBGT

11513; 30 Jul 2008, TBGT 11554; 16 Dec 2009, TBGT 13058; Wayanap District,

Nadavayal: 27 Sep 2007, TBGT 10575 (all at TBGT).

Discussion — Entoloma suaveolens is characterized by the unique

combination of medium to large bluish terrestrial tricholomatoid basidiomes,

convex umbonate appressed squamulose pileus and stipe, adnate pink

lamellae, heterodiametric ovate basidiospores, absence of hymenial cystidia,

trichodermial pileipellis, abundant caulocystidia, clamp connections present

in all hyphae, and strong fragrant odor.

The present collections invite comparison with a number of blue Entoloma

species described from various regions of the world by different workers.

Entoloma rugosopruinatum Corner & E. Horak, originally described from Sabah

(Horak 1980) and recently recorded from Kerala (Manimohan et al. 1995),

differs by its violaceous purple finely tomentose pileus, presence of cylindro-

clavate cheilocystidia, and subfarinaceous odor. Entoloma divum Corner &

E. Horak is distinguished by a deep blue pruinose pileus, palisadic pileipellis,

and lack of any odor. Three other comparable blue species —E. simillimum

Corner & E. Horak, E. egregium E. Horak, E. marinum Corner & E. Horak— are

easily separated by their velutinous pileus, presence of cheilocystidia, and lack

of any distinctive odor. The European taxa E. chalybeum (Pers.) Noordel. and

E. chalybeum var. lazulinum (Fr.) Noordel. (Noordeloos 1992) are distinguished

by their depressed velutinous pileus, heterodiametrical 5-9-angled spores, sterile

lamella edge, a cutis pileipellis, and absence of clamps. Entoloma bloxamii (Berk.

& Broome) Sacc. and E. nitidum Quél. (Horak 1973, 1978, 1980; Noordeloos

1992), though comparable in gross morphology and pigmentation, vary

markedly in their smooth glabrous subviscid pileus (pileipellis an ixocutis) and

distinctive farinaceous odor. Entoloma dichroum (Pers.) P. Kumm. (Noordeloos

1992) differs in its violaceous purple pileus, heterogeneous lamella edge, and

unpleasant odor. The recently described E. eugenei Noordel. & O. Morozova

(Noordeloos & Morozova 2010) from Russia differs in its velvety deep blue

pileus, sterile lamellae edge with cylindrical, narrowly lageniform or irregularly

shaped cheilocystidia, and mild spicy odor.

Leptonia carnea Largent, a rather uncommon species restricted to coastal

and northern California, is characterized by a blue black densely appressed

fibrillose squamulose pileus and stipe, blue—violet-black lamella edge, cuticular

pileipellis, and distinctive farinaceous odor and taste (Largent 1977); L. carnea

is further distinguished by the insoluble blue pigment, contrasting with the

readily KOH-soluble pigment in E. suaveolens.

Entoloma spp. nov. (India) ... 335

Two recently described blue species from Kerala State, E. griseolazulinum

Manim. & Noordel. and E. indoviolaceum Manim. & Noordel. (Manimohan

et al 2006), clearly differ in their velutinous pileus, presence of cheilocystidia,

large heterodiametric spores, and lack of any distinct odor.

Tricholomatoid habit, convex umbonate bluish pileus, trichodermial

pileipellis, and presence of clamp connections place this species in subg.

Leptonia sect. Leptonia (Noordeloos 1981a).

Entoloma crassum C.K. Pradeep & K.B. Vrinda, sp. nov. PLATE 1C-D; Fic. 2

MycoBank MB 564455

Differs from Entoloma flavidum in its glabrous creamy to yellow white pileus, lack of

cheilocystidia, and strong, nauseating odor.

Type — India, Kerala State, Trivandrum District, Palode, TBGRI campus: 7 Oct 2004,

Pradeep 8154 (Holotype, K(M) 172394).

ETYMOLOGY —crassum refers to the thick, stout basidiomes

Basip1oMsEs thick, fleshy, robust, tricholomatoid. PiLeus 45-160 mm diam.,

convex to planoconvex with a broad obtuse umbo, becoming uplifted in old

ones; surface uniformly creamy white to yellowish white (442/443) when young

and yellowish white to brownish orange (442/443/3B3/3C3/4B2/ 4B3/4B4) with

brownish disc (48B3/4B4/4C4/5C2/5C3/5D3/5D4/5E3/5E4/6E4) in mature ones,

moist, smooth, glabrous, not hygrophanous; margin incurved when young

becoming straight, wavy, folded, entire to incised. LAMELLAE broadly adnate to

subdecurrent, yellowish white to greyish red (442/5A2/6A2/5B3/6C6/7B4), < 20

mm wide, close to crowded with lamellulae of 3-4 lengths; edge concolourous,

entire. STIPE 35-150 x 10-25 mm, central (often excentric in some specimens),

cylindric (rarely compressed), twisted, thick, fibrous, solid, equal, with a slightly

broad base; surface white, cream, ivory (483) turning brownish orange (5c3),

hair brown (584), greyish brown (6D3), brownish grey (6£2) on handling,

fibrillose striate, smooth. Basal white mycelial mat present in some specimens.

CONTEXT white, < 11 mm thick, soft, fleshy. ODor strong, unpleasant, pungent,

nauseating. SPORE PRINT cinnamon to reddish brown (6D6/8D6).

BASIDIOSPORES (8—)9-10 x (6-)6.5-7.5 um, avL = 9.3, avW = 6.8, Q = 1.25-1.50,

avQ = 1.34, heterodiametric ovate with 5-6 facets in profile. Basidia 45-58.5

x 7.5-10 um, clavate, 4-spored. LAMELLA EDGE fertile, CHEILOCYSTIDIA and

pleurocystidia absent. HYMENOPHORAL TRAMA regular; hyphae 2.5-14 um

wide, thin-walled, hyaline. PILEAL TRAMA interwoven; hyphae similar to that of

hymenophoral trama. PILEIPELLIs a cutis passing to a trichoderm at the centre,

elements 15-38 x 4-8 um, clavate, cylindro-clavate to strangulated, thin-

walled, hyaline. STIPITIPELLIs a cutis of longitudinally parallel hyphae, 2.5-5

uum wide, thin-walled, hyaline. CauLocystip1a abundant throughout the stipe

surface, 25-80 x 6-9 um, lageniform or narrowly utriform, cylindric, cylindro-

336 ... Pradeep & al.

clavate, subutriform, strangulated, flexuous, often with a short subobtuse apex,

thin-walled with intracellular granular contents. CLAMP CONNECTIONS and

OLEIFEROUS HYPHAE present.

HasitT & Hasitat — Solitary, scattered or in groups on soil among litter in

evergreen forest or under reeds in sandy soil on riverbanks, April-December.

ADDITIONAL SPECIMENS EXAMINED — INDIA, KERALA STATE, TRIVANDRUM DISTRICT,

Palode, TBGRI campus: 21 Oct 1995, TBGT 2758; 19 May 1997, TBGT 3858; 13 May

1999, TBGT 4659; 19 Jun 2000, TBGT 5072; 21 Jun 2000, TBGT 5081; 27 Jun 2001,

TBGT 5351; 29 Apr 2002, TBGT 5502; 7 Jul 2005, TBGT 9064; 23 Sep 2005, TBGT

9278; 17 Jun 2008, TBGT 11122; 25 Jun 2008, TBGT 11237; 30 Sep 2008, TBGT 12064;

4 Nov 2008, TBGT 12189; 11 Jun 2009, TBGT 12577; 16 Jun 2009, TBGT 12606; 29

Jun 2009, TBGT 12685; 16 Sep 2009, TBGT 12918; 23 Sep 2009, TBGT 12940; 14 Oct

2009, TBGT 12980; 23 Oct 2009, TBGT 13012; 25 Nov 2009, TBGT 13110; 11 Dec 2009,

TBGT 13128; 6 Jul 2010, TBGT 13373; 4 Aug 2011, TBGT 13768; Kallar: 10 Jul 2007,

TBGT 10411 (all at TBGT).

Discuss1on — Entoloma crassum is a very remarkable species characterized by

large robust tricholomatoid basidiomata, a convex umbonate smooth glabrous

yellowish white pileus with a brownish disc, broadly adnate to subdecurrent

lamellae, a fibrillose striate white stipe that bruises brown, strong unpleasant

nauseating odor, heterodiametric spores, fertile lamella edge, trichodermial

pileipellis, caulocystidia that are abundant over the entire stipe surface, and

presence of clamp connections in all hyphae.

The most closely related entolomas with large tricholomatoid yellowish

white basidiomes are compared and discussed here. Despite its macroscopic

and microscopic similarities, E. flavidum (Massee) Corner & E. Horak (Horak

1980) differs from E. crassum in its white subtomentose to floccose pileus that

yellows with age, hollow fibrillose-striate non-yellowing white stipe, cylindric

to subclavate cheilocystidia arrayed on a sterile lamella edge, a cuticular

pileipellis, sour odor, and absence of caulocystidia. Among similar species that

differ by possessing an ixocuticular pileipellis, E. sinuatum (Bull.) P. Kumm.

(Noordeloos 1981b) is further distinguished by its viscid pale ochraceous

pileus, characteristically yellow lamellae, and small isodiametric spores;

E. niphoides Romagn. ex Noordel. (Largent 1994) is separated by its snow-white

pileus and stipe and fruity odor; E. moserianum Noordel. (Noordeloos 1983)

is distinguished by its typically yellow spotted pileus, lamellae, and stipe and

sterile lamella edge; E. albomagnum G.M. Gates & Noordel. and E. cretaceum

G.M. Gates & Noordel. (Gates & Noordeloos 2007) disagree by their pure white

pileus and small spores; and E. sepium (Noulet & Dass.) Richon & Roze and

E. saundersii (Fr.) Sacc. (Breitenbach & Kranzlin 1995) differ mainly in their

silky white pileus, farinaceous odor, and lack of caulocystidia.

The large tricholomatoid habit, nonhygrophanous convex umbonate smooth

pileus, adnate decurrent lamellae, regular hymenophoral trama, and abundant

Entoloma spp. nov. (India) ... 337

Fic. 2: Entoloma crassum.

A, basidia; B, basidiospores; C, trichodermial elements; D, caulocystidia. Scale bars = 10 um.

338 ... Pradeep & al.

clamps indicate that the species belongs to subg. Entoloma sect. Entoloma

(Noordeloos 1981a).

Entoloma aurantioquadratum C.K. Pradeep & K.B. Vrinda, sp. nov.

MycoBAank MB 564456 PLATE 1E-F; FIG. 3

Differs from Entoloma talisporum and E. gracilius in the absence of clamp connections,

from E. hyalodepas in its large quadrate basidiospores, and from E. albogracile in the

absence of hymenial cystidia.

Type — India, Kerala State, Trivandrum District, Palode, TBGRI campus: 11 Nov 2009,

Shibu 13048 (Holotype, K(M) 172395).

ETYMOLOGY — aurantioquadratum refers to the orange basidiome with quadrate

spores

PrLEus 10-30 mm diam., convex to applanate with a small shallow depression

at centre, often uplifted in old ones; surface orange white to brownish orange

(5A2/5A3/5B2/5B3/5C3), with hair brown to greyish brown (5£4/7D3/7E3)

centre or even paler towards margin, dry, velvety at disc (under a lens), elsewhere

smooth, glabrous, nonhygrophanous; margin straight to uplifted in old ones,

wavy, entire to incised, pellucid striate when wet. LAMELLAE adnexed, orange

white to pinkish white (542/7A2), up to 4 mm wide, close with lamellulae of

different lengths; edge concolourous to the sides, entire. STIPE 20-50 x 1-3

mm, central, cylindric, twisted, hollow, equal, curved, often narrowly tapering

up from a slightly broad base; surface white turning brown on handling or

bruising, pruinose in the upper part (under the lens) elsewhere smooth and

glabrous. CONTEXT thin, concolourous to pileus, soft. Odor none.

BASIDIOSPORES (9-)9.5-12.5(-13) x (6-)6.5-8.5(-9.5) um, avL = 10.8, avW

= 7.6, Q = 1.05-1.7, avQ = 1.4, quadrate in profile. Basidia 23-36 x 9-12.5 um,

clavate, 4-spored. LAMELLA EDGE fertile, CHEILOCystTID1A and pleurocystidia

absent. HYMENOPHORAL TRAMA regular, 2-12 um wide, thin-walled, hyaline.

PILEAL TRAMA interwoven; hyphae similar to hymenophoral trama. PILEIPELLIS

a cutis with transitions towards a trichoderm at centre, elements 19.5-65 x

7.5-11 um, clavate to cylindro-clavate, thin-walled with brown plasmatic

intracellular contents. STIPITIPELLIS a cutis of loosely arranged, septate, thin-

walled, 2-4 um wide, hyaline hyphae. CAULOCysTIDIA in groups on the upper

part, 14-32 x 4.5-10.5 um, clavate, cylindro-clavate with septate base, thin-

walled, hyaline. CLAMP CONNECTIONS absent. OLEIFEROUS HYPHAE present.

HaBiT & HaBiTaT — Scattered on forest floor in tropical evergreen forest,

July-November.

ADDITIONAL SPECIMENS EXAMINED — INDIA, KERALA STATE, TRIVANDRUM DISTRICT,

TBGRI campus: 10 Nov 2009, TBGT 13043; 15 Jul 2011, TBGT 13685; 11 Aug 2011,

TBGT 13781 (all at TBGT).

Discussion — Entoloma aurantioquadratum is a typical member of subg.

Inocephalus due to its nonhygrophanous nonstriate pileus and quadrate spores.

Entoloma spp. nov. (India) ... 339

Fic. 3: Entoloma aurantioquadratum.

A, basidia; B, basidiospores; C, trichodermial elements; D, caulocystidia. Scale bars = 10 um.

340 ... Pradeep & al.

PLATE 1: A-B, Entoloma suaveolens; C-D, E. crassum;

E-E, E. aurantioquadratum. Scale bars: A-B, E-F = 10 mm; C-D = 20 mm.

Entoloma spp. nov. (India) ... 341

The combination of small collybioid basidiomes with pale orange pileus, white

stipe turning brownish, large spores, fertile lamellae edge, cuticular pileipellis

with transitions towards a trichoderm, presence of caulocystidia, and lack of

clamp connections are diagnostic for this species.

A comprehensive literature search (Horak 1976, 1977, Hesler 1967, Pegler

1986, Noordeloos 1987, 1992, Noordeloos & Hausknecht 2007, Manimohan

et al. 1995, 2006) revealed that although some species are comparable, none

matches E. aurantioquadratum exactly. Entoloma talisporum Corner & E.

Horak differs in its yellowing white fibrillose pileus, smaller cuboid spores, and

presence of clamp connections. Entoloma hyalodepas (Berk. & Broome) E. Horak

is distinguished by its depressed to infundibuliform ivory yellow or ochraceous

buff pileus, decurrent lamellae, large pentagonal spores, and cuticular pileipellis

(Baroni & Lodge 1998). Entoloma albogracile E. Horak has a convex-umbonate

papillate white to yellowish pileus, cylindrical to subclavate cheilocystidia, and

cuboid to tetrahedral spores. Entoloma gracilius E. Horak can be separated

by its small fragile basidiomes, yellowish green tinted pileus, broadly adnate

lamellae, smaller cuboid spores, and presence of clamp connections.

Acknowledgments

The authors are grateful to Drs. Alfredo Justo (Biology Department, Clark

University, Worcester, USA) and Laura Guzman-Davalos (Departamanto de Botanica

y Zoologia, Universidad de Guadalajara, Mexico) for their pre-submission reviews and

suggestions.

Literature cited

Baroni TJ, Lodge DJ. 1998. Alboleptonia from the Greater Antilles. Mycologia 90: 680-696.

http://dx.doi.org/10.2307/3761227

Breitenbach J, Kranzlin F 1995. Fungi of Switzerland 4: Agarics 2nd part. Mykologia Lucerne,

Switzerland.

Gates GM, Noordeloos ME. 2007. Preliminary studies in the genus Entoloma in Tasmania - I.

Persoonia 19: 157-226.

Hesler LR. 1967. Entoloma in southeastern North America. Beihefte Nova Hedwigia 23: 1-196.

Horak E. 1973. Fungi Agaricini Novazeylandiae. 1. Entoloma Fr. and related genera. Beihefte Nova

Hedwigia 43: 1-86.

Horak E. 1976. On cuboid species of Entoloma. Sydowia 28: 171-236.

Horak E. 1977. Additions to “On cuboid-spored species of Entoloma”. Sydowia 29: 289-299.

Horak E. 1978. Entoloma in South America. 1. Sydowia 30: 40-111.

Horak E. 1980. Entoloma (Agaricales) in Indomalaya and Australasia. Beihefte Nova Hedwigia 65:

1-352;

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3rd ed. Methuen, London.

Largent DL. 1977. The genus Leptonia on the Pacific coast of the United States. Bibliotheca

Mycologica 55: 1-309.

Largent DL. 1994. Entolomatoid fungi of the Western United States and Alaska. Mad River Press,

Eureka, CA.

342 ... Pradeep & al.

Manimohan P, Vijaya Joseph A, Leelavathy KM. 1995. The genus Entoloma in Kerala State, India.

Mycological Research 99: 1083-1097. http://dx.doi.org/10.1016/S0953-7562(09)80777-6

Manimohan P, Leelavathy KM, Noordeloos ME. 2002. Three new species of Entoloma from Kerala

State, India. Persoonia 17: 625-630.

Manimohan P, Noordeloos ME, Dhanya AM. 2006. Studies on the genus Entoloma (Basidiomycetes,

Agaricales) in Kerala State, India. Persoonia 19: 45-93.

Noordeloos ME. 1981a. Introduction to the taxonomy of the genus Entoloma sensu lato (Agaricales).

Persoonia 11: 121-151.

Noordeloos ME. 1981b. Entoloma subgenera Entoloma and Allocybe in the Netherlands and adjacent

regions with a reconnaissance of their remaining taxa in Europe. Persoonia 11: 153-256.

Noordeloos ME. 1983. Studies in Entoloma — 9. On two new European species in section Entoloma.

Sydowia 36: 208-212.

Noordeloos ME. 1987. Entoloma (Agaricales) in Europe. Beihefte Nova Hedwigia 91: 1-417.

Noordeloos ME. 1992. Entoloma s.l. Fungi Europaei, Saronno, Italy.

Noordeloos ME, Hausknecht A. 2007. The genus Entoloma (Basidiomycetes, Agaricales) of the

Mascarenes and Seychelles. Fungal Diversity 27: 111-144.

Noordeloos ME, Morozova OV. 2010. New and noteworthy Entoloma species from the Primorsky

Territory, Russian Far East. Mycotaxon 112: 231-255. http://dx.doi.org/10.5248/112.231.

Pegler DN. 1986. Agaric flora of Sri Lanka. Kew Bulletin Additional Series 12: 1-519.

MY COTAXON

http://dx.doi.org/10.5248/120.343

Volume 120, pp. 343-352 April-June 2012

A new species of Infundichalara from pine litter

ONDREJ KOUKOL

Department of Botany, Charles University in Prague,

Bendtska 2, CZ-128 01 Prague, Czech Republic

CORRESPONDENCE TO: ondrej.koukol@natur.cuni.cz

ABSTRACT — A new species of Infundichalara (anamorphic Helotiales), a hitherto monotypic

genus, is described from pine needle litter. Infundichalara minuta sp. nov. forms two

conidiophore types. Erect penicillate conidiophores producing catenulate non-septate

hyaline conidia form more frequently than Chalara-like conidiophores with funnel-shaped

collarettes produce wedge-shaped phialoconidia. Although its morphological characters

correspond more with Xenopolyscytalum, a three-region DNA analyses (ITS, 28S rDNA and

EF-1la) of Infundichalara, Xenopolyscytalum, and related Chalara species in the Helotiales

confirm the phylogenetic position of the new species within Infundichalara. It differs from

I. microchona by smaller phialides and wedge-shaped conidia.

Key worps — hyphomycete, dimorphic conidiophores, litter saprotroph

Introduction

Scots pine (Pinus sylvestris L.) needles in litter host several microfungal

species producing straight erect penicillate conidiophores with catenulate

hyaline conidia. Polyscytalum fecundissimum Riess and P. pini P.M. Kirk &

Minter produce uniseptate conidia from denticulate conidiogenous cells

growing on a straight, erect conidiophore (Kirk 1983). Hormiactella asetosa

Hol.-Jech., differing from Polyscytalum by conidia with rounded ends and dark

grey colonies (Holubova-Jechova 1978), predominantly colonizes bark but

may also be found on needles in the litter (Koukol 2007). A monotypic genus

Xenopolyscytalum Crous was described for X. pinea Crous to accommodate

two strains isolated from pine needles (Crous & Groenewald 2010). Unlike

Polyscytalum and Hormiactella representatives, X. pinea forms white tufts on

needles and is characterized by aseptate conidia with somewhat darkened hila

and a Chalara-like synanamorph. The synanamorph produces phialides with a

flaring collarette that differs from the tubular collarette found in Chalara (Corda)

344 ... Koukol

Rabenh. (Nag Raj & Kendrick 1975). The only Chalara species characterized by

a funnel-shaped collarette, C. microchona W. Gams, was recently placed into a

new genus, Infundichalara Réblova & W. Gams (Réblova et al. 2011).

I have isolated one fungus with dimorphic (penicillate/Chalara-like)

conidiophores from needles of several pine species from various parts of Europe.

These isolates are morphologically similar to both Xenopolyscytalum (abundant

production of penicillate conidiophores) and Infundichalara (Chalara-like

conidiophores with funnel-shaped collarettes) but obviously represent a

distinct species. Both morphological and molecular criteria were considered to

accommodate the species in a genus reflecting evolutionary relationships.

Materials & methods

Collection and isolation

Cultures were isolated from needles of Scots pine (Pinus sylvestris), Swiss stone pine

(P. cembra L.), Siberian pine (P. sibirica Du Tour), and Siberian dwarf pine (P pumila

(Pallas) Regel) in the litter. Needles were sampled in pure or mixed pine forests in the

NP Bohemian Switzerland (Czech Republic), along the rivers Timpton (Republica

Sakha) and Vyerchni Sakujan (Zabaykalskaya Oblast) in Russia, and in the Windachtal

Valley, Tyrol (Austria). Surface sterilisation of needles and cultivation conditions were

identical to those described in Koukol (2010). Culture characteristics were observed on

2% malt agar (2MA, final sucrose content 2% w/v, 18 g agar, 1 | distilled water) prepared

from brewer's wort (Staropramen Brewery, Prague, Czech Republic), potato carrot agar

(PCA), oatmeal agar (OA), and potato-dextrose agar (PDA), all prepared from fresh

ingredients according to Fassatiova (1986). Agar plates were maintained at laboratory

temperature (22-25°C). For microscopy and measurements, conidiophores with conidia

were mounted in Melzer’s reagent and examined with phase or differential interference

contrast (using an Olympus BX51 microscope with digital camera, measured using

Quick Photo software). Pictures were further edited in Adobe Photoshop®. Microscopic

measurements are reported as the mean + standard deviation of 30-50 measurements

with the extremes given in parentheses.

The holotype specimen is deposited in the PRM (National Museum, Prague, Czech

Republic) and living cultures are maintained in the CCF (Culture Collection of Fungi,

Prague, Czech Republic).

DNA extraction, amplification and sequencing

DNA was isolated from cultures grown on 2MA using the ZR Fungal/Bacterial DNA

kit (Zymo Research, Orange, CA, USA). DNA analyses were made with sequences from

the regions ITS1+5.8S+ITS2 (referred to it as the ITS rDNA) and 28S rDNA together

with the gene coding for the translation elongation factor 1 a (EF-la). Amplification

and sequencing were performed according to Koukol (2011) with the exception that

only the shorter part of EF-1a was amplified with primer pairs 983F a 1567R (Rehner

& Buckley 2005). New sequences generated in this study from newly isolated strains

are listed in TaBLE 1. Sequences of ITS and 28S rDNA of X. pinea were obtained from

Infundichalara minuta sp. nov. ... 345

TABLE 1. Accession numbers of new sequences generated in this study.

SPECIES / CULTURE ITS 28S RDNA EF-1a

Infundichalara minuta

CCF4156, ex-type HE603986 HE603981 HE603978

CCF4157 HE603987 HE603982 HE603977

CCF4158 HE603988 HE603983 HE603976

CCF4159 HE603989 HE603984 —

CCF4160 HE603990 HE603985 —

Xenopolyscytalum pinea

CPC14225, ex-type — _— HE603980

CPC14234 — — HE603979

GenBank. The remaining sequences of Chalara spp., Cistella acuum (Alb. & Schwein.)

Svréek, and Infundichalara microchona (W. Gams) Réblova & W. Gams originated in

Koukol (2011).

Phylogenetic analyses

All markers were aligned using MUSCLE (Edgar 2004) and adjusted manually

in BioEdit, v. 4.7.1 (Hall 1999) to maximize similarities. The dataset consisted of 32

sequences and comprised 1322 characters with 272 parsimony-informative sites.

Phylogenetic analyses were conducted using MEGA v. 5 (Tamura et al. 2011), PhyML

v. 3.0 (Guindon et al. 2010) and MrBayes v. 3.1.2 (Ronquist & Huelsenbeck 2003). For

the Bayesian analyses, General Time Reversible model with invariant sites and a gamma

rate distribution (GTR+I+G) was used due to its maximum complexity. The dataset was

divided to three partitions (ITS, 28S rDNA and EF-1a). The proportion of variable sites,

rate frequencies and the gamma shape were estimated separately for the three partitions.

Markov chains were initiated from a random tree and were run for 6,000,000 generations;

samples were taken every 100th generation. Posterior probabilities (PP) were used as a

Bayesian branch support on the consensus trees. In addition, bootstrap branch support

(BS) values were estimated using the Maximum-Likelihood (ML) analyzes with 10,000

bootstrap replicates (MEGA 5.0, PhyML v. 3.0). The tree was rooted with Tryblidiopsis

pinastri (Pers.) P. Karst. (AFTOL-ID 1319, sequences of 28S rDNA DQ470983, EF-1la

DQ471106).

Results

Five strains isolated during this study (CCF4156-4160) formed a strongly

supported group sister to I. microchona (CBS175.74 and CBS889.73). The isolate

B282 obtained from pine needles in Tyrol (Austria) clustered with X. pinea

(Fic. 1). It most probably belonged to X. pinea, but was not morphologically

observed, because it was lost due to contamination after DNA extraction.

Xenopolyscytalum pinea (CPC14225 and CPC14234) clustered with low support

with Cistella acuum (CCF3970).

346 ... Koukol

100/99 , C. hyalocuspica CCF3976

C. hyalocuspica CCF3975

C. affinis CBS562.77

C. pseudoaffinis CBS261.75

159 100/- | C. pseudoaffinis CCF3979

C. microspora CBS131.74

C. microspora CCF3980

135+ C. microspora CCF3981

100/98 - C. holubovae CCF3978

C. holubovae ZK68

Cistella acuum CCF3970

99/100

100/100

61/- 86/74, X. pinea CPC14225

an 99/62! | X pinea CPC14234

X pinea B2S2

100/71 C. recta CCF 3936

C. recta S17-2

79/82 | C. longipes CCF3973

C. longipes CCF3972

92/88 C. longipes CBS264.94

56/- C. longipes CBS411.76

100/99 C. longipes CCF3974

70184 | ©. longipes ZK66

C. piceae-abietis CCF3983

100/84 C. piceae-abietis CCF3982

81/100 — |. microchona CBS175.74

| microchona CBS889.73

/5A |. minuta CCF4160

|. minuta CCF4159

I. minuta CCF4156

|. minuta CCF4157

100/67 L |. minuta CCF4158

Tryblidiopsis pinastri AFTOL-ID 1319

96/-

100/63 100/-

0.1

Fic. 1. Phylogenetic relationships of Infundichalara minuta and selected members of Helotiales

derived from ITS, 28S rDNA and EF-la gene regions using ML and BA analyses. Genera are

abbreviated as C. = Chalara, X. = Xenopolyscytalum, and I. = Infundichalara. Ex-type sequences are

in bold. Bootstrap values > 50% are indicated along nodes. Thick lines show PP > 0.95. The tree was

rooted with Tryblidiopsis pinastri.

Taxonomy

Infundichalara minuta Koukol, sp. nov. Figs 2, 3 a,b

MycoBank MB 563538

Differs from Infundichalara microchona by its smaller Chalara-like phialides

producing wedge-shaped conidia with truncate base and synanamorph with penicillate

conidiophores, ramoconidia, and subcylindrical conidia.

Infundichalara minuta sp. nov. ... 347

Fic. 2. Infundichalara minuta: colonial and microscopic morphology. a-d, Morphological

variation on 2MA after 30 d incubation at 22-25°C; e, penicillate conidiophore with ramoconidia.

f-h, Chalara-like conidiophores (f, intercalary; h, terminal); i, conidia produced on penicillate

conidiophores; j, conidia produced from Chalara-like conidiophores—single, short chain,

and cluster. (a,e,g,i,j = ex-type strain CCF4156; b = CCF4160; c,fjh = CCF4158; d = CCF4159.

Bars = 20 um.)

TyPE: Czech Republic, Bohemian Switzerland National Park, Doubice, Tokan, forest

of P. sylvestris on sandstone rocks, 50°53.3'N 14°24.8'E, 450 m asl., on needles in the

litter of P. sylvestris, 11 Dec 2006, leg. O. Koukol (Holotype, PRM899344; ex-type strain

CCF4156).

ETYMOLOGY: minutus = referring to the minute Chalara-like conidiophores.

VEGETATIVE MYCELIUM consisting of smooth or warted, branched, septate,

hyaline to pale brown hyphae 1.5-3 um diam. ConipiopHoREs dimorphic.

PENICILLATE CONIDIOPHORES erect, producing branched, catenulate conidia

visible as white tufts on needles and in culture, pale brown, 3-5-septate, < 40

um tall, 3-3.5 um wide. CONIDIOGENOUS cells apical, hyaline to pale brown,

smooth, 9.5-13.5(-15.5) x 1.5-3.5 um, proliferating sympodially, usually giving

rise to 3 ramoconidia. RaMoconrpzI¢ hyaline, smooth, aseptate, cylindrical to

subcylindrical, (8—)9-13(-16) x 1.5-3 um. Conip1A subcylindrical, hyaline,

smooth, aseptate, occurring in unbranched dispersible chains, ends with a

flattened, slightly protruding scar, 0.8 um wide, (5-)7-9.5(-11.5) x 1.5-2.5 um.

CHALARA-LIKE CONIDIOPHORES produced less frequently, formed either by

conidiogenous cell on the mycelium or by conidiogenous cell on 1-4 basal cells,

348 ... Koukol

that are erect, cylindrical, unbranched, pale brown, smooth, 4-10 um tall (rarely

< 50 um), 2.5-4 um wide. CONIDIOGENOUS CELLS phialidic, hyaline to slightly

pigmented, mostly formed terminally or intercalary directly on mycelium or

on one supporting cell, subconical to lageniform, (8-)9.5-13.5(-20) x 3-4.5

um, collarette mostly funnel-shaped, rarely cylindrical, 2.5-5 x 1.5-2.5 um.

ConipiA wedge-shaped with truncate base, hyaline, smooth, 3-4(-5) x 2-3

um, forming either short chains or clusters on top of the phialide.

Cotonts variable, different strains produced different morphologies in

the dark, after 1 mo at 22-25°C (Fic. 2a-d). Colonies on 2MA erumpent in

the centre, with dense aerial mycelium, surface dark gray to brown with pale

margin; reverse pale gray to dark brown, reaching 8-28 mm diam. On PDA

aerial mycelium developed, colonies gray to pale brown, protuberant in the

centre, reaching 10-26 mm diam. On OA dendritic, slimy, aerial mycelium

only at margins, surface dark gray to brown, reaching 17-22 mm diam. On

PCA gray to pale brown, immersed, with protuberant centre, reaching 8-33

mm diam. The strain CCF4157 produced pale white immersed mycelium on

OA, 2MA and PCA and diffused violet pigment into agar on PDA and reached

< 40 mm diam. on each agar medium tested. Most intensive sporulation of all

strains was observed on 2MA.

EcoLocy & DISTRIBUTION — colonizes needles in the litter and humus of

P. sylvestris, P. cembra, P. sibirica, and P. pumila, in Czech Republic, Austria,

Russia (this study) and Sweden (concluded from Lindahl et al. 2010).

ADDITIONAL SPECIMENS EXAMINED: AUSTRIA, TyRroi, Windachtal Valley, sparse P.

cembra forest above river, 46°57.57'N 11°02.61'E, 1828 m asl., on P. cembra needles

in litter, 25 Jun 2010, leg. O. Koukol (PRM899346, living culture CCF4160). RUSSIA,

REPUBLICA SAKHA, sparse growth of P sibirica within Larix gmelinii (Rupr.) Rupr. on

the right bank of river Timpton near the branching with Ojumrak, 57°11.3'N 126°6.6'E,

550 m asl., on P. sibirica needles in litter, 4 Aug 2010, leg. D. Svoboda (living culture

CCF4157); sparse growth of P sibirica within L. gmelinii on the right bank of river

Timpton near the branching with Bolshoy Ulimakh 58°37.6’N 127°1.1'E, 270 m asl.,

P. sibirica needles in litter, 10 Aug 2010, leg. D. Svoboda (PRM899345, living culture

CCF4158); ZABAYKALSKAYA OBLAST, P. pumila forest on the right bank of river

Vyerchni Sakujan, 56°48.5'N 118°6.9’E, 766 m asl., on P pumila needles in litter, 12 Aug

2010, leg. D. Svoboda (living culture CCF4159).

Discussion

Infundichalara minuta shares morphological characteristics with both

Infundichalara and Xenopolyscytalum. It produces hyaline to slightly

pigmented phialides with funnel shaped collarettes and conidia that are

truncate on the base and rounded at the apex (Fic. 2f-h,j), which is consistent

with Infundichalara (Réblova et al. 2011). On the other hand, Chalara-like

phialides and a Polyscytalum-like synanamorph with penicillate conidiophores

(Fra. 2e,i) forming white tufts on the substrate are typical for Xenopolyscytalum

Infundichalara minuta sp. nov. ... 349

Fic. 3. Infundichalara minuta (ex-type strain CCF4156): a, Penicillate conidiophore and conidia;

b, Chalara-like conidiophores and conidia. Xenopolyscytalum pinea (ex-type strain CPC14225):

c, penicillate conidiophore and conidia. (Bar = 20 um).

(Crous & Groenewald 2010). Molecular data confirmed placement of the new

species in Infundichalara, despite the closer morphological similarity with

Xenopolyscytalum. Obviously, the presence of a Polyscytalum-like synanamorph

is not phylogenetically significant at the genus level but may be used for

recognizing the species (e.g., C. holubovae Koukol) that form typical Chalara

phialides and produce a synanamorph with fusiform conidia (Koukol 2011).

The previously monotypic Infundichalara and Xenopolyscytalum represented

by I. microchona and X. pinea respectively may be distinguished from I. minuta

350 ... Koukol

based on morphology. Infundichalara microchona produces larger phialides

(18-35 x 2.5-4.0 um) but with shorter collarettes. The similarly sized

phialoconidia of I. minuta and I. microchona differ in shape — wedge-shaped

in I. minuta but clavate in I. microchona (Gams & Holubova-Jechova 1976).

Infundichalara minuta phialides with the rather narrow collarette typical of

Chalara s.str. are considered exceptional. Such variation has been noted as well

for I. microchona (strain CBS125.74) with slightly longer collarettes (to 4 um

deep; Gams & Holubova-Jechova 1976). Vice versa, molecular data support

C. longipes (strain CBS867.73), which produces phialides with both types of

collarettes, in Chalara (Koukol 2011). Xenopolyscytalum pinea is distinguished

by shorter conidia formed on penicillate conidiophores (3-4(-7) x 1.5(-2)

um (Fic. 3c), larger Chalara-like phialides (15-25 x 2-3 um), and cylindrical

phialoconidia. Culture characteristics also differ, for X. pinea grew more quickly

on all tested media (Crous & Groenewald 2010).

In this study, only freshly isolated cultures of I. minuta were surveyed. Pure

cultures were obtained by picking conidia from penicillate conidiophores

(white tufts on the surface-sterilised needles) in primary isolations after several

weeks of cultivation. In pure cultures, this synanamorph also dominated and

Chalara-like phialides were formed relatively rarely. Penicillate conidiophores

and phialides were never observed growing from the same hyphae, but potential

contamination of surveyed strains may be disproved as individual isolates were

obtained independently from three distant localities.

It remains unknown whether I. minuta forms predominantly penicillate

synanamorph also on field substrates. Similarly, Crous & Groenewald (2010)

do not mention the prevalence of either form of X. pinea on natural substrates.

The assumption that penicillate conidiophores may dominate on the substrate

as well is indicated by the white colonies formed by an unknown Polyscytalum-

like fungus that have been repeatedly found on Scots pine needles sampled

close to the type locality of I. minuta and cultivated in damp chambers (Koukol

2007). Herbarium specimen “Polyscytalum sp. X07” (private herbarium of the

author) sampled in September 2000 and surveyed in this study contained a

fungus producing relatively dense growth of Polyscytalum-like conidiophores

(12-18 x 2 um) with chains of cylindrical, 0-1-septate, hyaline conidia (5-13

x 1.5 um). Except for the septate conidia, its description matches I. minuta,

but a fresh collection must first be isolated into pure culture and molecularly

analyzed before it can be regarded as conspecific with I. minuta.

All I. minuta strains were obtained from pine litter, but the species also

colonizes Scots pine humus according to 99% similarity (505/507 bp) of the ITS

rDNA region with sequence GU559086 obtained in the study of Lindahl et al.

(2010). Infundichalara minuta seems to have a negligible effect on degradation

of structural polysaccharides in the litter. In a parallel study, strain CCF4156,

Infundichalara minuta sp. nov.... 351

analyzed for the production of ten hydrolytic enzymes during submersed

cultivation in a low-nitrogen medium, expressed only very low amounts of

cellobiohydrolase and B-glucosidase (Koukol & Baldrian 2012).

Acknowledgments

Iam very grateful to Dr. David Svoboda, who sampled litter needles on his journey

across Russia, to Dr. Pedro Crous for sending me two strains of X. pinea. Dr. Pedro

Crous and Dr. Roland Kirschner significantly contributed to the improvement of the

manuscript as pre-submission reviewers. This study was supported by the Grant Agency

of the Czech Republic, project No. 206/09/P295 and by the Ministry of Education, Youth

and Sports of the Czech Republic (MSM0021620828).

Literature cited

Crous PW, Groenewald JZ. 2010. Fungal planet 55. Xenopolyscytalum pinea. Persoonia 25:

130-131.

Crous PW, Schubert K, Braun U, Hoog GS de, Hocking AD, Shin H-D, Groenewald JZ. 2007.

Opportunistic, human-pathogenic species in the Herpotrichiellaceae are phenotypically similar

to saprobic or phytopathogenic species in the Venturiaceae. Studies in Mycology 58: 185-217.

Edgar RC. 2004. MUSCLE: a multiple sequence alignment method with reduced time and space

complexity. BMC Bioinformatics 5: 113. http://dx.doi.org/10.1186/1471-2105-5-113

Fassatiova O. 1986. Moulds and filamentous fungi in technical microbiology. Amsterdam,

Elsevier.

Gams W, Holubova-Jechova V. 1976. Chloridium and some other dematiaceous hyphomycetes

growing on decaying wood. Studies in Mycology 13: 1-99.

Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O. 2010. New algorithms and

methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML

3.0. Systematic Biology 59: 307-321. http://dx.doi.org/10.1093/sysbio/syq010

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NTT. Nucleic Acids Symposium Series 41: 95-98.

Holubova-Jechova V. 1978. Lignicolous hyphomycetes from Czechoslovakia 5. Septonema,

Hormiactella and Lylea. Folia Geobotanica Phytotaxonomica 13: 421-442.

Kirk PM. 1983. New or interesting microfungi. IX. Dematiaceous hyphomycetes from Esher

Common. Transactions British Mycological Society 80: 449-467.

http://dx.doi.org/10.1016/S0007-1536(83)80041-2

Koukol O. 2007. Effect of Pinus strobus L. invasion on the mycoflora of pine litter needles in the

Bohemian Switzerland National Park (Czech Republic). 229-233, in: H Hartel et al. (eds).

Sandstone Landscapes. Academia, Prague.

Koukol O. 2010. Revision of “Septonema ochraceum” revealed three new species of Venturiaceae

and Herpotrichiellaceae. Mycological Progress 9: 369-378.

http://dx.doi.org/10.1007/s11557-009-0645-x

Koukol O. 2011. New species of Chalara occupying coniferous needles. Fungal Diversity 49: 75-91.

http://dx.doi.org/10.1007/s13225-011-0092-2

Koukol O, Baldrian P. 2012. Intergeneric variability in enzyme production of microfungi from pine

litter. Soil Biology & Biochemistry 49: 1-3. http://dx.doi.org/10.1016/j.soilbio.2012.02.004

Nag Raj TR, Kendrick WB. 1975. A monograph of Chalara and allied genera. Wilfr. Laurier Univ

Press, Waterloo

352 ... Koukol

Lindahl BD, de Boer W, Finlay RD. 2010. Disruption of root carbon transport into forest humus

stimulates fungal opportunists at the expense of mycorrhizal fungi. ISME Journal 4: 872-881.

http://dx.doi.org/10.1038/ismej.2010.19

Réblova M, Gams W, Stépanek V. 2011. Thenewhyphomycete genera Brachyalara and Infundichalara,

the similar Exochalara and species of ‘Phialophora sect. Catenulatae’ (Leotiomycetes). Fungal

Diversity 46: 67-86. http://dx.doi.org/10.1007/s13225-010-0077-6

Rehner SA, Buckley E. 2005. A Beauveria phylogeny inferred from nuclear ITS and EF1-a sequences:

evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97: 84-98.

http://dx.doi.org/10.3852/mycologia.97.1.84

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. http://dx.doi.org/10.1093/bioinformatics/btg180

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGAS5: Molecular

Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and

Maximum Parsimony Methods. Molecular Biology and Evolution.

http://dx.doi.org/10.1093/molbev/msr121

MY COTAXON

http://dx.doi.org/10.5248/120.353

Volume 120, pp. 353-355 April-June 2012

Clavulicium hallenbergii, a new corticioid species

from India

AVNEET P. SINGH, JASPREET KAUR & G.S. DHINGRA*

Department of Botany, Punjabi University

Patiala 147 002, India

*CORRESPONDENCE TO: dhingragurpaul@gmail.com

ABSTRACT - A new corticioid species, Clavulicium hallenbergii, is described from Chamba,

in Himachal Pradesh.

Key worps -— Manimahesh, Pinus wallichiana

During the fungal forays conducted in Dibri-Dhansho area in district Chamba

of Himachal Pradesh, India, Avneet P. Singh collected a corticioid fungus on a

trunk of Pinus wallichiana. Oil-rich clavate basidia and oil-rich basidiospores

pointed in the direction of the genus Clavulicium (Agaricomycetes). After

detailed comparison of macroscopic and microscopic characters with the

relevant literature (Burt 1926, Parmasto 1968, Eriksson & Ryvarden 1973) we

concluded that the fungus represents a new species in the genus Clavulicium,

which herewith is described.

Clavulicium hallenbergii Avneet P. Singh, J. Kaur & Dhingra sp. nov.

MycoBAnk MB 560931 FIGs 1-5

Differs from Clavulicium macounii by its narrower basidia, smaller basidiospores, and

lack of gloeocystidia.

Type: India, Himachal Pradesh: Chamba, Manimahesh, on way from Dibri towards

Dhansho, on the trunk of Pinus wallichiana A.B. Jacks., 20 September 2005, Avneet 3683

(PUN, holotype)

Erymo.ocy: In honour of the mycologist Prof. Nils Hallenberg, University of

Gothenburg, Gothenburg, Sweden.

Basidiocarp resupinate, adnate, effused, up to 175 um thick in section;

hymenial surface smooth, brownish red to light brown; margins whitish,

thinning, fibrillose, to indeterminate, loosening upon drying in mature

354 ...Singh, Kaur & Dhingra

10 poo at

BOCOO0

20 ym

pole

\ iN a

Fics 1-4. Clavulicium hallenbergii, microscopic structures.

1. basidiospores; 2. basidia; 3. generative hyphae; 4. section through basidiocarp.

basidiocarp. Hyphal system monomitic; generative hyphae up to 3.0 um wide,

branched, septate, clamped; basal hyphae somewhat thick-walled, parallel to

the substrate; subhymenial hyphae thin-walled, vertically directed. Basidia

39.0-47.0 x 3.0-5.0 um, clavate, sinuous, 4-sterigmate, with oily contents and

basal clamp; sterigmata up to 3.0 um long. Basidiospores 6.2-7.8 x 4.5-5.5 um,

broadly ellipsoid to obovate, smooth, thin-walled, inamyloid, acyanophilous,

with oil droplets.

REMARKS—C. hallenbergii resembles C. macounii (Burt) J. Erikss. & Boidin

ex Parmasto in terms of morphology of the basidiocarp, clamped generative

Clavulicium hallenbergii sp. nov. (India) ... 355

Fic. 5. Clavulicium hallenbergii, basidiocarp (holotype).

hyphae and abundant oily contents in basidia and basidiospores. However,

C. macounii differs in having gloeocystidia, wider basidia, and larger

basidiospores (9-12 (-13) x 6.5-8 um; Eriksson and Ryvarden, 1973).

Acknowledgements

Authors thank Prof. Nils Hallenberg (Evolutionary Botany, Botanical Institute,

Gothenburg, Sweden) and Prof. B.M. Sharma (Department of Plant Pathology, COA,

CSKHPAU, Palampur, H.P., India) for peer review and Prof. Joost Stalpers (CBS

Fungal Biodiversity Centre, Netherlands) for providing the Mycobank number. Head,

Department of Botany, Punjabi University, Patiala, is thanked for providing research

facilities.

Literature cited

Burt EA. 1926. The Thelephoraceae of North America: XV. Corticium and supplement to the whole

series. Ann. Mo. Bot. Gard. 13: 173-357.

Parmasto E. 1968. Conspectus systematis Corticiacearum. Tartu. 262 pp.

Eriksson J, Ryvarden L. 1973. Corticiaceae of North Europe - II. Fungiflora, Oslo. pp 59-286.

MY COTAXON

http://dx.doi.org/10.5248/120.357

Volume 120, pp. 357-360 April-June 2012

Vararia longicystidiata sp. nov. (Agaricomycetes) from India

SAMITA, S.K. SANYAL’, G.S. DHINGRA & AVNEET P. SINGH

Department of Botany, Punjabi University, Patiala 147 002, India

*CORRESPONDENCE TO: skskumar731@gmail.com

ABSTRACT - A new corticioid species, Vararia longicystidiata, is described on decaying wood

of Quercus incana from Uttarakhand state in India.

Key worps - Basidiomycota, Chaurangi Khal, Uttarakashi

While conducting fungal forays in Chaurangi Khal area of district Uttarakashi,

Uttarakhand (India), Samita collected an unknown corticioid fungus on

decaying wood of Quercus incana. The presence of dichohyphidia and

inamyloid smooth basidiospores indicated that the species belonged to Vararia,

but it could not be assigned to any previously described species (Welden 1965,

Parmasto 1970, Boidin & Lanquetin 1976, Rattan 1977, Boidin et al. 1980,

Hallenberg & Eriksson 1985, Pegler & Young 1993). A detailed description of

the specimen along with the microphotographs and line diagrams was sent

to Prof. Nils Hallenberg (Sweden), who supported the description of a new

species in the genus Vararia.

Vararia longicystidiata Samita, Sanyal, Dhingra & Avneet P. Singh, sp. nov.

MycoBAnk 562081 Fics 1-7

Differs from Vararia gomezii in its larger cystidia, basidia, and basidiospores.

Type: India, Uttarakhand: Uttarakashi, Chaurangi Khal, on decaying wood of Quercus

incana Bartram, 29 September 2011, Samita 4413 (PUN, holotype).

Erymo.oey: The epithet refers to the long cylindrical cystidia.

Basidiocarp resupinate, adnate, effused, up to 230 um thick in section,

hymenial surface smooth to somewhat tuberculate, light orange; margins

thinning, fibrillose, paler concolorous to whitish. Hyphal system monomitic;

generative hyphae up to 5.5 um wide, branched, septate, generally without

clamps; basal hyphae loosely arranged, more or less parallel to the substrate,

interspersed with some thick-walled hyphae with oily contents, some cells

358 ... Samita & al.

10nm

—

-—

~——_ ae

microscopic structures.

Fics 1-6. Vararia longicystidiata,

1. Basidiospores; 2. basidia; 3. generative hyphae; 4. cystidia;

5. dichohyphidia; 6.section through basidiocarp.

Vararia longicystidiata sp. nov. (India) ... 359

Fic. 7. Vararia longicystidiata, basidiocarp (holotype).

inflated; subhymenial hyphae vertical, compactly arranged. Dichohyphidia

more common in subhymenium than hymenium, abundant, coralloid,

branches irregular with blunt endings, dextrinoid. Cystidia 100.0-188.0 x

8.8-17.5 um, abundant, subcylindrical to sinuous, thick-walled, without basal

clamp, negative to sulphovanillin. Basidia 70.0-110.0 x 9.0-13.0 um, clavate,

4-sterigmate, frequently with secondary septa, without basal clamp, with or

without oily contents; sterigmata up to 12.5 um long; basidioles rich in oily

contents. Basidiospores 15.0-20.0 x 6.0-7.5 um, sub-fusiform to navicular,

with numerous oil-drops, thin- to somewhat thick-walled, inamyloid,

acyanophilous.

REMARKS—Vararia longicystidiata resembles V. gomezii Boidin. & Lanq.

(reported from South America and Africa) in having similar basidiocarps and

basidiospore shape. However, V. gomezii differs in having ovoid unthickened

cystidia (14-50 x 8-14 um), smaller basidia (34.0-45.0- x 7.5), and smaller

basidiospores (13.0-14.8 x 4.8-6. 25 um; Boidin and Lanquetin; 1977).

Acknowledgements

The authors thank Head, Department of Botany, Punjabi University, Patiala,

for providing research facilities; Prof. Nils Hallenberg (University of Gothenburg,

360 ... Samita & al.

Gothenburg, Sweden) for expert comments and peer review; Prof. B.M. Sharma

(Department of Plant Pathology, COA, CSKHPAU, Palampur, H.P., India) for peer

review.

Literature cited

Boidin J, Lanquetin P. 1976 [“1975”]. Vararia subgenus Vararia (Basidiomycetes Lachnocladiaceae):

étude spéciale des espéces d'Afrique intertropicale. Bull. Soc. Mycol. France 91(4): 457-513.

Boidin J, Lanquetin P. 1977. Les genres Dichostereum et Vararia en Guadeloupe (Basidiomycetes,

Lachnocladiaceae). Mycotaxon 6: 277-336

Boidin J, Lanquetin P, Gilles G. 1980. Application du concept biologique de Tespéces aux

basidiomycetes: le genre Vararia (section Vararia) au Gabon. Cryptog. Mycol. 1: 265-384.

Hallenberg N, Eriksson J. 1985. The Lachnocladiaceae and Coniophoraceae of North Europe.

Fungiflora, Oslo. pp. 1-96.

Pegler DN, Young TWK. 1993. Basidiome structure in Lachnodiales sensu lato. Kew Bullein 48:

oye.

Parmasto E. 1970. The Lachnocladiaceae of the Soviet Union with a key to boreal species. Scripta

Mycologia 2: 1-168

Rattan SS. 1977. The resupinate Aphyllophorales of the northwestern Himalayas. Bibliotheca

Mycologica 60: 1-427.

Welden AL. 1965. West Indian species of Vararia with notes on extralimital species. Mycologia 57:

502-520.

MY COTAXON

http://dx.doi.org/10.5248/120.361

Volume 120, pp. 361-371 April-June 2012

A new cystidiate variety of Omphalina pyxidata (Basidiomycota,

tricholomatoid clade) from Italy

ALFREDO VIZZINI’*, MARIANO CurRTI’?,, MARCO CONTU? & ENRICO ERCOLE!

‘Dipartimento di Scienze della Vita e Biologia dei Sistemi - Universita degli Studi di Torino

Viale Mattioli 25, I-10125, Torino, Italy

? Via Tito Nicolini 12, Pozzaglia Sabina, I-02030 Rieti, Italy

> Via Marmilla 12 (I Gioielli 2), I-07026 Olbia (OT), Italy

*CORRESPONDENCE TO: alfredo. vizzini@unito. it

ABSTRACT — A new variety of Omphalina pyxidata, var. cystidiata, is here described and

illustrated based on morphological and molecular data. The new combination Infundibulicybe

lateritia is introduced.

Key worps — Agaricomycetes, Agaricales, omphalinoid fungi, Contumyces, taxonomy

Introduction

Within the omphalinoid fungi — small agarics with a convex to deeply

umbilicate pileus, central stipe, thin context, decurrent lamellae, white spore-

print, and thin-walled inamyloid smooth spores (Norvell et al. 1994, Lutzoni

1997, Redhead et al. 2002a,b) — taxa with cystidiate basidiomata are thus far

known only in the hymenochaetoid clade (Blasiphalia Redhead, Contumyces

Redhead et al., Rickenella Raithelh.; Moncalvo et al. 2000, 2002, Redhead et al.

2002b, Larsson et al. 2006, Larsson 2007).

During fieldwork focused on bryophilous Galerina species, we collected an

omphalinoid fungus growing on mosses close to Galerina discreta E. Horak

et al. We initially believed that these specimens represented a new species

of Contumyces (Contu 1997, Redhead et al. 2002b) based on the presence of

well-differentiated pileo-, caulo-, cheilo-, and pleurocystidia and an irregular

hymenophoral trama. However an ITS-rDNA analysis implied they might

represent an unknown as-yet undescribed variety of Omphalina pyxidata (Bull.)

Quél., the type species of Omphalina Quél. (Omphalina s.s. = tricholomatoid

clade sensu Matheny et al. 2006, Binder et al. 2010 and Vizzini et al. 201 1a). This

genus is characterized by bryophilous basidiomata with a reddish brown tinged

362 ... Vizzini & al.

pileus and stipe, paler and well-developed lamellae, a smooth (not scaly) pileus,

absence of hymenial and pileal cystidia, and presence of clamp-connections

(Redhead et al. 2002a, Elborne 2008). We fully describe and illustrate the new

taxon below.

Materials & methods

Morphology

The macromorphological descriptions follow the detailed field notes taken on fresh

material. The micromorphological descriptions are based both upon study of fresh

and herbarium material. Dried material was revived in 5% KOH and stained in Congo

Red, Cotton Blue, and Melzer’s reagent. The basidiospore range and means are based

on measurements of 25 spores. In the macro- and microscopic descriptions Q = the

quotient of spore length and width and Qm = the average quotient; L = number of entire

lamellae; | = number of lamellulae between each pair of entire lamellae. The basidial

width was measured at the broadest part, and the length was measured from the apex

(sterigmata excluded) to the basal septum. Colour codes in brackets (e.g., Se 45) follow

Séguy (1936), hereafter referred to as (Se). Author citations follow the IPNI Authors

and the Index Fungorum Authors of Fungal Names websites. Herbarium abbreviations

follow Thiers (2011) with the exception of “EM” and “GT”, referring to the personal

herbaria of Enzo Musumeci and Gérard Trichies, respectively. The type collection is

housed at TO (Herbarium of the Department of Plant Biology, University of Turin, Italy),

and both name and Latin diagnosis of the new variety as well as the new combination

are deposited in MycoBank (http://www.mycobank.org).

DNA extraction, PCR amplification, DNA sequencing

Genomic DNA was isolated from 1 mg of 3 herbarium specimens (TABLE 1) using the

DNeasy Plant Mini Kit (Qiagen, Milan Italy). Universal primers ITS1£/ITS4 were used

for the ITS region amplification (White et al. 1990; Gardes & Bruns 1993). Amplification

reactions were performed in PE9700 thermal cycler (Perkin-Elmer, Applied Biosystems)

in a 25 ul reaction mixture using the following final concentrations or total amounts:

5 ng DNA, 1x PCR buffer (20 mM Tris/HCl pH 8.4, 50 mM KCl), 1 uM of each primer,

2.5 mM MgCl2, 0.25 mM of each dNTP, 0.5 unit of Taq polymerase (Promega). The

PCR program was as follows: 3 min at 95°C for 1 cycle; 30 s at 94°C, 45 s at 50°C,

2 min at 72°C for 35 cycles, 10 min at 72°C for 1 cycle. PCR products were resolved on a

1.0% agarose gel and visualized by staining with ethidium bromide. PCR products were

purified and sequenced by DiNAMYCODE srl (Turin, Italy). Sequence assembly and

editing were performed using Geneious v5.3 (Drummond et al. 2010). The sequences

are deposited in GenBank under the accession numbers given in TABLE 1.

Sequence alignment and phylogenetic analysis

Sequences included in the phylogenetic analyses were either generated in this study

(TABLE 1) or retrieved from GenBank. Multiple sequence alignments for ITS fragments

were generated using MAFFT (Katoh et al., 2002) with default conditions for gap opening

and gap extension penalty. The alignment was slightly edited using MEGA 5.0 (Tamura

et al. 2011). Phylogenetic analyses were performed using both Bayesian Inference (BI)

Omphalina pyxidata var. cystidiata var. nov. (Italy) ... 363

TABLE 1. Omphalina pyxidata collections used in this study.

COLLECTIONS ! COLL. NO., COUNTRY, DATE, COLLECTOR ! ITS Acc. No.

. pyxidata , France, 12/1 , G. Trichies 1

*O. pyxidata 1 GT99398, F 02/12/1999, G. Trich JQ671000

*O. pyxidata var. cystidiata : TO HG2512, Italy, 11/11/2010, M. Curti : JQ671002

* = collections newly sequenced in this study.

and Maximum Likelihood (ML) approaches. The BI was performed with MrBayes

3.1.2 (Huelsenbeck & Ronquist 2001) with four incrementally heated simultaneous

Monte Carlo Markov Chains (MCMC) ran over 10 millions generations, under GTR+I

evolutionary model. Trees were sampled every 1000 generations resulting in an overall

sampling of 10,001 trees; the first 2500 trees were discarded as “burn-in” (25%). For the

remaining trees, a majority rule consensus tree showing all compatible partitions was

computed to obtain estimates for Bayesian Posterior Probabilities (BPP). Branch lengths

were estimated as mean values over the sampled trees. ML was performed with RAXML

(Stamatakis 2006) under GTRGAMMA model and using thorough bootstrap with

20 runs and 1000 replicates. In both analyses a Pseudoclitocybe cyathiformis sequence

(Genbank HM191730) was used as outgroup taxon, according to Binder et al. (2010)

and Vizzini et al. (2011a). The ML consensus tree was used merely for comparison with

the Bayesian tree and to support the analysis. However the BPP (Bayesian posterior

probability) over 0.75 and the ML bootstrap (MLB) over 50% values are reported in the

resulting tree. Pairwise % identity values of ITS sequences were calculated using MEGA

5.0 (Tamura et al. 2011).

Results

Molecular results

Bayesian and Maximum likelihood inferences were performed on a total

of 18 samples, including 15 sequences available from GenBank and three

newly sequenced specimens (TABLE 1). Final alignment length was 731 bp.

Both Bayesian and Maximum likelihood analyses produced the same topology

(Fic. 1). In both analyses our cystidiate collection clusters with four O. pyxidata

sequences clearly forming a strongly supported clade (BPP 0.92 and MLB 85%).

The five O. pyxidata sequences show a pairwise % identity value of 99.8 and

could be considered conspecific.

The O. pyxidata sequence deposited in Genbank under the accession number

JF908502 (MCVE 15669) was not used in the analysis because a Blastn search

364 ... Vizzini & al.

Omphalina pyxidata 3 JN944402

0.77/90

Omphalina pyxidata 4 OPU66450

Omphalina pyxidata 1 JQ671000

0.92/85

Omphalina pyxidata var. cystidiata JQ671002

Ompbhalina pyxidata 2 JQ671001

Omphalina rivulicola GU234117

diss Omphalina cf. rivulicola HQ445617

Omphalina rivulicola ORU66451

Omphalina chionophila GU234144

Omphalina rivulicola JF908501

Clitocybe lateritia CLU66431

Omphalina cf. rivulicola HQ445603

0.76/84 Omphalina rivulicola GU234109

Infundibulicybe gibba DQ490635

Infundibulicybe gibba HM631716

Infundibulicybe mediterranea HM631724

Clitocybe alkaliviolascens JF907807

Pseudoclitocybe cyathiformis HM191730

0.03 expected changes per site

FicuRe 1. Bayesian phylogram obtained from the ITS sequence alignment. Support values for clades

that are supported in either the Bayesian (Posterior Probabilities values - BPP) and Maximum

likelihood (ML Bootstrap percentage - MLB) analyses are indicated. BPP above 0.75 and MLB

above 50% are given above branches. Taxa in bold represent newly sequenced collections. Numbers

(1-4) refer to the collections reported in TABLE I.

implies it represents a non-omphalinoid species in the Pluteus cinereofuscus/

P. nanus complex.

Clitocybe lateritia, together with two O. rivulicola (J. Favre) Lamoure

collections, clearly falls within Infundibulicybe. Clitocybe alkaliviolascens Bellu

clusters close to Infundibulicybe mediterranea Vizzini et al.

Taxonomy

Omphalina pyxidata var. cystidiata M. Curti, Contu & Vizzini, var. nov. Fics 2-3

MycoBank MB564484

A varietate typica differt praesentia cystidiorum in lamella, pileo, et stipite. Habitat:

graegatim, ad muscos.

Type — Italy, Latium, prov. Rieti, com. Pozzaglia Sabina, Valle del Turano, 42.1596°N

12.9652°E, 878 m, 11.X1.2010, leg. M. Curti (TO HG2512 holotype).

EryMoLocy — the specific epithet refers to the presence of hymenial as well as of pileo-

and caulocystidia.

Omphalina pyxidata var. cystidiata var. nov. (Italy) ... 365

FiGuRE 2. Omphalina pyxidata var. cystidiata. Basidiomata. Bar = 20 mm.

Piteus 3-15 mm broad, at first convex to applanate, later with a slightly

depressed centre, not or slightly hygrophanous, minutely pubescent, dark

brown-fulvous (Se 96, 121, 126, 146, 251, 252) fading to light brown (Se 133,

134, 173); margin at first involute then expanding, smooth to slightly striate in

old basidiomes, whitish in dried basidiomes. LAMELLAE (L = 18-25; ] = 1-2)

decurrent, thick, distant with lamellulae occasionally interspersed, strongly

forked and intervenose, whitish to light ochre (Se 199-200), with entire

concolorous edge. Stripe 15-40 x 1-3 mm, cylindrical, slightly broadened at

the base, sinuous-flexuous, solid then fistulous, cartilaginous, paler than pileus,

light ochre-brown (Se 133, 174, 203-204), minutely pruinose to subglabrous.

CoNnTEXT elastic, thin, whitish-ochre to light ochre-brown (Se 133, 134) in

surface; smell and taste not distinctive. SPORE PRINT white.

SPORES (7-)7.5-9(-10) x 4.5-6(-6.5) um, Q = 1.3-1.7, Qm = 1.51, largely

ellipsoid to oblong in frontal view, lacrymoid to slightly amygdaliform in side-

view, smooth, thin-walled, hyaline, inamyloid, mono-biguttulate, with a thick

and evident hilum. Basrp1a 28-46(-60) x 6-9 um, clavate, (two-) four-spored,

sterigmata 3-7 um long, usually clamped. CHEILOCysTIDIA 23-45 x 3-6 x

2.5-5 um, abundant, cylindrical, slender, apically tapered or slightly swollen,

some laterally ventricose, hyaline, thin-walled, often with inner microguttules.

PLEUROCYSTIDIA 25-50(-65) x 3.5-5.5 x 2.5-5(—7) um, similar to cheilocystidia,

but slightly longer. HYMENOPHORAL TRAMA irregular, consisting of 3.3-7 um

wide hyphae. PILEIPELLIS a cutis consisting of cylindrical, interwoven, 6-10

366 ... Vizzini & al.

FIGURE 3. Omphalina pyxidata var. cystidiata. Microscopic features. a. Spores showing protruding

hilum. b. Basidium with basal clamp. c. Pileipellis and pileocystidia. d. Encrusting pigment of

pileipellis elements. e. Cheilocystidia (with inner microguttules). f. Pleurocystidia. g. Caulocystidia.

h. Irregular hymenophoral trama. Bars: a~d = 10 um; e—h = 20 um.

Omphalina pyxidata var. cystidiata var. nov. (Italy) ... 367

um wide hyphae; terminal elements with a subclavate 7-13 um wide apex;

subcutis made up of 3-7 um wide hyphae; PIGMENT epiparietal, minutely to

strongly encrusting; PILEOCYSTIDIA numerous, fusiform to lageniform, 35-60

x 3.5-9 x 3.5-5 um. STIPITIPELLIS a cutis made up, in 5% KOH, of ochre-yellow

3-8 um wide hyphae bearing minute incrustations; CAULOCYSTIDIA similar to

hymenial cystidia (30-65 x 4-5 x 3-5 um). STIPITITRAMA non-sarcodimitic,

consisting of hyphae up to 16 um wide. CLAMP CONNECTIONS present at all

septa. THROMBOPLEROUS HYPHAE (= oOleiferous hyphae sensu Clémencon

2004) present. APPRESSORIA on host surface not observed.

HaBItatT — Gregarious, rarely subcaespitose, on Tortula muralis

gametophytes (Bryophyta).

ADDITIONAL MATERIAL STUDIED — Omphalina pyxidata var. pyxidata: FRANCE,

LORRAINE, MOSELLE, Moyeuvre-Petite, 02 Dec 1999, on mosses, leg. G. Trichies

(GT99398); SWITZERLAND, CANTON BasEL StTapT, Lange Erlen, 06 Jul 2005,

on mosses, 250 m asl, leg. E. Musumeci (EM0434-05); ITALY, PrEDMONT, CHISONE

VALLEY, Pinerolo, Pra Martino, 10 Oct 2010, on mosses, 1000 m asl, leg. A. Vizzini (TO

AV98).

Discussion

As highlighted by Vizzini et al. (201la), Omphalina s.s. is sister to

Infundibulicybe Harmaja, a genus recently segregated from Clitocybe (Fr.)

Staude (Harmaja 2003). In the six-gene region sequence analyses by Matheny &

al. (2006), Infundibulicybe falls outside the tricholomatoid clade, occupying an

isolated position in the Agaricales (incertae sedis). However, recent analyses by

Binder et al. (2010), Vizzini et al. (2011a) and Matheny (pers. comm.) place it

with robust support as sister to the rest of the tricholomatoid clade. Omphalina

and Infundibulicybe share the cream-reddish brown tinges of pileus and stipe,

the long-decurrent lamellae, and strongly encrusting pigment (Harmaja 2003,

Vizzini et al. 2011b).

Our molecular analysis (Fic. 1) and morphological comparison support

the new taxon as a cystidiate variant of O. pyxidata thus far never described.

Omphalina pyxidata var. cystidiata is unique within Omphalina s.s., where it is

the only taxon thus far included that possesses well-developed cheilo-, pleuro-

, pileo-, and caulocystidia (Bigelow 1970, 1985, Singer 1970, Lamoure 1974,

1975 1982; Clémencon 1982, Bon 1997, Elborne 2008). Furthermore, we have

found no cystidia in three examined European collections of O. pyxidata (see

ADDITIONAL MATERIAL STUDIED). Morphologically, our new taxon resembles

the omphalinoid bryophilous genera belonging to the hymenochaetoid clade

(Redhead et al. 2002b, Antonin & Noordeloos 2004). However, Rickenella

differs in having a regular hymenophoral trama (Contu 1997, Redhead et al.

2002b) and Blasiphalia by growing specifically on the Blasia pusilla gametophyte

(Marchantiophyta) and forming appressoria on host surface (Larsson et al.

368 ... Vizzini & al.

2006). Apart from molecular data that clearly support our new variety within

the Agaricales, morphologically Contumyces (= Jacobia Contu, nom. illeg.;

Hymenochaetales) might accommodate our taxon. Contumyces so far comprises

only three species — C. rosellus (M.M. Moser) Redhead et al., C. vesuvianus

(F. Brig.) Redhead et al., and C. brunneolilacinus (Contu et al.) Redhead et al.

(Redhead et al. 2002b, Antonin & Noordeloos 2004) — but only the first named

has been sequenced. Contumyces rosellus differs from Omphalina pyxidata var.

cystidiata by the pink to pale salmon-pink colouration, white stipe, and longer

and larger cystidia. Contumyces vesuvianus has orange to fulvous basidiomata,

larger basidiospores, rarer differently shaped (mucronate/rostrate) cystidia

and intracellular pigment in the pileipellis hyphae. As originally described

(Contu 1989), C. brunneolilacinus differs in having a lilaceous-brown, sharply

tomentose, hygrophanous pileus, pink-lilac lamellae, a Pelargonium-like smell,

lageno-capitate cystidia, and intracellular pigment in the pileipellis hyphae.

Finally, C. brunneolilacinus sensu Antonin & Noordeloos (2004) differs in

having longer and larger cystidia and a non-tomentose pileal surface.

Our sequence analyses have also revealed that Clitocybe lateritia clusters

in Infundibulicybe with two collections clearly misdetermined as O. rivulicola

(Fic. 1). The three sequences are clearly conspecific (pairwise % identity

value = 99.7). The two “O. rivulicola” sequences were derived from soil or

ectomycorrhizal samples sampled from Dryas octopetala beds, as reported in two

recent papers dealing with alpine-arctic fungi (Bjorbeekmo et al. 2010, Geml et

al. 2012). Clitocybe lateritia, a rare alpine-arctic species strictly associated with

Dryas octopetala (Rosaceae), should be transferred to Infundibulicybe based on

both sequence analyses and its Infundibulicybe-like features (reddish-brown

colouration, strongly encrusting pigment in the pileipellis, partly lacrymoid

spores) as inferred from Favre (1955), Gulden & Jennsen (1988), and Bon

(1997). Consequently, we propose the following new combination:

Infundibulicybe lateritia (J. Favre) Vizzini & Contu, comb. nov.

MycoBank MB564485

= Clitocybe lateritia J. Favre, Ergebn. wiss. Unters. schweiz.

Natn. Parks, n.s. 5(33): 54, 199 (1955).

Type: Switzerland, Munt la Schera, 2400 m a.s.l., in Dryas beds, 21.08.1941, leg. J. Favre

(Lectotype designated here, G-K16433; specimen illustrated in Favre 1955: Fig. 34; Pl.

IV, Fig. 11).

Finally, Clitocybe alkaliviolascens, a species of the Infundibulicybe gibba complex

recently described from Mediterranean areas and characterized by a dark brown

pileus surface turning pink-violet with a 30% KOH drop, and spores reaching

10 um in length (Bellu 1995, 1996), also belongs in Infundibulicybe. Bellu will

propose the new combination in a forthcoming paper (Bellu, pers. comm.).

Omphalina pyxidata var. cystidiata var. nov. (Italy) ... 369

Acknowledgements

Our most sincere thanks are due to Scott Redhead (Ottawa, Ontario, Canada), Lorelei

Norvell (Portland, Oregon, USA), and Shaun Pennycook (Auckland, New Zealand) for

their pre-submission reviews. We would like also to thank Luca Miserere (Torino, Italy)

for identifying the moss species and Enzo Musumeci (Basel, Switzerland), Philippe

Clerc (Chambésy, Switzerland) and Gérard Trichies (Neufchef, France) for sending

herbarium specimens.

Literature cited

Antonin V, Noordeloos M. 2004. A monograph of Hemimycena, Delicatula, Fayodia, Gamundia,

Myxomphalia, Resinomycena, Rickenella, and Xeromphalina (Tribus Mycenae sensu Singer,

Mycena excluded) in Europe. IHW Verlag, Eching.

Bellu. F. 1995. Die Trichterlinge (Clitocybe) der Sekt. Infundibuliformes Fr. und ihre Reaktion

gegentiber Kalilauge. Sydowia Beih. 10: 28-34.

Bellu F 1996. Alcune specie mediterranee di recente identificazione con particolare riguardo al

genere Clitocybe. Riv. Micol. 39: 99-114.

Bigelow HE. 1970. Omphalina in North America. Mycologia 62: 1-32.

http://dx.doi.org/10.2307/3757709

Bigelow HE. 1985. North American species of Clitocybe. Part Il. Beih. Nova Hedwigia 81:

281-471.

Binder M, Larsson K.-H, Matheny PB, Hibbett DS. 2010. Amylocorticiales ord. nov. and Jaapiales

ord. nov.: early diverging clades of Agaricomycetidae were dominated by corticioid forms.

Mycologia 102: 865-880. http://dx.doi.org/10.3852/09-288

Bjorbekmo MFM, Carlsen T, Brysting A, Vralstad T, Hoiland H, Ugland KI, Geml J, Schumacher

T, Kauserud H. 2010. High diversity of root associated fungi in both alpine and arctic Dryas

octopetala. BMC Plant Biol. 2010, 10: 244. http://dx.doi.org/10.1186/1471-2229-10-244

Bon M. 1997. Flore mycologique d'Europe 4. Les Clitocybes, Omphales et ressemblants. Doc.

Mycol., Mém. Hors-Sér. 4: 1-181.

Clémencon H. 1982. Kompendium der Blatterpilze Europaische omphalinoide Tricholomataceae.

Zeitschr. F. Mykol. 48: 195-237.

Clémencon H. 2004. Cytology and plectology of the Hymenomycetes. Bibl. Mycol. 199: 1-488.

Contu M. 1989. Rickenella brunneolilacina sp. nov. con note sul genere Rickenella in Sardegna.

Micol. Veget. Medit. 4: 57-62.

Contu M. 1997. Jacobia, un nuovo genere di Tricholomataceae omfalinoidi. Boll. Gruppo Micol. G.

Bres. (n.s.) 40(2—3): 169-713.

Drummond AJ, Ashton B, Buxton $, Cheung M, Cooper A, Duran C, Field M, Heled J, Kearse M,

Markowitz S, Moir R, Stones-Havas S, Sturrock S, Thierer T, Wilson A. 2010. Geneious v5.3.

Available from http://www.geneious.com/.

Elborne SA. 2008. Omphalina Quél. 235-237, in: H Knudsen, J Vesterhold (eds). Funga Nordica

- Agaricoid, boletoid and cyphelloid genera. Nordsvamp, Copenhagen.

Favre J. 1955. Les champignons supérieurs de la zone alpine du Parc national Suisse. Ergebn. wiss.

Unters. schweiz. Natn. Parks, n.s. 5(33): 1-212.

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes - application

to the identification of mycorrhizae and rusts. Mol. Ecol. 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

370 ... Vizzini & al.

GemlG, Timling I, Robinson CH, Lennon N, Nusbaum HC, Brochmann C, Noordeloos ME, Taylor

DL. 2012. An arctic community of symbiotic fungi assembled by long-distance dispersers:

phylogenetic diversity of ectomycorrhizal basidiomycetes in Svalbard based on soil and

sporocarp DNA. J. Biogeogr. 39: 74-88. http://dx.doi.org/10.1111/j.1365-2699.2011.02588.x

Gulden G, Jenssen KM. 1988. Arctic and alpine fungi, 2. Soopkonsulenten, Oslo.

Harmaja H. 2003. Notes on Clitocybe s. lato (Agaricales). Ann. Bot. Fennici 40: 213-218.

Huelsenbeck JP, Ronquist FE 2001. MrBayes: Bayesian inference of phylogeny. Bioinformatics 17:

754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

Katoh K, Misawa K, Kuma K, Miyata T. 2002. MAFFT: a novel method for rapid multiple

sequence alignment based on fast Fourier transform. Nucl. Acids Res. 30: 3059-3066.

http://dx.doi.org/10.1093/nar/gkf436

Lamoure D. 1974. Agaricales de la zone alpine. Genre Omphalina (lere partie). Trav. Sci. Parc Natl.

Vanoise 5: 149-164.

Lamoure D. 1975. Agaricales de la zone alpine. Genre Omphalina (2e partie). Trav. Sci. Parc Natl.

Vanoise 6: 153-166.

Lamoure D. 1982. Alpine and circumpolar Omphalina species. 201-215, in: GA Laursen, JF

Ammirati (eds). Arctic and Alpine Mycology 1. University of Washington Press, Seattle.

Larsson K-H. 2007. Re-thinking the classification of corticioid fungi. Mycol. Res. 111: 1040-1063.

http://dx.doi.org/10.1016/j.mycres.2007.08.001

Larsson K-H, Parmasto E, Fischer M, Langer E, Nakasone KK, Redhead SA, 2006. Hymenochaetales:

a molecular phylogeny for the hymenochaetoid clade. Mycologia 98: 926-936.

http://dx.doi.org/10.3852/mycologia.98.6.926

Lutzoni F, 1997. Phylogeny of lichen- and non-lichen-forming omphalinoid mushrooms and

the utility of testing for combinability among multiple datasets. Syst. Biol. 46: 373-406.

http://dx.doi.org/10.1093/sysbio/46.3.373

Matheny PM, Curtis JM, Hofstetter V, Aime MC, Moncalvo J-M, Ge Z-W, Yang Z-L, Slot JC,

Ammirati JF, Baroni TJ, Bougher NL, Hughes KW, Lodge DJ, Kerrigan RW, Seidl MT, Aanen

DK, DeNitis M, Daniele GM, Desjardin DE, Kropp BR, Norvell LL, Parker A, Vellinga EC,

Vilgalys R, Hibbett DS. Mycologia 98: 982-995. http://dx.doi.org/10.3852/mycologia.98.6.982

Moncalvo J-M, Lutzoni F, Rehner SA, Johnson J, Vilgalys R. 2000. Phylogenetic relationships of

agaric fungi based on nuclear large subunit ribosomal DNA sequences. Syst. Biol. 49: 278-305.

http://dx.doi.org/10.1093/sysbio/49.2.278

Moncalvo J-M, Vilgalys R, Redhead SA, Johnson JE, James TY, Aime MC, Hofstetter V,

Verduin AJW, Larsson E, Baroni TJ, Thorn RG, Jacobsson S, Clémencon H, Miller OK.

2002. One hundred and seventeen clades of euagarics. Mol. Phylog. Evol. 23: 357-400.

http://dx.doi.org/10.1016/S1055-7903(02)00027-1

Norvell LL, Redhead SA, Ammirati JE 1994. Omphalina sensu lato in North America 1-2. 1:

Omphalina wynniae and the genus Chrysomphalina. 2: Omphalina sensu Bigelow. Mycotaxon

50: 379-407.

Redhead SA, Lutzoni F, Moncalvo JM, Vilgalys R, 2002a. Phylogeny of agarics: partial systematics

solutions for core omphalinoid genera in the Agaricales (Euagarics). Mycotaxon 83: 19-57.

Redhead SA, Moncalvo J-M, Vilgalys R, Lutzoni F, 2002b. Phylogeny of agarics: partial systematics

solutions for bryophilous omphalinoid agarics outside of the Agaricales. Mycotaxon 82:

151-168.

Séguy E. 1936. Code universel des couleurs. Paul Chevalier, ed. Paris.

Singer R. 1970. Omphalinae (Clitocybeae-Tricholomataceae Basidiomycetes). Flora Neotropica 3:

1-84.

Omphalina pyxidata var. cystidiata var. nov. (Italy) ... 371

Stamatakis A. 2006. RAxML-VI-HPC: Maximum Likelihood-based phylogenetic analyses with

thousands of taxa and mixed models. Bioinformatics 22: 2688-2690.

http://dx.doi.org/10.1093/bioinformatics/btl446

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGAS5: Molecular

Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and

Maximum Parsimony Methods. Mol. Biol. Evol. 28(10): 2731-2739.

http://dx.doi.org/10.1093/molbev/msr121

Thiers B. 2011. [continuously updated] Index Herbariorum: A global directory of public herbaria

and associated staff. New York Botanical Garden's Virtual Herbarium.

http://sweetgum.nybg.org/ih/

Vizzini A, Contu M, Ercole E. 201la. Musumecia gen. nov. in the tricholomatoid clade

(Basidiomycota, Agaricales) related to Pseudoclitocybe. Nord. J. Bot. 29(6): 734-740.

http://dx.doi.org/10.1111/j.1756-1051.2011.01169.x

Vizzini A, Contu M, Musumeci E, Ercole E. 2011b. A new taxon in the Infundibulicybe gibba

complex (Basidiomycota, Agaricales, Tricholomataceae) from Sardinia (Italy). Mycologia 103:

203-208. http://dx.doi.org/10.3852/10-137

White TJ, Bruns TD, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR Protocols. Academic

Press, London.

MY COTAXON

http://dx.doi.org/10.5248/120.373

Volume 120, pp. 373-384 April-June 2012

Four new records of Aspergillus sect. Usti

from Shandong Province, China

LonG WANG *

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing

100101, China

* CORRESPONDENCE TO: wl_dgk@yahoo.com.cn

ABSTRACT—Four new Chinese records of Aspergillus species: Emericella heterothallica,

Aspergillus calidoustus, A. keveii, and A. pseudodeflectus are reported from Shandong

Province. Emericella heterothallica, despite being uncommonly reported, should be regarded

as geographically widely distributed.

Key worps — morphology, sequence analysis, taxonomy

Introduction

Raper & Fennell (1965) recognized five species in the Aspergillus ustus

group for which Gams et al. (1985) later established subgen. Nidulantes sect.

Usti. Kozakiewicz (1989) added two species to this section and removed one

species, while Klich (1993) transferred A. granulosus Raper & Thom from sect.

Usti to sect. Versicolores. The phenotypic evidence spawned different opinions

on phylogenetic relationships.

Peterson (2000) rejected sect. Usti based on Lsu rDNA sequence data (with

low bootstrap support) and placed the species in sect. Nidulantes. Houbraken

et al. (2007), who revived A. insuetus (Bainier) Thom & Church based on

multilocus DNA data along with chemical and phenotypic evidence, accepted

eight species in sect. Usti; Peterson (2008), accepting the species Houbraken et

al. included, added seven additional species to the section. Recently, Samson

et al. (2011) added five new taxa to this section, enlarging to twenty-one but

without including A. ochraceoroseus and A. versicolor.

Only three species representing sect. Usti have thus far been recorded in

China: A. deflectus Fennell & Raper, A. puniceus Kwon-Chung & Fennell, and

A. ustus (Qi et al. 1997). In the present paper, we report the recovery of four

additional members of this section: E. heterothallica, A. calidoustus, A. keveii,

374 ... Wang

TABLE 1. Strains included in molecular phylogenetic analyses.

GENBANK ACCESSION NUMBERS “

SPECIES STRAINS *

ITS1-5.8S-ITS2 BenA CaM

Aspergillus amylovorus Panas. ex Samson NRRL5813 T EF652503 EF652327 EF652415

A. calidoustus CBS 114380 EF591741 EF591729 EF591716

CBS 113228 EF591739 EF591730 EF591715

AS3.15302 JN982696 JN982686 JN982676

A. deflectus NRRL2206 T EF652437 EF652261 EF652349

A. egyptiacus Moub. & Mustafa NRRL5920 T EF652504 EF652328 EF652416

A. elongatus J.N. Rai & S.C. Agarwal NRRL5176 T EF652502 EF652326 EF652414

A. granulosus NRRL 1932 T EF652430 EF652254 EF652342

A. insuetus CBS 107.25 T EU076356 EU076371 EU076366

CBS 119.27 EU076355 EU076372 EU076367

A. keveii NRRL 1974 EF652432 EF652256 EF652344

CBS 561.65 EU076352 EU076375 EU076364

CBS 209.92 T EU076354 EU076376 EU076365

AS3.15305 JN982704 JN982694 JN982684

A. lucknowensis J.N. Rai et al. NRRL 3491 T EF652459 EF652283 EF652371

A. minutus Abbott NRRL 4876 T EF652481 EF652305 EF652393

NRRL 279 EF652457 EF652281 EF652369

A. pseudodeflectus NRRL6135 T EF652507 EF652331 EF652419

NRRL278 EF652456 EF652280 EF652368

AS3.15306 JN982697 JN982687 JN982677

AS3.15307 JN982700 JN982690 JN982680

AS3.15308 JN982699 JN982689 JN982679

AS3.15309 JN982701 JN982691 JN982681

AS3.15310 JN982703 JN982693 JN982683

A. puniceus NRRL 5077 T EF652498 EF652322 EF652410

A. ustus NRRL 4991 EF652492 EF652316 EF652404

NRRL 275 T EF652455 EF652279 EF652367

AS3.15311 JN982695 JN982685 JN982675

AS3.15312 JN982702 JN982692 JN982682

A. versicolor (Vuill.) Tirab. NRRL238 T EF652442 EF652266 EF652354

Emericella heterothallica NRRL 5096 T EF652449 EF652323 EF652411

NRRL 5097 EF652500 EF652324 EF652412

AS3.15313 JN982698 JN982688 JN982678

* Ex-type strains are indicated with “T”.

A Sequences JN982675-JN982704 were obtained in the present study.

and A. pseudodeflectus. All were isolated from soil samples collected at the foot

of Mount Tai in Shandong Province, located in the monsoon area of moderate-

temperate zone of China (36°15'17"N 117°06'15"E) in late July 2011. The

average altitude of that area is 134 m with an atmospheric pressure of 1004.1

Aspergillus spp. new to China... 375

hPa; the annual average temperature is 12.8 °C, with the monthly average of

—1.4 °C in January and 26.5 °C in July; the soil is typically frozen from late

October to late March; the frost-free period is about 200 days from late March

to late September; the annual precipitation is 700 mm (http://www.weather.

com.cn).

Materials & methods

Soil samples were collected underneath leaf litter and kept in sterilized plastic bags.

The fungi were isolated through dilution plating (Malloch 1981) using Dichloran Rose

Bengal Chlortetracycline agar (King et al. 1979) as the selective medium. Ten sect. Usti

strains were obtained and deposited in the China General Microbiological Culture

Collection Center, Institute of Microbiology, Chinese Academy of Sciences, Beijing

(CGMCC).

Morphological characters were assessed according to Houbraken et al. (2007), Klich

(2002), and Raper & Fennell (1965). Color names (shown here in title case) follow

Ridgway (1912). Wet mounts were prepared using material from colonies grown on

Czapek yeast autolysate agar (CYA) at 25 °C after 14 days mounted in lactophenol

without dye (Raper & Fennell 1965). Optical microscopic examination and photography

were performed with a Nikon Eclipse 80i and Nikon Digital Sight DS-L1 Microscope

(Nikon Co. Ltd., Japan).

Total DNA extraction followed the method of Scott et al. (2000). For amplification

of a portion of the B-tubulin gene, we used the primers described by Glass & Donaldson

(1995); to amplify the ITS1-5.8S-ITS2 region of the Nuc rRNA gene, we used the primers

the following primers: cmdAD1 5'-Gcc GAC TCT TTG ACT GAA GAG C-3’, cmd AD2: 5'-

GCC GAT TCT TTG ACC GAG GAA C-3' and cmdAD3: 5’-GCc GAT TCT TTG ACC GAA GAA

c-3' (sense primers); cmdQ1: 5'-GCA TCA TGA GCT GGA CGA ACT C-3’ and cmdQ2?: 5’-

GCA TCA TGA GCT GGA CGA ATT C-3’ (antisense primers). Polymerase chain reaction

(PCR) protocols for amplification of the above three gene regions, and the purification

and sequencing of PCR products followed the methods of Wang & Zhuang (2007).

Raw sequences were assembled and edited manually with BioEdit 5.0.9 (Hall

1999). Edited sequences were aligned using Clustal X 1.81 (Thompson et al. 1997), and

adjusted manually, as needed. Thirty-three strains of sect. Usti (TABLE 1) were analyzed

using the neighbor-joining (NJ) method and subjected to 1000 bootstrap replicates. All

the phylograms yielded the same results (e.g., Fic. 5, based on partial calmodulin gene

sequences).

Taxonomy

Emericella heterothallica (Kwon-Chung, Fennell & Raper) Malloch & Cain PL. 1

Colonies on Czapek agar (CA) at 25 °C growing rapidly, Colonies in 14

days reaching 40-43 mm in diam., densely floccose to velvety, radially sulcate,

margins entire; Hiille cells aggregated into small white masses visible to the

unaided eye, distributed sparsely on the colony surfaces; conidiogenesis sparse,

only present in the adjacent areas of colonies, brown in color, similar to Verona

376 ... Wang

PratE 1. Emericella heterothallica (AS3.15313): A-B. colonies on CA, CYA at 25 °C after 14 days;

C-D. conidiophores; E. conidia; F. hiille cells. Bar = 10 um

Brown, mycelia Orange-Cinnamon to Vinaceous-Cinnamon, white at margins;

exudate abundant, Brazil Red to Strawberry Pink; soluble pigment abundant,

Congo Pink; reverse Indian Red.

Aspergillus spp. new to China... 377

Colonies on CYA at 25 °C growing very rapidly, attaining 50-53 mm in

diam. in 14 days; texture velvet, radially sulcate with lightly discernable annular

plications, margins entire; Hiille cells moderately abundant, aggregated

into conspicuous, small white masses, distributed sparsely on the surfaces;

conidiogenesis moderate, in marginal areas, Verona Brown; mycelia Cinnamon

to Vinaceous-Cinnamon, white at colony peripheries; exudate light yellow

brown, moderately abundant; soluble pigment Empire Yellow, moderately

abundant; reverse Mars Orange to Orange Rufous.

Conidial heads globose when young, splitting at maturity into 2-3 loosely

divergent, short columns, (100-) 120-180 um long; conidiophores arising from

substratum or surface hyphae, stipes colorless becoming light brown in upper

portion, thin-walled, 240-360(-420) x (5-)7-9(-11) um; vesicles subglobose

to ellipsoidal, thin-walled, 14-18(-—22) x 11-14 um, biseriate; metulae 5-6.5 x

2-3 um; phialides ampuliform, 6.5-7 um, with short collula; conidia globose

(3-)3.5-5.5 um, echinulate, brown-colored en masse; Hiille cells ellipsoidal to

very elongate, or joined into botuliform chains, thick-walled, commonly 36-45

x 9-16 um, smaller ones 16-27 um, longer ones up to 54-63 um.

Colonies on CYA with 20% sucrose (CY20S) at 25 °C growing very rapidly,

reaching 52-55 mm in diam. in 14 days, densely floccose to velvety, radially

sulcate with annular plications, margins entire; conidiogenesis absent; mycelia

Ochraceous-Buff, white at margins; exudate and soluble pigment absent;

reverse Nopal Red.

Colonies on CYA at 37 °C growing moderately, in 14 days attaining 36-38

mm diam.; texture velvety, flat, margins entire; conidiogenesis moderately

heavy in central areas, brown-colored near Bister, with a Mikado tinge; mycelia

white at margins to Baryta Yellow in submarginal areas; exudate and soluble

pigment absent; reverse Lemon Chrome, with Buff Yellow tinge in centers.

ISOLATE EXAMINED: China, Shandong Province, foot of Mount Tai, from soil, 23 July

2011, Z. Yang (CGMCC AS3.15313).

Aspergillus calidoustus Varga, Houbraken & Samson PL. 2

Colonies on CA at 25 °C growing moderately, colonies attaining 35-40 mm

in diam. in 14 days, flat, sparsely floccose, margins fimbriate; conidiogenesis

sparse, Light Drab; mycelia white; exudate and soluble pigment absent; reverse

white.

Colonies on CYA at 25 °C growing rapidly, attaining 48-49 mm in diam. in

14 days, moderately deep, margins entire, floccose; conidiogenesis heavy, Drab

Gray to Light Drab; mycelia white; exudate and soluble pigment absent; reverse

Pale Yellow—Green to Light Viridine Green.

Conidial heads small, radiate; conidiophores borne from aerial hyphae,

stipes straight, with light yellow tinge, smooth-walled, (130—)150-250(-300)

378 ... Wang

PLATE 2. Aspergillus calidoustus (AS3.15302): A—B. colonies on CYA, CY20S at 25 °C after 14 days;

C-D. conidiophores; E. conidia. Bar = 10 um

x 5-7 um; vesicles subglobose, light brown, 9-18(-24) um, biseriate; metulae

very short, 3.5 x 2-3 um; phialides ampuliform, 7-7.5 x 2-3 um, with short

collula; conidia spheroidal, conspicuously echinulate, 2.5-3.6(-4) um; Hiille

cells not found.

Colonies on CY208S at 25 °C growing rapidly, attaining 46-49 mm in diam.

in 14 days, moderately deep, depressed at centers, margins entire, floccose;

conidiogenesis heavy, Drab Gray to Light Drab; mycelia white; exudate and

soluble pigment absent; reverse Pale Yellow-Green.

Aspergillus spp. new to China... 379

Colonies on CYA at 37 °C after 14 days: growing fast, 48-50 mm in diam.,

plane, centrally umbonate, margins entire, compactly floccose; conidiogenesis

heavy, Drab Gray; mycelia white, only conspicuous at margins; exudate and

soluble pigment absent; reverse Pale Yellow-Green to Light Viridine Green.

ISOLATE EXAMINED: China, Shandong Province, foot of Mount Tai, from soil, 23 July

2011, Z. Yang (CGMCC AS3.15302).

PLATE 3. Aspergillus keveii (AS3.15305): A-B. colonies on CYA, CY20S at 25 °C after 14 days;

C-D. conidiophores; E. conidia. Bar = 10 um

380 ... Wang

Aspergillus keveii Varga, Houbraken & Samson PE 3

Colonies on CA at 25 °C growing moderately, reaching 38-40 mm in diam.

in 14 days, flat, sparse, floccose, margins fimbriate; conidiogenesis sparse;

mycelia white; exudate and soluble pigment absent; reverse white.

Colonies on CYA at 25 °C growing rapidly, reaching 44-47 mm in diam. in

14 days, deep, margins fimbriate, floccose; conidiogenesis sparse, mostly in the

central areas, Light Drab; mycelia white with Pale Brownish Vinaceous tinge;

exudate and soluble pigment absent; reverse Pale Ochraceous-Buff.

Conidial heads small, radiate; conidiophores borne from aerial hyphae,

stipes smooth-walled, brown in color, 200-280 x 5-7.5 um; vesicles subglobose,

brown-colored, 10-18 um in diam., biseriate; metulae short, 3.5 x 2-3 um;

phialides ampuliform, 7-7.5 x 2-3 um, with short collula; conidia spheroidal,

conspicuously roughened, 3-3.6(-4) um; Hiille cells not observed.

Colonies on CY20S at 25 °C growing moderately, reaching 35-38 mm in

diam. in 14 days, deep, margins fimbriate, floccose; conidiogenesis sparse;

mycelia white with Pale Grayish Vinaceous tinge; exudate and soluble pigment

absent; reverse Light Ochraceous-Salmon.

Colonies on CYA at 37 °C showing no growth after 14 days.

ISOLATE EXAMINED: China, Shandong Province, foot of Mount Tai, from soil, 23 July

2011, Z. Yang (CGMCC AS3.15305).

Aspergillus pseudodeflectus Samson & Mouch. PL. 4

Colonies on CA at 25 °C growing moderately, attaining 28-30 mm in diam.

in 14 days, deep, flat, floccose; conidiogenesis absent; mycelia white; exudate

and soluble pigment absent; reverse white.

Colonies on CYA at 25 °C fast-growing, reaching 43-45 mm in diam. in

14 days, deep, convex, floccose; conidiogenesis moderate, Light Drab; mycelia

white; exudate and soluble pigment absent; reverse Marguerite Yellow to Reed

Yellow.

Conidial heads small, radiate; conidiophores borne on aerial hyphae, stipes

curved and slender, smooth-walled but with occasional concretions, light

brown in color, (50-)100-180 x 2-3.5 um; vesicles hemispherical, brown-

colored, 4-7.5 um in diam., biseriate; metulae very short, 3.5 x 2-3 um;

phialides ampuliform, 7-7.5 x 2-3 um, with long collula up to 3.5 um; conidia

spheroidal, warty, 2.5-3.6(—4) um; Hille cells not observed.

Colonies on CY20S at 25 °C growing rapidly, reaching 42-43 mm in diam.

in 14 days, deep, convex with depression at centers, floccose; conidiogenesis

moderate, Light Drab to Drab Gray; mycelia white; exudate and soluble pigment

absent; reverse Reed Yellow.

Colonies on CYA at 37 °C growing rapidly, reaching 48-50 mm in diam. in 14

days, flat, annularly and radiately plicate, umbonate lightly in centers, floccose;

Aspergillus spp. new to China... 381

ae!

PiaTE 4. Aspergillus pseudodeflectus (AS3.15306): A-B. colonies on CYA, CY20S at 25 °C after

14 days; C-E. conidiophores; F. conidia. Bar = 10 um

382 ... Wang

conidiogenesis heavy, Light Cinnamon-Drab to Light Drab; mycelia white;

exudate and soluble pigment absent; reverse Dark Olive to Buffy Brown.

ISOLATE EXAMINED: China, Shandong Province, foot of Mount Tai, from soil, 23 July

2011, Z. Yang (CGMCC AS3.1506-3.15310).

Discussion

Emericella heterothallica was the first reported heterothallic teleomorph

associated with the aspergilli. Raper & Fennell (1965) reported the production

of cleistothecia following the cross of yellow series and reddish orange series.

Houbraken et al. (2007), however, were unable to reproduce this result.

A. pseudodeflectus AS 3.15310

A. pseudodeflectus AS 3.15309

A. pseudodeflectus AS 3.15307

0.02 A. pseudodeflectus AS 3.15308

ie 4 A. pseudodeflectus AS 3.15306

844. pseudodeflectus NRRL 278

100}"4. pseudodeflectus NRRL 6135 T

A. calidoustus CBS 113228

7) A. calidoustus AS 3.15302

A. calidoustus CBS 114380

A. insuetus CBS 107.25 T

100} 4. minutus NRRL 4876 T

8884. minutus NRRL 279

100 A. insuetus CBS 119.27

99f A. keveii CBS 209.92 T

99 A. keveii CBS 561.65

94 A. keveii NRRL 1974

A. keveii AS 3.15305

100] A. ustus AS 3.15312

A. ustus AS 3.15311

100 A. ustus NRRL 257 T

A. ustus NRRL 4991

99 89 A, puniceus NRRL 5077 T

89 E. heterothallica NRRL 5096 T

100}"F. heterothallica NRRL 5097

E. heterothallica AS 3.15313

‘A. granulosus NRRL 1932 T

100 ‘A. lucknowensis NRRL 3491 T

fe A. deflectus NRRL 2206 T

‘A. elongatus NRRL 5176 T

‘A. versicolor NRRL 238 T

‘A. egyptiacus NRRL 5920 T

A. amylovorus NRRL 5813 T

100

PLaTE 5. The NJ tree inferred from the partial sequences of calmodulin gene data set. Bootstrap

percentages > 70% derived from 1000 replicates are indicated at the nodes. Bar = 0.02 substitutions

per nucleotide position.

Aspergillus spp. new to China ... 383

According to the characters presented, isolate AS3.15313 is a member of the

reddish orange series. Although E. heterothallica has only been reported rarely,

our observation of this species in China suggests that, despite its relative rarity,

it is geographically widely distributed.

Raper & Fennell (1965) cited six isolates —WB275 (type culture), WB278,

WB280, WB281, WB1974, WB4876— as typical of the six morphotypes they

recognized from among the hundreds of A. ustus isolates they had examined. It

is interesting how predictive of distinct species their observations were.

Based on partial 6-tubulin, calmodulin, actin genes, and ITS2-5.8S-ITS2

rDNA sequences, Houbraken et al. (2007) restricted A. ustus to isolates WB275

and WB280 and designated WB1974 as a new species, A. keveii. Varga et

al. (2008) included WB281 in their new taxon, A. calidoustus. We observed

much variation among our isolates under the name of A. ustus sensu Raper

& Fennell (1965), with prominent differences in the stipe and vesicle shapes

and dimensions. The growth rate at 37 °C varied greatly, some strains rapid,

others moderate, still others with no growth at all. Conidiation was also highly

variable, ranging from abundant to none depending on the isolates and culture

conditions, and conidial color ranged from grey-green to drab and brown.

These morphological differences suggest that multiple cryptic taxa may remain

embedded in A. ustus sensu Raper & Fennell (1965). Phylogenetic analyses of

three genetic markers (the Nuc ITS1-5.8S-ITS2 rRNA region and the partial

6-tubulin and calmodulin gene sequences) affiliate our isolates with the four taxa

included in sect. Usti by Houbraken et al. (2007) and Samson et al. (2011).

Acknowledgments

The author would like to thank Dr. J.A. Scott and S.W. Peterson for pre-submission

review and valuable suggestions. This work was supported by the National Natural

Foundation of China (no. 31070015).

Literature cited

Gams W, Christensen M, Onions AH, Pitt JI, Samson RA. 1985. Infrageneric taxa of Aspergillus.

55-61, in RA Samson, JI Pitt (eds). Advances in Penicillium and Aspergillus sytematics. Plenum

Press, New York.

Glass NL, Donaldson GC. 1995. Development of primer sets designed for use with the PCR to

amplify conserved genes from filamentous Ascomycetes. Appl. Environ. Microbiol. 61:

1323-1330.

Hall TA. 1999. Bioedit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NTT. Nucl. Acids Symp. Ser. 41: 95-98.

Houbraken J, Due M, Warga J, Meijer M, Frisvad JC, Samson RA. 2007. Polyphasic taxonomy of

Aspergillus section Usti. Stud. Mycol. 59: 107-128. http://dx.doi.org/10.3114/sim.2007.59.12

King DA, Hocking AD, Pitt JI. 1979. Dichloran-rose bengal medium for enumeration and isolation

of molds from foods. J. Appl. Environ. Microbiol. 37: 959-964.

Klich MA. 1993. Morphological studies of Aspergillus section Versicolores and related species.

Mycologia 85: 100-107

384 ... Wang

Klich MA. 2002. Identification of common Aspergillus species. CBS Press, Utrecht.

http://dx.doi.org/10.2307/3760484

Kozakiewicz Z. 1989. Aspergillus species in stored products. Mycol. Pap., 161: 1-188

Malloch D. 1981. Moulds their isolation, cultivation and identification. University of Toronto Press,

Toronto.

Peterson SW. 2000. Phylogenetic relationships in Aspergillus based on rDNA sequence analysis.

323-355, in RA Samson, JI Pitt (eds). Integration of modern taxonomic methods for Penicillium

and Aspergillus classification. Harwood Academic Publishers, Amsterdam.

Peterson SW. 2008. Phylogenetic analysis of Aspergillus species using DNA sequences from four

loci. Mycologia 100: 205-226. http://dx.doi.org/10.3852/mycologia.100.2.205

Qi Z-T, Kong H-Z, Sun Z-M. 1997. Flora fungorum sinicorum vol. 5: Aspergillus et teleomorphi

cognati. Science Press, Beijing. (in Chinese)

Raper KB, Fennell DI. 1965. The genus Aspergillus. Williams & Wilkins, Baltimore.

Ridgway R. 1912. Color standards and color nomenclature. Publ. by author, Washington D C.

Samson RA, Varga J, Meijer M, Frisvad JC. 2011. New taxa in Aspergillus section Usti. Stud Mycol.,

69: 81-97. http://dx.doi.org/10.3114/sim.2011.69.06

Scott J, Malloch D, Wong B, Sawa T, Straus N. 2000. DNA heteroduplex fingerprinting in Penicillium.

225-236, in RA Samson, JI Pitt (eds). Integration of modern taxonomic methods for Penicillium

and Aspergillus classification. Harwood Academic Publishers, Amsterdam.

Thompson JD, Gibbson TJ, Plewniak F, Jeanmougi F, Higgins DG. 1997. The CLUSTAL-X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucl. Acids Res. 25: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Varga J, Houbraken J, Van Der Lee HAL, Verweij PE, Samson RA. 2008. Aspergillus calidoustus sp.

nov., causative agent of human infections previously assigned to Aspergillus ustus. Eukaryot.

Cell 7: 630-638. http://dx.doi.org/10.1128/EC.00425-07

Wang L, Zhuang W-Y. 2007. Phylogenetic analyses of penicillia based on partial calmodulin gene

sequences. BioSystems 88: 113-126. http://dx.doi.org/10.1016/j.biosystems.2006.04.008

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in MA Innis et al. (eds). PCR Protocols: a guide to

methods and applications. Academic Press, San Diego.

MY COTAXON

http://dx.doi.org/10.5248/120.385

Volume 120, pp. 385-400 April-June 2012

Systematics of the Gomphales:

the genus Gomphus sensu stricto

ADMIR J. GIACHINI’*, CARLA M. CAMELINI’, MArcrio J. Rossr’,

CLAUDIO R. F. S. SOARES’ & JAMES M. TRAPPE?

Universidade Federal de Santa Catarina, Departamento de Microbiologia,

Imunologia e Parasitologia, Florianopolis, Santa Catarina 88040-970, Brazil

Oregon State University, Department of Forest Ecosystems and Society,

Corvallis, Oregon 97331-5752, USA

*CORRESPONDENCE TO: admir.giachini@ufsc. br

ABSTRACT — Gomphus sensu lato (Gomphales) was described to include species of

cantharelloid-gomphoid fungi that had ‘merulioid’ (wrinkled) hymenia and verrucose spores.

Gomphus sensu stricto is currently characterized by unipileate to merismatoid (composed

of several pilei) basidiomata, depressed funnel- to fan-shaped pilei, presence of clamp

connections, and verrucose spores; it has three described species: G. brunneus, G. clavatus,

and G. crassipes. Basidiomata and spore features are reliable identifiers of Gomphus sensu

stricto that distinguishes the species from other genera in the Gomphales.

Key worps — Gloeocantharellus, Phaeoclavulina, Turbinellus

Introduction

The genus Gomphus was originally proposed as a segregate from the genera

Clavaria, Geoglossum, Mitrula, and Spathularia and described as “thickened,

truncate, smooth, laterally plicate-venose, the pileus weakly developed”

(Persoon 1797). No species were assigned to the genus when described. The

first citation of a species for Gomphus came only when Gray (1821) described

G. clavatus based on Merulius clavatus Pers., a species known to have

merismatoid (composed of several pilei) basidiomata, orangish brown to violet

pilei, violet hymenia, and verrucose non-anastomosed ornamented spores.

From the earliest citations of Persoon (1797) to the latest descriptions of

Petersen (1971), the systematics and nomenclature of Gomphus have had a

checkered history. Before Giachini (2004) analyzed the molecular phylogeny

of 320+ collections of Gomphus sensu lato and related genera in the Gomphales

and reviewed the systematics and nomenclatural history for this group, species

of Gomphus sensu stricto were assigned to three different genera: Cantharellus

386 ... Giachini & al.

(Fries 1821), Craterellus (Fries 1838), and Neurophyllum (Doassans & Patouillard

1886).

The reduced number of distinctive morphological features and (more

importantly) the lack of molecular data to clarify the systematics of this group

have contributed to its confused classification. Nevertheless, the significant

contributions by many taxonomists have led to a better understanding of the

overall placement of gomphoid fungi.

Recently Giachini & Castellano (2011) presented a new classification

for Gomphus sensu lato. Giachini (2004) and Giachini & Castellano

(2011) emphasized that Gomphus sensu stricto is the only genus in the

Gomphaceae with strictly violet, lavender-brown, or milky-coffee colored

hymenia, distinguishing it from other representatives of Gomphus sensu lato

(Gloeocantharellus, Phaeoclavulina, Turbinellus) characterized by orange,

brown or greenish olive hymenia. Furthermore, all Gomphus species produce

clamp connections and verrucose spores. The unique combination of these

morphological characteristics separates Gomphus from other genera within the

Gomphales.

Below we review the important diagnostic characters of Gomphus sensu

stricto and provide a dichotomous key and complete descriptions of the three

currently accepted species: G. brunneus, G. clavatus, and G. crassipes. For

discussion on the nomenclatural history of Gomphus sensu lato see Giachini

(2004), Giachini et al. (2010), and Giachini & Castellano (2011).

Materials & methods

Collections examined

Collections of Gomphus were obtained from BR, DAOM, FH, K, MICH, NYS, OSC,

S, SFSU, TENN, and WTU (http://www.nybg.org/bsci/ih/ih.html). Since Dufour (1889)

did not assign a type for G. crassipes and the only known collections (two) deposited at

the herbarium RAB were not granted for loan, its description and further notes were

taken from the relevant literature.

In addition to dried herbarium material, we examined fresh specimens of G. clavatus

from northern California, Oregon, and Washington. These were collected mostly in late

September to mid-December, the usual fruiting season. Therefore, colors for G. clavatus

are based on fresh and dried material while for G. brunneus are based on dried material

only.

After selected collections were photographed and macroscopic features recorded,

they were dried within 24 h of collection with a forced-air food dehydrator set at

38-40°C, and deposited at OSC.

Habitat and fruiting patterns are based on information from the original descriptions

and/or field observations.

Macroscopic characterization

Macroscopic features are based on original descriptions and herbarium notes

supplemented by our own data on fresh or dried specimens. Features of the pileus,

Gomphus sensu stricto ... 387

hymenium, stipe, flesh, odor and taste were recorded whenever possible. Size of

basidiomata may vary depending on conditions of the fruiting season, so the ranges of

minimum and maximum sizes are presented as the range for 5-10 collections. Several

specimens were cut longitudinally through the vertical axis to examine the context

structure of pileus and stipe. Application of chemicals often used and referred to in the

literature (Corner 1966, Petersen 1971) in describing species of Gomphus sensu lato

proved of little value and so are not reported here.

Microscopic characterization

Features observed with a compound microscope were described from free-hand

sections mounted in Melzer’s reagent, 5% KOH, cotton blue, or H,O. Structures were

measured with an optical micrometer at x1000 magnification, mostly in mountants.

Hyphal wall thicknesses are recorded as ‘thin’ <0.5 um and ‘thick’ >0.5 um. The pileipellis

and stipitipellis were examined in surface view. When material was available, dimensions

were recorded from at least 3 specimens from each of at least 5 collections. Both mature

and immature specimens were examined for developmental changes. Maturity was

judged by the relative abundance of fully ornamented spores.

Spore shape was determined by the length-width ratio of 20-30 randomly selected

spores (Kirk et al. 2008). Dimensions are given as: minlength-maxlength x minwidth-

maxwidth, excluding ornamentation and apiculus. Immature or oversized spores were

not included in the measurements, though variation was noted.

Taxonomy

Species are presented in alphabetical order. Descriptions include citation of

all examined material and the herbarium where the type and other collections

are deposited. Descriptions are based on notes available with collections, the

observations made in this study, and literature reports.

Gomphus Pers., Tent. Disp. Meth. Fung.: 74 (1797) [non Gomphus (Fr.) Weinm.1826].

(For synonymy, see Giachini & Castellano 2011:186.)

TYPE SPECIES: Gomphus clavatus (Pers.) Gray 1821.

BASIDIOMATA <18 cm tall, erect, unipileate to merismatoid, when merismatoid

with two to several pilei arising from a single stipe. PILEUs 4-18 cm wide,

plane to depressed, fan- to funnel-shaped, flexuous, fleshy, dry, glabrous

(G. brunneus) to subpruinose, covered with brown hyphae that form minute

separate and distinct patches toward the margin but merge into a continuous

felty tomentum over the disc (G. clavatus), rosaceous, sordid yellow to orange-

brown, creamy violet, violet, brown; margin lacerate to crenate. HYMENIUM

decurrent, wrinkled, generally longitudinally oriented, dichotomous, often

to rarely anastomosing, reticulate to almost poroid (G. brunneus), especially

near the stipe apex, more lamellate toward the margin of the pileus, violet,

vinaceous brown to milky-coffee colored. Stipe <80 mm long, solid, cylindrical

to tapering downward, generally dilating into the pileus, pale violet to brown,

nearly black in G. crassipes, often pale red-brown where handled. CONTEXT

388 ... Giachini & al.

firm, pliable, often with anastomosing cavities, especially in G. clavatus, white,

off-white, violet, pale rose, brownish pink on exposure. OpDorR faint to sweet.

TasTE mild to bitter. SPORE PRINT brown for G. brunneus and G. clavatus, not

recorded for G. crassipes.

PILEIPELLIS of scattered to fasciculate, simple or branched, rarely slightly

inflated hyphae, in G. clavatus with pileocystidia; clamp connections present.

STIPITIPELLIS of parallel hyphae at surface, interwoven beneath, hyaline, <2 um

wide; clamp connections present. PILEUS and STIPE CONTEXT of interwoven,

hyaline hyphae 2.5-6 um wide, generally uninflated except adjacent to the

clamp connections. HYMENIAL TRAMA of thin- to thick-walled hyphae; clamp

connections present. SUBHYMENIAL TRAMA of interwoven, hyaline hyphae

<8 um wide; clamp connections present. Bastp1A <100 um long, 12 um wide,

clavate, in general with (2-)4 slightly divergent, slightly incurved sterigmata;

clamp connections present. HYMENIAL CYSTIDIA absent. BASIDIOSPORES 7.5-17

x 3.5-7.5 um, orange, yellow-brown to dark olive in mass, ellipsoid to obovoid;

ornamentation verrucose, cyanophilic; apiculus eccentric (not recorded for

G. crassipes).

ECOLOGY & DISTRIBUTION: epigeous, solitary to gregarious or caespitose, in

leaf litter or terrestrial. When gregarious, basidiomata are often separated by a

few centimeters. Encountered in Africa, Asia, Europe, and North America.

REMARKS: Defining boundaries for species within Gomphus sensu lato has been

a constant challenge because of the lack of consistent morphological features.

Only recently have morphology and molecular techniques combined to provide

a natural classification for the Gomphaceae. Giachini et al. (2010) and Giachini

& Castellano (2011) verified by molecular and morphological data that species

of Gomphus sensu stricto (G. brunneus, G. clavatus, and G. crassipes) are in

a lineage separate from all other species of Gomphus sensu lato. Accordingly,

Gomphus sensu stricto is reduced to three species vs. the 35 formerly assigned

to Gomphus sensu lato. Main features diagnosing Gomphus are the smooth to

subpruinose pileus, wrinkled or poroid-like hymenium, and clamp connections.

Species of Gomphus sensu stricto occur across the northern hemisphere and as

far south in Africa as the Democratic Republic of the Congo.

Gomphus brunneus (Heinem.) Corner, Ann. Bot. Mem. 2: 116. 1966. Fic. 1

(For synonymy, see Giachini & Castellano 2011:186.)

Type: DEMOCRATIC REPUBLIC OF CONGO. Equateur, Binga, April 1928, M.

Goosens-Fontana 683 (BR A253).

BASIDIOMATA <9 x 4 cm, unipileate, erect, clavate, truncate at apex. PILEUS

<4 cm wide, depressed to funnel-shaped, thick in the center, thin toward the

ascending and somewhat lacerate margin, brown or occasionally more or

less rosaceous, with paler tones in the center. HYMENIUM wrinkled, reticulate

Gomphus sensu stricto ... 389

Fic. 1. Gomphus brunneus. A. Basidioma (bar = 2 cm). B. Basidiospores (bar = 5 um).

to almost poroid, pale violet to milky-coffee colored. StrPE <80 mm long,

tapered downward, dilating obconically into the pileus, brown, the base white

tomentose. CONTEXT pliable, white, then brownish pink on exposure. ODOR

not recorded. TasTE bitter. SPORE PRINT brown.

PILEIPELLIS of scattered to fasciculate, simple hyphae; clamp connections

present. STIPITIPELLIS of hyaline hyphae <2 um wide; clamp connections

present. PILEUs and STIPE CONTEXT of interwoven, hyaline hyphae 2.5-6 um

wide; clamp connections present. HYMENIAL TRAMA of thin-walled hyphae;

clamp connections present. SUBHYMENIAL TRAMA Of slightly thickened, hyaline

hyphae 3-7 um wide, with refractive walls; clamp connections present. BASIDIA

50 x 8 um, with 2-4 sterigmata. HYMENIAL CyYSTIDIA absent. BASIDIOSPORES

7.5-10 x 3.5-5 um, yellow-brown in mass, ellipsoid to obovoid; ornamentation

of cyanophilic warts arranged more or less in longitudinal rows; apiculus

rounded, eccentric.

ECOLOGY & DISTRIBUTION terrestrial, in leaf litter, on floor of dry forests of

Cameroon (Roberts 1999), the Democratic Republic of Congo, and Uganda

(Corner 1966, Roberts 1999).

ADDITIONAL SPECIMENS EXAMINED: UGANDA: Gunda Forest, May 1918, T:D. Maitland

275 (as Gomphus clavatus var. parvisporus; K 57307).

390 ... Giachini & al.

REMARKS: Gomphus brunneus is distinguished by its simple basidiomata with

smooth pilei, weakly wrinkled hymenia, clamped hyphae, and comparatively

small basidiospores. Its morphology, distribution, and molecular profile make

it unique (Giachini 2004, Giachini et al. 2010, Giachini & Castellano 2011).

Only two G. brunneus herbarium collections were available for examination.

After studying the type, Roberts (1999) considered G. clavatus var. parvisporus

from Uganda (described by Corner 1966 and said to occur in Europe and the

western USA) a synonym of G. brunneus. Both species are macroscopically

similar with unusually small verrucose spores and are known only from

equatorial Africa. Heinemann (1958, 1959) reported two collections from

forests of the Democratic Republic of Congo, Africa. Corner (1966) reported

one collection (as Gomphus clavatus var. parvisporus) from Cameroon, while

Roberts (1999) cited two collections from Cameroon obtained in 1996 and

reported abundant basidiomata from the same location on the following year.

Otherwise, the species appears to be restricted to the Democratic Republic of

Congo and Uganda.

For several decades Cantharellus pseudoclavatus A.H. Sm. (Smith & Morse

1947) was classified under Gomphus sensu lato (Corner 1966). After analyzing

the type collection (MICH 6916), however, Giachini (2004) concluded that it

better fit Cantharellus. The main morphological character justifying its removal

from Gomphus sensu lato was the presence of 8-spored basidia, a feature not

found elsewhere in Gomphus but present in Cantharellus. Petersen (1971) had

previously transferred the species to Pseudocraterellus, a genus which also lacks

8-spored sterigmata. Although Corner (1966) suggested Gomphus clavatus var.

parvisporus occurs in Europe and is common in western USA, he mentioned

only specimens obtained in Africa in his original description of that variety.

Cantharellus pseudoclavatus is rare under conifers of the western USA and is

definitely distinct from G. brunneus.

Gomphus clavatus (Pers.) Gray, Nat. Arr. Brit. Pl. 1: 638. 1821. Fig, 2

(For synonymy, see Giachini & Castellano 2011:187.)

BASIDIOMATA S17 cm tall, unipileate at first and then merismatoid with

<15 subpilei. Prteus <15 cm wide, the surface flat, subundulate, fan-shaped,

glabrous to covered with brown hyphae that form minute separate and distinct

patches toward the margin but merge into a continuous felty tomentum

over the disc, orangish brown to overall creamy violet to dark violet; margin

crenate. HyMENIUM surface wrinkled, generally longitudinally oriented, with

or without discrete folds or pits, bright violet at the margin and junction with

stipe and overall when immature, at maturity covered with spores and then

paler (vinaceous brown). STIPE tomentose to hispid toward the base, glabrous

above and there blending to pale violet, white at the base and where covered

Gomphus sensu stricto ... 391

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Dr. Robert Thoma$ anMargarett

Fic. 2. Gomphus clavatus. A-B. Basidiomata. C. Spores (bar = 10 um).

392 ... Giachini & al.

(soil, debris), often pale red-brown where handled. CoNnTEXT often with

anastomosing cavities, violet around those, off-white to pale rose elsewhere.

Opor faint or none. TasTE mild. SPORE PRINT brown.

PILEIPELLIS often crowded, thin-walled, scattered to fasciculate, with simple

or branched hyphae, usually incrusted apically, with pileocystidia 3-4.5 um

wide, rarely slightly inflated, <5.5 um, protruding 50-120 um from the surface;

clamp connections present. Hyphae and bases of pileocystidia often with

brown amorphous deposits. STIPITIPELLIS of periclinal and parallel, hyaline

hyphae 1.5-2 um wide at the surface, becoming interwoven beneath, with few

projecting hyphae where stipe is glabrous, or fascicles of crowded hyphae where

stipe is hispid; clamp connections present. PILEUS and STIPE CONTEXT of thin-

to slightly thick-walled, generally oriented on the long axis of the basidioma

but profusely interwoven, hyaline hyphae 2.5-6 um wide, generally uninflated

except adjacent to the clamp connections, where it is slightly thick-walled

(<1 um thick) and <12 um wide. HyMENIAL TRAMA hyphae more loosely

interwoven than the rest of the context, undifferentiated from the sUBHYMENIAL

TRAMA. BASIDIA 60-95 x 8.5-12 um, clavate, collapsing after spore discharge,

hyaline, with (2-)4 slightly divergent, slightly incurved sterigmata <10 um

long; clamp connections present at base. HYMENIAL CYSTIDIA not observed.

BASIDIOSPORES (9-)10-15(-17) x 4-7.5 um, orange in Melzer’s reagent,

dark olive in mass (KOH), ellipsoid to obovoid, contents weakly cyanophilic;

ornamentation of cyanophilic warts, usually in discrete, raised patches that give

an undulate appearance to the spore surface; apiculus prominent, eccentric.

ECOLOGY & DISTRIBUTION: solitary, gregarious to caespitose, when gregarious

often separated by a few centimeters, terrestrial. Generally in conifer forests and

suggested to form mycorrhizae with species of Abies (Pantidou 1980) and Picea

(Agerer et al. 1998). Known from Austria (Petersen 1971), Canada (East and

West, Petersen 1971), China (Corner 1966), Czech Republic (Kluzak 1994),

France (Doassans & Patouillard 1886), Greece (Petersen 1971), Italy (Petersen

1971), Japan (Corner 1966), Lithuania (Urbonas et al. 1990), Mexico (Petersen

1971), Pakistan (Corner 1966), Poland (Adamczyk 1996), Russia (Bulakh 1978,

Bulakh & Govorova 2000), Spain (Fernando Sanchez, pers. comm., 10 Oct.

1999), Sweden, Switzerland (Petersen 1971), Turkey (Sesli 1997), and the USA

(especially northwestern/northeastern and into the Appalachian Mountains).

Loans were requested from herbaria located in most of these countries but

several did not respond, so our examination of specimens is restricted to those

listed under collections examined.

ADDITIONAL SPECIMENS EXAMINED: CANADA. BRITISH COLUMBIA, Vancouver

Island, 15 October 1962, M. Pantidou (DAOM 91369). Quebec, Duchesnay, 26 Aug.

1938, H.S. Jackson (DAOM 8624). SWEDEN: Uppsala, Sédermanland, Handen, 4

Sep. 1998, L. Kerwién (S); Fiby Urskog, 20 Sep. 1998, A. Taylor (S). UNITED STATES.

CALIFORNIA, DEL NorTE Co., Del Norte Coast Redwoods State Park, 9 Dec. 1999, M.A.

Gomphus sensu stricto ... 393

Castellano (OSC 97685); Jedediah Smith Redwoods State Park, 17 Nov. 1961, D. Isely

1479 (WTU), and 12 Nov. 1966, H.D. Thiers (SFSU 17740); Siskiyou Nat. Forest, Smith

River, 30 Nov. 1937, A.H. Smith 9243 (MICH); EL Dorapo Co., Pleasant Valley, 22

October 1967, D.E. Stuntz 14410 (WTU); MENDocINo Co., 5 Nov. 1961, D. Largent

71 (SFSU); Aleuria Glenn, 14 Nov. 1967, R.H. Petersen (TENN 33256); Aleuria Glenn,

along Rd. 404, 25 October 1986, M. Seidl 2120 (WTU)); Jackson State Forest, 5 Nov.

1967, R.H. Petersen and H.D. Thiers (TENN 33174), and 8 Dec. 1999, A.J. Giachini, M.A.

Castellano and E. Nouhra (OSC 97679, 97680, 97681, 97682, 97683, 97684). IDAHO,

BONNER Co., Priest River, Binarch Creek, 9 Sep. 1966, R.H. Petersen (TENN 32166);

Priest River Experimental Forest, Canyon Creek Road, 11 Nov. 1939, A.H. Slipp 621

(MICH). MassAcHusETTS, Essex Co., Gloucester, Aug. 1877, unnamed collector

(FH). NEw YorK, SARATOGA Co., Ballston Lake, Branch, 1879, C.H. Peck (holotype of

Cantharellus brevipes; NYS). OREGON, BENTON Co., Siuslaw Nat. Forest, Mary’s River,

24 October 1999, S. Ashkannejhad (OSC 97622); CLacKAMAS Co., Wemme, 23 Sep.

1946, A.H. Smith 23644 (MICH); Coos Co., South Slough, 23 Sep. 1985, C. Ardrey 800

(WTU), and 3 Sep. 1992, C. Ardrey 1806 (WTU); LANE Co., Cougar Reservoir, Road

19, 27 Nov. 1999, A.J. Giachini (OSC 97671, 97672, 97673, 97674, 97675, 97676, 97677);

Eugene, 30 October 1999, unnamed collector (OSC 97646); H. J. Andrews Experimental

Forest, Lookout Mt., Rd. 1506, 23 October 1999, A.J. Giachini (OSC 97616, 97620, 97669,

97670, 97678); H. J. Andrews Experimental Forest, 23 October 1999, A.J. Giachini (OSC

97617), 5 Nov. 1999, A.J. Giachini (OSC 97656), and 7 Nov. 2000, Y. Yano (OSC 97694);

Oakridge, Road 1934, 15 Nov. 1999, A.J. Giachini (OSC 97668); Willamette Nat. Forest,

25 October 1999, C. Lefevre (OSC 97623); LINN Co., Sweet Home, 4 Nov. 2000, A.J.

Giachini (OSC 97696, 97698); Willamette Nat. Forest, Pamela Lake Trail head, 2 Nov.

1999, A.J. Giachini (OSC 97624); MARION Co., Detroit Lake, Road 46, 15 Nov. 1999, A.J.

Giachini (OSC 97667). WASHINGTON, CHELAN Co., Lake Wenatchee, 7 Nov. 1980, D.E.

Stuntz 21131 (WTU); CLALLAM Co., Morse Creek camp, 17 October 1942, D.E. Stuntz

1245 (WTU); Olympic Mountains, 30 October 1931, H.S. Hotson (WTU); KinGs Co.,

Snoqualmie Nat. Forest, Asahel Curtis Nature Trail, 4 Aug. 1992, G.R. Walker 10600

(WTU); Lewis Co., Mt. Rainier Nat. Park, Bumping Lake, October 1937, D.E. Stuntz

759 (WTU); PreRcE Co., Mt. Rainier Nat. Park, Lower Tahoma Lake, 23 Sep. 1948,

D.E. Stuntz 4732 (WTU); Old Tahoma campground, 23 Sep. 1960, D.E. Stuntz 12000

(WTU); Mt. Rainier National Park, Tahoma Creek, 18 Aug. 1948, D.E. Stuntz and A.H.

Smith 4003 (WTU); Mt. Rainier Nat. Park, Tahoma Creek, 23 Sep. 1960, D. Isely 1257

(WTU).

REMARKS: Gomphus clavatus has traditionally been placed in a monotypic

subgenus (Gomphus subg. Gomphus) based on morphological features (Corner

1966). However, molecular studies by Giachini (2004) and Giachini et al. (2010)

support its placement together with G. brunneus and G. crassipes in Gomphus

sensu stricto and suggest it to be phylogenetically closely related to species of

Gloeocantharellus. Petersen (1971) noted that these two genera share several

morphological similarities, the most striking of which is that both exhibit the

same reaction to 10% KOH applied to the basidioma pileus. In reference to

G. clavatus Petersen (1971) stated, “the normal color immediately becomes a

most unique salmon-yellow, almost precisely the normal color of the pileus

and stipe of Gloeocantharellus purpurascens (Hesler) Singer.” The pileus surface

tomentum of G. clavatus is composed of pileocystidia, just as in G. purpurascens,

394 ... Giachini & al.

and, even though the pileocystidia of the two species differ in shape and

orientation, they have a similar brown agglutinating substance. Despite these

similarities, the molecular evidence shows Gomphus and Gloeocantharellus to

be in distinctly different, but related lineages (Giachini et al. 2010).

Other taxa in the Gomphaceae share the merismatoid habit of G. clavatus,

notably Phaeoclavulina grandis (Corner) Giachini, P. guadelupensis (Pat.)

Giachini, P. subclaviformis (Berk.) Giachini, and P. viridis (Pat.) Giachini and

Gloeocantharellus dingleyae (Segedin) Giachini, G. novae-zelandiae (Segedin)

Giachini, and G. pallidus (Yasuda) Giachini.

Gomphus crassipes (L.M. Dufour) Maire, in Maire & Werner, Mém. Soc. Sci. Nat.

Maroc 45: 81 (1937). FIG. 3

(For synonymy, see Giachini & Castellano 2011:187.)

Lectotype designated here, Pl. 13, in L.M. Dufour, Rev. Gen. Bot. 1: 357-358. 1889.

BASIDIOMATA <18 cm tall, unipileate or occasionally merismatoid. PILEUs 4-18

cm wide, plane to slightly depressed in the center, funnel-shaped to fan-shaped,

non hygrophanous, sordid yellow to orangish brown; margin subundulate.

HyMENIumM decurrent, wrinkled, occasionally anastomosing, dichotomous,

violet. Stipe 40-70 x 25-45 mm, cylindrical to slightly tapering downward,

almost entirely underground, central or lateral, nearly black. CONTEXT firm,

pale violet, marbled in the stipe with violet tones. ODOR sweet. TASTE not

recorded. SPORE PRINT color not recorded.

All HyPHAE 5-8 um wide; clamp connections present. Basrp1a 70-100 x

8-10(-12) um, clavate, with (2-)4 straight sterigmata. HYMENIAL CYSTIDIA

absent. Bastip1osporEs (11-)13-15(-17) x 5.5-6(—7) um, pale orange in mass,

ellipsoid to obovoid; ornamentation of cyanophilic, fine warts; data on apiculus

not recorded (Dufour 1889, Maire 1914).

ECOLOGY & DISTRIBUTION: on the ground, generally grouped, in conifer

forests in Algeria (Dufour 1889, Maire 1914, Maire & Werner 1937), Morocco

(Malencon 1958), and Spain (Miguel A. Pérez-De-Gregorio i Capella, pers.

comm., 5 Jan 2004).

REMARKS: As the only known collections are at herbarium RAB in Morocco,

and our loan request was denied due to the material’s fragile condition, our

description is based on Dufour (1889), Maire (1914), and Maire & Werner

(1937). Dufour (1889), who did not cite any collection or herbarium for the

type species, stated that “cette espéce est assez voisine du Cantharellus brevipes

Peck” As C. brevipes is a synonym of G. clavatus, in effect Dufour related

G. crassipes to G. clavatus, with similar macro- and microscopic characteristics,

differing only by the violet to marbled violet color of the context, which is

violet, off-white to pale rose in G. clavatus. However, its overall spore size and

Gomphus sensu stricto ... 395

Fic. 3. Gomphus crassipes.

A. Basidiomata [from Maire (1914: Pl. VII.2)].

B. Dried basidiomata [modified from Dufour (1889; Pl. 13), with permission].

396 ... Giachini & al.

geographic distribution suggest that G. crassipes is distinct from G. clavatus

(Giachini et al. 2010).

Only a few other references to G. crassipes are observed in the literature.

Maire (1914), who transferred Dufour’s C. crassipes to Neurophyllum, provided

a detailed description and excellent drawing of its basidiomata and spores

(Maire 1914) and subsequently (Maire & Werner 1937) transferred the species

to Gomphus. Later Malencon (1958) called attention to the putative relationship

of G. crassipes to the coral fungi, especially to Ramaria, but observed that Maire’s

drawings were not good representations of what Dufour (1889) had described

as C. crassipes and that in particular the were not those suggested by Dufour.

This species “pulls too much into the yellows and reds” according to Malencon

(1958), who described G. crassipes as having a pale ocher to rose pileus surface.

However, as seen for other species of Gomphus sensu lato, (e.g., G. clavatus)

the color of both pileus and hymenium surface can vary considerably. For

G. clavatus, for example, the pileus surface can be orangish brown to overall

creamy violet to dark violet. It is possible that Maire (Maire & Werner 1937)

observed a color variant of G. crassipes.

Because the designated lectotype does not adequately characterize the

species, an epitype is desirable. However, we refrain from designating an epitype

since no herbarium specimen is available and Malencon (1937) challenged

the Maire’s (1914) excellent illustration. A fresh collection (preferably from

Morocco) that can provide DNA sequences would be an ideal candidate for

epitypification.

Key to species

la. Basidiomata initially unipileate, then becoming merismatoid with <15 subpilei;

pileus fan-shaped, orangish brown, creamy violet to dark violet; hymenium

wrinkled, violet, vinaceous brown, lavender-brown;

ASTAENITO PSs aN OFT AMTICRICAS. 5 jc a-seslin bee hart wast ecare wat siaet ea otsee eee one G. clavatus

1b. Basidiomata unipileate to merismatoid; pileus depressed, funnel-shaped,

occasionally fan-shaped, brown to rosaceous, sordid yellow to orangish brown;

hymenium wrinkled, reticulate to almost poroid, violet to milky-coffee colored;

PNT CAs DAIS Jap ctrae tons ee oa escalate ted et A Ie as mete doses Ape datetents Z

2a. Spores 7.5-10 x 3.5-5 uum, verrucose; basidiomata unipileate; pileus brown or

occasionally rosaceous; hymenium slightly wrinkled, reticulate to almost poroid,

pale violet to milky-coffee colored;

Cameroon, Democratic Republic of Congo, Uganda .............. G. brunneus

2b. Spores (11-)13-15(-17) x 5.5-6(-7) um, finely verrucose; basidiomata

occasionally merismatoid; pileus sordid yellow to orangish brown; hymenium

wrinkled, violet;

I SETIa, -WIOKOCES, ANC SPallics J.:.8 ce Tank rath a tise asrablatstoreaaek eh earasen G. crassipes

Gomphus sensu stricto ... 397

Discussion

Gomphus was originally described to include cantharelloid-gomphoid

species that produced basidiomata resembling those of Cantharellus. The recent

molecular analyses of Giachini et al. (2010), which sharpened generic and

species concepts within the Gomphaceae and revealed Gomphus sensu lato as

non-monophyletic, support redistribution of those into Gomphus sensu stricto,

Gloeocantharellus, Phaeoclavulina, and Turbinellus. Only three original species

are retained within Gomphus sensu stricto — G. brunneus characterized by

somewhat funnel-shaped pilei and G. clavatus and G. crassipes, characterized

by fan- to funnel-shaped pilei.

Gomphus clavatus, widely distributed in the northern hemisphere, is

believed to form ectomycorrhizae with species of Abies (Pantidou 1980) and

Picea (Agerer et al. 1998). It is the most easily identified of the three species,

due primarily to the size and color of its basidiomata. Since the development

and implementation of the Northwest Forest Plan in the United States (USDA/

USDI 1994a,b, 2000, 2001), G. clavatus has been considered rare and potentially

restricted to old-growth forests of the Pacific Northwest (PNW), requiring all

federal and state land to be surveyed for the species before any management

activity could be approved.

Both G. brunneus and G. crassipes are known only from a few collections

from Africa and Spain, as noted above.

Morphologically Gomphus sensu stricto is readily separated from the other

gomphalean genera based on the violet, violet-brown to orangish brown colors

and fan- to somewhat funnel-shaped basidiomata pilei.

Species of Gomphus sensu stricto most resemble those of the resurrected

genus Phaeoclavulina (Giachini & Castellano 2011), which share fan- to funnel

shaped pilei and clamp connections. However, they differ based on spore

ornamentation, with most Phaeoclavulina species diagnosed by echinulate,

subreticulate, or reticulate spores in contrast to the strictly verrucose spores of

Gomphus.

Even though all Turbinellus species were assigned to Gomphus until

recently, they share only few morphological characters, most notably spore

ornamentation. They differ in basidiomata shape, color, and presence of clamp

connections (absent in Turbinellus).

Gomphus and Gloeocantharellus occupy an important position in the

evolution of the fungi in the Gomphales. Molecular analyses (Giachini et al.

2001, Giachini et al. 2010, Humpert et al. 2001), indicate Gloeocantharellus and

Gomphus as ancestral to all other genera within the Gomphales. The two genera

share spore shape and ornamentation and (for some Gloeocantharellus species)

clamp connections.

Unavailability of samples restricted more in-depth evolutionary inferences

of Gomphus to other genera in the Gomphales. Additional sampling, especially

398 ... Giachini & al.

for G. crassipes, is necessary to determine whether subgeneric arrangements

exist within Gomphus sensu stricto and to infer higher-level relationships to

other members of the Gomphales. A complete discussion on the evolutionary

relationships and implications of this taxonomic classification in relation to

other members of the Gomphales can be found in Giachini (2004), Hosaka et

al. (2006), and Giachini et al. (2010).

Acknowledgments

This research was partially supported by the Forest Mycology Team (Pacific Northwest

Research Station, Forest Service, US Department of Agriculture, Corvallis, Oregon).

AJG thanks Conselho Nacional de Desenvolvimento Cientifico e Tecnolégico (CNPq)

of the Brazilian Ministry of Education for the doctorate fellowship. Special thanks to

the following herbaria for providing specimens: Herbarium of the Jardin Botanique

National de Belgique - Meise (BR), National Mycological Herbarium —- Ottawa (DAOM),

Farlow Herbarium of Cryptogamic Botany - Cambridge (FH), Royal Botanic Gardens

Herbarium - Kew (K), University of Michigan Fungus Collection - Ann Arbor (MICH),

Herbarium of the New York State Museum - Albany (NYS), Oregon State University

Herbarium - Corvallis (OSC), Herbarium of the Swedish Museum of Natural History

— Stockholm (S), Harry D. Thiers Herbarium at San Francisco State University - San

Francisco (SFSU), University of Tennessee Herbarium - Knoxville (TENN), and the

University of Washington Herbarium - Seattle (WTU). Also thanks to Dr. Andy Taylor

for specimens of G. clavatus from Sweden. Boleslaw Kuznik, Michael Wood, Robert

Thomas, and Margaret Orr are acknowledged for permission to reproduce photographs

and drawings. We are grateful to Drs. Richard E. Baird, Efrén G. Cazares, and Shaun

Pennycook for critically reading the manuscript.

Literature cited

Adamczyk J. 1996. Les champignons supérieurs des hétrais du Nord du plateau de Czestochowa

(Pologne méridionale). Rev Bot 150: 1-83.

Agerer R, Beenken L, Christian J. 1998. Gomphus clavatus (Pers.: Fr.) S. F Gray + Picea abies (L.)

Karst. In: R Agerer et al. (eds). Descriptions of ectomycorrhizae 3: 25-29.

Bulakh EM. 1978. [Macromycetes of fir forests]. Biocenotic studies at the Berkhneussuriysk station,

Akademia Nauk SSSR Far Eastern Branch, Biology-Soils Institute. p 73-81.

Bulakh EM, Govorova OK. 2000. Rare and new for Russia basidiomycetes from Primorsky territory.

Mycol Phytopathol 34: 21-25.

Corner EJH. 1966. A monograph of the cantharelloid fungi. Ann Bot Mem 2: 1-255.

Doassans MME, Patouillard NT. 1886. Champignons du Béarn (2° liste). Revue Mycologique 8(29):

25-28 (Reprint from ‘Collected Mycological papers’ chronologically arranged and edited by

L. Vogelenzang, Rijksherbarium, Leiden, Amsterdam, vols. 1-3, 1978).

Dufour ML. 1889. Une nouvelle espéce de chanterelle. Rev Gen Bot 1: 357-358.

Fries EM. 1821. Systema mycologicum, vols. 1-3 (Reprint 1952, Johnson Reprint Corporation,

New York).

Fries EM.1838. Epicrisis systematis mycologici. typographia academica, Uppsala.

Giachini AJ. 2004. Systematics of the Gomphales: the genus Gomphus Pers. sensu lato. PhD

Dissertation, Department of Forest Science, Oregon State University, Corvallis.

Gomphus sensu stricto ... 399

Giachini AJ, Castellano MA. 2011. A new taxonomic classification for species in Gomphus sensu

lato. Mycotaxon 115: 183-201. http://dx.doi.org/10.5248/115.183

Giachini AJ, Spatafora JW, Cazares E, Trappe JM. 2001. Molecular phylogenetics of Gomphus

and related genera inferred from nuclear large and mitochondrial small subunits ribosomal

DNA sequences. Abstracts of the 3rd International Conference on Mycorrhizas (ICOM III).

Adelaide, Australia. p. 95.

Giachini AJ, Hosaka K, Nouhra ER, Spatafora JW, Trappe JM. 2010. Phylogenetic relationships

of the Gomphales based on nuc-25S-rDNA, mit-12S-rDNA and mit-atrp6-DNA combined

sequences. Fungal Biology 114: 224-234. http://dx.doi.org/10.1016/j.funbio.2010.01.002

Gray SF. 1821. Natural arrangement of British plants, vol. 1. Baldwin, Craddock and Joy, London.

Heinemann P. 1958. Champignons récoltes au Congo Belge par Madame M. Goosens-Fontana. III.

Cantharellineae. Bull Jard Bot Etat Brux 28: 385-438.

Heinemann P. 1959. Cantharellineae. Fl Icon Champ Congo, Fasc. 8: 153-165.

Hosaka K, Bates ST, Beever RE, Castellano MA, Colgan W, Dominguez LS, Nouhra ER, Gem J,

Giachini AJ, Kenney SR, Simpson NB, Spatafora JW, Trappe JM. 2006. Molecular phylogenetics

of the gomphoid-phalloid fungi with an establishment of the new subclass Phallomycetidae and

two new orders. Mycologia 98: 949-959. http://dx.doi.org/10.3852/mycologia.98.6.949

Humpert AJ, Muench EL, Giachini AJ, Castellano MA, Spatafora JW. 2001. Molecular

phylogenetics of Ramaria (Gomphales) and related genera: evidence from nuclear large

subunit and mitochondrial small subunit rDNA sequences. Mycologia 93: 465-477.

http://dx.doi.org/10.2307/3761733

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi, 10th ed.. CAB

International, Wallingford, UK. 771 p.

Kluzak Z. 1994. Gomphus clavatus, a seriously endangered species in the Czech Republic. Zeit

Mykol 60: 113-116.

Maire R. 1914. La flore mycologique des foréts de cédres de I’Atlas. Bull Soc Mycol Fr 30: 199-220.

Maire R, Werner RG. 1937. Fungi Maroccani. Mém Soc Sci Nat Maroc 45: 1-147.

Malencon G. 1958 [“1957”]. Prodrome d’une flore mycologique du Moyen Atlas. Bull Soc Mycol

Fr 73: 289-330.

Pantidou ME. 1980. Macrofungi in forests of Abies cephalonica in Greece. Nova Hedwigia 32:

709-723.

Persoon CH. 1797. Tentamen dispositionis methodicae fungorum in classes, ordines, genera et

familias. Leipzig.

Petersen RH. 1971. The genera Gomphus and Gloeocantharellus in North America. Nova Hedwigia

21: 1-118.

Roberts P. 1999. Clavarioid fungi from Korup National Park, Cameroon. Kew Bull 54: 517-539.

http://dx.doi.org/10.2307/4110853

Sesli E. 1997. Two new records of cantharelloid fungi for Turkey. Israel J Plant Science 45: 71-74.

Smith AH, Morse EE. 1947. The genus Cantharellus in western United States. Mycologia 39:

497-534. http://dx.doi.org/10.2307/3755192

Urbonas VA, Matyalis AA, Gritsyus AI. 1990. Trends of variability of macromycetes, extinguishing

species and principles of their protection in Lithuania. Mycol and Phytophatol 24: 385-388.

USDA/USDI [U.S.D.A. Forest Service; U.S.D.I. Bureau of Land Management]. 1994a. Final

supplemental environmental impact statement on management of habitat for late-successional

and old-growth forest related species within the range of the northern spotted owl (northwest

forest plan). Portland, OR.

AOO ... Giachini & al.

USDA/USDI [U.S.D.A. Forest Service; U.S.D.I. Bureau of Land Management]. 1994b. Record of

decision on management of habitat for late-successional and old-growth forest related species

within the range of the northern spotted owl (northwest forest plan). Portland, OR.

USDA/USDI [U.S.D.A. Forest Service; U.S.D.I. Bureau of Land Management]. 2000. Final

supplemental environmental impact statement for amendment to the survey and manage,

protection buffer, and other mitigation measures standard and guidelines. Volume 1, chapters

1-4. Portland, OR.

USDA/USDI [U.S.D.A. Forest Service; U.S.D.I. Bureau of Land Management]. 2001. Record of

decision and standards and guidelines for amendments to the survey and manage, protection

buffer, and other mitigation measures standards and guidelines. Portland, OR.

MYCOTAXON

http://dx.doi.org/10.5248/120.401

Volume 120, pp. 401-405 April-June 2012

Myxomycetes from China 15:

Arcyria galericulata sp. nov.

Bo ZHANG’, T1AN-Hao LI’, QI WANG’ & Yu LI’*

‘Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi &

?Institute of Agricultural Modernization,

Jilin Agricultural University, 2888 Xincheng Street, Changchun City, P. R. China

* CORRESPONDENCE TO: yuli966@126.com

ABSTRACT — A new species, Arcyria galericulata, is described and illustrated with scanning

electron micrographs. This species is characterized by a helmet-like (galeiform) structure at

the top of the sporocarp. Holotype and isotype specimens are deposited in the Herbarium of

Mycological Institute of Jilin Agricultural University (HMJAU), Changchun, China.

Key worps — SEM, taxonomy, Trichiales

Introduction

Arcyria is a common and important genus of Trichiaceae. Since Wiggers

established the genus in 1780, more than 55 species have been reported (Kirk

et al. 2008, Lado 2001, 2005-12), of which 29 species have been reported in

China (Chen & Li 1999; Chou 1937; Li & Li 1989; Li et al. 1993; Liu et al. 2002;

Wang & Li 1995, 1996, 2006). A new species found on bark surface of dead

described and illustrated below.

Materials & methods

The fruiting bodies and microscopic structures of the new species were examined

by light and scanning electron microscopes (Martin & Alexopoulos 1969; Li et al.

1993) and compared with other morphologically similar Arcyria species. Permanent

slides are mounted in Hoyer’s medium (Martin & Alexopoulos 1969). Coloured slides

were prepared according to Robbrecht (1974) by spreading capillitium in a drop of

94% alcohol, determining colour after one minute, and then mounting in Hoyer’s.

Colour terms are given according to Anonymous (1969). Voucher specimens are

deposited in the Herbarium of Mycological Institute, Jilin Agricultural University

(HMJAU).

402 ... Zhang & al.

AccV Spot Maqn Det WD Exp /-——-+{ 500m

150kV30 38x SE 1001

AccV Spot Maqn Det bs = :

15.0KY 3.0 308K SE~, 1 OmIaRRIARS

& J o

Acc.V Spot Maan." Det pe . AA. AccV Spot Maqn- ‘Det WD Exp

15.0 kV 3:0 5000x., | =e y § Rats i a 2 1bO kV 3.0 20000x SE 10.1 1,

eee —_ avis head! ed " i si Pa

PiaTE 1. Arcyria galericulata (Isotype): A, fruiting bodies. B, a complete, expanded sporocarp.

C, galeiform structure at the top of the sporocarp and part of expansion of the capillitium.

D, part of the stalk, showing longitudinal striations, and part of calyculus and sporocarp. E, outer

surface of a galeiform structure marked with regular circular depressions. F, warted interior of a

depression.

Taxonomy

Arcyria galericulata B. Zhang & Yu Li, sp. nov. PLATE 1-2

MycoBank MB564122

Differs from Arcyria papilla by its smaller sporocarp with a larger helmet-like

protuberance and larger spores.

TypE—China, Jilin province, Jingyuetan National Forestry Park, on the bark of a dead

log, 26 July 2009, Zhang Bo 0628 (Holotype HMJAU10244; isotype HMJAU10245).

EryMoLocy—galericulata (Latin) = with a helmet-like covering, in reference to the

galeiform protuberance at the top of the sporocarp.

Arcyria galericulata sp. nov. (China) ... 403

SPOROCARPS aggregated or united in clusters of 3-8 (with fused stalks), stalked,

erect, 1-1.5 mm in total height, 2-2.2 mm after expansion of the capillitium.

Sporocarp obovoid or shortly cylindrical, fawn to snuff brown, fading to brown.

HyYPOTHALLUS common to a group of sporocarps. STALK 0.8-1.0 mm long, dark

brown by transmitted light, filled with spore-like cells, cells 18-20 um diam. at

the base of stalk, subglobose. PERrp1uM single, membranous, persistent, with a

helmet-like structure at the top of each sporocarp, outer surface of the structure

marked with regular circular depressions with warted interiors. CALYCULUS

translucent, saucer-shaped, light, yellow to colourless by transmitted light,

inner surface with many prominent warts, low irregular ridges forming a net

with small protuberances. COLUMELLA absent. CAPILLITIUM tubular, elastic,

branched and anastomosed, pale orange yellow by transmitted light, 3 um diam.

without ornamentations, 5-6 um diam. with ornamentations, firmly attached

to the calyculus, decorated with many cogs, half-rings and rings with irregular

edges, sometimes with a faint reticulation. Sporgs free, yellowish brown in

mass, yellowish pale to colourless by transmitted light, 6-8 um diam., densely

warted, with scattered groups of more prominent warts.

ComMENTSs: Arcyria galericulata has a galeiform structure at the top of the

sporocarp. Only one other Arcyria, A. papilla Ejale & L.S. Gill (Ejale & Gill

1992), has a similar papillate structure at the top of the sporocarp but the

Acc Spot Magn~ Det WD Pe —— ¥ AccV Spot Magn

15.0kV 3.0 10000x SE 10.3 1 15.0 kV 3.0 @500x

Acc.V Spot Maqn ID —_— - a , Ace.V Spot M

15.0kV 3.0 10000x SiERRIGaie of y 1b.0kKV 3.0 10) 80

4 =.

PLaTE 2. Arcyria galericulata (Isotype): A, part of calyculus, showing many prominent warts and

low irregular ridges forming a net with small protuberances. B-C, part of capillitium, showing the

ornamentation. D, spores.

404 ... Zhang & al.

papilla is smaller, the sporocarp is larger (about 4.4 x 1.0 mm after capillitial

expansion), the capillitium is marked with spines occasionally interrupted

by rings that form characteristic triangular loops, and the spores are smaller

(~5 um diam.) and double-walled.

The capillitial ornamentation of A. galericulata resembles that of A. affinis

Rostaf., A. helvetica (Meyl.) H. Neubert et al., and A. ferruginea Saut. Arcyria

affinis is distinguished by its reddish brown sporocarps (about 3-4 mm after

capillitial expansion), a capillitium broken away leaving only a few ends attached

to the stalk apex, and larger spores (about 7-9 um in diam.) (Rostafinsky

1875). Arcyria ferruginea differs in its shorter stalk (about 0.2-0.6 mm long),

capillitium tube 5-8(-10) um in diam., and larger spores (9-12 um in diam.)

(Sauter 1841), while A. helvetica is differentiated by a deeply funnel-shaped

calyculus and capillitium only attached to the calyculus at the center (Neubert

et al. 1989).

Acknowledgments

We express our deep appreciation to Prof. Guozhong Lit (Dalian Nationalities

University, P.R. China) and Prof. Uno Eliasson (University of Gothenburg, Sweden) for

their valuable suggestions in peer-reviewing this manuscript. We thank Dr. Pu Liu, Jilin

Agricultural University for correcting the manuscript. The study was supported by the

fund from Ministry of Agriculture of China.

Literature cited

Anonymous. 1969. Flora of British Fungi: colour identification chart. Royal Botanic Garden

Edinburgh, H.M. Stationery Off. 6 p

Chen SL, Li Y. 1999. A preliminary report on the myxomycetes from Zhangjiajie, Hunan Province.

J. Wuhan. Bot. Res. 17(3): 217-219.

Chou ZH. 1937. Notes on myxomycetes from China. Bull. Fan. Men. Inst. Biol. 7: 257-278.

Ejale UA, Gill LS. 1992. Two new species of myxomycetes from southern Nigeria. Acta Mycologica

27(2): 267-269.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008 Ainsworth & Bisby’s dictionary of the fungi.

10th Ed. CAB International, Wallingford 771 p.

Lado C. 2001. Nomenmyx: a nomenclatural taxabase of myxomycetes. Cuadernos de Trabajo de

Flora Micoldgica Ibérica 16. 224 p.

Lado C. 2005-2012. An online nomenclatural information system of Eumycetozoa.

http://www.nomen.eumycetozoa.com (Consulted April 15, 2012).

Li Y, Li HZ. 1989. Myxomycetes from China I: A checklist of Myxomycetes from China. Mycotaxon

35(2): 429-436.

Li Y, Chen SL, Li HZ. 1993. Myxomycetes from China X: Additions and notes to Trichiaceae from

China. Mycosystema 6: 107-112.

Liu CH, Yang FH, Chang JH. 2002. Myxomycetes of Taiwan XIV: Three new records of Trichiales.

Tainwainia 47(2): 97-105.

Martin GM, Alexopoulos CJ. 1969. The myxomycetes. University of Iowa Press. Iowa. 561 p.

http://dx.doi.org/10.2307/1218569

Arcyria galericulata sp. nov. (China) ... 405

Neubert H, Nowotny W, Baumann K. 1989. Myxomyceten aus der Bundesrepublik Deutschland

V. Carolinea 47: 25-46.

Robbrecht E. 1974. The genus Arcyria Wiggers in Belgium. Bull. Jard. Bot. Nat. Belg. 44: 303-353

http://dx.doi.org/10.2307/3667676

Rostafinsky JT. 1875. Sluzowce monografia. Pamietn. Towarz.Nauk. Sci. Paryzu. 6(1): 216-432.

Sauter AE. 1841. Beitrage zur Kenntnis der Pilz-Vegetation des Ober-Pinzgaues in Herzogthume

Salzburg. Flora 24: 305-320.

Wang Q, Li Y. 1995. Two new varieties of Arcyria. J. Jilin Agri. Univ. 17(4): 83-85.

Wang Q, Li Y. 1996. A new species of Arcyria myxomycetes. Bull. Bot. Res. 16(2): 179-181.

Wang Q, Li Y. 2006. Trichiales in China. Institute of Science Publish. 134p.

MY COTAXON

http://dx.doi.org/10.5248/120.407

Volume 120, pp. 407-413 April-June 2012

New records of Puccinia species on Poaceae

from Fairy Meadows, Pakistan

N.S. AFSHAN”™, A.N. KHALID? & A.R. NIAZI’

™Centre for Undergraduate Studies & *Department of Botany,

University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

*CORRESPONDENCE TO: pakrust@gmail.com

AsBsTRACT — During a survey of rust fungi of Fairy Meadows, Puccinia brachypodii var.

major on Poa attenuata and P. substriata var. indica on Pennisetum orientale were reported

for the first time. These new rust fungi records bring to 70 the number of Puccinia species

reported on Poaceae from Pakistan. Puccinia brachypodii var. poae-nemoralis and P. poarum

are also reported for the first time from Fairy Meadows.

Key worps — Anthoxanthum odoratum, graminicolous rust, Hunza

Introduction

This paper continues our study of graminicolous rust fungi from Pakistan.

Previously, about 100 species of graminicolous rust fungi including 68 taxa of

Puccinia have been reported from Pakistan (Afshan et al. 2010, 2011a,b). During

a 2007 survey of the rust flora of Fairy Meadows, four members of Poaceae

infected with rust fungi were collected. Among these, Puccinia brachypodii var.

major on Poa attenuata and P. substriata var. indica on Pennisetum orientale

represent new records for Pakistan, while P brachypodii var. poae-nemoralis on

Anthoxanthum odoratum and P. poarum on Poa pratensis are additions to the

rust flora of Fairy Meadows. Anthoxanthum odoratum represents a new host for

rust fungi from Pakistan.

Materials & methods

Freehand sections of infected tissue and spores were mounted in lactophenol and

gently heated to boiling. The preparations were observed under a NIKON YS 100

microscope. Drawings of spores and paraphyses were made using a Camera Lucida

(Ernst Leitz Wetzlar, Germany). Spores were measured using an ocular micrometer.

At least 25 measurements per fungal structure were taken. Both light microscope and

scanning electron microscope (SEM) images were obtained of the spores. Collections

AO8 ... Afshan, Khalid & Niazi

have been deposited in the herbarium of the Botany Department, at the University of

the Punjab, Lahore (LAH).

Enumeration of taxa

Puccinia brachypodii var. major Cummins & H.C. Greene, Mycologia 58: 711

(1966) Fics A-B

SPERMOGONIA and AEcIA unknown. UREDINIA amphigenous, subepidermal,

light brown, 0.6-0.8 x 1.9-2.0 mm. UREDINIOSPORES globose to subglobose

or obovoid, (16-)21-24(-29) x (23-)27-35(-39) um (mean 22 x 30 um);

wall 1.5-2 um thick, light yellow to pale brown, echinulate; germ pores (6-)

Fics. A-B: Puccinia brachypodii var. major.

A: Urediniospores and paraphyses. B: Teliospores. Scale bars = 10 um.

Puccinia species on Poaceae (Pakistan) ... 409

8-11, scattered, obscure; pedicel hyaline, 6-8 um wide and 32-50 um long.

PARAPHYSES 70-95 um long, hyaline to pale yellow, capitate, head 15-17 um

wide; wall = 2 um thick. TELIA amphigenous, sub-epidermal, loculate, with

stromatic paraphyses, dark brown, 0.4-0.7 x 0.6-0.8 mm. TELIOsPORES 1-2

celled, oblong to clavate, golden brown to chestnut brown but paler basally,

smooth; 20-26 x (27—)39-43(-55) um (mean 26 x 34 um); wall 1.5-2 um thick;

apex truncate or rounded, sometimes conical, 3-5 um thick; germ pore 1 per

cell, obscure; pedicel short, hyaline, 6-10 x 31-45 um.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Fairy Meadows, at 3036 ma.s.l.,

stages II + III, on Poa attenuata Trin., 12 August, 2007, NSA # 52. (LAH NSA1034).

ComMENTSs: Puccinia brachypodii var. major is a new record for Pakistan, and

Poa attenuata is a new host record for this fungus. Previously this rust has been

reported on Poa horridula and P. candamoana from Peru (Cummins 1971).

In Pakistan, Puccinia brachypodii var. brachypodii has been reported on

Brachypodium sp. from KPK and Kaghan Valley (Kakishima et al. 1993b);

and P. brachypodii var. poae-nemoralis on Agrostis munroana, Poa nemoralis,

P. pratensis, and P. sterilis from Kaghan Valley, Sharan, Swat, and Azad Jammu

& Kashmir (Ahmad 1956a; Kakishima et al. 1993a,b; Masood et al. 1995).

Puccinia substriata var. indica Ramachar & Cummins, Mycopath. Mycol. appl. 25:

30 (1965) Figs C-D

SPERMOGONIA and AEcIA unknown. UREDINIAamphigenous, sub-epidermal,

yellowish-brown to golden-brown, 0.09-0.1 x 0.1-2.0 mm. UREDINIOSPORES

variable in shape, globose to subglobose or ovoid to ellipsoid, 18-24(-29) x

21-24(-30) um, pale brown to golden brown, echinulate; wall 1-1.5 um thick;

germ pores 3-4(-5), equatorials; pedicel hyaline, 4-5 um wide and = 15 um

long. TeL1a amphigenous, mostly adaxial, sub-epidermal, erumpent, dark

brown to blackish brown, 0.09-0.5 x 0.2-0.6 mm. TELIOsPORES 1-2-celled,

oblong to clavate, 17-24(-29) x (29-)34-52(-60) um, cinnamon brown to

golden brown, paler basally, smooth; wall 2-3 um thick; apex mostly truncate,

sometimes rounded, 4-9 um thick; germ pores obscure; pedicel hyaline to light

brown, 6-8 x 8-12 um.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Fairy Meadows, at 3036 m

a.s.l., stages II + III, on Pennisetum orientale Rich., 12" August, 2007, NSA # 70. (LAH

NSA1095).

ComMENTS: Puccinia substriata var. indica is a new record for Pakistan. The

variety has been reported from India on Pennisetum typhoides (Cummins 1971),

and Afshan et al. (2008) reported another variety, P. substriata var. insolita (on

Panicum antidotale) from Munchinabad in Pakistan. Puccinia penniseti-lanati

(on P lanatum) and Uromyces penniseti (on P americanum from Kaghan

Valley, Tandojam, and Karachi are two other poaceous rusts known to occur

410 ... Afshan, Khalid & Niazi

Fics. C-D: Puccinia substriata var. indica.

C: Urediniospores. D: 1-2-celled teliospores. Scale bars = 10 um.

on Pennisetum species in Pakistan (Ahmad 1969; Hasnain et al. 1959; Khan &

Kamal 1968).

Puccinia brachypodii var. poae-nemoralis (G.H. Otth) Cummins & H.C. Greene,

Mycologia 58: 705 (1966) Fics E-F

SPERMOGONIA and AECIA unknown. UREDINIA on leaves, mostly adaxial,

yellowish or yellowish brown, 0.09-0.2 x 0.2-0.4 mm. UREDINIOSPORES

obovoid or ellipsoid to broadly ellipsoid, 19-26 x 23-36 um (23.2 x 28.6 um);

wall 2-3 um thick, light brown to cinnamon brown, closely echinulate; germ

pores 5-9, obscure; pedicel hyaline, short, 7-8 x 38-45 um. PARAPHYSES

cylindric to capitate, hyaline or yellowish, mostly 80-100 um long and 14-18

um wide, usually geniculata, wall 2-4 um thick throughout or to 6 um thick in

the head. TeL1a on leaves, mostly abaxial, sub-epidermal, 0.06-0.1 x 0.1-0.2

mm, blackish, loculate with a few brown paraphyses surrounding the sori.

TELIOSPORES 1-2 celled (few one-celled spores also observed), oblong to clavate,

13-24 x 40-59(-65) um (mean 17.7 x 50.6 um), not or slightly constricted at

Puccinia species on Poaceae (Pakistan) ... 411

Fics. E-F: Puccinia brachypodii var. poae-nemoralis.

E: Urediniospores and paraphyses. F: Teliospores. Scale bars = 10 um.

the septum; wall 1-1.5 um thick, smooth, brown to chestnut brown or paler

basally; apex 5-9 um thick, truncate or conical; germ pore 1 per cell, upper

sub-apical, lower at the equator or near the septum; pedicel short, light brown,

5-6 x 9-15 um, not collapsing, thick walled.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Karimabad, Hunza, at 2438 m

a.s.l., stages I] + III, on Anthoxanthum odoratum L., 13 August, 2007; Fairy Meadows,

at 3036 m a.s.l., 12 August, 2007, NSA # Gr 58 (G06). (LAH NSA1035a & 1035b).

ComMENTs: Puccinia brachypodii var. poae-nemoralis has been reported on

leaves of Agrostis munroana, Poa nemoralis, P. pratensis, and P. sterilis from

Kaghan Valley, Sharhan, Swat and Azad Jammu & Kashmir (Ahmad et al.

1997).

412 ... Afshan, Khalid & Niazi

P. brachypodii var. poae-nemoralis is reported for the first time from Fairy

Meadows and Hunza, Northern Areas of Pakistan; Anthoxanthum odoratum is

a new host for rust fungi from Pakistan.

Puccinia poarum P. Nielsen, Bot. Tidsskr., ser. 3, 2: 34 (1877) Fics G-H

SPERMOGONIA and AEcIA unknown. UREDINIA amphigenous, subepidermal,

light yellow to light brown, 0.6-0.8 x 1.9-2.0 mm. UREDINIOSPORES globose to

subglobose or obovoid, 20-26 x 23-30 um (mean 22.92 x 26.71 um); wall 1.5-2

uum thick, light yellow to pale brown, echinulate; germ pores 4—-5(-8), obscure;

pedicel hyaline, 5-6 um wide and = 15 um long. PARAPHysEs few, clavate to

capitate, peripheral, hyaline to pale yellow, head 19-21 um wide, wall = 2 um

thick, 94-100 um long. TELIA amphigenous, covered by the epidermis, rarely

Fics. G-H: Puccinia poarum.

G. Urediniospores and paraphyses. H: Teliospores. Scale bars = 10 um.

Puccinia species on Poaceae (Pakistan) ... 413

loculate, dark brown, 0.4-0.7 x 0.6-0.8 mm. TELIOSPORES 1-2 celled, oblong to

clavate; wall 2-3 um thick, golden brown to chestnut brown but paler basally,

smooth; 11-19(-22) x 37-58(-67) um (mean 15.7 x 47.9 um); apex truncate

or conical to obliquely conical, 3-5 um thick; germ pore 1 per cell, obscure;

pedicel short, brown, 6-7 x 9-11 um.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Fairy Meadows, at 3,036 ma.s.L.,

stages II + III, on Poa pratensis L., 12 August, 2007, NSA # G 371 (LAH NSA1077).

COMMENTS: Puccinia poarum has been reported on Poa annua from Lahore

by Ahmad (1956a,b), on Agrostis sp. from Kaghan valley and Azad Jammu &

Kashmir by Kakishima et al. (1993b). Puccinia poarum is newly recorded from

Fairy Meadows, Northern areas of Pakistan.

Acknowledgements

We sincerely thank Dr. José R. Hernandez, Risk Manager (Plant Pathology

Regulations, Permits & Manuals, USDA) and Dr. Omar Paino Perdomo (Dominican

Society of Mycology Santo Domingo, Dominican Republic) for their valuable

suggestions to improve the manuscript and acting as presubmission reviewers. We are

highly obliged to the Higher Education Commission (HEC) of Pakistan for providing

financial support for this research work.

Literature cited

Afshan NS, Khalid AN, Javed H. 2008. Further additions to the rust flora of Pakistan. Pakistan

Journal of Botany 40(3): 1285-1289.

Afshan NS, Khalid AN, Niazi AR. 2010. Three new species of rust fungi from Pakistan. Mycological

Progress 9: 485-490. http://dx.doi.org/10.1007/s11557-010-0655-8

Afshan NS, Khalid AN, Niazi AR, Iqbal SH. 2011a. New records of Uredinales from Fairy Meadows,

Pakistan. Mycotaxon 115: 203-213. http://dx.doi.org/10.5248/115.203

Afshan NS, Khalid AN, Iqbal SH, Niazi AR. 2011b. Puccinia species new to Azad Jammu & Kashmir,

Pakistan. Mycotaxon 116: 175-182. http://dx.doi.org/10.5248/116.175

Ahmad S. 1956a. Uredinales of West Pakistan. Biologia 2(1): 29-101.

Ahmad S. 1956b. Fungi of Pakistan. Biological Society of Pakistan, Lahore Monograph 1: 1-126.

Ahmad S. 1969. Fungi of Pakistan. Biological Society of Pakistan, Lahore, Monograph 5(Suppl.

1): 110.

Ahmad §, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Nabiza Printing Press, Karachi, Pakistan.

Cummins GB. 1971. The rust fungi of cereals, grasses and bamboos. Springer Verlag, Berlin-

Heidelberg—New York.

Hasnain SZ, Khan A, Zaidi AJ. 1959. Rust and smut of Karachi. Department of Botany, University

of Karachi, Monograph 2. 36 p.

Kakishima M, Izumi O, Ono Y. 1993a. Rust fungi (Uredinales) of Pakistan collected in 1991.

Cryptogamic Flora of Pakistan 2: 169-179.

Kakishima M, Izumi O, Ono Y. 1993b. Graminicolous rust fungi (Uredinales) from Pakistan.

Cryptogamic Flora of Pakistan 2: 181-186.

Khan SA, Kamal M. 1968. The fungi of South West Pakistan. Part 1. Pak. J. Sci. & Ind. Res. 11:

61-80.

Masood A, Khalid AN, Iqbal SH. 1995. New records of graminicolous rust fungi (Uredinales) from

Pakistan. Sci. Int. 7(3): 415-416.

MY COTAXON

http://dx.doi.org/10.5248/120.415

Volume 120, pp. 415-422 April-June 2012

Lectotypification and characterization of the natural phenotype

of Fusarium bactridioides'

KEITH A. SEIFERT & TOM GRAFENHAN

Biodiversity (Mycology & Botany), Agriculture & Agri-Food Canada,

Ottawa, ON, K1A 0C6 Canada

Grain Research Laboratory, Canadian Grain Commission,

Winnipeg, MB, R3C 3G8 Canada

* CORRESPONDENCE TO: keith. seifert@agr.gc.ca

ABSTRACT — Specimens of Fusarium bactridioides deposited in the US National Fungus

Collections and New York Botanical Garden were examined and a lectotype is selected based

on naturally infected galls of Cronartium conigenum collected in Arizona. This species was

previously only described from culture, and its natural phenotype is presented and illustrated

here. Historical experiments involving the attempted use of F. bactridioides as a biocontrol of

pine blister rusts in Oregon and New Hampshire are reviewed, but the unpublished records

of the ultimate fate of these experiments could not be located.

Key worps — Hypocreales, Nectriaceae, anamorph taxonomy, biological control

Introduction

Fusarium bactridioides was the last species of the genus to be formally

described in the august pages of the journal SciENcE. The single page outlined the

circumstances of the discovery of the fungus by the American botanist Arthur

Hinckley and forest pathologist L.N. Goodding. The Fusarium “thoroughly

parasitized” a cone of Pinus leiophylla var. chihuahuana (Chihuahua pine)

attacked by the cone blister rust, Cronartium conigenum Hedgc. & N.R. Hunt.

It was originally collected in the remote Chiricahua Mountains in Arizona (ca.

31°50'N 109°17'W), a so-called ‘sky island’ range of eroded volcanic rhyolite

rock arising from the surrounding grassland desert.

' This article, the first of a projected series on the typification and nomenclature of species of

Fusarium and allied genera, is dedicated to our friend and mentor Emory G. Simmons, whose

erudite and articulate ‘Alternaria: Themes and Variations, published in this journal, inspired

this pale imitation.

416 ... Seifert & Grafenhan

Following the formal description is a remarkable story of what must be among

the earliest deliberate releases of one fungus as a possible biological control of

another. The Cronartium host is one of several rusts infecting Pinus species in

the southwestern United States and adjacent Mexico, with uredinia and telia

developing on various Quercus spp. (Cannon 2007). In July 1932, only three

months after the discovery of the species, conidia of F. bactridioides were sprayed

onto living C. conigenum cankers on Pinus monticola in Clackamas County

Oregon. The following July, all the inoculated cankers were dead. Inoculations

were then attempted on galls of C. harknessii E. Meinecke and C. filamentosum

Hedge. on P. contorta in two locations in Oregon, where sporulating Fusarium

colonies were observed on the galls during a survey four months later. Field

inoculations in four localities in Idaho were reported, but only sketchy details

were provided of observations of the Fusarium sporulating on the cankers

afterwards and there are no subsequent literature reports. Unfortunately, we

were unable to locate any unpublished notes of L.N. Goodding that might give

more information on the ultimate fate of these inoculations.

Fusarium bactridioides was classified in Fusarium sect. Discolor by

Wollenweber (1934), Wollenweber & Reinking (1935), and Gerlach &

Nirenberg (1982). It was considered a synonym of F. sambucinum Fuckel (Booth

1971, Nelson et al. 1983) and E bactridioides vaguely fits the broad concept of

F. sambucinum adopted by these authors. However, the macroconidia lack

the asymmetrical, ‘dolphin-nose’ apical cell typical of the latter species. Of the

species segregated from F. sambucinum by Nirenberg (1985), the macroconidia

of F. bactridioides have some morphological similarity to those of the potato

pathogen E venenatum Nirenberg, but the latter species lacks microconidia. The

phylogenetic and taxonomic studies of O’Donnell et al. (1998) and Nirenberg

& O'Donnell (1998) accept F. bactridioides as a distinct species in the American

clade of the Fusarium fujikuroi complex.

Wollenweber (1934) did not illustrate the species with the protologue,

but later published a drawing as Fusarium autographice delineata no. 1153

(Wollenweber 1935), which was excerpted in his subsequent monographs. All

published descriptions and illustrations of this species are probably based on

one culture, exemplified by the modern species description and microscopic

photographs of Gerlach & Nirenberg (1982: 267). This in vitro characterization

is not repeated here, but the natural ‘wild type’ of this species on pine rust galls

is illustrated and described. No holotype was designated in the protologue, and

this shortcoming is remedied below by the selection of a lectotype.

Typification

The protologue of F. bactridioides explicitly mentions three specimens, the

first probably the original collection from Arizona, and two from the inoculation

Fusarium bactridioides lectotypified ... 417

Fics. 1-2. Fusarium bactridioides, habit photographs. 1. Naturally infected Cronartium conigenum

gall (BPI 451322). 2. Detail of sporodochia (lectotype, composite photograph assembled with

CombineZ, Hadley 2006).

experiments in Oregon. There is no designation of a holotype. The description

was based on a culture, and it is unlikely that Wollenweber ever saw any of the

specimens that he listed.

Eight specimens of E bactridioides are deposited in BPI and one in NY. Apart

from one specimen collected and identified by J.R. Hansbrough in Waterville,

New Hampshire (BPI US0451805) after the publication of the protologue, these

collections are authentic and represent; a) material collected by A. Hinckley

in the Chiricahua Mountains (NY 00936830 from 1932 on a host identified as

Cronartium quercuum (Berk.) Miyabe; BPI US0451322 from 1933 on a host

identified as C. conigenum); b) dried cultures from “cones collected by Arthur,

Mar. 15, 1934” (BPI US0451803) and four specimens collected in 1933 from

the inoculated locations in Oregon. It seems reasonable to designate the galls

of C. quercuum collected by Hinckley in 1932 as the lectotype, despite the

absence of the variety name for the host. The dates and locality match precisely

and the identification of the host probably was changed between the times

of specimen deposit and publication. This is presumably the specimen from

which the cultures and subsequent specimens from inoculated localities were

derived. The 1933 specimen in BPI appears also to be a natural infection from

the type locality and although it predates publication, it is not mentioned in the

protologue.

An argument could be made to designate the dried cultures in US0451803 as

the lectotype, because they probably represent the strain(s), but not the actual

transfers, that Wollenweber described. Unfortunately, the six dried PDA slants

have no evident sporodochia or macroconidia, although there are abundant

418 ... Seifert & Grafenhan

microconidia in the cottony, white aerial mycelium. The cultures are heavily

contaminated with coccoid bacteria. The date on the package containing the

dried cultures is “Mar. 15, 1934”, a scant three months before Wollenweber’s

description was published. The notes inside this packet suggest that the 1934

date is actually when the preserved cultures were dried. Only one strain of this

fungus now exists, i.e. Wollenweber 4748 > CBS 177.35 (> BBA 63602, CBS

> NRRL 22201, NRRL > DAOM 225115, NRRL > CBS 100057). The data for

CBS 100057 record P. leiophylla as the plant host and the location as Arizona,

suggesting that this strain, which was regarded as an ‘ex-type culture’ by

Gerlach & Nirenberg (1982) and Nirenberg & O'Donnell (1998), was derived

from material from the Chiricahua Mountains. The protologue is ambiguous

about how many strains were originally isolated and how many were sent to

Wollenweber by his American colleagues. One of these strains was illustrated as

Fusarium autographice delineata no. 1153, and we must assume that the culture

now represented by CBS 100057 is the same strain. We consider this to be the

ex-lectotype strain, and see no need to formalize this by epitypification.

For formal typification, we prefer to emphasize the concept of the species as

a parasite of Cronartium, and designate the lectotype accordingly below.

Taxonomy

Fusarium bactridioides Wollenw., Science 79: 572. 1934. FIGS 1-5

MycoBank MB 258078

Type: On Cronartium quercuum on Pinus chichuahuana (sic), USA, Arizona, Chiricahua

Mt., Cave Creek, HI.1932, Arthur Hinckley (lectotype designated here, NY 00936830).

Sporodochia are erumpent from host tissue and visible as Orange-White

(5A2, Kornerup & Wanscher 1978) to Light Orange (5A5) masses of conidial

slime. On the naturally infected specimens, they are irregular in outline and

form lesions up to 1 cm x 5 mm on the galls. On inoculated specimens, the

lesions are scattered and smaller, about 200-1000 um diam. The stroma is

poorly developed, about 100 um thick, and in optical section appears as a

textura angularis of thin- to slightly thick-walled cells about 3.5-7 um wide, but

its hyphal character is evident with changes of focus. A hymenium-like layer of

conidiophores and phialides arises from the stroma, with the conidiophores

more or less biverticillate, but with phialides also often arising at the first level

of branching. Metulae are cylindrical, doliiform to slightly clavate, 8.5-11.5 x

3.5-7 um at the broadest part. Phialides are 9.5-15 x 3-5 um at the broadest

part, narrowly doliiform, sometimes with a central constriction, in terminal

pairs or whorls of 3, or arising singly, in pairs or in whorls with metulae, with

a flared collarette about 1-1.5 um long and periclinal thickening sometimes

visible; conidiogenous aperture about 2 um wide.

Fusarium bactridioides lectotypified ... 419

+ - : 1 ™ _ 4

Fics 3-4. Microphotographs of Fusarium bactridioides, BPI 451804. 3. Macroconidia. (composite

image). 4. Hand section of the sporodochial stroma, showing conidiophores and phialides.

Scale bars = 10 um.

There is no obvious distinct difference between ‘microconidial’ and

« *4°_,p . ‘i .

macroconidial’ sporodochia and there is more or less a continuum between

the two conidial types. Few macroconidia occur on the naturally infected

420 ... Seifert & Grafenhan

specimens, but they are more abundant on the lectotype and inoculated

specimens. Macroconidia are 3-6 septate; 3-septate predominate and are

30.5-45.5 x 4.5-6 um (mean + SE = 38.3 + 0.8 x 5.3 + 0.1, n = 20), I/w = 5.5-9;

4-septate 27-36.5 x 5-6 um (mean + SE = 31.9+0.8 x 5.4+0.1,n= 10), l/w=5-7;

5-septate 29-39.5 x 5.5-6.5 um (mean + SE = 34.6 + 0.9 x 5.8 + 0.1, n = 13),

I/w = 5-7; 6-septate 35.5-41 x 5-6 um (n = 3), I/w = 6-7. In side view, the

ventral surface is more or less straight or gently curved, and the dorsal surface

is moderately curved, with the walls more or less parallel in the central two cells

of the conidia, with the widest point near the middle or above the middle. The

apical cell is bluntly rounded, and roughly the same length as the penultimate

cell. The basal cell is tapered more acutely than apical cell; the base is rounded,

flat, or has a slight indentation on the dorsal side or central papilla, indicating a

foot cell. In front view, the macroconidia appear somewhat clavate. 4-6 septate

macroconidia tend to have unequal lengths of cells, with the additional septa

dividing one but not all of the original four cells. Microconidia are abundant

in the sporodochia, are 0-3 septate, and vary in shape and size from small,

ellipsoidal, oblong-ellipsoidal or allantoid cells that are obviously microconidia

to fusiform to clavate, septate spores that intergrade with macroconidia.

Aseptate conidia 6-18 x 3.5-6.5 um (mean + SE = 11.8 + 0.6 x 4.9 + 0.1, n = 25),

I/w = 1.5-4. 1-septate conidia 9.5-19.5 x 4-6.5 um (mean + SE = 14.9 + 0.5

x 5.1 + 0.1, n = 25), l/w = 2-4. 2-septate conidia are infrequent and 15-24 x

5-6.5 uum. 3-septate microconidia also occur and can be distinguished from

macroconidia by their shorter length and their more clavate shape; they are

16-31 x 5-7 um (mean + SE = 24.0 + 0.7 x 6.1 + 0.1, n = 25), I/w = 3-5. In

general, the bases of microconidia are conical, symmetrical or asymmetrical,

with a flat secession scar or relatively conspicuous papilla; the apical cell or part

of the conidium is rounded.

ADDITIONAL SPECIMENS EXAMINED: USA, Arizona, Chiricahua Mt. (as Chricicalma),

Cave Creek, on Cronartium conigenum on Pinus leiophylla, 25.V1I1.1933, Arthur Hinckley

(BPI US0451322); Oregon, Mt. Hood National Forest, On Cronartium harknessii on

Pinus contorta, 8.V1I.1933, L.N. Goodding (BPI US0451323); on Cronartium ribicola,

24.X.1933, L.N. Goodding (BPI US0451325); Eagle Creek, on Cronartium ribicola on

Pinus monticola, 24.X.1933, L.N. Goodding (BPI US0451326); six dried slant cultures on

PDA from “cones collected by Arthur” (BPI US0451803); Hood River Co., Eagle Creek,

on Cronartium ribicola on Pinus sp., 24.X.1933, L.N. Goodding (BPI US0451804);

New Hampshire, Waterville, on Pinus strobus, 1.[X.1935, J.R. Hansbrough (BPI

US0451805).

Discussion

The micromorphology of F. bactridioides reported here from natural

material is comparable to that reported from cultures of this species by

Wollenweber (1934) and Gerlach & Nirenberg (1982). The main distinction

is that on the natural substrate the microconidia are produced from the

Fusarium bactridioides lectotypified ... 421

Fic. 5. Microphotograph of microconidia of Fusarium bactridioides, lectotype, NY 00936830

(composite image). Scale bar = 10 um.

same sporodochia as the macroconidia, whereas in culture microconidia are

produced independently. In nature, there is a continuum of shape and size

between macro- and microconidia, but in culture the two kinds seem to be

morphologically distinct. Gerlach & Nirenberg (1982) reported macroconidia

with up to 11 septa in culture, with 3-5 septa being the typical condition. On

the specimens, the macroconidia with three septa predominated, with a small

number of 4-6 septate macroconidia seen on some specimens. Morphologically,

Fusarium bactridioides has more robust macroconidia than is typical for the F.

fujikuroi complex, and the 0-1 septate oval macroconidia are produced on the

agar surface in vitro, rather than in the aerial mycelium as they are in related

species. The mycoparasitic habit is also unusual in this group.

To our knowledge, Fusarium bactridioides has not been collected after

1935 and no natural infections have been reported outside of Arizona. The

Hansbrough specimens (USO451805) collected in 1935 are the result of

inoculation experiments in New Hampshire in 1934, about which apparently

nothing was ever published. We were unsuccessful in locating the typewritten

report by Hansbrough cited in a note included with the specimens. Whether

EF. bactridioides is indigenous and perhaps restricted to the remote Chiricahua

Mountains, and whether the inoculations of blister rust cankers in Oregon,

422 ... Seifert & Grafenhan

Idaho and New Hampshire still persist, seem questions worthy of investigation.

Such inoculation experiments would not easily be conducted today, with the

need for risk assessments and legal permits from regulatory agencies. Fusarium

bactridioides may provide a unique opportunity to search for the lingering

fingerprints of innovative scientific activities of a more innocent time.

Acknowledgments

We are grateful to Helgard Nirenberg and Walter Gams for their insights into the

practices of H.W. Wollenweber in Berlin, the curators of BPI and NY for the kind

loans of specimens discussed in this article, and Drs T. Aoki and B. Summerell for pre-

submission reviews. We appreciate the efforts of staff at the herbarium (S. Hunkins)

and library (M. Carpenter) of the University of Arizona to find archival material of L.N.

Goodding, and staff at the Connecticut Agricultural Experiment Station (D.W. Li, S.

Anagnostakis, V. Bomba-Lewandoski, W. Elmer) and USDA Forestry Lab in Hamden,

CT (M. Keena) for searching for archives of J.R. Hansbrough.

Literature cited

Booth C. 1971. The genus Fusarium. Commonwealth Mycological Institute, Kew.

Cannon PE. 2007. Cronartium conigenum. IMI descriptions of fungi and bacteria No. 1722. 4 p.

Gerlach W, Nirenberg HI. 1982. The genus Fusarium - a pictorial atlas. Mitt. Biol. Bundesanst.

Ld.- u. Forstw. 209: 1-406.

Hadley A. 2006. CombineZ, version 5. Published by the author. www.hadleyweb.pwp

Kornerup A, Wanscher JH. 1978. Methuen handbook of color, 3rd ed. Denmark, Sankt Jorgen

Tryk. 243 p.

Nelson PE, Toussoun TA, Marasas WFO. 1983. Fusarium species, an illustrated manual for

identification. Pennsylvania State Univ. Press, University Park, London. 193 p.

Nirenberg HI, O'Donnell K. 1998. New Fusarium species and combinations within the Gibberella

fujikuroi species complex. Mycologia 90: 434-458. http://dx.doi.org/10.2307/3761403

O’Donnell K, Cigelnik E, Nirenberg HI. 1998. Molecular systematics and phylogeography of the

Gibberella fujikuroi species complex. Mycologia 90: 465-493.

http://dx.doi.org/10.2307/3761407

Wollenweber HW. 1934. Fusarium bactridioides sp. nov., associated with Cronartium. Science 79:

572. http://dx.doi.org/10.1126/science.79.2060.572

Wollenweber HW. 1935. Fusaria autographice delineata [4]: nos 1101-1200. Self published, Berlin.

1200 p.

Wollenweber HW. 1943. Fusarium—Monographie IJ. Fungi parasitici et saprophytici. ZenBl. Bakt,,

Abt. 2, 106: 171-202.

Wollenweber HW, Reinking OA. 1935. Die Fusarien, ihre Beschreibung, Schadwirkung und

Bekampfung. Paul Parey, Berlin. 355 p.

MY COTAXON

http://dx.doi.org/10.5248/120.423

Volume 120, pp. 423-426 April-June 2012

Lectotypification of Crepidotus variabilis var. subsphaerosporus

SONA JANCOVICOVA' & SHAUN R. PENNYCOOK?’

' Comenius University in Bratislava, Faculty of Natural Sciences,

Department of Botany, Révovd 39, 811 02 Bratislava, Slovakia

? Manaaki Whenua Landcare Research,

Private Bag 92 170, Auckland 1142, New Zealand

CORRESPONDENCE TO: ' jancovicova@fns.uniba.sk & * PennycookS@LandcareResearch.co.nz

ABSTRACT — An original watercolour painting by J.E. Lange is designated as the lectotype

of Crepidotus variabilis var. subsphaerosporus. This painting was cited in two descriptions

published invalidly by Lange prior to his validation of the name; it therefore constitutes part

of the original material on which he based his description. As a lectotype, it supersedes the

neotype previously designated by Senn-Irlet, which is here designated as epitype.

Key worps — Afbildninger af Danmarks Agaricaceer, Basidiomycota, Crepidotus cesatii var.

subsphaerosporus, Crepidotus kubickae, Flora Agaricina Danica

Introduction

The name Crepidotus variabilis var. subsphaerosporus was first proposed by

J.E. Lange (1938: 52, as ‘subsphaerospora’) as a nomen non rite publicatum,

lacking a Latin description (McNeill et al. 2006: Art. 36.1). No type specimen

was designated; according to M. Lange (1969: 131), J.E. Lange “never tried to

make up a set of herbarium specimens.’ However, an illustration was cited

(as “D.A. pl. 533”), together with details of collection locality, habitat, and

date. The abbreviation “D.A.” refers to the extensive series of watercolours

painted by J.E. Lange during the years 1893-1910 and compiled under the

title “Afbildninger af Danmarks Agaricaceer” (J.E. Lange 1914: 1; M. Lange

1969: 122-123). The original paintings (initially held by J.E. Lange) are now

conserved in the Herbarium, Natural History Museum of Denmark, University

of Copenhagen (C), and are reproduced on the Flora Agaricina Danica

website (http://130.225.211.158/agaricina/flagar-search.htm; H. Knudsen,

pers. comm.). Subsequently, J.E. Lange painted a duplicate set for the Museum

on better quality paper (M. Lange 1969: 123); this set is now held in the

private collection of his granddaughter, Dr. Lene Lange, Aalborg University

(H. Knudsen, pers. comm.).

424 ... Jan¢éovitova & Pennycook

J.E. Lange (1939: 46, pl. 133 fig. E) republished the description with minor

changes accompanied by a printed reproduction of the original painting (with

some repositioning of component elements), but the name remained invalid

for lack of a Latin description.

J.E. Lange (1940: IV) finally validated the name in a set of Latin diagnoses

of names invalidly published in “Flora Agaricina Danica.” Although there is no

direct reference to the earlier publication pages, the trinomial is marked with

an asterisk, indicating that the name had originally been published in “Studies

in the Agarics of Denmark” during the period “1935-40” (J.E. Lange 1940: IJ),

i.e., in the series of papers that included J.E. Lange (1938).

When Senn-Irlet (1995: 53) recombined the name as Crepidotus cesatii var.

subsphaerosporus (J.E. Lange) Senn-Irlet, she designated a Swiss collection as

neotype. However, this designation ignored the availability of original material

(the watercolour painting) that should have been selected as lectotype (McNeill

et al. 2006: Art. 9.10, 9.11).

Here we designate the original watercolour painting as lectotype, and the

superseded Swiss “neotype” as epitype.

Taxonomy and typification

The name Crepidotus variabilis var. subsphaerosporus has priority at variety

rank (e.g., as treated in Senn-Irlet 1995). Ripkova (2009: 272) summarized the

nomenclatural history of the taxon under Crepidotus kubickae, the synonym

that has priority at species rank. Two widely cited purported recombinations of

J.E. Lange's basionym at species rank were not validly published (McNeill et al.

2006: Art. 33.4): “Crepidotus subsphaerosporus’ (J.E. Lange) Kuhner & Romagn.

(Kihner & Romagnesi 1953: 76) lacked a full and direct basionym reference,

and “Crepidotus subsphaerosporus” (J.E. Lange) Kithner & Romagn. ex Hesler &

A.H. Sm. (Hesler & Smith 1965: 121) gave a full and direct reference to the 1938

nomen non rite publicatum, a non-correctable nomenclatural error (McNeill

et al. 2006: Art. 33.7).

J.E. Lange preserved no herbarium specimens other than an occasional

spore print. There are no J.E. Lange collections or spore prints of C. variabilis

var. subsphaerosporus at C; the watercolour painting, conserved at C (viewable

on http://130.225.211.158/agaricina/flagar-search.htm under “Crepidotus sub-

sphaerosporus’), is apparently the only extant original material. (J.E. Lange's

duplicate painting is extant, but it is debatable whether this can be considered

to be “original material.”) The original painting is on a 26.5 x 17.7 cm sheet of

thick, cardboard-like, aquarelle paper, which has now yellowed considerably.

The images comprise a drawing of two basidiospores, a painted representation of

the spore print colour (easily overlooked because of its resemblance to adjacent

blotches of foxing discoloration), and three habit paintings of the basidiomata.

Crepidotus variabilis var. subsphaerosporus, lecto- & epitypified ... 425

(The three habit paintings and the basidiospore drawing, but not the spore print

colour representation, are also reproduced in J.E. Lange 1939: pl. 133 fig. E.)

The original painting bears various handwritten ink and pencil annotations in

Danish: in the upper right corner “Tavle 533”; below the basidiospore drawing

“sp x 1000”; and along the lower margin “Crepidotus sessilis (Bull.) Schroet.

f. subsphaerospora. Sporestov blegt okkerbrunt. Sp spheerisk-ellips, 6 x 4 %.

Heesbjerg, pa pinde af eg o.a. lovtreer. Okt. 1901” Hzesbjerg is an old spelling

for the modern Hesbjerg, the name of a number of localities on Funen, the

Danish island where J.E. Lange lived and did most of his collecting; the most

likely collection locality is at 55°22'N 10°13’E, the only forested Hesbjerg on

Funen (H. Knudsen, pers. comm.).

We propose the original painting as lectotype of C. variabilis var. subsphaero-

sporus. We also propose Senn-Irlet’s superseded “neotype’” as epitype, providing

micromorphological data to supplement the macromorphology illustrated in

the lectotype.

Crepidotus kubickae Pilat, Stud. Bot. Cechoslov. 10: 150, 1949, as ‘Kubickae’.

TyPE (Pilat 1949, Senn-Irlet 1992, Ripkova 2009): CZECH REPUBLIC. Central Bohemia:

“Poricko nad Sazavou, ad terram inter muscis, 29.V.1949, legit J. Kubicka, det. Pilat,

Typus.” (PRM 665190, holotype).

= Crepidotus variabilis var. subsphaerosporus J.E. Lange, Fl. Agaric Danic. 5: IV, 1940.

Types: DENMARK. Funen: Hesbjerg, “Crepidotus sessilis (Bull.) Schroet. f.

subsphaerospora. Sporeprint pale ochre brown. Sp sphaerical-ellipsoid, 6 x 4%. Heesbjerg,

on sticks of Quercus and other deciduous trees. Oct. 1901.” [in Danish]. (C, Danmarks

Agaricaceer Tavle 533, original watercolour painting by J.E. Lange, lectotype designated

here). SWITZERLAND. Bern: Rothenbach, Schineggschwand am Schallberg, alt. 1000

m, Abieti-Fagetum, on fallen Picea twigs, 11 Oct. 1989, Senn-Irlet 89/240 (G, epitype

designated here).

Acknowledgments

We thank Henning Knudsen (the Herbarium, Natural History Museum, University

of Copenhagen), without whose willingness to answer numerous questions concerning

J.E. Lange’s paintings and their curation this paper could not have been written, and

Henning Knudsen and Scott Redhead (Systematic Mycology and Botany, Agriculture

and Agri-Food Canada, Ottawa) for refereeing the manuscript.

Literature cited

Hesler LR, Smith AH. 1965. North American species of Crepidotus. Hafner Publishing Company,

New York. 168 p.

Kiihner R, Romagnesi H. 1953. Flore analytique des champignons supérieurs (agarics, bolets,

chanterelles). Masson et Cie, Paris. 556 p.

Lange JE. 1914. Studies in the agarics of Denmark. I. General Introduction. The genus Mycena.

Dansk Botanisk Arkiv 1(5): 1-40.

426 ... Jan¢éoviéova & Pennycook

Lange JE. 1938. Studies in the agarics of Denmark. Part XII. Hebeloma, Naucoria, Tubaria,

Galera, Bolbitius, Pluteolus, Crepidotus, Pseudopaxillus, Paxillus. Additional descriptions and

supplementary notes to part I-XI. Dansk Botanisk Arkiv 9(6): 1-104.

Lange JE. 1939. Flora Agaricina Danica, Vol. 4. The Society for the Advancement of Mycology in

Denmark and the Danish Botanical Society, Copenhagen. pp. 1-119, pl. 121-160.

Lange JE. 1940. Flora Agaricina Danica, Vol. 5. The Society for the Advancement of Mycology in

Denmark and the Danish Botanical Society, Copenhagen. pp. 1-103 + I-XXIV, pl. 161-200.

Lange M. 1969. Jakob E. Lange and the creation of “Flora Agaricina Danica.’ Friesia 9(1-2):

121-132.

McNeill J, Barrie FR, Burdet HM, Demoulin V, Hawksworth DL, Marhold K, Nicolson DH,

Prado J, Silva PC, Skog JE, Wiersema JH, Turland NJ. 2006. International Code of Botanical

Nomenclature (Vienna Code) adopted by the Seventeenth International Botanical Congress,

Vienna, Austria, July 2005. Regnum Vegetabile 146. 568 p.

Pilat A. 1949. Ad monographiam Crepidotorum europaeorum. Supplementum I. Studia Botanica

Cechoslovaca 10(4): 149-154.

Ripkova S. 2009. Crepidotus kubickae - a forgotten name. Mycotaxon 110: 271-281.

http://dx.doi.org/10.5248/110.271

Senn-Irlet B. 1992. Type studies in Crepidotus — I. Persoonia 14(4): 615-623.

Senn-Irlet B. 1995. The genus Crepidotus (Fr.) Staude in Europe. Persoonia 16(1): 1-80.

MY COTAXON

http://dx.doi.org/10.5248/120.427

Volume 120, pp. 427-435 April-June 2012

A new species of Conidiobolus (Ancylistaceae)

from Anhui, China

YONG Nig?, Cul-ZHu Yu', XIAO- YONG Liu?’ & Bo HUANG"

‘Anhui Provincial Key Laboratory of Microbial Control, Anhui Agricultural University,

West Changjiang Road 130, Hefei, Anhui 230036, China

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

* CORRESPONDENCE TO: bhuang@ahau.edu.cn, liuxiaoyong@im.ac.cn

ABSTRACT —Conidiobolus sinensis was isolated from plant detritus in Huoshan, Anhui

Province, eastern China. It produces primary conidiophores from cushion mycelium,

which is distinct from all other species in the genus except C. stromoideus and C. lichenicola.

Morphologically C. sinensis differs from C. stromoideus in the shape of the mycelia at the

colony edge and conidiophore length and from C. lichenicola by colony color and mycelial

form. A phylogram based on partial 28S rDNA and EF-1a sequences from 14 Conidiobolus

species shows C. sinensis most closely related to C. stromoideus, forming a clade of sister

taxa with a 100% bootstrap. DNA similarity levels between these two species were 94% (28S

rDNA) and 96% (EF-1a). Based on the morphological and molecular evidence, C. sinensis is

considered a new species.

Key worps —Entomophthorales, hyphal knots, taxonomy

Introduction

Species belonging to Conidiobolus can be easily isolated from soil, decaying

leaf litter, rotten vegetables and some dead insects, although the type of the

genus, C. utriculosus Bref., was first isolated from the decaying fleshy fruitbodies

of Exidia and Hirneola. The genus is diagnosed by (i) nuclei that do not stain in

aceto-orcein and lacking obviously granular contents, (ii) simple conidiophores,

(iii) globose to pyriform multinucleate conidia, (iv) resting spores formed in the

axis of hypha (mostly as zygospores), and (v) walled vegetative cells (Humber

1997). After Huang et al. (2007) recognized 30 species within Conidiobolus, only

two additional species — C. margaritatus (Huang et al. 2007), C. thermophilus

(Waingankar et al. 2008) — have been added.

While many phylogenetic studies of entomogenous fungi have been

conducted in recent years, there is little information regarding the phylogeny

A428 ... Nie & al.

and molecular taxonomy of Conidiobolus. The limited SSU analysis of Jensen

et al. (1998) suggested that Conidiobolus may be polyphyletic. Vilela et al.

(2010) were the first to detail the taxonomic and phylogenetic features of three

pathogenic Conidiobolus: C. coronatus (Costantin) A. Batko, C. lamprauges

Drechsler, and C. incongruus Drechsler.

Anhui Province is located at 29°04'-34°06'N 114°09'-119°06’E, eastern

China, and most parts of the province have subtropical vegetation, but some

temperate vegetation occurs in mountains. Recently, several new fungal species

on decaying wood have been reported from the area (Dai 2010; Cui et al. 2011).

In the course of studies on Conidiobolus species from China, one strain with

cushion mycelia was isolated from decaying plant material in Anhui and is

described in this report as a new species, based on both morphological and

molecular data.

Materials & methods

Morphological studies

Plant detritus was sampled on 17 June 2010 near a reservoir in Huoshan, Anhui,

China. The sample was screened for saprotrophic Conidiobolus by canopying moistened

detritus on agar plates following Drechsler (1952) and King (1976a) to exploit the

forcible discharge of Conidiobolus conidia. These isolation plates were incubated at 21°C

and examined daily for one week. Once Conidiobolus cultures were detected on the PDA

canopy, they were transferred to new PDA plates for purification and morphological

study. The measurements of different fungal structures followed King (1976a).

Molecular studies

The Conidiobolus strains used in the molecular study are shown in TABLE 1. Ten

ex-type strains of Conidiobolus spp. were purchased from American Type Culture

Collection (ATCC; Manassas, VA, USA), and the remaining Conidiobolus strains were

obtained from the Research Center for Entomogenous Fungi (RCEF; Anhui Agricultural

University, Hefei, China). Genomic DNA was extracted using the CTAB method (Yi et

al. 2003). The extracted DNA was stored in 50-100 uL of HPLC-H,O at -20°C, and

was diluted 10-fold with HPLC-H,O for use in PCR reactions. Regions of two genes

were amplified by PCR: 1) nuclear ribosomal large subunit (LSU rDNA) by primers

LROR and LRS (Vilgalys & Hester 1990) and 2) elongation factor 1-alpha (EF-1a) by

primers EF983 and EF1aZ-1R (http://www.aftol.org/primers.php). All procedures used

in this study for LSU amplification have been described previously (Liu et al. 2005). The

PCR reaction mixture for amplifying EF-1a contained 200 uM each dNTP, 1x Mg-free

buffer, 2.5 mM MgCl, 0.5 uM each primers, 1 ng/uL genomic DNA, and 0.04 Unit/L

Taq polymerase. The cycle program included initial denaturation at 100°C for 5 min

followed by 95°C for 5 min (during which time Taq polymerase was added to each tube),

34 cycles of 94°C for 1 min, 55°C for 2 min, and 72°C for 2 min, and a final extension

at 72°C for 10 min. The nucleotide sequences of the PCR products were determined on

both strands by using dideoxy-nucleotide chain termination on an ABI 3700 automated

sequencer at Shanghai Genecore Biotechnologies Company. Sequence data of the 19

Conidiobolus sinensis sp. nov. (China) ... 429

TABLE 1. Conidiobolus and Entomophthora cultures and sequences used in

phylogenetic analyses.*

FUNGAL TAXON STRAIN # 28S RDNA EF-1a

C. chlamydosporus Drechsler ATCC12242 (T) JF816212 JF816234

C. denaeosporus Drechsler ATCC12940 (T) JF816215 JF816228

C. firmipilleus Drechsler RCEF4429 JF816222 JF816237

C. gonimodes Drechsler ATCC14445 (T) JF816221 JF816226

C. coronatus RCEF5598 JQO004791 JQ004795

RCEF5599 JQ004792 JQ004796

RCEF5600 JQ004793 JQ004797

RCEF5601 JQ004794 JQ004798

AFTOL-ID137 AY546691 DQ275337

C. heterosporus Drechsler RCEF4430 JF816225 JF816239

C. humicola M.C. Sriniv. & Thirum. ATCC28849 (T) JF816220 JF816231

C. lichenicola ATCC16200 (T) JF816216 JF816232

C. lobatus M.C. Sriniv. & Thirum. ATCC18153 (T) JF816218 JF816233

C. nodosus M.C. Sriniv. & Thirum. ATCC16577 (T) JF816217 JF816235

C. polytocus Drechsler ATCC12244 (T) JF816213 JF816227

C. stromoideus ATCC15430 (T) JF816219 JF816229

C. sinensis RCEF4952 (T) JF816224 JF816238

C. thromboides Drechsler ATCC12587 (T) JF816214 JF816230

RCEF4492 JF816223 JF8 16236

E. muscae (Cohn) Fresen. ARSEF3074 DQ273772 DQ275343

* The fungal taxonomy follows that of King (1976a, b, 1977). ARSEF = ARS Entomopathogenic

Fungus Collection (Ithaca, USA). arcc = American Type Culture Collection (Manassas,

USA). RCEF = Research Center for Entomogenous Fungi (Hefei, China). AFTOL-ID =

Assembling the Fungal Tree of Life Identity. T = ex type.

strains of Conidiobolus have been deposited in the GenBank database under the access

numbers shown in TABLE 1.

Sequences were aligned with Clustal X (Thompson et al. 1997). The combined data

of the two loci, partial 28S rDNA and EF-1a, were analyzed with Maximum Parsimony

(MP) in PAUP* 4.0b10 (Swofford 2003), by using 1000 replicates of heuristic search of

random sequence additions, branch swapping algorithm by tree bisection-reconnection

(TBR) and MULTrees in effect. Gaps were treated as missing data and all characters were

equally weighted. Branch support was estimated by 1000 bootstraps of 10 replicates of

heuristic search with the same options as the parsimony search (Felsenstein 1985). The

alignments were fed to DNAMAN software package (Version 5.2.2, Lynnon Biosoft,

Canada) for calculating genetic similarities.

Results

The combined alignment of partial 28S rDNA and the EF-1a dataset was

1468 bp in length, including 981bp from the LROR/LR5 region of 28S rDNA

and 487 bp from the EF983/EF1aZ-1R region of EF-1la. 108 sites in 28S rDNA

and 24 sites in EF-la with ambiguous alignment were excluded from the

analysis and the final alignment contained 643 parsimony-informative sites.

Maximum parsimony analysis of 20-taxon dataset resulted in a single tree

(TL = 1942, CI = 0.6130, RI = 0.7826, HI = 0.3553) shown in PLaTE 1, and

430 ... Nie & al.

: €. coronatus RCEF5598

€. coronatus RCEF5599

"'F ¢. coronatus RCEFS6O0

C. coronaius AF TOL-137

€. coronaius RCEF5601

€. lichenicola ATCC16200

2 C. gonimodes ATCC 14445

7 C. polytocus ATCC12244

100) C. chlampdosporus ATCC12242

94 a C. fiemipilleus RCEF4429

C. Augnicolus ATCC28849

% C. nodosus ATCC16577

C. sinensis. RCEF4952

100

C. thromboides RCEF4492

€. denaeosporus ATCC12940

8 C. heterosporus. RCEF4430

C. dobaius ATCC18153

Entomophihora muscae ARSEF3074

10 changes

PraTE 1. The single most parsimonious tree (TL = 1942, CI = 0.6130, RI = 0.7826, HI = 0.3553)

showing phylogenetic relationships among species of Conidiobolus inferred from a combined

dataset of partial 28S rDNA and EF-la sequences. Bootstrap values =>50 % are labeled above

relevant branches. Entomophthora muscae served as the outgroup. The bar at the lower left corner

represents 10 changes.

Conidiobolus sinensis sp. nov. (China) ... 431

TABLE 2. Similarities of partial 28S rDNA and EF-1a sequences

from Conidiobolus strains.*

% SIMILARITY

SPECIES— STRAIN sss OO eae.

1 2 3 4 5 6 7 8 9 10 11 12 13

1 C. chlamydosporus 99 85 85 67 67 67 65 85 85 85 85 84

—ATCC12242

Po ecinipilletts 98 8 85 68 68 68 66 85 85 85 85 85

— RCEF4429

3 C. gonimodes

91 91 97 68 68 66 66 9% 9% %% 9% 96

—ATCC14445

SIE CON 90 89 93 68 68 66 66 % 9% 9% £96 9%

—ATCC16200

SG faromboiges Feet G34 Fe 76 99 88 86 67 68 68 68 67

—ATCC12587

ae ds ats 73 73 75 76 99 88 86 67 68 68 68 66

— RCEF4492

Te TMI EUs 7595 75y 77 1 9D 94 65 66 66 65 65

—ATCC15430

Ars de 73 73 74 75 91 91 96 65 65 65 65 65

—RCEF4952

P CacerenHts 92 92 94 95 76 76 77 75 99 99 98 99

—RCEF5598

BOR conan 92 91 94 95 76 76 77 75 99 99 98 99

—RCEF5599

pera 91 91 94 9% 76 76 77 «+75 99 99 99 99

—RCEF5600

inp giherecs 91 91 93 95 76 76 77 75 98 98 99 98

—RCEF5601

13 C. coronatus

—AFTOL-ID137

91 91 94 96 76 76 $77 #7 99 99 100 99

* Data refer to the overall similarities of the partial 288 rDNA (above the diagonal) and

EF-1a sequences (below the diagonal)

the DNA similarities among 13 representing strains are listed in TABLE 2. The

phylogenetic tree shows that the isolate RCEF4952 clustered with the ex-

type strain of Conidiobolus stromoideus with 100% bootstrap support (PLATE

1), but the similarities between C. stromoideus and C. sinensis were only 94%

(28S rDNA) and 96% (EF-1a) (TABLE 2). Higher intraspecific DNA similarities

were measured from partial 28S rDNA and EF-1a sequence. For example, the

ranges of DNA similarities within C. coronatus have been found to be 98-99%

(28S rDNA) and 98-100% (EF-1a), and those within C. thromboides were 99%

for both genes. On the other hand, the Conidiobolus partial 28S rDNA and EF-

la showed high genetic divergence among species. The highest similarity of

partial 28S rDNA (97%) was recorded between C. gonimodes and C. lichenicola,

and the lowest (65%) between C. coronatus and C. stromoideus. The range of

similarities in EF-la among species was 73-96%. Although the similarities

between C. chlamydosporus and C. firmipilleus were 99% (28S rDNA) and 98%

(EF-1la), C. chlamydosporus was placed in synonymy with C. firmipilleus in

King’s classification (King 1977).

432 ... Nie & al.

Taxonomy

Conidiobolus sinensis Y. Nie, X.Y. Liu & B. Huang, sp. nov. PLATES 2-3

MycoBAnk MB563665

Differs from Conidiobolus stromoideus by its much longer conidiophores and rarely

branching mycelia at the colony edge.

Type: China, Anhui Province, Huoshan County, isolated from leaf litter, 17 June 2010,

[Yong Nie] (Holotype, RCEF4952; GenBank JF816224, jF816238).

EryMo_oey: sinensis (Lat.) = China, referring to the geographic origin of the strain.

Colonies grown on PDA for 3 days at 21°C, white, reaching ca 21 mm diameter.

Numerous hyphal knots giving the colony a coarse appearance with aging.

Mycelium colorless, tubular, filamentous, 5-10 um wide, forming hyphal

segments in older regions. Apical cells 80-450 um long, often unbranched

before cell division. Conidiophores colorless, unbranched and producing a

single conidium, arising as upward branches from hyphal knots formed by

irregular mycelium interweaving, 32.5-110 x 10-15 um. Primary conidia

colorless, globose to pyriform 17.5-25 um wide, 22.5-32.5um long including

a basal papilla 7.5-10 um high and 2.5-7.5 um wide. Primary conidia forcibly

discharged, on water agar forming globose secondary conidia resembling the

primary spore, 17.5-22.5 x 15-20 um. Zygospores formed between adjacent

conjugating cells of a hyphal body. Mature zygospores smooth, globose or

subglobose, 25-31 tm in diameter with wall 1-2 um thick.

Discussion

In comparing the morphological characteristics of primary conidiophores

from cushion mycelium with the known Conidiobolus species, C. sinensis

resembles C. lichenicola M.C. Sriniv. & Thirum. and C. stromoideus M.C. Sriniv.

& Thirum. (Srinivasan & Thirumalachar 1962, 1968). Colonies of C. lichenicola

are distinguished by a pale brownish mycelium with sinuous, lobate hyphae.

Conidiobolus stromoideus differs from the new species producing edge mycelia

that are usually branched (rarely branched in C. sinensis) and much shorter

(12-40um) conidiophores (PLATE 3).

The phylogenetic tree places the C. sinensis—C. stromoideus clade distant

from the C. lichenicola clade (PLATE 1), thus reinforcing the morphological

difference between C. sinensis and C. lichenicola. If C. chlamydosporus is accepted

as synonymous with C. firmipilleus in accordance with King (1977), there is a

clear-cut line between intraspecific and interspecific sequence similarity levels:

98-100% within species and 64-97% among species (TABLE 2). Although

C. sinensis groups with C. stromoideus, DNA similarity levels between the two

species fall within the interspecific range (94% (28S rDNA) and 96% (EF-1a)).

Thus, the phylogenetic analysis supports the morphological identification of

C. sinensis as a new species differing from C. stromoideus and C. lichenicola.

Conidiobolus sinensis sp. nov. (China) ... 433

PLATE 2. Conidiobolus sinensis. A. Colony on PDA after 3 days at 21°C. B. Rarely branched mycelia

at the margin of colony. C. Primary conidia. D. Primary conidiophores produced from hyphal

knots. E. Primary conidiophores. FE. Secondary conidia produced singly from the primary conidia.

G. Mature zygospores. Bars: A = 10 mm, B = 100 um, C-H = 20 um.

PiaTE 3. A. Conidiobolus sinensis: rarely branched mycelia at the colony edge. B. C. stromoideus:

moderately branched mycelia at the colony edge. C. C. sinensis: long conidiophores. D. C. stromoideus:

short conidiophores. Bars: A-B = 100 um, C-D = 20 um.

434 ... Nie & al.

Acknowledgments

We are grateful to Dr. Kathryn E. Bushley (Oregon State University, USA) for

improving manuscript writing and Dr. Paul Kirk (CABI UK) and Dr. Yu-Cheng Dai

(Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, China) for

reviewing this manuscript. This project was supported by the National Natural Science

Foundation of China (No. 30770008, No. 31070009, No.31070019) and the Key Science

Research Project of Anhui Province (No. TD200708).

Literature cited

Cui BK, Li HJ, Dai YC. 2011. Wood-rotting fungi in eastern China 6. Two new species of

Antrodia (Basidiomycota) from Yellow Mountain, Anhui Province. Mycotaxon 116: 13-20.

http://dx.doi.org/10.5248/116.13

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 131-343.

http://dx.doi.org/10.1007/s13225-010-0066-9

Drechsler C. 1952. Widespread distribution of Delacroixia coronata and other saprophytic

Entomophthoraceae in plant detritus. Science 115: 575-576.

http://dx.doi.org/10.1126/science.115.2995.575

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution

38: 783-791. http://dx.doi.org/10.2307/2408678

Huang B, Humber RA, Hodge KT. 2007. A new species of Conidiobolus from Great Smoky

Mountains National Park. Mycotaxon 100: 227-233.

Humber RA. 1997. Fungi: identification. In: LA Lacey (ed.). Manual of Techniques in Insect

Pathology. London, Academic Press.

Jensen AB, Gargas A, Eilenberg J, Rosendahl S. 1998. Relationships of the insect-pathogenic

order Entomophthorales (Zygomycota, Fungi) based on phylogenetic analyses of nuclear small

subunit ribosomal DNA sequences (SSU rDNA). Fungal Genetics and Biology 24: 325-334.

http://dx.doi.org/10.1006/fgbi.1998.1063

King DS. 1976a. Systematics of Conidiobolus (Entomophthorales) using numerical taxonomy I.

Biology and cluster analysis. Canadian Journal of Botany 54: 45-65.

http://dx.doi.org/10.1139/ b76-008

King DS. 1976b. Systematics of Conidiobolus (Entomophthorales) using numerical

taxonomy II. Taxonomic considerations. Canadian Journal of Botany 54: 1285-1296.

http://dx.doi.org/10.1139/b76-141

King DS. 1977. Systematics of Conidiobolus (Entomophthorales) using numerical taxonomy

III. Descriptions of recognized species. Canadian Journal of Botany 55: 718-729.

http://dx.doi.org/10.1139/b77-086

Liu M, Rombach MC, Humber RA, Hodge KT. 2005. What's in a name? Aschersonia insperata: a

new pleoanamorphic fungus with characteristics of Aschersonia and Hirsutella. Mycologia. 97:

249-256. http://dx.doi.org/10.3852/mycologia.97.1.246

Srinivasan MC, Thirumalachar MJ. 1962. Studies on species of Conidiobolus from India-II.

Sydowia, Annales Mycologici 16: 60-66.

Srinivasan MC, Thirumalachar MJ. 1968. Studies on species of Conidiobolus from India-V.

Mycopathologica et Mycologia Applicata 36: 341-346. http://dx.doi.org/10.1007/BF02050380.

Swofford DL. 2003. PAUP*. Phylogenetic analysis using parsimony and other methods. Version 4.

Sunderland, Massachusetts: Sinensisuer Associates.

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The Clustal-X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acid Research 63: 215-228. http://dx.doi.org/10.1093/nar/25.24.4876.

Conidiobolus sinensis sp. nov. (China) ... 435

Vilela R, Silva SMS, Correa FR, Dominguez E, Mendoza L. 2010. Morphologic and Phylogenetic

characterization of Conidiobolus lamprauges recovered from infected sheep. Journal of Clinical

Microbiology 48: 427-432. http://dx.doi.org/10.1128/JCM.01589-09

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238-4246.

Waingankar VM, Singh SK, Srinivasan MC. 2008. A new thermophilic species of Conidiobolus

from India. Mycopathologia 165: 173-177. http://dx.doi.org/10.1007/s11046-007-9088-6

Yi RH, Zhu XR, Zhou RX. 2003. Simplified the CTAB DNA extraction from filamentous fungi.

Journal of Zhanjiang Ocean University 23: 72-73.

MY COTAXON

http://dx.doi.org/10.5248/120.437

Volume 120, pp. 437-441 April-June 2012

First record of the sequestrate fungus Neosecotium macrosporum

(Agaricales, Lepiotaceae) from Mexico

Marcos LIZARRAGA', MARTIN ESQUEDA’ ,

MARIO VARGAS-LUNA’ & GABRIEL MORENO?

"Dpto. de Ciencias Quimico-Bioldgicas, Instituto de Ciencias Biomédicas,

Universidad Auténoma de Ciudad Juarez, Anillo Envolvente Pronaf y Estocolmo s/n,

Cd. Juarez, Chihuahua 32300, México

*Centro de Investigacién en Alimentacion y Desarrollo,

A.C. Apartado Postal 1735, Hermosillo, Sonora 83000, México

°Dpto. de Biologia Vegetal, Facultad de Biologia, Universidad de Alcala,

Alcala de Henares, Madrid 28871, Spain

* CORRESPONDENCE TO: esqueda@ciad.mx

ABSTRACT — Neosecotium macrosporum, previously known only from two USA records, is

reported for the first time in Mexico. Observations of macro- and microscopic characters

including for the holotype and the Chihuahua collection are presented. SEM photomicrographs

illustrating spore ornamentation are included.

Key worps — Basidiomycota, taxonomy, hypogeous fungi, secotiaceous fungi

Introduction

Sequestrate fungi are considered a polyphyletic group with no evolutionary

relationships whose morphological similarity comes from the adaptation

to extreme habitats and environmental conditions (Albee-Scott 2007). The

approximately 1200 species are distributed among eleven orders representing

the phyla Zygomycota, Ascomycota, and Basidiomycota. Distinguishing

characteristics are a fleshy, cartilaginous or elastic fruiting body with a persistent

peridium that encloses the spore producing tissue and thus prevents the easy

release of the spores (Kendrick 1992). Most sequestrates are mycorrhizogenic

and serve as a source of food for a number of mammals (Trappe & Claridge

2003).

There are few studies of this group in Mexico. The first sequestrate fungus

recorded for Mexico was collected by Lumholtz (1902) and later designated

as the holotype of Melanogaster umbriniglebus Trappe & Guzman. Trappe &

438 ... Lizarraga & al.

Guzman (1971) described three other new hypogeous species and 13 new

records for Mexico, Cazares et al. (1992) cited 17 new records, and Cazares et

al. (2008) described M. minisporus Cazares et al. as a new species and reported

the first Mexican records of three species of Hysterangium Vittad.

Neosecotium Singer & A.H. Sm. is a genus characterized by a dry firm

whitish stipe-columella that extends below a powdery gleba containing few

fibulae and strongly verrucose spores (Miller & Miller 1988). In comparison,

Secotium Kunze is characterized by the presence of a volva, a less powdery

gleba with abundant fibulae, and smooth ellipsoid large spores. Neosecotium is

represented by two species: N. africanum and N. macrosporum.

Materials & methods

The collections were studied according to the standard mycological techniques

(Cifuentes et al. 1986, Moreno & Manjén 2010). The basidia, spores, peridial hyphae,

gleba, and columella of specimens mounted in Hoyer’s medium were measured using

a Nikon Eclipse E200 contrast phase microscope, with some samples examined using a

Zeiss DSM 950 scanning electron microscope after processing in a Polaron E-2000 for

1 min at 1.2 Kv and 20 mA, in an argon atmosphere to obtain a 500 A thick deposition

of gold. The specimens are deposited in the macromycetes collection of the Centro de

Estudios Superiores de Estado de Sonora (CESUES) and the Universidad Auténoma

de Ciudad Juarez (UACJ). The holotype was examined using the New York Botanical

Garden's Virtual Herbarium (NY). Herbarium acronyms follow Thiers (2011).

Species studied

Neosecotium macrosporum (Lloyd) Singer & A.H. Sm.,

Madrofio 15(5): 154 (1960). Fics 1-6

= Secotium macrosporum Lloyd Mycol. Writ. 1: 139. (1903).

Basip1oMatTa subglobose to pyriform, measuring 11-15 x 7-10 mm. PERIDIUM

thick, 0.5-0.8 mm, membranaceous, yellowish brown when mature. GLEBA

composed of tightly packed lamellae, compact or forming pseudo-chambers,

labyrinthiform in young specimens and powdery in old specimens; yellowish

and dark brown when young and mature respectively. STIPE-COLUMELLA

percurrent, whitish, measuring 7-15 x 3-9 mm. PERIDIUM of septate hyphae,

branched, 2.5-4 um in diam. CoLUMELLA with hyphae similar to the peridium,

septate, branched, 5-7.5 um in diam. Basrp1a clavate, (25—)28-32(-35) x 3-5

uum at the base and 9-12 um in the middle and upper part, yellowish brown in

water; evanescent, leaving amorphous remains when mature. BASIDIOSPORES

globose, subglobose to ovoid, pseudoamyloid, 10-13(-14) x 8-11 um including

ornamentation, which is strongly spiny with flat distal end; episporium 2-3 um

thick, covered by a membrane, which covers the depressions between the spines,

giving a cracked appearance to the wall as observed under the SEM. PEDICEL

straight to slightly curved, 2-3 x 2-4 um, narrowing toward the apex.

Neosecotium macrosporum in Mexico ... 439

Fics 1-6: Neosecotium macrosporum (CESUES 5600).

1. Basidiome. 2. Peridium hyphae (LM). 3. Basidium (LM). 4. Spores (LM). 5-6. Spore (SEM).

SPECIMENS EXAMINED: MEXICO, CurHuanua: Municipality of Juarez, 31°13'53.1"N

106°30'36.8" W, 1120 masl, in sandy soil near Prosopis glandulosa Torr., 24.V1I.2009, M.

Lizarraga, C. Salazar, D. Lopez-Pefia, D. Saenz, A.H. Gutiérrez, E. Navarrete (CESUES

5550); 31°17'54.35"N 106°31'55.45"W, 1303 masl, 17.[X.2010, M. Lizarraga, M. Vargas,

C. Salazar, D. Saenz (UAC] 1560, CESUES 5600); 19.III.2011, M. Vargas, M. Lizarraga

(UAC) 1683); 26.11.2011 (UACJ 1684).

OBSERVATIONS— The Mexican materials were compared with the type and other

authentic collections of Neosecotium macrosporum in the Virtual Herbarium

of the New York Botanical Garden (VHNYBG 2011), which allowed us to

440 ... Lizarraga & al.

confirm this determination. Although the holotype (NY 809166, from Texas)

is in a state of deterioration, there are two collections of authentic material

(NY 809164 and 809165, from North Dakota) identified as N. macrosporum by

S.M. Zeller.

Basidiomes of the type and authentic material, in immature stages with

chambers in an obscure lamellar orientation, wood brown to cinnamon color;

stipe-columella percurrent, whitish, little projection of the stipe (2-3 mm); spores

globose, subglobose to ovate, 13-18 or 14-18 x 12-15 um, pseudoamyloid, with

sterigmal appendage, exosporium smooth becoming cracked into an areolate

pattern and the fissures produce a warty to echinulate effect; basidia clavate to

subelliptic-pedicellate, 25-37 x 14-17 um (Lloyd 1903; Singer & Smith 1960).

The Mexican collections differ in the smaller spore size compared with those

previously reported. This high variability appears to be inherent to the species

and has also been reported for taxa close to N. macrosporum.

According to descriptions by Lloyd (1903) and Singer & Smith (1960), this

species is easily recognized by its stipe-columella, which extends from the

base of the gleba to the apex of the peridium, lamelliform gleba and strongly

verrucose pigmented spores. Although unopened fruiting bodies of Geastrum

spp. could be confused macroscopically with Neosecotium, the Geastrum is

easily distinguished by violet spores, a two-layered peridium, and capillitium.

Based on similar habitat and morphology, Neosecotium might also be confused

with Endoptychum Czern., which can be distinguished by its smooth spores

and somewhat larger fruiting bodies (Arora 1986).

Singer & Smith (1960) regarded Neosecotium as intermediate between the

Agaricaceae and Lycoperdaceae, based on the variation in color and spore

ornamentation in some genera in both families. Castellano et al. (2004)

placed Neosecotium into Lepiotaceae based on recent molecular-supported

classifications.

Neosecotium macrosporum has been recorded from two localities in the

United States of America (Lloyd 1903, Singer & Smith 1960); the present study

represents a new record for Mexico, and the third for the world.

Acknowledgments

The authors are grateful to CONABIO (project GT016) for funding this study. Our

gratitude to Dr. H. Kreisel and Dr. R. Valenzuela for reviewing the manuscript and their

useful comments. MVL thanks CONACYT (Mexico) for the award of a fellowship to

carry out his M.Sc. studies and Aldo Gutierrez (CIAD) for preparing the plates and

formatting the text. Bianca Delfosse translated the text from the original in Spanish.

Literature cited

Albee-Scott S. 2007. Does secotioid inertia drive the evolution of false-truffles? Mycological

Research 111: 1030-1039. http://dx.doi.org/10.1016/j.mycres.2007.08.008

Neosecotium macrosporum in Mexico ... 441

Arora D. 1986. Mushrooms demystified: a comprehensive guide to the fleshy mushrooms. Ten

Speed Press, Berkeley. 959 p.

Castellano MA, Trappe JM, Luoma DL. 2004. Sequestrate fungi. 197-213, in: GM Mueller et al.

(eds). Biodiversity of fungi: inventory and monitoring methods. Elsevier Academic Press, San

Diego.

Cazares E, Garcia J, Castillo J, Trappe JM. 1992. Hypogeous fungi from northern Mexico. Mycologia

84: 341-359. http://dx.doi.org/10.2307/3760186

Cazares E, Guevara G, Garcia J, Trappe JM. 2008. Melanogaster minisporus sp. nov., a new

sequestrate member of the Boletales from Mexico. Revista Mexicana de Micologia 28: 67-69.

Cifuentes J, Villegas M, Pérez-Ramirez L, Sierra S. 1986. Hongos. 55-64, in: A Lot, F Chiang (eds).

Manual de herbario. Consejo Nacional de la Flora de México, A.C., México.

Guevara G, Castellano MA, Garcia J, Cazares E, Trappe JM. 2008. Hysterangium (Hysterangiales,

Hysterangiaceae) from northern Mexico. Revista Mexicana de Micologia 28: 95-100.

Kendrick B. 1992. The fifth kingdom. Focus Publishing, Newburyport. 406 p.

Lumholtz C. 1902. Unknown Mexico. Vol. 1. Charles Scribner’s Sons, New York. 530 p.

Miller OK Jr, Miller HH. 1988. Gasteromycetes. Morphological and developmental features with

keys to the orders, families, and genera. Mad River Press, Eureka. 157 p.

Moreno G, Manjén JL. 2010. Guia de hongos de la Peninsula Ibérica. Ediciones Omega, Madrid.

1417 p.

Singer R, Smith AH. 1960. Studies on secotiaceous fungi VII. Secotium and Neosecotium. Madrofio

15: 152-158.

Thiers B. 2011. (continuously updated). Index Herbariorum. Part I: the herbaria of the world. New

York Botanical Garden's Virtual Herbarium. http://sweetgum.nybg.org/ih/

Trappe JM, Claridge AW. 2003. Australasian sequestrate (truffle-like) fungi. 15. New species from

tree line in the Australian Alps. Australasian Mycologist 22: 27-38.

Trappe JM, Guzman G. 1971. Notes on some hypogeous fungi from Mexico. Mycologia 63:

317-332. http://dx.doi.org/10.2307/3757764

VHNYBG. 2011. The C.V. Starr Virtual Herbarium. New York Botanical Garden, International

Plant Science Center. http://sciweb.nybg.org/science2/vii2.asp [accessed September 2011].

MYCOTAXON

http://dx.doi.org/10.5248/120.443

Volume 120, pp. 443-450 April-June 2012

New combinations in Lactifluus. 3.

L. subgenera Lactifluus and Piperati

A. VERBEKEN’, K. VAN DE PUTTE' & E. DE CRopP’

'Ghent University, Department of Biology, Research Group Mycology,

K.L. Ledeganckstraat 35, B-9000 Gent, Belgium

CORRESPONDENCE TO: mieke. verbeken@ugent.be

ABSTRACT — In this last of a series of three papers, new combinations in the genus Lactifluus

are proposed. This paper treats Lactifluus subg. Lactifluus (an autonymous subgenus) and

Lactifluus subg. Piperati (proposed as a new subgenus). In Lactifluus subg. Lactifluus,

six sections are recognized (five of them as new combinations) and 46 new combinations

are proposed at species level. In Lactifluus subg. Piperati, two sections are recognized (as

new combinations) and nine new species combinations are proposed. In addition, new

combinations are proposed for an unassigned section and its single species as well as for

three unassigned species.

Key worps — milkcaps, nomenclature, Lactarius, Russulaceae

Introduction

This is the third treatment of species formerly treated in Lactarius Pers.

and now classified in Lactifluus (Pers.) Roussel. Lactifluus subgenera Edules,

Lactariopsis, and Russulopsis were discussed in the first article (Verbeken et al.

2012) and Lactifluus subg. Gerardii in the second (Stubbe et al. 2012).

Taxonomy

Lactifluus subg. Lactifluus

This large and diverse subgenus comprises 6 sections. Lactifluus sect.

Rubroviolascentini, with 2 species, is endemic to tropical Africa. Two sections,

L. sect. Polysphaerophori and L. sect. Pseudogymnocarpi (each with 8 species)

are almost completely African, except for one South American species in

L. sect. Polysphaerophori and one Chinese species in L. sect. Pseudogymnocarpi.

L. sect. Phlebonemi (11 species) and L. sect. Tomentosi (9 species) also have

their major distribution in Africa, but contain some species from North and

Central America, Australia, Europe and Asia. Lactifluus sect. Lactifluus with

444 ... Verbeken, Van de Putte & de Crop

Asian, American, and European species is the only section not represented in

tropical Africa.

Five species in Lactifluus subg. Lactifluus have already been recombined:

L. corrugis, L. oedematopus, L. princeps and L. volemus in L. sect. Lactifluus; and

L. hygrophoroides in L. sect. Tomentosi.

Lactifluus (Pers.) Roussel, Fl. Calvados, Ed. 2: 66. 1806, subg. Lactifluus

Type: Agaricus lactifluus L. [= L. volemus (Fr. : Fr.) Kuntze] (see Buyck et al. 2010).

= Lactarius subg. Lactifluus (Burl.) Hesler & A.H. Sm., N. Am. Species Lactarius: 158. 1979.

TYPE: Agaricus volemus Fr. : Fr.

Lactifluus (Pers.) Roussel, Fl. Calvados, Ed. 2: 66. 1806, sect. Lactifluus

Type: Agaricus lactifluus L.

= Lactarius subsect. Lactifluini (Burl.) Singer, Ann. Mycol. 40: 114. 1942.

= Lactarius subsect. Volemi Pacioni & Lalli, Mycotaxon 44: 190. 1992, nom. superfl.

TYPE: Agaricus volemus Fr. : Fr.

Lactifluus acicularis (Van de Putte & Verbeken) Van de Putte, comb. nov.

MycoBank MB 564580

= Lactarius acicularis Van de Putte & Verbeken, Fungal Diversity 45: 108. 2010.

Lactifluus austrovolemus (Hongo) Verbeken, comb. nov.

MycoBaAnk MB 564581

= Lactarius austrovolemus Hongo, Rep. Tottori Mycol. Inst. 10: 362. 1973.

Lactifluus corrugis (Peck) Kuntze, Revis. Gen. Pl. 2: 856. 1891.

= Lactarius corrugis Peck, Annual Rep. New York State Mus. 32: 31. 1880 (“1878”).

Lactifluus crocatus (Van de Putte & Verbeken) Van de Putte, comb. nov.

MycoBank MB 564582

= Lactarius crocatus Van de Putte & Verbeken, Fungal Diversity 45: 112. 2010..

Lactifluus distantifolius (Van de Putte, Stubbe & Verbeken) Van de Putte,

comb. nov.

MycoBank MB 564583

= Lactarius distantifolius Van de Putte, Stubbe & Verbeken,

Fungal Diversity 45: 115. 2010.

Lactifluus lamprocystidiatus (Verbeken & E. Horak) Verbeken, comb. nov.

MyYcoBANK MB 564584

= Lactarius lamprocystidiatus Verbeken & E. Horak, Austr. Syst. Bot. 13: 674. 2000.

Lactifluus longipilus (Van de Putte, H.T. Le & Verbeken) Van de Putte, comb. nov.

MycoBank MB 564585

= Lactarius longipilus Van de Putte, H.T. Le & Verbeken, Fungal Diversity 45: 117. 2010.

Lactifluus oedematopus (Scop.) Kuntze, Revis. Gen. Pl. 2: 857. 1891.

= Agaricus oedematopus Scop., Fl. Carniol., Ed. 2, 2: 453. 1772.

Lactifluus pallidilamellatus (Montoya & Bandala) Van de Putte, comb. nov.

MycoBank MB 564586

= Lactarius pallidilamellatus Montoya & Bandala, Cryptog. Mycol. 25: 16. 2004.

Lactifluus subg. Lactifluus & Piperati ...

Lactifluus pinguis (Van de Putte & Verbeken) Van de Putte, comb. nov.

MycosBank MB 564587

= Lactarius pinguis Van de Putte & Verbeken, Fungal Diversity 45: 119. 2010.

Lactifluus princeps (Berk.) Kuntze, Revis. Gen. Pl. 2: 857. 1891.

= Lactarius princeps Berk., Hooker's J. Bot. Kew Gard. Misc. 4: 135. 1852.

Lactifluus vitellinus (Van de Putte & Verbeken) Van de Putte, comb. nov.

MycoBank MB 564588

= Lactarius vitellinus Van de Putte & Verbeken, Fungal Diversity 45: 121. 2010.

Lactifluus volemus (Fr.: Fr.) Kuntze, Revis. Gen. Pl. 2: 857. 1891.

= Agaricus volemus Fr.: Fr., Syst. Mycol. 1: 69. 1821.

Lactifluus sect. Polysphaerophori (Singer) Verbeken, comb. nov.

MycoBank MB 564589

= Lactarius sect. Polysphaerophori Singer, Beih. Sydowia 7: 106. 1973.

Type: Lactarius veraecrucis Singer.

= Lactarius sect. Gymnocarpi R. Heim ex Verbeken, Mycotaxon 66: 374. 1998.

Type: Lactarius gymnocarpus R. Heim ex Singer

Lactifluus albocinctus (Verbeken) Verbeken, comb. nov.

MycoBank MB 564590

= Lactarius albocinctus Verbeken, Syst. Geogr. Pl. 70: 182. 2000.

Lactifluus brunnescens (Verbeken) Verbeken, comb. nov.

MycoBank MB 564591

= Lactarius brunnescens Verbeken, Bull. Jard. Bot. Belg. 65: 199. 1996.

Lactifluus flammans (Verbeken) Verbeken, comb. nov.

MycoBank MB 564592

= Lactarius flammans Verbeken, Mycotaxon 55: 539. 1995.

Lactifluus foetens (Verbeken & Van Rooij) Verbeken, comb. nov.

MycoBank MB 564593

= Lactarius foetens Verbeken & Van Rooij, Nova Hedwigia 77: 230. 2003.

Lactifluus goossensiae (Beeli) Verbeken, comb. nov.

MycoBank MB 564594

= Lactarius goossensiae Beeli, Bull. Soc. Roy. Bot. Belgique 60: 165. 1928.

Lactifluus gymnocarpus (R. Heim ex Singer) Verbeken, comb. nov.

MycoBank MB 564595

= Lactarius gymnocarpus R. Heim ex Singer, Pap. Michigan Acad. Sci. 32: 107. 1946.

Lactifluus tanzanicus (Karhula & Verbeken) Verbeken, comb. nov.

MycoBank MB 564596

= Lactarius tanzanicus Karhula & Verbeken, Karstenia 38: 50. 1998.

Lactifluus veraecrucis (Singer) Verbeken, comb. nov.

MycosBank MB 564597

= Lactarius veraecrucis Singer, Beih. Sydowia 7: 104. 1973.

445

446 ... Verbeken, Van de Putte & de Crop

Lactifluus sect. Phlebonemi (R. Heim ex Verbeken) Verbeken, comb. nov.

MycoBank MB 564598

= Lactarius sect. Phlebonemi R. Heim ex Verbeken, Mycotaxon 66: 378. 1998.

Type: Lactarius phlebonemus R. Heim & Gooss.-Font.

= Lactarius subsect. Luteoli Pacioni & Lalli, Mycotaxon 44: 190. 1992.

= Lactarius sect. Luteoli (Pacioni & Lalli) Pierotti, Boll. Gruppo Micol. Bres. 48: 54. 2007.

Tye: Lactarius luteolus Peck

Lactifluus angustus (R. Heim & Gooss.-Font.) Verbeken, comb. nov.

MycosBank MB 564599

= Lactarius angustus R. Heim & Gooss.-Font., Bull. Jard. Bot. Etat 25: 67. 1955.

Lactifluus arsenei (R. Heim) Verbeken, comb. nov.

Mycosank MB 564600

= Lactarius arsenei R. Heim, Candollea 7: 380. 1938, as “arsenii”.

Lactifluus brunneoviolascens (Bon) Verbeken, comb. nov.

MycoBank MB 564601

= Lactarius brunneoviolascens Bon, Doc. Mycol. 1 (2): 45. 1971.

Lactifluus caribaeus (Pegler) Verbeken, comb. nov.

MycoBank MB 564602

= Lactarius caribaeus Pegler, Kew Bull. 33: 617. 1979.

Lactifluus longivelutinus (X.H. Wang & Verbeken) X.H. Wang, comb. nov.

MycosBank MB 564603

= Lactarius longivelutinus X.H. Wang & Verbeken, Nova Hedwigia 83 (1-2): 168, 2006.

Lactifluus luteolus (Peck) Verbeken, comb. nov.

MycoBank MB 564604

= Lactarius luteolus Peck, Bull. Torrey Bot. Club 23: 412. 1896.

Lactifluus nonpiscis (Verbeken) Verbeken, comb. nov.

MycoBank MB 564605

= Lactarius nonpiscis Verbeken, Bull. Jard. Bot. Belg. 65: 204. 1996.

Lactifluus phlebonemus (R. Heim & Gooss.-Font.) Verbeken, comb. nov.

MycoBank MB 564606

= Lactarius phlebonemus R. Heim & Gooss.-Font., Bull. Jard. Bot. Etat 25: 38. 1955.

Lactifluus pisciodorus (R. Heim) Verbeken, comb. nov.

MycoBank MB 564607

= Lactarius pisciodorus R. Heim, Candollea 7: 380. 1938.

Lactifluus putidus (Pegler) Verbeken, comb. nov.

MycoBank MB 564608

= Lactarius putidus Pegler, Kew Bull. 33: 620. 1979.

Lactifluus rubrobrunnescens (Verbeken, E. Horak & Desjardin) Verbeken, comb.

nov.

MycoBank MB 564609

= Lactarius rubrobrunnescens Verbeken, E. Horak & Desjardin, Sydowia 53: 274. 2001.

Lactifluus subg. Lactifluus & Piperati ...

Lactifluus sect. Pseudogymnocarpi (Verbeken) Verbeken, comb. nov.

MycoBank MB 564610

= Lactarius sect. Pseudogymnocarpi Verbeken, Mycotaxon 66: 376. 1998.

Type: Lactarius gymnocarpoides Verbeken

Lactifluus carmineus (Verbeken & Walleyn) Verbeken, comb. nov.

MycoBank MB 564611

= Lactarius carmineus Verbeken & Walleyn, Syst. Geogr. Pl. 70: 190. 2000.

Lactifluus gymnocarpoides (Verbeken) Verbeken, comb. nov.

MycoBank MB 564612

= Lactarius gymnocarpoides Verbeken, Mycotaxon 55: 530. 1995.

Lactifluus longisporus (Verbeken) Verbeken, comb. nov.

MycoBank MB 564613

= Lactarius longisporus Verbeken, Mycotaxon 55: 527. 1995.

Lactifluus luteopus (Verbeken) Verbeken, comb. nov.

MycoBank MB 564614

= Lactarius luteopus Verbeken, Mycotaxon 55: 536. 1995.

Lactifluus medusae (Verbeken) Verbeken, comb. nov.

MycoBank MB 564615

= Lactarius medusae Verbeken, Mycotaxon 55: 532. 1995.

Lactifluus pseudogymnocarpus (Verbeken) Verbeken, comb. nov.

MycoBank MB 564616

= Lactarius pseudogymnocarpus Verbeken, Mycotaxon 55: 523. 1995.

Lactifluus pumilus (Verbeken) Verbeken, comb. nov.

MycoBank MB 564617

= Lactarius pumilus Verbeken, Bull. Jard. Bot. Belg. 65: 205. 1996.

Lactifluus tenuicystidiatus (X.H. Wang & Verbeken) X.H. Wang, comb. nov.

MycoBank MB 564618

= Lactarius tenuicystidiatus X.H. Wang & Verbeken, Nova Hedwigia 83: 173, 2006.

Lactifluus sect. Rubroviolascentini (Singer) Verbeken, comb. nov.

MycoBank MB 564619

= Lactarius subsect. Rubroviolascentini Singer, Ann. Mycol. 40: 114. 1942.

= Lactarius sect. Rubroviolascentini (Singer) Verbeken,

Mycotaxon 66: 380. 1998, as “Rubroviolascentes”.

Type: Lactarius rubroviolascens R. Heim

Lactifluus denigricans (Verbeken & Karhula) Verbeken, comb. nov.

MycoBank MB 564620

= Lactarius denigricans Verbeken & Karhula, Persoonia 16: 219. 1996.

Lactifluus rubroviolascens (R. Heim) Verbeken, comb. nov.

MycoBank MB 564621

= Lactarius rubroviolascens R. Heim, Candollea 7: 377. 1938.

447

448 ... Verbeken, Van de Putte & de Crop

Lactifluus sect. Tomentosi (McNabb) Verbeken, comb. nov.

MycoBank MB 564622

= Lactarius sect. Tomentosi McNabb, New Zealand J. Bot. 9: 59. 1971.

= Lactarius subsect. Clarkeina McNabb, New Zealand J. Bot. 9: 59. 1971.

Type: Lactarius clarkeae Cleland

= Lactarius subsect. Rugati Pacioni & Lalli, Mycotaxon 44: 190. 1992, nom. superfl.

= Lactarius sect. Rugati Verbeken, Mycotaxon 66: 372. 998, 1998, nom. superfl.

Tye: Lactarius rugatus Kiihner & Romagn.

Lactifluus clarkeae (Cleland) Verbeken, comb. nov.

MycoBank MB 564623

= Lactarius clarkeae Cleland, Trans. & Proc. Roy. Soc.

S. Australia 51: 302. 1927, as “clarkei”.

Lactifluus hygrophoroides (Berk. & M.A. Curtis) Kuntze, Revis. Gen. Pl. 2: 857.

1891.

= Lactarius hygrophoroides Berk. & M.A. Curtis, Ann. Mag. Nat. Hist., Ser. 3, 4: 293. 1859.

Lactifluus kivuensis (Verbeken) Verbeken, comb. nov.

MycoBank MB 564624

= Lactarius kivuensis Verbeken, Bull. Jard. Bot. Belg. 65: 202. 1996.

Lactifluus pseudoluteopus (X.H. Wang & Verbeken) X.H. Wang, comb. nov.

MycoBaAnk MB 564625

= Lactarius pseudoluteopus X.H. Wang & Verbeken, Nova Hedwigia 83: 171. 2006.

Lactifluus pseudovolemus (R. Heim) Verbeken, comb. nov.

MycoBank MB 564626

= Lactarius pseudovolemus R. Heim, Candollea 7: 378. 1938.

Lactifluus rubiginosus (Verbeken) Verbeken, comb. nov.

MycoBank MB 564627

= Lactarius rubiginosus Verbeken, Bull. Jard. Bot. Belg. 65: 207. 1996.

Lactifluus rugatus (Kihner & Romagn.) Verbeken, comb. nov.

MycosBank MB 564628

= Lactarius rugatus Kihner & Romagn., Bull. Soc. Mycol. France 69: 362. 1954 (“1953”).

Lactifluus xerampelinus (Karhula & Verbeken) Verbeken, comb. nov.

MycoBank MB 564629

= Lactarius xerampelinus Karhula & Verbeken, Karstenia 38: 59. 1998.

Lactifluus volemoides (Karhula) Verbeken, comb. nov.

MycoBank MB 564630

= Lactarius volemoides Karhula, Karstenia 38: 53. 1998.

Lactifluus subg. Piperati

This group consists of two sections, one with 9 species described from

Europe and Asia (but also with records from North America), and the other

with one American species. The combination Lactifluus piperatus has already

been proposed.

Lactifluus subg. Lactifluus & Piperati ... 449

Lactifluus subg. Piperati Verbeken, subg. nov.

MycoBank MB 564631

Pileus pallidus, saepe albus vel albidus, siccus. Pileipellis hyphoepithelium, tenue stratum

hyphis hyalinis super cellullis globosis. Dermatocystidia interdum presentia.

TyPE: Agaricus piperatus L.: Fr.

Lactifluus sect. Piperati (Fr.) Verbeken, comb. nov.

MycoBank MB 564632

= Agaricus sect. Piperati Fr., Syst. Mycol. 1: 73. 1821.

= Lactarius sect. Piperati (Fr.: Fr.) Fr., Epicr. Syst. Mycol.: 338. 1838.

TyPE: Agaricus piperatus L.: Fr.

Lactifluus dwaliensis (K. Das, J.R. Sharma & Verbeken) K. Das, comb. nov.

MycoBank MB 564633

= Lactarius dwaliensis K. Das, J.R. Sharma & Verbeken, Mycotaxon 88: 334. 2003.

Lactifluus glaucescens (Crossl.) Verbeken, comb. nov.

MyYcoBANK MB 564634

= Lactarius glaucescens Crossl., Naturalist, J. Nat. Hist. N. England 1900(516): 5. 1900.

= Lactarius piperatus var. glaucescens (Crossl.) Hesler & A.H.

Sm., N. Amer. Species Lactarius: 186. 1979

Lactifluus leuacophaeus (Verbeken & E. Horak) Verbeken, comb. nov.

MycoBank MB 564635

= Lactarius leucophaeus Verbeken & E. Horak, Austr. Syst. Bot. 12: 768. 1999.

Lactifluus novoguineensis (Henn.) Verbeken, comb. nov.

MycoBank MB 564636

= Lactarius novoguineensis Henn., Bot. Jahrb. Syst. 25: 503. 1898.

Lactifluus olivescens (Verbeken & E. Horak) Verbeken, comb. nov.

MycoBank MB 564637

= Lactarius olivescens Verbeken & E. Horak, Austr. Syst. Bot. 13: 678. 2000.

Lactifluus paleus (Verbeken & E. Horak) Verbeken, comb. nov.

MycoBank MB 564638

= Lactarius paleus Verbeken & E. Horak, Austr. Syst. Bot. 12: 771. 1999.

Lactifluus piperatus (L. : Fr.) Kuntze, Revis. Gen. Pl. 2: 857. 1891.

= Agaricus piperatus L.: Fr., Sp. Pl.: 1173. 1753.

= Lactarius piperatus (L. : Fr.) Pers., Tent. Disp. Meth. Fung.: 64. 1797.

= Galorrheus piperatus (L.: Fr.) Fr., Stirp. Agri Femsion. 3: 57. 1825.

Lactifluus roseophyllus (R. Heim) De Crop, comb. nov.

MycoBank MB 564639

= Lactarius roseophyllus R. Heim, Rev. Mycol. (Paris) 30: 237. 1966 (“1965”).

Lactifluus subpiperatus (Hongo) Verbeken, comb. nov.

MyYcoBANK MB 564647

= Lactarius subpiperatus Hongo, Mem. Fac. Liberal Arts

Shiga Univ., Nat. Sci. 15: 46. 1964.

450 ... Verbeken, Van de Putte & de Crop

Lactifluus sect. Allardii (Hesler & A.H. Sm.) De Crop, comb. nov.

MycosBank MB 564640

= Lactarius sect. Allardii Hesler & A.H. Sm., N. Amer. Species Lactarius: 207. 1979.

Type: Lactarius allardii Coker

Lactifluus allardii (Coker) De Crop, comb. nov.

MycoBank MB 564641

= Lactarius allardii Coker, J. Elisha Mitchell Sci. Soc. 34: 12. 1918.

Unassigned taxa

The following section has not yet been assigned to a subgenus. Only one

species, described from tropical Africa, is known.

Lactifluus sect. Aurantiifolii (Verbeken) Verbeken, comb. nov.

MycoBank MB 564642

= Lactarius sect. Aurantiifolii Verbeken, Mycotaxon 77: 441. 2001.

Tye: Lactarius aurantiifolius Verbeken

Lactifluus aurantiifolius (Verbeken) Verbeken, comb. nov.

MyYcoBANK MB 564643

= Lactarius aurantiifolius Verbeken, Bull. Jard. Bot. Belg. 65: 197. 1996.

The following species have an uncertain systematic position, but morphological

and/or molecular data support their placement in Lactifluus.

Lactifluus caperatus (R. Heim & Gooss.-Font.) Verbeken, comb. nov.

MyYcoBANK MB 564644

= Lactarius caperatus R. Heim & Gooss.-Font., Bull. Jard. Bot. Etat 25: 36. 1955.

Lactifluus cocosmus (Van de Putte & De Kesel) Van de Putte, comb. nov.

MyYcoBANK MB 564645

= Lactarius cocosmus Van de Putte & De Kesel, Cryptog. Mycol. 30: 40. 2009.

Lactifluus subclarkeae (Grgur.) Verbeken, comb. nov.

MycoBank MB 564646

= Lactarius subclarkeae Grgur., Larger Fungi S. Australia: 63, 1997.

Acknowledgments

The authors acknowledge Scott Redhead and Shaun Pennycook for valuable

comments and for reviewing the manuscript.

Literature cited

Buyck B, Hofstetter V, Verbeken A, Walleyn R. 2010. Proposal to conserve Lactarius nom. cons.

(Basidiomycota) with a conserved type. Taxon 59: 295-296.

Stubbe D, Wang X-H, Verbeken A. 2012. New combinations in Lactifluus. 2. L. subg. Gerardii.

Mycotaxon 119: 483-485. http://dx.doi.org/10.5248/119.483

Verbeken A, Nuytinck J, Buyck B. 2012. New combinations in Lactifluus. 1. L. subgenera Edules,

Lactariopsis, and Russulopsis. Mycotaxon 118: 455-458. http://dx.doi.org/10.5248/118.447

MY COTAXON

http://dx.doi.org/10.5248/120.451

Volume 120, pp. 451-460 April-June 2012

Exobasidium ferrugineae sp. nov., associated with hypertrophied

flowers of Lyonia ferruginea in the southeastern USA

AARON H. KENNEDY’, NISSE A. GOLDBERG? & ANDREW M. MINNIS?

‘National Identification Services, USDA-APHIS-PPQ-PHP,

10300 Baltimore Ave., B 580, Beltsville, MD, 20705, USA

Jacksonville University, Dept. of Biology and Marine Science,

2800 University Blvd. North, Jacksonville, FL 32211, USA

°Center for Forest Mycology Research, Northern Research Station,

USDA-Forest Service, One Gifford Pinochet Drive, Madison, WI 53726, USA

* CORRESPONDENCE TO: amminnis@fs.fed.us

Apstract — Exobasidium ferrugineae, associated with hypertrophied flowers and less

commonly leaves of Lyonia ferruginea (rusty staggerbush), is formally described here as a

new species. Morphological and DNA sequence (ITS, nLSU) data are provided. Phylogenetic

analyses confirm that it is not conspecific with any species of Exobasidium represented by

existing DNA sequence data. A key to North American species of Exobasidium on Lyonia is

presented.

Key worps — Basidiomycota, Ericaceae, Exobasidiales, Exobasidiomycetes, plant pathogen

Introduction

Exobasidium Woronin (Exobasidiales, Exobasidiomycetes) is a basidio-

mycetous genus associated with diseases of ericaceous plants commonly

characterized by formation of galls on leaves, shoots, and flowers (Burt 1915,

Savile 1959, Nannfeldt 1981). Early authors named species on the basis of

symptomatology and host association, whereas monographers, including Burt

(1915) and Savile (1959), advocated broader taxonomic concepts. These authors

suggested that symptoms were variable, overlapping, and dependent on time and

environmental conditions. Furthermore, fungal morphology was not definitive

for species recognition and usually poorly known, and host associations are

not supported by inoculation and cross-inoculation experiments. According to

these authorities, only a small number of species were accepted in Exobasidium

and the citation of numerous synonyms was deemed unreliable (Burt 1915,

Savile 1959). Nannfeldt (1981) reviewed a large amount of data and literature

452 ... Kennedy, Goldberg & Minnis

with special emphasis on life cycles and symptoms. He concluded that species

of Exobasidium, though often quite similar morphologically, are rigidly host-

limited taxa. Recent molecular studies including those by Begerow et al. (2002)

and Piatek et al. (2012) support Nannfeldt’s hypotheses on the importance of

host association and host specificity.

In April 2011, an unusual flower of Lyonia ferruginea (Walter) Nutt. (rusty

staggerbush; Ericaceae) that was infected and highly transformed by a fungus

was found in Florida. Microscopic examination confirmed that a species

of Exobasidium was growing on the hypertrophied flower. This species was

subsequently characterized with morphological and DNA sequence (ITS,

nLSU) data. Careful comparison with known species using historical literature,

herbarium specimens, and phylogenetic analyses revealed that it represents an

undescribed taxon that is described herein. A key to North American species

of Exobasidium on Lyonia is presented in order to facilitate identification and

promote further study of Exobasidium from North America.

Materials & methods

Morphology

Observations and measurements of microscopic characters were made from crush

mounts of material scraped from the hymenial surface that covered the flower. Material

was rehydrated and viewed in 3% KOH, and occasionally phloxine solution was used

to improve the visibility of fungal structures (Largent et al. 1977). Sterigmata are not

included in length measurements of basidia. Voucher specimens were deposited into the

US. National Fungus Collections (BPI).

Molecular data collection & phylogenetic analysis

DNA was extracted from scraped hymenial surface material from the infected flower

of L. ferruginea (N.A. Goldberg, s.n.; BPI 882571) using Qiagen’s DNeasy Plant Mini Kit

(Germantown, MD). Ribosomal DNA sequence data were generated from the internal

transcribed spacer region (ITS; ITS1, 5.88, ITS2) and the nuclear encoded large subunit

(nLSU) using the primer pairs ITS5/ITS4 (White et al. 1990) and NL1/NL4 (O’Donnell

1992), respectively. GoTaq (Promega, Madison, WI) and associated standard reagents

were used for PCR following the manufacturer recommendations including 2.0 mM

MgCl. Standard thermal cycling was used including an annealing temperature of 55°C

for the ITS and 50°C for the nLSU. PCR products were fluorescently labeled using

respective forward and reverse PCR primers and the BigDye v3.1 dye terminator kit

(Applied Biosystems; Foster City, CA). The purified products were then sequenced on

an ABI 3730 automated DNA sequencer. Electropherograms were edited in Geneious

Pro v.5 (Drummond et al. 2010). Resulting consensus sequences were submitted to

GenBank (http://www.ncbi.nlm.nih.gov).

BLAST searches were conducted independently with the ITS and nLSU sequences

against GenBank using the megablast algorithm. Taxon sampling for phylogenetic

analyses was primarily determined by selecting the top hits from each search, sorted

by Max Score; the phylogenetic results of Pigtek et al. (2012); and morphological and

Exobasidium ferrugineae sp. nov. (USA) ... 453

host association similarity. The ITS and nLSU sequences for each taxonomic unit were

derived from the same isolate.

Multiple sequence alignments of the ITS and nLSU were conducted using

MUSCLE v3.6 (Edgar 2004) within Geneious Pro v5 (Drummond et al. 2010) and

then concatenated. Although nLSU data were available for all taxa included in

the concatenated alignment, ITS data were unavailable for Exobasidium karstenii,

E. oxycocci, E. pieridis, and E. sundstroemii and therefore treated as missing data. The

best-fit model of DNA sequence evolution was independently determined for ITS

and nLSU in MrModeltest v2.2 (Nylander 2004) according to the Akaike Information

Criterion (AIC; Posada & Buckley 2004). The GTR+I+G model was selected for both

data sets and implemented in the following analysis. The concatenated alignment was

analyzed phylogenetically with Bayesian Inference (BI) in MrBayes v3.1.2 (Huelsenbeck

& Ronquist 2001; Ronquist & Huelsenbeck 2003) leaving all other parameters as default.

The posterior probability (pp) distribution of trees was estimated from two independent

Markov Chain Monte Carlo (MCMC) simulations of 3 million generations, sampling

trees every 100 generations until the standard deviation of split frequencies reached 0.01.

The burn-in was determined using the program Tracer v1.5 (Rambaut & Drummond

2007). A 50% majority rule consensus tree was constructed from the remaining trees in

FigTree v1.3.1 (Rambaut 2009).

The concatenated alignment was also analyzed using Maximum Likelihood (ML).

This analysis was conducted using RAxML (Stamatakis 2006) via the on-line CIPRES

(Cyberinfrastructure for Phylogenetic Research) Science Gateway V. 3.1 (http://www.

phylo.org/sub_sections/portal/; Miller et al. 2010) with the RAxML-HPC BlackBox

(7.2.8) tool. One thousand bootstrap (bs) replicates were run and all other parameters

were left as default.

Results

Alignment of the ITS resulted in a matrix of 595 nucleotide positions, 296

of which were variable (49.7%) and 156 of which were parsimony informative

(26.2%). Alignment of the nLSU resulted in a matrix of 555 nucleotide

positions, 146 of which were variable (26.3%) and 71 of which were parsimony

informative (12.8%).

The phylogenies resulting from the BI and ML analyses were entirely

congruent with the exception of the E. pieridis/E. ferrugineae clade, which

occupied a different position in the most optimal ML tree vs. the BI consensus

tree (Fic. 1). This is an insignificant observation considering the position of

this clade is entirely unsupported in the ML bootstrap tree and received very

low posterior probability in the BI tree. Each analysis revealed high support

(pp = 0.99, bs = 99%) for the E. pieridis/E. ferrugineae clade, revealing a close

relationship between these species and demonstrating that E. ferrugineae is a

species distinct from E. vaccinii (Fic. 1).

The relationship between Exobasidium ferrugineae and E. karstenii was

of particular interest because E. karstenii was among the top hits for nLSU.

454 ... Kennedy, Goldberg & Minnis

Tilletiopsis pallescens AY259059/AY 272004

Laurobasidium hachijoense AB180359/AB 177562

E. sundstroemii N.A./AF487396

E. pieridis-ovalifoliae AB180367/AB 1177601

0.67/* ¥ p= E. bisporum AB180364/AB177598

i a AB180368/AB177596

E. darwinii FJ896133/FJ896134

eR . pachysporum AB180352/AB177573

c E. arescens FJ896135*/FJ896136

E. rostrupii FJ896132*/FJ896137

E. inconspicuum AB180347/AB177556

0.96/97 E. cylindrosporum AB245089/AB 176713

0.62/*}* E. pulchrum EU692776*/EU692795

0.67/"t_— E, pentasporium AB180334/AB177581

0.75/L E. nobeyamense AB180331/AB177582

O.BRIe E. otanianum AB180345/AB177593

E. miyabei AB180330/AB177579

E. japonicum AB180315/AB177548*

0 Yoyo E. shiraianum AB180336/AB177549

E. rhododendri DQ667153/DQ667151*

0.9/72L_ E. woronichinii AB180348/AB177557

1/100 E. camelliae AB176712/AB180317

1/100 E. gracile DQ663700/DQ663699

0.99/82 E. reticulatum AB180377/AB 180381

0.56/** E. symploci-japonicae AB180678/AB 178255

OY p> E. ferrugineae JQ611710/IQ611711_ <<——

E. pieridis N.A./AB177575

OVE. kishianum AB180353/AB177577

, E. kishianum AB180354/AB177555

099° L 1098/68. Fe vaccinii AB180362/AB177560

0.63/* E. karstenii N.A./AF487389*

E. oxycocci N.A./AF487391

0.9

0.94/84

0.69/*

0.07

Fic. 1. A phylogenetic summary of relationships among Exobasidium species resulting from

Bayesian Inference (BI) and Maximum Likelihood (ML) analyses of a combined ITS/nLSU data

matrix. This phylogram is the resulting 50% majority rule consensus tree from the BI analysis

with corresponding posterior probabilities (pp) and appended bootstrap (bs) values from the

ML analysis on each branch (pp/bs). One asterisk indicates a bs value < 50%, while two asterisks

indicates a clade that was not recovered in the most optimal ML tree. The hatch marks on the

“Laurobasidium hachijoense” nom. prov. in GenBank [= E. hachijoense] branch indicate that it was

shortened for presentation purposes from 8.5 times the scale bar. GenBank accession numbers

are provided after each taxon name (ITS/nLSU). Asterisks here indicate top BLAST hits for the

associated marker. The arrow indicates the position of E. ferrugineae (holotype, BPI 882571).

However, our phylogenetic results suggested, albeit with very weak support

(pp=0.53, bs = 68), that E. karstenii is most closely related to E. vaccinii. Again,

the relationship of this clade to others was unsupported.

Exobasidium ferrugineae sp. nov. (USA) ... 455

Taxonomy

Exobasidium ferrugineae Minnis, A.H. Kenn. & N.A. Goldberg, sp. nov. Fics. 2-4

MycoBANnk 564773

Characterized by its association with hypertrophied flowers of Lyonia ferruginea.

TYPE: USA. Florida: Jacksonville, Jacksonville Arboretum, Rosemary Ridge Trail, on

flower of Lyonia ferruginea, 18.IV.2011, leg. N.A. Goldberg, s.n., (Holotype BPI 882571;

GenBank accession nos., ITS JQ611710, nLSU JQ611711).

Erymo oey: The species epithet is derived from the host, Lyonia ferruginea.

HYMENIUM consisting of a thin white layer on hypertrophied flowers having

all parts proportionally enlarged, infected flowers up to approx. 6 cm long, or

less commonly on leaves that may also be hypertrophied. Basip1a 37-54.5 x

6.5-8.5 um, cylindrical to cylindrical-clavate, 3-5 sterigmate, thin walled,

hyaline, often with granular contents. STERIGMATA approx. 4.5-6.5 x 1-2

um, typically slightly wider towards base. BAsipriospoREs 13.5-19 x 4-6.5

um, ellipsoid to musiform, with a prominent apiculus, walls thin and smooth,

initially with one more or less median septum at maturity, later with up to six

septa, hyaline. GERMINATION via germ tubes at both ends of basidiospores.

ConipiA 4-9.5 x 1-2.5 um, bacilliform, ellipsoid or slightly clavate, walls thin

and smooth, aseptate, hyaline, contents often with a few scattered guttules.

HABITAT & DISTRIBUTION— ‘This species is known from hypertrophied

flowers and less commonly hypertrophied leaves of Lyonia ferruginea

(Ericaceae). It is known from Florida and Georgia, and is predicted to occur

in South Carolina, and throughout the narrow, native range of its host (USDA

2011).

ADDITIONAL SPECIMENS EXAMINED— USA. FLoripa: on flower of Lyonia ferruginea,

leg. Chapman, s.n., Mo. Bot. Gard. Herb. 44409 (BPI 292003 as Exobasidium vaccinii);

Dunedin, on flower of Lyonia ferruginea, 16.1V.1900, leg. S.M. Tracy, SMT 7159, Mo.

Bot. Gard. Herb. 4962 (BPI 292004 as E. vaccinii); Jacksonville, Jacksonville Arboretum,

Rosemary Ridge Trail, on leaves of Lyonia ferruginea, 06.V.2011, leg. N.A. Goldberg, s.n.

(BPI 882572); additional collections on Lyonia ferruginea made by N.A. Goldberg from

the area of the type locality, Rosemary Ridge Trail: 18.IV.2011 (BPI 882566, BPI 882567,

BPI 882568, BPI 882569, BPI 882570, BPI 882580, BPI 882582, BPI 882583), 06.V.2011

(BPI 882573, BPI 882574, BPI 882575, BPI 882576, BPI 882577, BPI 882578, BPI 882579,

BPI 882581). Georetra: Brunswick, on flower of Lyonia ferruginea, 01.V1.1901, leg.

W. Trelease, s.n., Mo. Bot. Gard. Herb. 4955 (BPI 292005 as E. vaccinii).

CoMMENTS— Burt (1915) studied collections of hypertrophied flowers of Lyonia

ferruginea caused by Exobasidium ferrugineae and concluded that the species

was conspecific with E. vaccinii. Burt (1915) also commented on the fact that

the hypertrophied flowers were much larger than those of L. mariana found

in association with E. peckii Halst., which he also concluded was a synonym

of E. vaccinii. However, the results presented here show that E. ferrugineae is

distinct from E. vaccinii. Exobasidium vaccinii has been shown to be restricted

456 ... Kennedy, Goldberg & Minnis

Fr, - i

\\ ry at ry WWW

ae Hl

my ‘aig ape

Fic. 2. Exobasidium ferrugineae associated with hypertrophied flowers of Lyonia ferruginea.

Note uninfected flowers in upper image for comparison. All flower parts are proportionally

enlarged.

to Vaccinium, likely just Vaccinium vitis-idaea (Nannfeldt 1981, Begerow et al.

2002, Piatek et al. 2012).

Exobasidium ferrugineae sp. nov. (USA) ... 457

Fic. 3. Exobasidium ferrugineae associated with hypertrophied leaves of Lyonia ferruginea.

Several species of Exobasidium are known from Lyonia (Farr & Rossman

2011) and eight of these were described originally from this host genus.

Five Asian species, E. kunmingense Zhen Ying Li & L. Guo (Li & Guo 2009),

E. lyoniae Zhen Ying Li & L. Guo (Li & Guo 2006), E. ovalifoliae Zhen Ying Li

& L. Guo (Li & Guo 2008), E. pieridis Henn. (Hennings 1903), and E. pieridis-

ovalifoliae Sawada (Sawada 1931), are distinguished from E. ferrugineae by

their occurrence on L. ovalifolia and their association that is confined to leaf

diseases. There are three previously described species on Lyonia from North

America. Exobasidium andromedae Peck (Peck 1874) and E. fawcettii Massee

(Massee 1908) occur on different hosts than E. ferrugineae and are associated

with leaf diseases. At times, E. andromedae may produce unique bag galls where

developing leaves are transformed into hollow bags (Peck 1874, Burt 1915).

Exobasidium peckii is similar to E. ferrugineae as it infects both flowers and

leaves (Halstead 1893, Stewart 1896), but it is distinguished by its occurrence

on Lyonia mariana and a lesser amount of hypertrophy on flowers. Based

on our morphological examination of a collection of E. peckii (USA: New

York, Hicksville, Long Island, on flower of Lyonia mariana, 25.V1.1919, leg.

G.P. Clinton, s.n. (BPI 292023 as E. vaccinii)), this species is nearly

indistinguishable morphologically from E. ferrugineae. However, E. peckii

differs in the basidiospores with none having been observed with more than 5

septa along with typically fewer septa per basidiospore and the conidia having

a tendency towards an allantoid shape.

Our phylogenetic analyses indicate that E. ferrugineae is distinct from all

species with existing ITS and nLSU sequence data in GenBank (data were

458 ... Kennedy, Goldberg & Minnis

Fic. 4. Exobasidium ferrugineae (holotype, BPI 882571).

A-B. Basidia. C. Basidiospore. D. Conidia. Scale bars are 20 um for A-B, 10 um for C-D.

unavailable for species on Lyonia including E. andromedae, E. fawcettii, E.

kunmingense, E. lyoniae, E. ovalifoliae, E. peckii) including the isolates that

are labeled E. pieridis and E. pieridis-ovalifoliae from Lyonia neziki. ‘The close

relationship between E. ferrugineae and E. pieridis is noteworthy because of

their highly discontinuous geographic ranges. We suggest that this geographic

isolation has led to reproductive isolation and that the relationship between

E. ferrugineae and E. pieridis is a product of the tendency for host specificity

in Exobasidium, in this case with Lyonia, and divergence in host range

relative to their recent common ancestor. Therefore, their well-supported

sister-relationship found here suggests recent common ancestry, but not

conspecificity. Unfortunately, ITS data were unavailable for E. pieridis,

resulting in missing data for combined analysis. This is significant because the

ITS region is more informative than nLSU primarily because it more rapidly

accumulates substitutions among species. This is evidenced by the fact that the

Exobasidium ferrugineae sp. nov. (USA) ... 459

ITS region contains twice the number of variable and informative characters

among Exobasidium species. Therefore, support for this relationship may be

artificially high in this combined analysis due to missing ITS data for E. pieridis.

Further sampling will be needed to address the complex patterns of speciation

including potential co-speciation in Exobasidium as discussed by Begerow et al.

(2002) and Piatek et al. (2012).

Key to Exobasidium on Lyonia of North America

1. Onhligustring, with bag Sallss ea ghee gba 2g heer Ba bee Babar 3 ghee E. andromedae

}- On. other species of: Lyvonta, bac.ealls absent oats gta! edna ads a ddanaie « enwictas. date 2

2. On L. jamaicensis, with hypertrophied leaves ...................0000- E. fawcettii

2. On other species of Lyonia, associated with flowers & occasionally leaves ........ 3

3. On L. mariana, with hypertrophied flowers relatively small (up to approx.

2-<longet);some- conidia allantoid tos .% 06 wea a ole bia ew eget w a E. peckii

3. On L. ferruginea, with hypertrophied flowers relatively large (up to approx.

5. xlonger);conidia not, allantoid. we. 62 west wate s wees 2 wanes 2 E. ferrugineae

Acknowledgments

The authors thank Dr. Lori M. Carris of Washington State University and Dr. Alistair

R. McTaggart of Louisiana State University for their expertise and time in reviewing

this work. Dr. Amy Y. Rossman of Systematic Mycology and Microbiology Laboratory

kindly provided additional helpful suggestions.

Literature cited

Begerow D, Bauer R, Oberwinkler F. 2002. The Exobasidiales: an evolutionary hypothesis. Mycol.

Progr. 1: 187-199. http://dx.doi.org/10.1007/s11557-006-0018-7

Burt EA. 1915. The Thelephoraceae of North America. IV. Exobasidium. Ann. Missouri Bot. Gard.

2: 627-658. http://dx.doi.org/10.2307/2990117

Drummond AJ, Ashton B, Buxton $, Cheung M, Cooper A, Heled J, Kearse M, Moir R, Stones-

Havas S, Sturrock S, Thierer T, Wilson A. 2010. Geneious v5.1, from http://www.geneious.com.

Edgar RC. 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput.

Nucl. Acids Res. 32: 1792-1797. http://dx.doi.org/10.1093/nar/gkh340

Farr DF, Rossman AY. 2011. Fungal Databases, Systematic Mycology and Microbiology Laboratory,

ARS, USDA. Retrieved November 26, 2011,

from http://nt.ars-grin.gov/fungaldatabases/fungushost/fungushost.cfm

Halsted BD. 1893. Notes upon a new Exobasidium. Bull. Torrey Bot. Club 20: 437-440.

http://dx.doi.org/10.2307/2996716

Hennings P. 1903. Fungi japonica. III. Bot. Jahrb. Syst. 32: 34-46.

Huelsenbeck JP, Ronquist FE. 2001. MrBayes: Bayesian inference of phylogeny. Bioinformatics 17:

754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

Largent D, Johnson D, Watling R. 1977. How to identify mushrooms to genus III: Microscopic

features. Mad River Press, Eureka, CA.

Li Z, Guo L. 2006. A new species and a new Chinese record of Exobasidium (Exobasidiales) from

China. Mycotaxon 97: 379-384.

460 ... Kennedy, Goldberg & Minnis

Li Z, Guo L. 2008. Two new species of Exobasidium (Exobasidiales) from China. Mycotaxon 104:

331-336.

Li Z, Guo L. 2009. Three new species of Exobasidium (Exobasidiales) from China. Mycotaxon 107:

215-220. http://dx.doi.org/10.5248/107.215

Massee GE. 1908. Fungi exotici: VII. Bull. Misc. Inform. Kew 1908: 216-219.

Miller MA, Pfeiffer W, Schwartz T. 2010. Creating the CIPRES Science Gateway for inference

of large phylogenetic trees. 1-8, in Proceedings of the Gateway Computing Environments

Workshop (GCE), 14 Nov., New Orleans, LA.

Nannfeldt JA. 1981. Exobasidium, a taxonomic reassessment applied to the European species.

Symb. Bot. Upsal. 23 (2): 1-72.

Nylander JAA. 2004. MrModeltest, v. 2. Program distributed by the author. Evolutionary Biology

Centre, Uppsala University.

O'Donnell KL. 1992. Ribosomal DNA internal transcribed spacers are highly divergent in the

phytopathogenic ascomycete Fusarium sambucinum (Gibberella pulicaris). Curr. Genet. 22:

213-220. http://dx.doi.org/10.1007/BF00351728

Peck CH. 1874 (1872). Report of the Botanist (1872). Rep. (Annual) New York State Mus. Nat.

Hist. 26: 35-91.

Piatek M, Lutz M, Welton P. 2012. Exobasidium darwinii, a new Hawaiian species infecting

endemic Vaccinium reticulatum in Haleakala National Park. Mycol. Progr. 11: 361-371.

http://dx.doi.org/10.1007/s11557-011-0751-4

Posada D, Buckley TR. 2004. Model selection and model averaging in phylogenetics: advantages of

akaike information criterion and Bayesian approaches over likelihood ratio tests. Syst. Biol. 53:

793-808. http://dx.doi.org/10.1080/10635150490522304

Rambaut A, Drummond AJ. 2007. Tracer v1.5. Computer program and documentation distributed

by the authors at http://beast.bio.ed.ac.uk/Tracer.

Rambaut A. 2009. FigTree v1.3.1. Computer program and documentation distributed by the author

at http://tree.bio.ed.ac.uk/ software/.

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. http://dx.doi.org/10.1093/bioinformatics/btg180

Savile DBO. 1959. Notes on Exobasidium. Canad. J. Bot 37: 641-656.

http://dx.doi.org/10.1139/b59-052

Sawada K. 1931. Descriptive catalogue of the Formosan fungi V. Rep. Dept. Agric, Gov. Res. Inst.

Formosa 51: 1-131.

Stamatakis A. 2006. RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with

thousands of taxa and mixed models. Bioinformatics 22: 2688-2690.

Stewart FC. 1896. Report of the mycologist. New York Agric. Exp. Sta. Annual Rep. 14: 519-546.

USDA, ARS, National Genetic Resources Program. 2011. Germplasm Resources Information

Network - (GRIN) [Online Database]. National Germplasm Resources Laboratory, Beltsville,

Maryland. URL: http://www.ars-grin.gov/cgi-bin/npgs/html/taxon.pl?404852 (26 November

2011).

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds.), PCR Protocols: a guide to

methods and applications. Academic Press, New York, NY.

MY COTAXON

http://dx.doi.org/10.5248/120.461

Volume 120, pp. 461-463 April-June 2012

Passalora aseptata, a new cercosporoid fungus

from northeastern Uttar Pradesh, India

RAGHVENDRA SINGH , BALMUKUND CHAURASIA,

KALAWATI SHUKLA & PARMATMA PRASAD UPADHYAYA

Department of Botany, D.D.U. Gorakhpur University, Gorakhpur, U.P. - 273 009, India

*CORRESPONDENCE TO: drsinghtaxon@gmail.com

AxsstRact — The anamorphic fungus Passalora aseptata sp. nov., found on Terminalia

bellirica (Combretaceae), in the Chowk Forest, Mahrajganj, U.P., India, is described and

illustrated.

Key worps — biodiversity, foliar diseases, phytopathogenic fungi, taxonomy

Introduction

During an investigation of plant diseases, fungi were collected on living

leaves from forests in northeastern Uttar Pradesh during 2008. Among the

collections an undescribed cercosporoid species was found. Based on pigmented

conidia and conidiophores as well as conidiogenous cells with thickened scars,

this fungus can be assigned to Passalora as circumscribed by Crous & Braun

(2003).

Materials & methods

Specimens with disease symptoms of cercosporoid fungi on living leaves were

collected during the course of field trips. Detailed observations of morphological

characters were carried out by means of an Olympus CX31 light microscope using oil

immersion (1000x). Specimens for microscopic observation were prepared by hand

sectioning. Water and lactophenol were used as mounting media. Measurements were

made of 30 conidia, hila, and conidiophores and of 15 stromata. Line drawings were

prepared at a magnification of 1000x. Morphotaxonomic determinations were made

with the help of current literature and available resident expertise. The holotype has been

deposited in Herbarium Cryptogamiae Indiae Orientalis (HCIO), Indian Agricultural

Research Institute, New Delhi, India; and an isotype was retained in the herbarium of

Department of Botany, D.D.U. Gorakhpur University (GPU) for further reference.

462 ... Singh & al.

Taxonomy

Passalora aseptata R. Singh, Chaurasia, K. Shukla & Upadhyaya, sp. nov.

MycoBank MB 519057 FIG. 1

Differs from all other Passalora species in aseptate conidia and conidiophores.

Type: India, Uttar Pradesh, Mahrajganj, Chowk Forest, on living leaves of Terminalia

bellirica (Gaertn.) Roxb. (Combretaceae), Jan. 2008, coll. Raghvendra Singh, (holotype,

HCIO 48785; isotype, GPU KSR-395).

ErymMo_oey: Latin, aseptata refers to aseptate conidia and conidiophores.

Infection spots hypogenous, discrete and initially vein-limited but later

coalescing to become irregular and more or less necrotic, light brown,

spreading over entire lower leaf surface. Colonies hypophyllous, effuse,

velvety, light brown. Mycelium internal. Stromata well developed, erumpent,

pseudoparenchymatous, substomatal, 26-43 x 22-31 um. Conidiophores

arising from stromata, in fascicles of 10-12, macronematous, simple, erect to

procumbent, straight to flexuous, cylindrical, unbranched, smooth, thin-walled,

Fic. 1. Passalora aseptata (holotype).

a: symptoms; b: stroma, conidia, and conidiophores.

(Scale bars: a = 20 mm, b = 20 um).

Passalora aseptata sp. nov. (India) ... 463

light brown to olivaceous-brown, aseptate, 29-48 x 4-7 um. Conidiogenous

cells integrated, terminal, monoblastic, scars conspicuously thickened,

darkened, 0.7-1.5 um wide. Conidia solitary, dry, acropleurogenous, simple,

obclavate-cylindrical, erect, straight to curved, sometimes rostrate, aseptate,

thin-walled, smooth, light brown to olivaceous-brown, 40-70 x 4-9 um, hila

slightly thickened, 0.7-1.5 um wide.

REMARKS — No species of Passalora Fr. has previously been described with

aseptate conidia and conidiophores. A survey of literature also indicates that no

species of Passalora has previously been described on a host of genus Terminalia

or family Combretaceae.

Prathigada terminaliae (Syd.) B. Sutton (Sutton 1994), Pseudocercospora

brevis B. Sutton (Sutton 1994), P combretacearum var. minima B. Sutton

(Sutton 1994), P combreti A.K. Singh & Kamal (Singh & Kamal 1987), and

Scolecostigmina combreti (J. Kranz) U. Braun (Braun 1999) have been reported

on the same host species. Prathigada terminaliae is different due to the presence

of polyblastic, cicatrized, conidiogenous cells with thin, flat conidiogenous loci

and rostrate conidia having smooth or rugulose, multiseptate (septa often rather

thick and dark) and unequally pigmented cells. All Pseudocercospora species

differ from Passalora aseptata in having euseptate conidia and conidiogenous

cells without thickened scars. Scolecostigmina combreti is readily separated by

its formation of sporodochia, annellations on conidiophores and truncate base

of conidia.

Passalora bacilligera (Mont. & Fr.) Mont. & Fr., type species of Passalora, and

all other species of this genus are easily distinguished from the novel species in

having septate conidia.

Acknowledgments

The authors are grateful to Prof. Dr. Uwe Braun and Dr. Eric H.C. McKenzie for

reviewing the manuscript. We also express our deep thanks to Dr. Shaun Pennycook

for nomenclatural review. We are much obliged to Prof. Dr. Kamal for the identification

of host plant and editing the manuscript. Authors’ thanks are also due to the Curator,

HCIO, New Delhi, for accepting the holotype specimens and providing an accession

number thereof.

Literature cited

Braun U. 1999. Some additional species of Scolecostigmina. Schlechtendalia 3: 33-42.

Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs: 1. Names published in Cercospora

and Passalora. CBS, Utrecht, The Netherlands, Fungal Biodiversity Centre. 571 p.

Singh AK, Kamal. 1987 [“1986”]. Fungi of Gorakhpur. XXXIX. Pseudocercospora. Kavaka 14:

Pier ey

Sutton BC. 1994. IMI Descriptions of Fungi and Bacteria, Nos 1181-1190. Mycopathologia 125(1):

43-64. http://dx.doi.org/10.1007/BF01103975

MY COTAXON

http://dx.doi.org/10.5248/120.465

Volume 120, pp. 465-469 April-June 2012

First records of Craterium aureonucleatum,

Perichaena quadrata, and Physarum mutabile in Italy

RICCARDO COMPAGNO, ALFONSO LA ROSA, & GIUSEPPE VENTURELLA*

Dipartimento di Biologia Ambientale e Biodiversita, Via Archirafi 38, I-90123 Palermo, Italy

*CORRESPONDENCE TO: giuseppe.venturella@unipa. it

ABsTRACT — Three species of Mycetozoa, new for Italy, were identified in garrigues and

maquis ecosystems on the volcanic island of Pantelleria (Sicily). The finding of Craterium

aureonucleatum, Perichaena quadrata, and Physarum mutabile widens their distribution in

Europe and identifies new substrata.

Key worps — biodiversity, Mediterranean area, ecology

Introduction

Within a wide-scoped biodiversity census, field investigations were carried

out on the island of Pantelleria (Trapani province, southwest Sicily, Italy). In

his checklist of Mycetozoa in the Mediterranean area, Lado (1994) reported

421 taxa, of which 164 were recorded from Italy. To increase the knowledge

of mycetozoan diversity in Italy, we focused primarily on the collection

of Mycetozoa in Mediterranean forest ecosystems characterizing the plant

landscape of Pantelleria (Gianguzzi 1999). The Erico arboreae-Quercetum ilicis

typicum plant association and Myrtus communis facies (Brullo et al. 1977)

is dominated by Quercus ilex L. in the tree layer and shrubs such as Arbutus

unedo L., Erica arborea L., Pistacia lentiscus L., Phillyrea latifolia L., and Myrtus

communis L. The degradation stages of the association are characterized by

garrigues with Rubus ulmifolius Schott. and Euphorbia dendroides L. (Brullo

et al 1977).

The three myxomycete species collected — identified as Craterium

aureonucleatum (Physaraceae), Perichaena quadrata (Trichiaceae), and

Physarum mutabile (Physaraceae) — are new records for Italy.

Materials & methods

The investigation was carried out weekly on Pantelleria from October 2007 to May

2009 in Mediterranean maquis and garrigues ecosystems. The collected samples were

identified by observing sporocysts in a water solution under Leica DMLB microscope.

466 ... Compagno, La Rosa & Venturella

Nomenclature of the recorded taxa follows MycoBank (http://www.mycobank.org/). The

macro- and microscopic features of peridium, stipe (if present), hypothallus, capillitium,

columella and/or pseudocolumella, and spores were observed. The habitat type was

noted for each recorded species. Voucher specimens were deposited in Herbarium

Mediterraneum Panormitanum, Palermo (PAL).

Species recorded

Craterium aureonucleatum Nann.-Bremek. FIG. 1

Teh Le:

yam, =£=> a>

(¢- 3; ay ~f . .

( | Ss

\ _* = se

“= =

" =

~ = 4

Ss b

- - ——

Fic. 1: Craterium aureonucleatum. [PAL 001(Myz)]

a. Sporocysts. b. Spores. c. A net with yellow nodes in the capillitium.

Myxomycetes new to Italy ... 467

Sporocysts with a short stem, grouped, sub-conical, 0.3-0.5 mm in diameter

and height, orange-dull brown. Hypothallus like a wide membranous disk,

ridged, orange, stem not exceeding 0.2 mm in height. Peridium with two

appressed layers, translucent. The outer layer of peridium is rather uniform

while the inner layer is membranous. ‘The two layers are orange at the base,

colorless or yellow in the upper part. The dehiscence of the peridial apex is

circular, forming a cup-shaped structure. The capillitium consists of a net with

small meshes, pale yellow with numerous cells, often branched, calcareous

yellow knots, pseudocolumella orange-yellow. Spore print dark brown; spores

dark brown-lilac in transmitted light, 9-10 um diam., covered by light tiny

warts.

SPECIMEN EXAMINED: ITALY. SicILy, TRAPANI PROVINCE, Pantelleria, Cuddia di Mida

and Montagna Grande, on litter of Quercus ilex L. and Erica arborea L. 10 Nov 2008, coll.

R. Compagno [PAL 001 (Myz)].

Comments - The sporocyst morphology in our collected specimens differs

slightly from the original description of Ing (1999), where sporocysts are

described as sub-globose, not sub-conical. This difference may be due to

ecological conditions and infraspecific variability.

Craterium aureonucleatum has been recorded from Canary Islands,

Germany, Netherlands, and Scotland (Beltran & al. 2004, Ing 1999, Nannenga-

Bremekamp 1991).

Perichaena quadrata T. Macbr. FIG. 2

tam. .¢

o > ~—

a 8 ay "4

he aie} . 7

77 - ror!

north Ti ist 4 Bibi HE

i nalitte c TL VA Gk OC

eet sa aris

at fs

hes

‘gal

Fic. 2: Perichaena quadrata [PAL 002(Myz)]. a. Sporocysts. b. Spores and capillitium.

468 ... Compagno, La Rosa & Venturella

Sporocysts rarely single, usually appressed with polyhedral shape, discoid,

sometimes lengthened, purple-brown or entirely black, with exudates of calcium

oxalate, 0.1-0.5 mm in diameter. Hypothallus extended under the sporocysts,

dark brown. Peridium double, initially adherent, the other layer thick and

opaque, brownish with inclusion of the inner membrane and translucent, pale

yellow, smooth. The capillitium consists of thin tubules, usually elastic and

abundant, sometimes with rings that segment the tubules. Spore print yellow;

spores yellow in transmitted light, 9-12 um diam., lightly punctate.

SPECIMEN EXAMINED: ITALY. SICILY, TRAPANI PROVINCE, Pantelleria, Bugeber, on dead

stems of Rubus ulmifolius Schott, 15 Feb 2009, coll. R. Compagno [PAL 002 (Myz)].

ComMENTs - Perichaena quadrata has also been recorded from Africa, Asia,

Australasia, Europe (including France and Spain), and North America (Lado

1994, The Eumycetozoan Project — http://slimemold.uark.edu/index.htm).

Physarum mutabile (Rostaf.) G. Lister FIG. 3

a wy a ti

- aT ! ert - zg

patagbes as Bs i Lor Si ate ees

ye, . 2 e. basil os8he

ais 4 : ar : ara ate:

if 4 i iii er Soa - 35h, ug yee Oi i

7 zs ree oe

Fic. 3: Sana ree mutabile [PAL 003(Myz)]. a. Sporocysts. b. Soe

Tei

Sporocysts with a short stipe; sub-globose; 0.3-0.6 mm wide and 2 mm long;

white or pale grey. Hypothallus membranous, colorless, pale yellow or pale

brown, sometimes with exudates. If present, the stipe is short, yellow to ochre,

Myxomycetes new to Italy... 469

usually covered by exudates. Peridium single, thin, colorless, smooth or rarely

with calcium carbonate flakes, upper crack irregular. Capillitium persistent,

elastic, with small meshes and white nodules. Pseudocolumella located in the

center of sporocyst or at the base of plasmodiocarp. Spore print black; spores

purple-brown in transmitted light, 7-10 um diam., densely spinulose with

spines sometimes grouped.

SPECIMEN EXAMINED: ITALY. Sicity, TRAPANI PROVINCE, Pantelleria, Montagna

Grande, on litter of Q. ilex L. and E. arborea L., 18 Oct 2007, coll. R. Compagno [PAL

003 (Myz)].

CoMMENTS — P. mutabile has been recorded from Africa, Asia, Europe, North

America and South America (Emoto 1977, Ing 199, Lado 1994, Nannenga-

Bremekamp 1991, The Eumycetozoan Project — http://slimemold.uark.edu/index.

htm).

Conclusions

The new records of C. aureonucleatum, P. quadrata and P. mutabile extend

their known European distribution into Italy and add valuable information

regarding substrata.

Acknowledgements

The authors wish to thank Dr Franco Bersan (Trieste, Italy) and Dr Gabriel Moreno

(Alcala de Henares, Madrid, Spain) for critically reviewing the manuscript and Cassandra

Funsten (USA) for her help with linguistic revision.

Literature cited

Beltran E, Lado C, Barrera J, Gonzales E. 2004. Myxomycetes diversity in the laurel forest of

Garajonay National Park (Canary Islands, Spain). Systematics and Geography of Plants 74:

159= 173:

Brullo S, Di Martino A, Marceno C. 1977. La vegetazione di Pantelleria (studio fitosociologico).

Pubblicazioni Istituto Botanico Universita di Catania, pp. 5-110. Tipografia Coniglione,

Catania.

Emoto Y. 1977. The myxomycetes of Japan. Sangyo Tosho Publishing Co., Ltd. Tokyo, Japan. 263

Gianguzzi L. 1999. Vegetazione e bioclimatologia dell’isola di Pantelleria (Canale di Sicilia). Braun-

Blanquetia 22: 3-70.

Ing B. 1999. The myxomycetes of Britain and Ireland: an identification handbook. The Richmond

Publishing Co. Ltd., England. 374 p.

Lado C. 1994. A checklist of myxomycetes of the Mediterranean countries. Mycotaxon 52:

117-185.

Nannenga-Bremekamp NE. 1991. A guide to temperate myxomycetes. Biopress Limited, England.

409 p.

MY COTAXON

http://dx.doi.org/10.5248/120.471

Volume 120, pp. 471-475 April-June 2012

Tuber microsphaerosporum and Paradoxa sinensis spp. nov.

Li FAN’, JIN-ZHONG CAO? & Yu Lr’

' College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

? Institute of Mycology, Jilin Agricultural University, Changchun 130118, China

* CORRESPONDENCE TO: fanli@mail.cnu.edu.cn

ABSTRACT — Two new truffle species from China are described and illustrated. Tuber

microsphaerosporum is recognized by its small ascospores and 1-7-spored asci. Paradoxa

sinensis is separated from the European P. monospora by its pale color and larger ascospores.

KEY worps — Ascomycota, mushroom market, Tuberaceae

Introduction

Knowledge about truffles in China has become richer since the first

publication on Tuber in China by Liu (Liu 1985). Of the many species found,

a high percentage has proved endemic (Wang 1988; Chen et al. 2005, 2007;

Fan et al. 2011, 2012a, b). Recently we obtained two truffle-like fungi from a

mushroom market in Kunming that we recognized as two new truffle species,

a Tuber and a Paradoxa, described and illustrated here.

Materials & methods

The fresh fruiting bodies were collected from a local mushroom market in Kunming,

China. Macro- and microscopic characters were described from fresh specimens.

Sections were cut by razor-blade and mounted in 3% KOH for observation or stained

in Melzer’s reagent, rinsed, and mounted in polyvinyl lactic glycerol for permanent

slides for archiving to be kept with dried specimens. For scanning electron microscopy

(SEM), ascospores were scraped from the dried gleba onto doubled-sided tape, which

was mounted directly on an SEM stub, coated with gold-palladium, and examined

and photographed with a Hitachi S-4800 SEM. The specimens are deposited in BJTC

(Herbarium Biology Department, Capital Normal University).

Taxonomy

Tuber microsphaerosporum L. Fan & Yu Li, sp. nov. Fic. 1

MycoBank MB 564356

472 ... Fan, Cao & Li

Fic. 1. Tuber microsphaerosporum (BJTC FAN152, holotype)

a. Ascoma. b-c. Asci and ascospores observed under light microscope.

d. Ascospores observed under SEM.

Differs from other Tuber species with globose ascospores by its smaller ascospores, up

to 7 per ascus.

Type: China. Yunnan Province, Kunming, from a mushroom market. 20 Dec. 2010, Jin-

Zhong Cao 115 (Holotype, BJTC FAN152).

ETYMOLOGY: microsphaerosporum (Lat.), referring to the small globose ascospores.

ASCOMA 2 cm in diam., subglobose, soft or slightly firm, solid, surface smooth,

poorly pubescent to nearly glabrous, yellowish white to light yellow-brown

at maturity. Odor slight, not pungent. PERIDIUM 250-350 um thick, two

layers; outer layer 100-150 um thick, pseudoparenchymatous, composed

of subangular or subglobose cells mostly 12.5-30 um in diam., with thin or

slightly thickened walls, pale yellowish, intermixed with larger swollen cells

of 70-80 x 40-50 um with thickened walls 2.5-3 um in diam.; inner layer

150-200 um thick, composed of intricately interwoven hyphae, thin-walled,

branching, hyaline, septate, 3-5 um in diam. The outermost cells giving rise a

few hyphae-like hairs, the hairs 20-50 x 2.5-3 um, septate, thin-walled, light

brown. GLeEBA light brown at maturity, marbled with large, rarely branching

and white veins continuous with inner peridium. Asc1 globose, subglobose or

broadly ellipsoid, hyaline, thin-walled, 67.5-105 x 60-70 um, mostly sessile,

sometimes with a short stalk, 1-7-spored, mostly 3-6-spored. AscosPORES

Two new truffle species from China ... 473

globose, brown at maturity, 17.5-27.5 um in 2-7-spored asci and 32.5-37.5 um

in 1-spored asci excluding ornamentation; ornamentation reticulate, meshes

regular or irregular, 3-6 sided, 2.5-3 um tall, the meshes generally 3-8 across

the spore width.

ComMENts — Tuber microsphaerosporum differs from the other Tuber species

with globose ascospores mainly by its smaller ascospores and the asci containing

1-7 ascospores; moreover, its peridium structure with large swollen cells is also

diagnostic for this new species.

The North American Tuber californicum Harkn. and T: sphaerosporum

Gilkey and European T. oligospermum (Tul. & C. Tul.) Trappe and T. borchii var.

sphaerosperma Malencon also produce perfectly globose ascospores, but all also

produce 1-4- or 1-3-spored asci. Tuber sphaerosporum is further distinguished

by large reticulum meshes numbering 3-4 across the spore diameter while

T. californicum has a dark-colored mature gleba and T: oligospermum has a one-

layer peridium.

Although we found no 8-spored asci in our holotype, 7-spored asci are not

uncommon, it is possible that 8-spored asci may occur in the new species.

Paradoxa sinensis L. Fan & J.Z. Cao, sp. nov. Fic. 2

MycoBank MB 564357

Differs from Paradoxa monospora by its yellowish to yellow-brown ascomata and larger

ascospores.

Type: China. Yunnan Province, Kunming, from the local mushroom market. 20 Dec.

2010, Jin-Zhong Cao 113 (Holotype, BJTC FAN150).

EryMo_oey: sinensis (Lat.), referring to the presumed host country.

ASCOMA 2.5 cm in diam., subglobose, firm, solid, surface smooth, glabrous,

yellowish to yellow-brown at maturity. Odor slight, not pungent. PERIDIUM

200-250 um thick, one layer, prosenchymatous, composed of intricately

Fic. 2. Paradoxa sinensis (BJTC FAN150, holotype)

a. Asci and ascospores observed under light microscope. b. Ascoma.

474 ... Fan, Cao & Li

interwoven hyphae, branching, septate, the hyphae are light yellow-brown,

slightly thick-walled and 5-7.5(-10) um broad towards the outer side of

peridium, and hyaline, thin-walled and 5-7.5 um broad towards the inner

side. GLEBA blackish at maturity, marbled with large, rarely branching and

greyish or grey-brown veins continuous with the peridium. Asc1 1-spored,

rarely 2-spored, globose or subglobose for 1-spored and long-ellipsoid for

2-spored, 77.5-92.5 x 80-85 um for 1-spored and 120 x 80 um for 2-spored,

hyaline, thick-walled, 5-7.5 um thick, sessile or with a short tail. AscosPORES

globose, red-brown to dark red-brown at maturity, (55-)62.5-72.5 um in diam.

excluding ornamentation, ornamentation reticulate, irregular, meshes varying

greatly in shape and size, 5-20 um in diam., 1 um high, averagely 6-10 across

the spore width.

ComMENTS — Paradoxa sinensis closely resembles the European P monospora

Mattir., but Ceruti (1960) describes the ascomata of P monospora as black

brown and the ascospores as somewhat smaller (50-60um in diam.). The third

Paradoxa species, P. gigantospora (Y. Wang & Z.P. Li) Y. Wang (Wang & Hu

2008), was originally described from China by Wang & Li (1991, as Tuber

gigantosporum) and is diagnosed by its large elliptic ascospores (<100 um).

Paradoxa, originally described from Italy in 1935, was monotypic until Wang

& Hu (2008) transferred T. gigantospora to it. Paradoxa sinensis represents

the third species in the genus, which differs from Tuber mainly by its single-

spored asci. Vizzini (2008), who treated Paradoxa as a synonym of Tuber,

transferred P. monospora to Tuber as T:! monosporum. Kinoshita et al. (2011)

seemingly accepted this classification when they listed Paradoxa gigantospora

as a synonym of Tuber gigantosporum. However, in all three Paradoxa species,

the dominant character of one larger ascospore per ascus is always connected

with peculiar reticulum on the spore surface, which is highly irregular and

greatly lower, suggesting a taxonomic difference between Paradoxa and most

typical Tuber species. More research is needed to elucidate whether the two

genera are independent or conspecific. We prefer to treat Paradoxa and Tuber

as independent genera for the time being.

Acknowledgments

We are grateful to Prof. Zhu-Liang Yang (Kunming Institute of Botany, Chinese

Academy of Sciences) and Prof. Anthony Whalley (School of Pharmacy and

Biotechnology, Liverpool John Moores University) for reviewing the manuscript.

The study was supported by the National Natural Science Foundation of China (No.

30770005, 30870008) and the Beijing Natural Science Foundation (No. 5072006).

Literature cited

Ceruti A. 1960. Elaphomycetales et Tuberales. 1-48, in: J Bresadola. Iconographia Mycologica. 28,

suppl. 2.

Two new truffle species from China... 475

Chen J, Liu PG. 2007. Tuber latisporum sp. nov. and related taxa, based on morphology and DNA

sequence data. Mycologia 99: 475-481. http://dx.doi: 10.3852/mycologia.99.3.475

Chen J, Liu PG, Wang Y. 2005. Tuber umbilicatum, a new species from China, with a key to the

spinose-reticulate spored Tuber species. Mycotaxon 94: 1-6.

Fan L, Cao JZ, Liu YY, Li Y. 2011. Two new species of the genus Tuber from China. Mycotaxon 116:

349-354. http://dx.doi: 10.5248/116.349

Fan L, Hou CL, Cao JZ. 2012a [“2011”]. Tuber sinoalbidum and T. polyspermum — new species

from China. Mycotaxon 118: 403-410. http://dx.doi.org/10.5248/118.403

Fan L, Cao JZ, Zheng ZH, Li Y. 2012b. Tuber in China: T: microspermum and T. microspiculatum

spp. nov. Mycotaxon 119: xxx-xxx. http://dx.doi.org/10.5248/119.xxx

Kinoshita A, Sasaki H, Nara K. 2011. Phylogeny and diversity of Japanese truffles (Tuber spp.)

inferred from sequences of four nuclear loci. Mycologia 103: 779-794.

http://dx.doi.org/10.3852/10-138

Liu B. 1985. New species and new records of hypogeous fungi from China (I). Acta Mycologica

Sinica 4(2): 84-89.

Vizzini A. 2008. Novitates: Tubariaceae fam. nov. Rivista di Micologia 51: 174.

Wang Y. 1988. First report of study on Tuber species from China. Atti del II Congresso Internazionale

sul Tartufo, Spoleto, Nov. 24-27: 45-50.

Wang Y, Hu HT. 2008. Paradoxa gigantospora comb. nov. from China. Mycotaxon 106: 199-202.

MYCOTAXON

http://dx.doi.org/10.5248/120.477

Volume 120, pp. 477-492 April-June 2012

Entomophthoromycota: a new phylum and

reclassification for entomophthoroid fungi

RICHARD A. HUMBER

USDA-ARS Biological Integrated Pest Management Research,

Robert W. Holley Center for Agriculture and Health,

Tower Road, Ithaca, New York 14853-2901, U.S.A.

CORRESPONDENCE TO: richard. humber@ars.usda.gov

ABSTRACT — One result of the recent phylogenetically based rejection of the phylum

Zygomycota was the description of the subphylum Entomophthoromycotina (not assigned to

any phylum) for fungi traditionally treated in the order Entomophthorales. More extensive

gene-based analyses of these fungi suggest that they represent a monophyletic lineage

distinct from all other fungi that deserves now to be recognized at the level of a new fungal

phylum. These molecular data and further analyses of more traditional taxonomic criteria

lead to this reclassification that still treats these fungi in six families but recognizes the new

classes Basidiobolomycetes, Neozygitomycetes, and Entomophthoromycetes as well as the new

order Neozygitales. Ballocephala and Zygnemomyces are excluded from Entomophthorales

(Meristacraceae) and should be reclassified among the Kickxellomycotina.

KEY worps — Zygomycetes, sexuality, homothallism

Introduction

The reclassification of fungi by Hibbett et al. (2007) as the complement

to a kaleidoscopic phylogenetic study all major fungal groups (James et al.

2006) validated the long-accepted sense that Zygomycota was polyphyletic,

and recognized five taxa to replace this phylum: The phylum Glomeromycota

accommodates arbuscular mycorrhizal fungi, and all other zygomycetes were

distributed among subphyla Entomophthoromycotina, Kickxellomycotina,

Mucoromycotina, and Zoopagomycotina without assignment to any phylum. It

was assumed that subsequent research would determine whether any of these

subphyla should be regrouped as part of an effort that would necessarily result

in the recognition of one to four new phyla for these fungi.

The major characters traditionally used to classify the Entomophthorales

have been thoroughly reviewed (summarized in Humber 1975, 1981, 1982,

478 ... Humber

1984) and were applied to six families in the last major reclassification of

Entomophthorales (Humber 1989); this taxonomy is widely accepted despite

a few minor differences in the treatments of some entomopathogenic genera

(Balazy 1993; Keller 1987, 1991, 1997; Keller & Petrini 2005). Until now,

however, there have not been phylogenetic studies on a sufficiently broad range

of their genes and taxa to propose a more contemporary revision.

The fungi in Entomophthoromycotina pose a few mycological puzzles

(discussed below) for refining their current classification: Both the ultrastructure

of the mitosis-associated organelle and early phylogenetic studies suggested that

Basidiobolus has affinities with chytrid fungi and might better be excluded from

Entomophthorales. Further, significant gaps in the gene-based understandings

of entomophthoroid fungi exist because many taxa are very rarely collected

and/or resist growth in vitro. Among these understudied taxa, Neozygites and

related species represent the largest and most important ‘black boxes’ for which

needed data remain unavailable.

The gene sequences now conceded to have taxonomic value for many

fungi (nuclear rDNA genes, translation-elongation factor, $-tubulin, etc.)

have been used in studies of a few entomophthoraleans in a more diverse set

of fungi (Nagahama et al. 1995, Jensen et al. 2001, White et al. 2006) and for

narrower studies of entomophthoralean species or species complexes (Jensen &

Eilenberg 2001, Nielsen et al. 2001). A markedly different molecular approach

comparing amino acid sequences for proteins (including some of the same

proteins whose DNA sequences are widely used) has placed Entomophthorales

outside of the true fungi (Einax & Voigt 2004, Liu & Voigt 2010, Voigt & Kirk

2011). While some skepticism about the meaning of such results based on

amino acid sequences must be maintained, these findings do demonstrate

the existence of distinct differences between all other groups of zygomycete

fungi and Entomophthoromycotina. The amino-acid sequence-based ‘exclusion’

of Entomophthorales from the fungi echoes the hypotheses about placing

Basidiobolus with watermolds but such ‘anomalous’ conclusions also suggest

that molecular analyses based on highly limited inputs can yield results that

have little sensibility from the broader perspective of the overall biology of the

organisms analyzed.

A series of phylogenetic studies on entomophthoraleans (being prepared

by A. Gryganskyi, R. Vilgalys, R. Humber, and other authors) incorporated

more genes and a much broader range of entomophthoroid taxa than any

earlier studies. These new analyses confirm the finding by James et al. (2006)

that entomophthoroid fungi are a monophyletic group and that this group

does include Basidiobolus and Basidiobolaceae. A reasonable integration of

all results of traditional and phylogenetic analyses of entomophthoroid fungi

suggests that they are distinct from all other fungi (including those in the other

Entomophthoromycota phyl. nov. ... 479

unaffiliated zygomycete subphyla; Hibbett et al. 2007) and may occupy the

most basal position among all non-flagellate fungi. The best-supported, most

appropriate conclusion about the status of these fungi is, therefore, to recognize

them as a new phylum in kingdom Fungi.

Materials & methods

Cultures and specimens used for the analyses discussed here are primarily from the

USDA-ARS Collection of Entomopathogenic Fungal Cultures (ARSEF; http://www.ars.

usda.gov/Main/docs.htm?docid=12125) and its associated herbarium. Unpublished

molecular results of analyses of various genes are based on sequences of cultures obtained

from ARSEF or isolated from nature and, in most cases, subsequently deposited in

ARSEF. Other results involving reports on fungi and analyses using other isolates and

specimens were completed at the Zoology Section of the Department of Agriculture and

Ecology, University of Copenhagen, or at Agroscope FAL Reckenholz Eidgenéssische

Forschungsanstalt fir Agrardkologie und Landbau (Ziirich).

Major taxonomic issues affecting this reclassification

‘Linkage’ of Basidiobolus with flagellate fungi

The nuclei of entomophthoralean fungi and the details of their mitoses

present a comparatively richer number and variety of characters than in most

other fungal groups, and these nuclear characters are taxonomically informative,

especially above the generic rank (Humber 1975, 1981, 1982, 1984, 1989).

Mitosis by the huge nuclei of Basidiobolus species is sufficiently unusual to

have been studied repeatedly (Olive 1907, Robinow 1963, Sun & Bowen 1972,

Tanaka 1978). Among other surprises, the location of the mitosis-associated

organelle in this genus is not fixed at the spindle poles and can even occur in the

plane of the metaphase plate (Sun & Bowen 1972). The real controversies about

this mitosis arose, however, when this organelle proved to be a short cylinder

with a ring of 11-12 singlet microtubules (McKerracher & Heath 1985), and

comparisons between this structure and centrioles were used to question the

phylogenetic placement of this fungus; Basidiobolus remains the ONLY organism

producing no flagellum in its life history for which microtubules are proven to

be present in a mitosis-associated organelle. Nonetheless, any hypothesis that

this organelle in Basidiobolus (but whose existence, location and ultrastructure

remain unconfirmed from other fungi in its family) is linked with or derived

from centrioles seems neither credible nor responsible: The 9x3 microtubular

arrangement in centrioles and kinetosomes is invariant among ALL phyla of

eukaryotes having flagella; conversely, no organelle with microtubules arranged

like those in Basidiobolus is known from any other organism.

Later findings of gene sequence similarities between Basidiobolus and

several chytrid and blastocladian watermolds (Nagahama et al. 1995, Jensen et

al. 1998, Tanabe et al. 2000, White et al. 2006) have also been used to suggest

480 ... Humber

that Basidiobolus might not belong in Entomophthorales. However, other studies

allied Basidiobolus with kickxelloid fungi from Harpellales (Keeling 2003,

Tanabe et al. 2004) and one placed Conidiobolus coronatus (whose inclusion in

Entomophthorales was never disputed) on a branch with the blastocladialean

genus Allomyces (Tanabe et al 2005). These divergent findings underscore the

peril of suggesting phylogenetic relationships among major fungal groups after

comparing very limited sequence data and very sparse samplings of taxa within

large and inherently diverse groups of fungi.

The recent survey of phylogenetic relationships within kingdom Fungi

(James et al. 2006) supported the continued placement of Basidiobolus in

Entomophthorales. Regardless of molecular or ultrastructural rationales to the

contrary, hypotheses that Basidiobolus is not entomophthoroid are nonsensical

if one considers the overall biology of these fungi. Basidiobolus and its relatives

share many novel features in common with other entomophthoroid taxa

despite the scant few pieces of data suggesting otherwise. It is necessary to

bring a broader perspective to the uncertainties about Basidiobolus: Despite

any and all evidence to the contrary, if this genus does not belong among

the entomophthoroid fungi, then jusT WHERE AMONG FLAGELLATE (OR ANY

OTHER) FUNGI SHOULD IT BE CLASSIFIED? The lack of any comparably well-

supported answers for this question should quash any residual doubts about

where Basidiobolus belongs.

Neozygitaceae: a special ‘problem’ in data gathering

The status of taxa in Neozygitaceae also presents a (temporary) problem

for reclassifying the Entomophthorales. Very few cultures of Neozygites species

are available in vitro, and, sadly, all current cultures of Neozygites are of mite

pathogens with rough-walled, nearly globose zygospores. No gene-based data

are available for N. fresenii [=N. aphidis, the type species] or other hemipteran

pathogens that form ovoid, smooth-walled resting spores. A further taxonomic

frustration is that these morphological and host differences suggest that

Neozygites may eventually be split into two or more genera, but molecular

data will be required to support such a decision. DNA-based evaluations of

fungi from Neozygitaceae is encumbered by difficulties experienced in multiple

laboratories to obtain clean DNA useful for amplifying and sequencing

the genes needed to determine their phylogenetic relationships. While the

recognition of Neozygitomycetes as a new class without supporting gene-based

evidence may be dismissed by some as premature, such a treatment is the

most reasonable for these fungi at this time, based on their known organismal

biology (that integrates and represents a vastly larger proportion of the gnome

than those few genes now so widely treated as sufficient to complete high-level,

phylogenetically sound reclassifications of virtually all other fungal groups).

Entomophthoromycota phy]. nov. ... 481

Distinct differences between neozygitoid fungi and either basidioboloid or

entomophthoroid taxa supports the description of three classes in this new

phylum: As in basidioboloid fungi, neozygitoid fungi exert strong control

in vegetative cells over nuclear number (usually four in Neozygites), have a

central mitotic metaphase plate (Butt & Humber 1989), and, perhaps most

significantly, a round of mitosis in gametangial cells precedes conjugation and

zygosporogenesis while only one nucleus from each gametangium enters each

zygospore (Keller 1997). As in entomophthoroid fungi, all neozygitoid taxa

are obligate pathogens of insects or mites, and the nuclear membrane remains

intact throughout mitosis. On the other hand, the chromosomes of neozygitoid

fungi differ from basidioboloid and entomophthoroid fungi in being vermiform

and of moderate size, condensing during mitosis but uncoiled (euchromatic)

during interphase. Neozygites mitoses (Butt & Humber 1989) resemble those

in animal or plant cells more closely than those in any other entomophthoroid

fungi.

The presence of many novel characteristics shared among all of the fungi

traditionally classified in order Entomophthorales underscores the need to keep

these fungi together in a phylogenetically supported, coherent group and to

pursue further studies to obtain more vital data about the genes, development,

pathobiology, and other aspects for a better understanding of these fungi that

can be very important naturally occurring biological control agents. Because

these fungi occupy a very ancient position of the fungal tree of life it is also

important to note that a better understanding should help to understand

more about the enigmatic transition of fungi (as also occurred with plants and

animals) from waterborne to terrestrial life forms.

Taxonomy

Entomophthoromycota Humber, phyl. nov. [TABLE 1]

MycoBank MB 564375

VEGETATIVE GROWTH AS hyphae, hyphal bodies, or yeast-like; cells broad, walled or

protoplastic. CONIDIOPHORES simple or digitate, each branch forming one conidiogenous

cell and one conidium. PRIMARY SPORES conidia, uni- to multinucleate, usually forcibly

discharged; usually forming one or more types of SECONDARY CONIDIA. RESTING SPORES

homothallic zygospores or azygospores. HaBit mostly as arthropod pathogens, but

some saprobes or specialized phytopathogens.

TyPE GENUS: Entomophthora Fresen. 1856.

CONIDIOPHORES rise from mycelium or from body of host, usually with positive

phototropic orientation, simple or apically (digitately) branched, with single

conidiogenous cell on each branch giving rise to a single conidium or simple

erect conidiophore becomes septate and each cell forms a single conidium.

PRIMARY CONIDIA (not sporangia) with wall layers continuous with those on

482 ... Humber

TABLE 1. Proposed new classification for Entomophthoromycota.

New taxa described here are listed in boldface italics.

Puyitum Entomophthoromycota Humber, phyl. nov.

Cass Basidiobolomycetes Humber, cl. nov.

ORDER Basidiobolales Caval.-Sm., Biol. Rev. 73: 246. 1998.

FamILy Basidiobolaceae Claussen, Syllab. Pflanzenfam., Edn 9 & 10: 45. 1924.

Basidiobolus Eidam, Beitr. Biol. Pflanz. 4: 194. 1886.

Other new, undescribed genera (R.A. Humber, B.

Huang & K. Hodge, unpublished).

Ciass Neozygitomycetes Humber, cl. nov.

ORDER Neozygitales Humber, ord. nov.

FamiLy Neozygitaceae Ben-Zeev, R.G. Kenneth &

Uziel, Mycotaxon 28: 321. 1987.

Apterivorax S. Keller, Sydowia 57: 47. 2005.

Neozygites Witlaczil, Arch. Mikr. Anat. 24: 601. 1885.

Thaxterosporium Ben-Zeev & R.G. Kenneth, Mycotaxon 28: 323. 1987.

Cxass Entomophthoromycetes Humber, cl. nov.

ORDER Entomophthorales G. Winter, Rabenh. Krypt.-Fl., Edn 2, 1(1): 74. 1880.

FaMILy Ancylistaceae J. Schrot., Nat. Pflanzenfam. 1(1): 92. 1893.

Ancylistes Pfitzer, Monatsb. Konigl. Preuss. Akad. Wiss. Berlin: 396. 1872.

Conidiobolus Bref., Untersuch. Gesammtgeb. Mykol. 6: 37. 1884.

Macrobiotophthora Reukauf, Centrabl. Bakt., Abt 1, 63: 390. 1912.

FamILy Completoriaceae Humber, Mycotaxon 34: 453. 1989.

Completoria Lohde, Tagebl. Versamml. Deutsch. Naturf. Aertze 47: 206. 1874.

FamiLy Entomophthoraceae Nowak., Bot. Ztg. 35: 35. 1877.

SUBFAMILY Entomophthoroideae S. Keller, Sydowia 57: 28. 2005.

Batkoa Humber, Mycotaxon 34: 446. 1989.

Entomophaga A. Batko, Bull. Polon. Acad. Sci. Sér. Biol. Sci. 12: 325. 1964.

Entomophthora Fresen., Bot. Ztg. 14: 883. 1856.

Eryniopsis Humber, Mycotaxon 21: 258. 1984, pro parte.

Massospora Peck, Rep. New York State Mus. 31: 44. 1879.

SUBFAMILY Erynioideae S. Keller, Sydowia 57: 33. 2005.

Erynia (Nowak. ex A. Batko) Remaud. &

Hennebert, Mycotaxon 11: 333. 1980.

Eryniopsis Humber, Mycotaxon 21: 258. 1984, pro parte.

Furia (Batko) Humber, Mycotaxon 34: 450. 1989.

Orthomyces Steinkraus, Humber & J.B. Oliv., J. Invertebr. Pathol. 72: 5. 1998.

Pandora Humber, Mycotaxon 34: 451. 1989.

Strongwellsea A. Batko & Weiser, J. Invertebr. Pathol. 7: 463. 1965.

Zoophthora A. Batko, Bull. Polon. Acad. Sci. Sér. Biol. Sci. 12: 323. 1964.

FamILy Meristacraceae Humber, Mycotaxon 34: 456. 1989.

Meristacrum Drechsler, J. Wash. Acad. Sci. 30: 250. 1940.

Tabanomyces Couch, RV Andrejeva, Laird & Nolan,

Proc. Natl. Acad. Sci. USA 76: 2302. 1979.

Entomophthoromycota phy]. nov. ... 483

conidiogenous cells, inner wall layer invaginating to form two-layered septum

between conidium and conidiogenous cell; almost always forcibly discharged

(several possible mechanisms are known). SECONDARY CONIDIA formed by

most taxa: if forcibly discharged from short secondary conidiophore then

usually similar in shape to primary conidium; if passively dispersed from long,

thin (capillary) secondary conidiophore then usually distinctly differing in

morphology from primary conidium. RESTING sPpoRES (when mature) with

thick, distinctly 2-layered walls, colored or hyaline, outer layer surface smooth

or variously decorated; formed as zygospores (after gametangial conjugation)

or azygospores (with no conjugation) either in the axis of the parental cells

or budded off laterally; nuclear number in mature spores varies from 2 (from

initiation or progressively reducing to 2) to multiple; germinating directly

by forming germ conidiophore and germ conidium (usually resembling a

secondary spore type) or indirectly by forming a small germ mycelium and

then germ conidia (usually like primary conidia). HasiTs: saprobes in soil or

litter, primary pathogens of arthropods (insect, mites, spiders) or other soil

invertebrates (nematodes, tardigrades), or highly specific phytopathogens

(e.g., of desmid algae or fern gametophytes). ARTHROPOD PATHOGENS may

form specialized organs: RHIZOIDs with or without differentiated holdfasts may

anchor host to substrate, and cysTip1A may perforate host cuticle and facilitate

emergence of conidiophores.

Primary and secondary conidia are the major spore forms in this phylum and

constitute the primary basis for the taxonomy of these fungi. The resting spores

are formed much less commonly than are conidia. ‘The majority of species

are pathogens of arthropods although pathogens of other soil invertebrates

(nematodes and tardigrades) or of plants (desmid algae or fern gametophytes)

are rare. The primary habit (especially in Basidiobolaceae and Ancylistaceae)

may be in soil and plant detritus, but some species in these groups are best

known as colonists of amphibian and reptile guts (Basidiobolus) or as facultative

or obligate entomopathogens (Conidiobolus).

Any continued use of subphylum Entomophthoromycotina Humber

(Hibbett et al. 2007: 517) is now superfluous until any future decision divides

Entomophthoromycota into subphyla. This reclassification does not take up

the phylum Basidiobolomycota Doweld (2001; LXXVII) because Doweld’s

name was proposed as part of a general reclassification of all fungi that does

not agree with current understandings of fungal biology and relationships

and, as circumscribed, Basidiobolomycota and the class Basidiobolomycetes

Doweld (2001: LXXVII) used fragmentary knowledge of characters that may

not apply to all taxa intended to be included while failing to account in any

way for most taxa specifically included in this circumscription of phylum

Entomophthoromycota; also see discussion below for class Basidiobolomycetes.

484 ... Humber

Basidiobolomycetes Humber, cl. nov.

MycoBank MB 564376

Differs from Entomophthoromycetes and Neozygitomycetes by unusually large nuclei

(often 210 um long) with a large central nucleolus that is the major feature of uninucleate

cells. Mitoses involve barrel-shaped spindles, mitotic organelles incorporating

microtubules (but not centrioles) but not always located at the spindle poles, and the

nuclear content isolated from the cytoplasm by a layer of nuclear and cytoplasmic

membrane fragments.

Type GENUS: Basidiobolus Eidam 1886.

VEGETATIVE CELLS uninucleate, as regularly septate mycelium or yeast-like cells

cleaved from contents of a parental cells (e.g., so-called ‘Darmform’ growth).

Mitosis begins with fragmentation of nuclear membrane and aggregation of

these and other membranes around a nuclear zone; chromosomes numerous,

tiny, condensed and aligned on central metaphase plate (usually embedded

inside the nucleolus) in association with a barrel-shaped spindle, chromosomes

uncoil during interphase. CONIDIOGENOUS CELL (CONIDIOPHORE) simple

but with bulbous apical swelling immediately below developing conidium.

CONIDIA uninucleate, globose, with small conical basal papilla (projecting

into spore body but everting during discharge), unitunicate (wall layers not

separable). CONIDIA DISCHARGE forcibly by rocket-like ejection when central

circumscissile weakness of the subconidial swelling ruptures; the upper portion

of the swelling discharges together with conidium but may detach during

flight. SECONDARY CONIDIA (if formed) usually elongate, often curved, with

or without an apical mucoid droplet, formed apically on an elongated capillary

conidiophore, passively dispersed. RESTING spoREs (RS) usually zygospores,

formed homothallically in axis of parental cells; gametangial nuclei undergo

mitosis before conjugation but only one nucleus from each cell enters the

zygospore. MATURE ZYGOSPORES have thick, bi-layered walls; RS GERMINATE

by direct formation of germ conidium (usually a secondary conidial type:

elongate, passively dispersed from a capillary conidiophore).

The foremost diagnostic character for basidioboloid fungi is their huge nucleus

(often =10 um in length) with a prominent central nucleolus that is the major

feature of uninucleate cells (either as a broad, septate mycelium or cells cleaving

internally in yeast-like growth mode). There is no staining of interphasic nuclei

(nor, in any obvious manner, of mitotic chromosomes) in Basidiobolomycetes

in aceto-orcein or other nuclear stains. The individual volumes of these nuclei

may be many times greater than the entire cells of most ascomycete yeasts),

and their mitoses are unusual for more than just the microtubular nucleus-

associated organelle (McKerracher & Heath 1985): As mitosis begins, the

nuclear envelope breaks down but the endoplasmic reticulum and other

membranous cell components cluster around the nuclear zone so that the

Entomophthoromycota phyl. nov. ... 485

spindle and chromosomes remain well isolated from the cytoplasm despite the

fragmentation of the nuclear envelope; the corollary effect of this membrane

organization is that mitotic nuclei ‘disappear’ when viewed with light microscopy

(Robinow 1963).

Zygosporogenesis in Basidiobolus (Eidam 1886) is also very distinctive as

short beak-like, lateral projections form at the septum between gametangial

cells; gametangial nuclei move into the beaks, undergo mitosis, and the

(uninucleate) beak cells are walled off before the septum dissolves and

zygosporogenesis proceeds; remnants of these ‘beaks’ often remain visible on

mature zygospores.

As noted in the discussion for the new phylum, two available names for

this new class were not adopted: Bolomycetes Cav.-Smith (Cavalier-Smith 1998:

243) was based mainly on the microtubular mitotic organelle and ‘beaked’

zygospores in Basidiobolus. This mitotic organelle is not confirmed as present

in all taxa in the Basidiobolaceae (including at least two still undescribed new

genera), and the zygospores of some basidiobolaceous fungi are not “beaked’ as

in Basidiobolus. Basidiobolomycetes Doweld (2001: LXXVII) was proposed as a

nomen novum for Bolomycetes and cited Cavalier-Smith’s description for this

class; Doweld neither placed nor mentioned other entomophthoralean fungi in

any rank in his general reclassification of fungi.

Neozygitomycetes Humber, cl. nov.

MycoBank MB 564377

Differs from Basidiobolomycetes and Entomophthoromycetes by vermiform, moderately

sized chromosomes that condense during mitosis on a central metaphase plate but

uncoil during interphase. Nuclear numbers in vegetative cells and conidia are low and

apparently controlled at (3)-4-(5).

TyPE GENUS: Neozygites Witlaczil 1885.

VEGETATIVE CELLS are rod-like hyphal bodies, walled or protoplastic, usually

with 4 (3-5) nuclei, elongating until + synchronous mitosis; daughter cells

separate by splitting of septum. NUCLEAR NUMBER in all cell types strongly

regulated; usually 4 (3-5) in vegetative cells and conidia, 2 in resting spores.

MiTOsEs intranuclear, + synchronous in any cell; nuclei fusoid at metaphase

with central, fusoid spindle; no nucleus-associated mitotic organelle observed;

chromosomes uncoil (euchromatic) during interphase. CONIDIOPHORES

simple; forming apical conidiogenous cell and one conidium. PRIMARY

conip1A subglobose to broadly ovoid, basal papilla short, comparatively flat;

forcibly discharged to short distance by papillar eversion. SECONDARY CONIDIA

usually form quickly after primary conidial discharge, most commonly form as

capilliconidia (that are the primary infective units). RESTING spoREs bud from

short conjugation bridge between rounded-up hyphal bodies (gametangia)

486 ... Humber

after preconjugational mitosis in contacting gametangia; zygospore receives

one nucleus from each gametangium; only outer wall layer is melanized.

Mature resting spores with two adjacent round fenestrae (‘holes’ through outer

wall layer) and raised ridge of gametangial wall remnants between them.

Melanization of all spore types is a major feature of Neozygitomycetes. Primary

and secondary conidia are pale, smoky gray; individual resting spores are much

more strongly colored, and dark gray to black in mass.

Neozygitales Humber, ord. nov.

MycoBank MB

ORDER having all characteristics of class Neozygitomycetes.

TYPE GENUS: Neozygites Witlaczil 1885.

This order has all characters of class Neozygitomycetes (which includes only a

single order and family).

Entomophthoromycetes Humber, cl. nov.

MycoBank MB 564381

Differs from Basidiobolomycetes by lack of uniformly uninucleate cells, nuclear

morphology, details of mitoses, and modes of zygosporogenesis; and from

Neozygitomycetes by cells not having uniformly small numbers of nuclei, details of

mitoses, and lack of melanization of all spore types.

TYPE GENUS: Entomophthora Fresen. 1856.

VEGETATIVE GROWTH as coenocytic mycelium or rod-like to variably shaped

hyphal bodies, walled or naturally protoplastic; if wall-less, rod-like to highly

variable in shape and/amoeboid. CONIDIOPHORES simple or digitately branched,

each branch with a single apical conidiogenous cell, or (in Meristacraceae) an

unbranched erect, septate conidiophore forming one conidium per cell. Conidia

unitunicate (wall layers not separating in liquid) or bitunicate (with separable

outer wall layer); variously shaped, uni- to multinucleate, with basal papilla flat,

conical or rounded; forcibly discharged by papillar eversion in most genera.

SECONDARY CONIDIA more or less similar in shape to primary conidia and

forcibly discharged if formed on short secondary conidiophore, or elongate and

passively dispersed if formed on elongated capillary secondary conidiophore.

NUCLEI (interphase) with small nucleolus, interphasic heterochromatin present

in Entomophthoraceae but absent in all other families; mitoses intranuclear,

with small lateral metaphase plate lateral; interphasic chromosomes are partly

condensed (heterochromatic) and stain readily in Entomophthoraceae but

euchromatic (uncoiled and nonstaining) in other families. RESTING SPORES

globose to subglobose, formed as zygospores or azygospores. HasiT obligately

pathogenic for invertebrates (Entomophthoraceae, Meristacraceae, some

Ancylistaceae), saprobic (some Ancylistaceae), or phytopathogenic (Completoria

[Completoriaceae| and Ancylistes [Ancylistaceae]).

Entomophthoromycota phyl. nov. ... 487

This class includes all members of Ancylistaceae, Entomophthoraceae,

Completoriaceae, and Meristacraceae but omits those entomophthoralean

taxa reassigned here to Basidiobolomycetes or Neozygitomycetes (TABLE 1) or

removed from Entomophthoromycota as noted below.

Genera incertae sedis:

Eryniopsis Humber, Mycotaxon 21: 258. 1984.

All species are in Entomophthoraceae but would appear to be a mix of taxa

representing both subfamilies Erynioideae and Entomophthoroideae; the type

species, E. lampyridarum, has morphological characters suggestive of both

subfamilies and cannot be placed in either without molecular studies.

Tarichium Cohn, Beitr. Biol. Pflanzen 1: 69. 1870.

This form genus for species known only from resting spores apparently represents

a mix of species attributable to Neozygitaceae (especially species pathogenic to

mites) and Entomophthoraceae. No new species should be added to this genus;

DNA-based studies and morphological re-evaluations should allow most species

to be recognized as synonyms of other species or transferred to other genera in

Entomophthoraceae and Neozygitaceae.

Taxa inadoptata vel excludenda:

Massosporoideae S. Keller, Sydowia 57: 44. 2005.

This subfamily (accommodating only the genus Massospora) seems not to be

supported by phylogenetic evidence and is treated as a synonym of subfamily

Entomophthoroideae.

Ballocephala Drechsler, Bull. Torrey Bot. Club 78: 199. 1951.

Zygnemomyces K. Miura, Rep. Tottori Mycol. Inst. 10: 520. 1973.

These two genera are excluded from Meristacraceae and reassigned to

Kickxellomycotina Benny (Hibbett et al. 2007) based on the bifurcate septa with

lenticular plugs in their vegetative hyphae (Saikawa 1989; Saikawa et al. 1997).

Discussion

The terms ‘mitospore’ and ‘meiospore’ are not used in characterizing taxa of

Entomophthoromycota. They were originally adopted to describe ascomycete

and basidiomycete spores, and are not applicable to entomophthoroid fungi

because the reproductive products and life histories of entomophthoroid

fungi are not strictly comparable with those of the Dikarya: The thin-walled

primary conidia (the basis for entomophthoroid taxonomy) are produced

by the vegetative cells of these fungi, usually forcibly discharged, and usually

able to produce one or more subsequent forms of secondary conidium if

conidia do not germinate by producing a germ tube. Entomophthoroid

resting spores may be conventionally sexual in nature (zygospores in which

it may be AssUMED, although not yet proven, that karyogamy and meiotic

488 ... Humber

divisions occur) in Basidiobolomycetes and Neozygitomycetes; for taxa in the

Entomophthoromycetes, and especially those in Entomophthoraceae, it was

noted that the MORPHOLOGICAL events of sexuality (the presence or absence

of gametangial conjugations define zygo- and azygosporogenesis, respectively)

and the GENETIC events of sexuality (karyogamy and meiosis, that presumably

happen during resting spore germination) may be completely independent

processes (Humber 1981, McCabe et al. 1984). Entomophthoroid fungi may be

the only fungi in which the morphological and genetic definitions of sexuality

(or their absences) are present in all possible permutations and without the

routine linkage between the morphological and genetic events of sexuality that

is taken for granted in virtually all other types of organisms.

No zygomycetous or flagellate fungi produce spores that can accurately be

referred to as meiospores in the sense of basidiospores and ascospores. The

proven or presumably ‘sexual’ spores of fungi below the subkingdom Dikarya

are thick-walled, environmentally resistant spores (zygospores, azygospores,

resistant sporangia, etc.) that go through a quiescent dormancy before

germinating to undergo a type of sporulation that is neither functionally nor

developmentally comparable to the basidiospores and ascospores that are the

direct and obligatory developmental products of the cells in which karyogamy

and meiosis occur.

To obtain clean DNA and good sequence data from entomophthoroid fungi

may be more difficult than for many, much more extensively studied fungal

groups. Part of this difficulty might involve the physical organization of the

genome in these fungi that might lead to overlapping but divergent sequences

for some ‘needed’ genes. Chromosomal counts for Basidiobolus (which was

long treated as including only one species, B. ranarum) have ranged from about

60 (Olive 1907) to several hundred (Sun & Bowen 1972) based on kinetochore

counts in serial sections for transmission electron microscopy. These high

numbers suggest that polyploidization events may have occurred repeatedly

in Basidiobolus. This possibility seems to be verified by genetic studies showing

multiple, genetically distinct allelic forms in B. ranarum for elongation-

translation factor genes that usually occur in single copies in the genome

(Henk & Fisher 2012). The few chromosome counts for entomophthoraceous

entomopathogens also suggest a tendency to towards polyploidy: While the

nature of chromosomes and mitoses may not facilitate chromosomal counts

in Entomophthoraceae (Olive 1906, Humber 1975), the few published numbers

in various taxa are 8, 12 (or more), 16, and 32 (see Humber 1982). No genetic

studies like those of Henk & Fisher (2012) are available for Entomophthoraceae

but no genetic studies of this family with techniques ranging from allozyme

polymorphisms (Hajek et al. 1990; B. May, personal communication) to the

latest gene sequencing efforts suggest that these fungi simultaneously harbor

Entomophthoromycota phyl. nov. ... 489

multiple allelic variants at single loci; there is no indication that vegetative nuclei

of these fungi are either diploid or include heterologous sets of chromosomes.

The interpretation of such seemingly numerous chromosomes in some

taxa in Entomophthoromycota becomes more problematic in the absence of

evidence suggesting that any putatively sexual reproduction in this phylum is

heterothallic rather than strictly homothallic. No data support the invocation

of heterothallic sexuality (even if outbreeding events were extremely rare) to

explain Henk & Fisher's (2012) conclusions about the Basidiobolus genome.

Except for gametangial fusions during zygosporogenesis, no cellular fusions

(even between the naturally protoplastic vegetative cells of some of the

pathogenic taxa) are known from cultures or natural collections of any

entomophthoroid fungus; such fastidious behavior by these fungi precludes

consideration of heterokaryosis and parasexuality as a mechanism to increase

or to sustain gene flow among taxa in Entomophthoromycota.

It is important to note again that the Entomophthorales as traditionally

recognized (Humber 1989) is the same group reclassified here except for the

removal of Ballocephala and Zygnemomyces to Kickxellomycotina based on the

bifurcate, plugged septa in their vegetative hyphae (Saikawa 1989, Saikawa et al.

1997). Despite earlier doubts about retaining Basidiobolus in Entomophthorales,

molecular studies of more genes and a broader spectrum of entomophthoraleans

(A. Gryganskyi, R. Vilgalys, and R. Humber, unpublished) confirm that this

order, as historically treated, is monophyletic. These fungi exemplify yet

another major group for which the traditional, pre-molecular classification

has been fundamentally confirmed (although amplified and adjusted) rather

than overturned by phylogenetic analyses. Phylogenetic techniques must not

be allowed to override or to supplant the existing knowledge about groups

of organisms despite the vital inputs, seductively authoritative-looking

dendrograms, and current pre-eminence among taxonomy methodologies.

The best role for phylogenetic techniques should be as partners with the much

broader (and usually older) perspectives gained by a thorough understanding

of the overall biology as the means to determine the most sensible and best

supported organismal classifications.

Acknowledgments

Special thanks are due to Andrii Gryganskyi, Annette Bruun Jensen, Kathie T. Hodge,

and Bo Huang for the molecular results that underpin this new classification, to Walter

Gams and Rytas Vilgalys for critical reviews, and to James L. Reveal for stimulating

discussion of nomenclatural issues.

Literature cited

Balazy S. 1993. Entomophthorales. Flora of Poland (Flora Polska), Fungi (Mycota) 24: 1-356. Polish

Acad Sci, W Szafer Inst Botany, Krakow.

490 ... Humber

Butt TM, Humber RA. 1989. An immunofluorescence study of mitosis in a mite-

pathogen, Neozygites sp. (Zygomycetes: Entomophthorales). Protoplasma 151: 115-123.

http://dx.doi.org/10.1007/BF01403448

Cavalier-Smith T. 1998. A revised six-kingdom system of Life. Biol. Rev. 73: 203-266.

http://dx.doi.org/10.1017/S0006323198005167

Doweld AB. 2001. Prosyllabus Tracheophytorum, Tentamen Systematis Plantarum Vascularium

(Tracheophyta). Geos, Moscow.

Eidam E. 1886. Basidiobolus, eine neue Gattung der Entomophthoraceen. Beitr. Biol. Pflanzen 4:

181-241. http://dx.doi.org/10.1016/0022-2011(86)90060-1

Einax E, Voigt K. 2004. Oligonucleotide primers for the universal amplification of 6-tubulin

genes facilitate phyogenetic analyses in the regnum Fungi. Org. Divers. Evol. 3: 185-194.

http://dx.doi.org/10.1078/1439-6092-00069

Fresenius G. 1856. Notiz, Insecten-Pilze betreffrend. Bot. Zeitg. 14: 882.

Hajek AE, Humber RA, Elkinton JS, May B, Walsh SRA, Silver JC. 1990. Allozymes and restriction

fragment length polymorphism analyses confirm Entomophaga maimaiga responsible for

1989 epizootics in North American gypsy moth populations. Proc. Natl. Acad. Sci. USA 87:

6979-6982. http://dx.doi.org/10.1073/pnas.87.18.6979

Henk DA, Fisher MC. 2012. The gut fungus Basidiobolus ranarum has a large genome and different

copy numbers of putatively functionally redundant elongation factor genes. PLoS ONE 7:

e31268. http://dx.plos.org/10.1371/journal.pone.0031268

Hibbett DS, Binder M, Bischoff JE, Blackwell M, Cannon PE, Eriksson OE, Huhndorf S, James

T, Kirk PM, Liicking R, Lumbsch HT, Lutzoni F, Matheny PB, McLaughlin DJ, Powell

MJ, Redhead S, Schoch CL, Spatafora JW, Stalpers JA, Vilgalys R, Aime MC, Aptroot A,

Bauer R, Begerow D, Benny GL, Castlebury LA, Crous PW, Dai YC, Gams W, Geiser DM,

Griffith GW, Gueidan C, Hawksworth DL, Hestmark G, Hosaka K, Humber RA, Hyde KD,

Ironside JE, Koljalg U, Kurtzman CP, Larsson KH, Lichtwardt R, Longcore J, Miadlikowska

J, Miller A, Moncalvo JM, Mozley-Standridge S$, Oberwinkler FE, Parmasto E, Reeb V,

Rogers JD, Roux C, Ryvarden L, Sampaio JP, Schiifler A, Sugiyama J, Thorn RG, Tibell L,

Untereiner WA, Walker C, Wang Z, Weir A, Weiss M, White MM, Winka K, Yao YJ, Zhang

N. 2007. A higher-level phylogenetic classification of the Fungi. Mycol. Res. 111: 509-547.

http://dx.doi.org/10.1016/ j.mycres.2007.03.004

Humber RA. 1975. Aspects of the biology of an insect-parasitic fungus, Strongwellsea magna

(Zygomycetes: Entomophthorales). PhD dissertation, University of Washington, Seattle.

Humber RA. 1981. An alternative view of certain taxonomic criteria used in the Entomophthorales

(Zygomycetes). Mycotaxon 13: 191-240.

Humber RA. 1982. Strongwellsea vs. Erynia: the case for a phylogenetic classification of the

Entomophthorales (Zygomycetes). Mycotaxon 15: 167-184.

Humber RA. 1984. Foundations for an evolutionary classification of the Entomophthorales

(Zygomycetes). 166-183, in: Q Wheeler, M Blackwell (eds). Fungus/insect relationships:

perspectives in ecology and evolution. Columbia University Press, New York.

Humber RA. 1989. Synopsis of a revised classification for the Entomophthorales (Zygomycotina).

Mycotaxon 34: 441-460.

James TY, Kauff FE, Schoch C, Matheny PB, Hofstetter V, Cox CJ, Celio G, Geuidan C, Fraker E,

Miadlikowska J, Lumbsch HT, Rauhut A, Reeb V, Arnold AE, Amtoft A, Stajich JE, Hosaka

K, Sung G-H, Johnson D, O’Rourke B, Crockett M, Binder M, Curtis JM, Slot JC, Wang Z,

Wilson AW, SchiiSler A, Longcore JE, O’Donnell K, Mozley-Standridge S, Porter D, Letcher

PM, Powell MJ, Taylor JW, White MW, Griffith GW, Davies DR, Humber RA, Morton JB,

Sugiyama J, Rossman A, Rogers JD, Pfister DH, Hewitt D, Hansen K, Hambleton S, Shoemaker

Entomophthoromycota phy]. nov. ... 491

RA, Kohlmeyer J, Volkmann-Kohlmeyer B, Spotts RA, Serdani M, Crous PW, Hughes KW,

Matsuura K, Langer E, Langer G, Untereiner WA, Liicking R, Biidel B, Geiser DM, Aptroot A,

Diederich P, Schmitt I, Schultz M, Yahr R, Hibbett DS, Lutzoni EF, McLaughlin DJ, Spatafora

JW, Vilgalys R. 2006. Reconstructing the early evolution of Fungi using a six-gene phylogeny.

Nature 443: 818-822. http://dx.doi.org/10.1038/nature05110

Jensen AB, Eilenberg J. 2001. Genetic variation with the insect-pathogenic genus Entomophthora,

focusing on the E. muscae complex, using PCR-RFLP of the ITS II and the LSU rDNA. Mycol.

Res. 105: 307-312. http://dx.doi.org/10.1017/S0953756201003434

Jensen AB, Gargas A, Eilenberg J, Rosendahl S. 1998. Relationships of the insect-pathogenic

order Entomophthorales (Zygomycota, Fungi) based on phylogenetic analyses of nuclear

small subunit ribosomal DNA sequences (SSU rDNA). Fung. Genet. Biol. 24: 325-334.

http://dx.doi.org/10.1006/fgbi.1998.1063

Keeling PJ. 2003. Congruent evidence from a-tubulin and 6-tubulin gene phylogenies for a

zygomycete origin of microsporidia. Fung. Genet. Biol. 38: 298-309.

http://dx.doi.org/10.1016/S1087-1845(02)00537-6

Keller S. 1987. Arthropod-pathogenic Entomophthorales of Switzerland. I. Conidiobolus,

Entomophaga, and Entomophthora. Sydowia 40: 122-167.

Keller S. 1991. Arthropod-pathogenic Entomophthorales of Switzerland. II. Erynia, Eryniopsis,

Neozygites, Zoophthora, and Tarichium. Sydowia 43: 39-122.

Keller S. 1997. The genus Neozygites (Zygomycetes, Entomophthorales) with special reference to

species found in tropical regions. Sydowia 49: 118-146.

Keller S, Petrini O. 2005. Keys to the identification of the arthopod pathogenic genera of the

families Entomophthoraceae and Neozygitaceae (Zygomycetes), with descriptions of three new

subfamilies and a new genus. Sydowia 57: 23-53.

Liu X-Y, Voigt K. 2010. Molecular characters of zygomycetous fungi. In: Molecular identification of

Fungi. Y Gherbawy, K. Voigt (eds). Springer-Verlag, Berlin.

http://dx.doi.org/10.1007/978-3-642-05042-8 20

McCabe DE, Humber RA, Soper RS. 1984. Observation and interpretation of nuclear reductions

during maturation and germination of entomophthoralean resting spores. Mycologia 76:

1104-1107. http://dx.doi.org/10.2307/3793025

McKerracher LJ, Heath IB. 1985. The structure and cycle of the nucleus-associated organelle in two

species of Basidiobolus. Mycologia 77: 412-417. http://dx.doi.org/10.2307/3793197

Nahaghama JT, Sato H, Shimazu M, Sugiyama J. 1995. Phylogenetic divergence of the

entomophthoralean fungi: Evidence from nuclear 18S ribosomal RNA gene sequences.

Mycologia 87: 203-209. http://dx.doi.org/10.2307/3760906

Nielsen C, Sommer C, Eilenberg J, Hansen KS, Humber RA. 2001. Characterization of aphid

pathogenic species in the genus Pandora by PCR techniques and digital image analysis.

Mycologia 93: 864-874. http://dx.doi.org/10.2307/3761752

Olive EW. 1906. Cytological sudies on the Entomphthoreae. II. Nuclear and cell division of Empusa.

Bot. Gaz. 41: 229-261. http://dx.doi.org/10.1086/328797

Olive EW. 1907. Cell and nuclear division in Basidiobolus Ann. Mycol. 5: 404-418.

Robinow CE. 1963. Observations on cell growth, mitosis, and division in the fungus Basidiobolus

ranarum. J. Cell Biol. 17: 123-152. http://dx.doi.org/10.1083/jcb.17.1.123

Saikawa M. 1989. Ultrastructure of the septum in Ballocephala verrucospora (Entomophthorales,

Zygomycetes). Can. J. Bot. 67: 2484-2488. http://dx.doi.org/10.1139/b89-318

Saikawa M, Oguchi M, Castafieda Ruiz RE 1997. Electron microscopy of two nematode-

destroying fungi, Meristacrum asterospermum and Zygnemomyces echinulatus (Meristacraceae,

Entomophthorales). Can. J. Bot. 75: 762-768. http://dx.doi.org/10.1139/b97-086

492 ... Humber

Sun NC, Bowen CC. 1972. Ultrastructural studies of nuclear division in Basidiobolus ranarum

Eidam. Caryologia 25: 471-494.

Tanabe Y, O'Donnell K, Saikawa M, Sugiyama J. 2000. Molecular phyogeny of parasitic Zygomycota

(Dimargaritales, Zoopagales) based on nuclear small subunit ribosomal DNA sequences. Mol.

Phylogenet. Evol. 16: 253-262. http://dx.doi.org/10.1006/mpev.2000.0775

Tanabe Y, Saikawa M, Watanabe MM, Sugiyama J. 2004. Molecular phylogeny of Zygomycota based

on EF-la and RPB1 sequences: limitations and utility of alternative markers to rDNA. Mol.

Phylogenet. Evol. 30: 438-449. http://dx.doi.org/10.1016/S1055-7903(03)00185-4

Tanabe Y, Watanabe MM, Sugiyama J. 2005. Evolutionary relationships among basal fungi

(Chytridiomycota and Zygomycota): Insights from molecular phylogenetics. J. Gen. Appl.

Microbiol. 51: 267-276. http://dx.doi.org/10.2323/jgam.51.267

Tanaka K. 1978. Mitosis in the fungus Basidiobolus ranarum revealed by electron microscopy.

Protoplasma 70: 423-440. http://dx.doi.org/10.1007/BF01275768

Voigt K, Kirk PM. 2011. Recent developments in the taxonomic affiliation and phylogenetic

positioning of fungi: impact in applied microbiology and environmental biotechnology. Appl.

Microbiol. Biotechnol. 90: 41-57. http://dx.doi.org/10.1007/s00253-011-3143-4

White MM, James TY, O'Donnell K, Cafaro MJ, Tanabe Y, Sugiyama J. 2006. Phylogeny

of the Zygomycota based on nuclear ribosomal sequence data. Mycologia 98: 885-895.

http://dx.doi.org/10.3852/mycologia.98.6.872

NOTE ADDED IN PROOF: Since the acceptance of this article, the bibliographic

citations for the two molecularly based papers that underpin and justify this new

classification of entomophthoroid fungi have become available:

Gryganskyi AP, Humber RA, Smith ME, Miadlikovska J., Wu S, Voigt K, Walther G, Anishchenko

IM, Vilgalys R. 2012. Molecular phylogeny of the Entomophthoromycota. Mol. Phylog. Evol.

65: 682-694. http://dx.doi.org/10.1016/j.ympev.2012.07.026

Gryganskyi AP, Humber RA, Smith ME, Hodge K, Huang B, Voigt K, Vilgalys R. 2012. Phylogenetic

lineages in Entomophthoromycota. Persoonia: in press.

MY COTAXON

http://dx.doi.org/10.5248/120.493

Volume 120, pp. 493-503 April-June 2012

Moniliophthora aurantiaca sp. nov.,

a Polynesian species occurring in littoral forests

BRADLEY R. Kropp?* & STEVEN ALBEE-SCOTT?”

‘Biology Department, 5305 Old Main Hill, Utah State University, Logan, Utah 84322 USA

?Wilbur L. Dungy Endowed Chair of the Sciences, Environmental Sciences,

Jackson Community College, 2111 Emmons Road, Jackson, MI USA

*CORRESPONDENCE TO: brad.kropp@usu.edu

ABSTRACT — A new species of Moniliophthora is described from the Samoan Islands. The

new species is characterized by its bright orange pileus and pale orange stipe and lamellae. It

occurs commonly on woody debris in moist littoral forests and has not been found in upland

forests. A phylogenetic analysis of nLSU and ITS sequences indicates that Moniliophthora

aurantiaca has an affinity with the Central and South American members of the genus.

Possible mechanisms for the dispersal of fungi from the Neotropics to the Samoan Islands

are discussed.

Key worps — Agaricales, American Samoa, Crinipellis, Oceania, phylogeny

Introduction

The mycobiota of the Samoan Islands has received very little attention.

What work that has been done consists of an inventory of the wood decay

fungi and plant pathogens present in American Samoa (Brooks 2004, 2006)

along with the recent description of an Inocybe species from the island of Tau

(Kropp & Albee-Scott 2010). Our preliminary work on Samoan fungi indicates

that the mycoflora of the islands is potentially quite rich and that a number of

undescribed species is present.

An attractive, bright orange species of Moniliophthora was recently

discovered in littoral forest on the islands of Tutuila and Tau, American Samoa.

In spite of its distinctive appearance and the fact that it appeared to be very

common at the time the material was collected, no name has been found for

this fungus among the published species of the Moniliophthora/Crinipellis

complex. In this article, we propose a new species of Moniliophthora from the

Samoan Islands and assess its phylogenetic position relative to other members

of Moniliophthora and to members of Crinipellis and Chaetocalathus.

494 ... Kropp & Albee-Scott

Materials & methods

Specimens were photographed in the field on natural substrates or using a gray

card as a background and field notes were taken while the material was still fresh. The

collections were then dried for herbarium specimens. Microscopic study of the dried

specimens was carried out in the laboratory using a light microscope after rehydrating

tissue sections in Melzer’s reagent. Microscopic characters were measured using oil

immersion at 1000x except for pileus hairs, which because of their length were made

at lower magnifications. All microscopical characters were illustrated with the aid of

a drawing tube. Spore measurements are given as an average with ranges, whereas

measurements of the other cells are given as ranges. Color notations were taken from

the digital photographs using the Munsell Soil Color Charts (Munsell Color 2000) and

Farver i Farver (Wanscher & Kornerup 1991). Formats for the Munsell and Farver i

Farver color notations are 5YR 8/8 and 54A respectively. All specimens examined,

including the holotype, have been accessioned into the Intermountain Herbarium

(UTC) at Utah State University.

DNA was extracted from dried herbarium material using standard protocols adapted

for use in our lab (Kropp et al. 1996). Amplified PCR products were obtained using

standard PCR protocols (White et al. 1990) for the internal transcribed spacer (ITS)

using primers ITS4 and ITS5 (White et al. 1990) and for the nuclear large ribosomal

subunit (nLSU) between primers LROR and LR5 (Moncalvo et al. 2000). Direct

sequencing of the purified PCR products was done for both the nLSU and the ITS using

the amplification primers. Both sequences for M. aurantiaca were deposited in GenBank

(ITS = JN692482, nLSU = JN692483).

Twenty-one species of Crinipellis representing Europe, Australasia, and North and

South America were sampled. Taxon sampling also included four Moniliophthora species

from Australasia, North America, and the American tropics. To identify apomorphies,

members of the closely related Chaetocalathus and representatives of the less closely

related Tetrapyrgos and Marasmius were used as outgroups in our analysis (TABLE I).

Clustal X (Thompson et al. 1997) was used to align the nLSU and ITS sequences

after they were concatenated in the phylogenetically uninformative terminal ends.

MrBayes 3.1 was used to analyze the aligned data set, 1613 bases in length (Ronquist &

Huelsenbeck 2003). Treespace was searched using a time reversible model of evolution

(Maddison 1994, Rodriguez et al. 1990) and a discrete gamma distribution with six

substitution types and some invariant sites (GTR+G+I). Every hundred trees were

sampled from the Bayesian simulation to approximate posterior probabilities. The

trees were simulated using the Markov Chain Monte Carlo Method (MCMC) and all

Bayesian simulations were conducted with eight active MCMC chains, heated at 0.2, and

started with a randomly chosen neighbor-joining tree. The first MCMC run was iterated

for 1,000,000 generations and three subsequent MCMC simulations were done using

1,000,000 generations, sampling every one hundredth tree. A majority consensus tree

was calculated from the last 7000 sampled trees from a 10,000 tree data set using all runs

to recover the posterior probabilities of the internal nodes using the sumT command in

MrBayes. All four consensus trees recovered from the four Bayesian simulations were

phylogenetically identical. The resulting statistics showed that the two independent

MCMC chains converged with a standard deviation <0.001 (Ronquist & Huelsenbeck

TABLE 1. Species, geographic origins, and GenBank accession numbers

TAXON

Chaetocalathus columellifer (Berk.) Singer

Ch. cf. columellifer

Ch. craterellus (Durieu & Lév.) Singer

Ch. fragilis (Pat.) Singer

Ch. galeatus (Berk. & M.A. Curtis) Singer

Ch. liliputianus (Mont.) Singer

Ch. magnus Halling

Crinipellis actinophora (Berk. & Broome) Singer

Cr. brasiliensis Arruda et al.

Cr. brunneipurpurea Corner

Cr. brunnescens Kerekes & Desjardin

Cr. campanella (Peck) Singer

Cr. cupreostipes Kerekes et al.

Cr. dipterocarpi Singer

Cr. furcata Kerekes et al.

Cr. cf. iopus Singer

Cr. malesiana Kerekes et al.

Cr. maxima A.H. Sm. & M.B. Walters

Cr. piceae Singer

Cr. procera G. Stev.

Cr. scabella (Alb. & Schwein.) Murrill

Cr. setipes (Peck) Singer

Cr. stipitaria (Fr.) Pat.

Crinipellis sp.

Cr. tabtim Kerekes et al.

Cr. trichialis (Lév.) Pat.

Cr. zonata (Peck) Sacc.

Marasmius apatelius Singer

M. leucorotalis Singer

M. rotula (Scop.) Fr.

Moniliophthora aurantiaca

M. canescens (Har. Takah.) Kerekes & Desjardin

M. perniciosa (Stahel) Aime & Phillips-Mora

M. roreri (Cif) H. C. Evans et al.

Moniliophthora sp.

Tetrapyrgos nigripes (Fr.) Horak

T. subcinerea (Berk. & Broome) Horak

T. subdendrophora (Redhead) Horak

ORIGIN

Malaysia

Ecuador

Italy

Thailand

Puerto Rico

Colombia

Malaysia

Brazil

Indonesia

Thailand

Indonesia

Papua NG

Malaysia

N. America

New Zealand

Europe

Thailand/USA

Germany

Thailand

Guyana

Thailand

Indonesia

Canada

Thailand

Malaysia

N. America

Samoa

Malaysia

Peru

Costa Rica

N. America

S. E. Asia

N. America

ITS

FJ167665

AY916686

FJ167664

FJ 167662

FJ167663

AY916682

FJ167666

FJ167626

AY317137

FJ167646

FJ 167627

FJ167641

FJ167651

FJ167658

FJ167639

FJ167629

FJ167633

FJ167660

FJ167635

FJ167634

AY571033

AY916698

AY916701

FJ167643

FJ167609

AY916692

EU935561

FJ431253

DQ182506

JN692482

FJ 167668

AF335590

AY916746

AY916754

DQ449942

EF175552

EF175521

* ITS sequence from a German collection, nLSU from an Estonian collection

**ITS sequence originated from a Thai collection, nLSU sequence from a North American collection.

Moniliophthora aurantiaca sp. nov. (Samoa) ... 495

nLSU

FJ167665

AY916680

AF042647

AF042630

AM946420*

AY916689**

AY207194

AY916690

DQ457686

JN692483

AY916744

AF261337

496 ... Kropp & Albee-Scott

2003). The potential scale reduction factors for all convergence statistics approached

1.001 for all parameters. Posterior probability support measures <50% are not shown.

Tetrapyrgos was used to root the consensus tree.

Results

The results of the Bayesian analysis show that the undescribed species

clusters with the Moniliophthora clade, which has very good posterior

probability support (Fic. 1). Certain other taxa, such as Crinipellis aff. iopus and

Crinipellis brasiliensis (which have not yet been transferred to Moniliophthora),

also clustered within this clade. Our analysis indicates that M. aurantiaca

has a stronger affinity with the neotropical members of the clade such as

M. roreri than it does with the Australasian representatives such as M.

canescens or Crinipellis aff. iopus. Moniliophthora aurantiaca is closely related to

M. roreri, which has a neotropical distribution. In addition, two other neotropical

taxa, M. perniciosa and Crinipellis brasiliensis, appear to be close relatives of

M. aurantiaca based on our analyses.

The species of Chaetocalathus and Marasmius formed two well-supported

clades basal to Crinipellis and Moniliophthora. In our analysis, the members of

Moniliophthora occupy a clade derived from within Crinipellis. Based on these

results, accepting Moniliophthora as a genus renders Crinipellis paraphyletic,

although the node separating Crinipellis clade 1 from Crinipellis clade 2 and

Moniliophthora is not well supported (Fie. 1).

Discussion

Moniliophthora aurantiaca is an unusually conspicuous element of the

mycobiota of American Samoa. It stands out primarily because of its bright

orange colors but also because it fruits commonly in littoral forests. ‘Thus far, it

has been found only in a narrow strip of damp littoral forest situated along the

shoreline on the islands of Tutuila and Ta’u, American Samoa. In one instance,

it was found among trees growing on a rocky outcrop a few meters above the

water but still at the shoreline. That M. aurantiaca was never found outside

the littoral zone in spite of an abundance of apparently suitable moist woody

substrates in nearby higher elevation forests suggests a halophilic or maritime

beach ecology. Known thus far only from American Samoa, M. aurantiaca may

be endemic to the Samoan Islands, but its distribution is still poorly known and

further work is needed to confirm whether it is really an endemic species.

The species belongs to the Moniliophthora/Crinipellis complex that currently

comprises five Moniliophthora species and 163 Crinipellis species (http.//www.

indexfungorum.org/). Of these, M. aurantiaca macroscopically most closely

resembles the South American species Crinipellis ticoi Halling, which is also

orange in color. Despite their macroscopic similarities, C. ticoi differs from

M. aurantiaca microscopically in larger spores (12.1-14.3 x 5-7.1 um) and

Moniliophthora aurantiaca sp. nov. (Samoa) ... 497

Tetrapyrgos nigripes

Tetrapyrgos subcinerea

Tetrapyrgos subdendrophora

Marasmius apatelius

Marasmius leucorotalis

0.78 L. Marasmuus rotula

0.94 Chaetocalathus cf columellifer

Chaetocalathus magnus

Chaetocalathus columellifer

1.00 7 Chaetocalathus craterellus

A068 Chaetocalathus galeatus

0.97 | - Chaetocalathus fragilis

1.00 | Chaetocalathus liliputianus

Crinipellis zonata (Canada)

0.99 | 1.00 Crinipellis procera (New Zealand)

sche Crinipellis furcata (Indonesia)

Crinipellis campanella

Crinipellis maxima

Moniliophthora sp (North America)

snyjepesojovyD

T sijedruna

0.63 Crinipellis aff iopus (Papua New Guinea) =

oe Moniliophthora canescens (Malaysia) =

ae Moniliophthora aurantiaca (Samoa) ep

1.00 ti Moniliophthora roreri (Costa Rica) a

1.00 Moniliophthora perniciosa (Central America) S

0.80 1.00 Crinipellis brasiliensis (Brazil) 4

1.00 + Crinipellis malesiana (Malaysia)

o99| | Crinipellis sp (Thailand)

Crinipellis trichialis (Indonesia)

100] 1 Crinipellis scabella (Europe)

1.00 § Crinipellis stipitaria (Germany)

1.00 |, oor Crinipellis piceae (North America)

Crinipellis setipes (Thailand)

Crinipellis brunneipurpurea (Indonesia)

098 |_| 1.00 r Crinipellis sp (Guyana)

Crinipellis actinophora (Malaysia)

Crinipellis brunnescens (Indonesia)

0.65 Crinipellis cupreostipes (Thailand)

Crinipellis dipterocarpi (Thailand)

1.00 L_ Crinipellis tabtim (Thailand)

Z siffediursD

1.00

0.4 1.00

FiGuRE 1. Phylogram from a Bayesian analysis of a combined ITS and nLSU dataset for Crinipellis,

Moniliophthora, and Chaetocalathus. Support measures are shown for nodes with 60 percent or

greater posterior probability support. Members of Tetrapyrgos were used to root the phylogram.

cheilocystidia that lack the apical appendages present on the cheilocystidia

of M. aurantiaca (Fic. 3). One other orange Crinipellis is the African

C. hygrocyboides (Henn.) Singer. Neither Hennings (1902) nor Singer (1989)

gives microscopic details or a detailed description for this fungus, making

comparisons with other taxa difficult. Since Halling (1993) reports that spores

and cystidia were not observed in the type material of C. hygrocyboides from

the Muséum National d’Histoire Naturelle, we have studied isotype material of

C. hygrocyboides (=Marasmius hygrocyboides Henn.) from the Royal Botanical

Gardens, Kew. The specimen resembles M. aurantiaca but differs by having a

mixture of long dextrinoid hairs and clavate, thin-walled dextrinoid cystidia

on the stipe surface and similar, but slightly smaller, spores. We were unable to

498 ... Kropp & Albee-Scott

reliably study cheilocystidia in the C. hygrocyboides material due to difficulty

reviving the material. However, all cells at the lamellar edges appeared to

be clavate and different from the cheilocystidia with apical appendages of

M. aurantiaca. It would be very useful to have molecular data for this species

to determine whether it belongs to the Moniliophthora clade. Another similar

taxon is M. canescens, which has orange hues when wet but is typically brownish;

additional differences include an initially white stipe that becomes brown and

narrower spores (Takahashi 2000). Crinipellis malesiana, which also develops

orange colors in age (Kerekes & Desjardin 2009), is phylogenetically separate

from M. aurantiaca (Fic. 1).

Based on our phylogenetic analysis, M. aurantiaca falls within the

Moniliophthora clade of Aime & Phillips-Mora (2005). These workers

found that fungi belonging to Moniliophthora, which had been used for an

anamorphic pathogen of cocoa (Evans et al. 1978), are not only closely allied to

certain members of Crinipellis but form a distinct lineage with Crinipellis in the

Marasmiaceae for which they proposed using Moniliophthora as a generic name.

The genus currently includes both conidial and agaricoid taxa. The agaricoid

members of Moniliophthora are usually relatively brightly colored species that

would otherwise have been placed in section lopodinae of Crinipellis.

In contrast, our analysis indicates that Moniliophthora does not occupy a

distinct lineage but is derived from within Crinipellis. In this case, accepting

Moniliophthora as a genus makes Crinipellis paraphyletic (Fic. 1). Based on

our results, one could abandon Moniliophthora and transfer everything in it,

including the anamorphic species, to Crinipellis. However, one cannot ignore

that the ITS analysis of Kerekes & Desjardin (2009) and the 5-gene phylogeny

of Aime & Phillips-Mora (2005) both agree that the Moniliophthora clade forms

a distinct lineage. In addition, the node separating the two Crinipellis clades

shown in our phylogram has weak posterior probability support (Fic. 1). Given

the discrepancy between our analysis and the other two studies, further work

is needed to clarify relationships in the Moniliophthora/Crinipellis complex.

For now, the preponderant information indicates that the Moniliophthora clade

forms a distinct lineage and until further work is done we retain Moniliophthora

as a genus in the sense of Aime & Phillips-Mora (2005).

Caution must be used when using our phylogram to identify biogeographical

patterns. Nonetheless, the close affinity of M. aurantiaca to the pathogenic South

and Central American members of the Moniliophthora clade is well supported

and implies that either M. aurantiaca or an ancestral taxon originated in the

neotropics (Fic. 1). This is somewhat unexpected, because we earlier found

that another Samoan fungus, Inocybe tauensis Kropp & Albee-Scott (2010), has

paleotropical roots and because Samoan plant communities have Australasian

ties (Van Balgooy et al. 1996).

Moniliophthora aurantiaca sp. nov. (Samoa) ... 499

Regardless of where M. aurantiaca originated, long-distance dispersal was

clearly involved because the islands are relatively young and have never been

connected to a land mass (McDougall 1985). In recent times, humans could have

carried M. aurantiaca to the Samoan Islands on plants or in woody materials

from almost any part of the world. However, long-distance dispersal might also

have occurred via other well-known mechanisms. Rafting is one potential means

of transport because M. aurantiaca grows in woody debris and could be carried

long distances by floating wood. Rafting is known to transport organisms and is

thought to have resulted in the dispersal and subsequent cladogenesis of snails

across large areas of the Pacific Ocean, including an endemic Samoan snail

(Donald et al. 2005). In another instance, an oyster species is thought to have

been carried via rafting from Chile to New Zealand (Foighil et al. 1999). Thus,

a plausible mechanism exists for the dispersal of fungi from the Neotropics to

Samoa. The mycelium of M. aurantiaca could have been carried within woody

material from South America to Samoa via the South Equatorial Current that

has a general westward flow across tropical portions of the South Pacific. Air-

borne spores are another means by which agarics could potentially cross the

Pacific and it is clear that fungal spores can travel long distances with air masses

(Brown & Hovmeller 2002). If M. aurantiaca has neotropical roots, then its

spores or those of an ancestral taxon would have needed to cross the Pacific

from the east to get to the Samoan Islands. ‘The easterly trade winds of the

equatorial South Pacific provide another plausible mechanism by which this

could have happened.

Taxonomy

Moniliophthora aurantiaca Kropp & Albee-Scott, sp. nov. Fics 2, 3

MyYcoBANK MB563345

Differs from Crinipellis ticoi by its smaller basidiospores and cheilocystidia with apical

appendages and from Crinipellis hygrocyboides by its short, moderately thick-walled

setae-like stipe hairs and the absence of clavate caulocystidia.

Type: American Samoa, National Park of American Samoa, Tau Unit, in littoral forest

along trail to Siu point, on woody debris, 17 May 2009, leg. B.R. Kropp BK17-May-

2009-20a (Holotype UTC253631; GenBank JN692482, JN692483).

ETYMOLOGy: aurantiaca refers to the bright orange coloration of the pileus.

PiLEus 3-15 mm diameter, convex with margin somewhat inrolled at first then

broadly convex to nearly plane, disc with a small umbo, sometimes slightly

depressed around the umbo, translucent-striate or slightly sulcate from the

margin nearly to the disk, surface matted fibrillose; bright red orange (78A,

2.5YR 7/12) at the center, becoming pale orange (54A, 5YR 8/8) toward

the margin especially in age; when dry, pale orange with the umbo darker.

LAMELLAE narrowly attached, subdistant, pale orange (65A, 5YR 8/8), very

500 ... Kropp & Albee-Scott

FiGuRE 2. Moniliophthora aurantiaca. Basidiomata: A. UTC253824; B. UTC253809. Scale = 1 cm.

pale orange when dry; lamellulae in 1-2 series. Stipe 5-8 x 0.25-1 mm,

central, terete, equal, densely pruinose, concolorous with the lamellae except

for the base which is often brown, extreme base on some specimens strigose,

stipe inserted for most specimens. PILEIPELLIs a cutis of fairly loosely woven

light brown, lightly encrusted hyphae 5-8 um wide, not well differentiated

Moniliophthora aurantiaca sp. nov. (Samoa) ... 501

FiGuRE 3. Moniliophthora aurantiaca. A. Basidiospores B. Basidia C. Stipe hairs D. Pileipellis hairs

E. Basidioles F. Cheilocystidia. Scales: D = 50 um; A-C, E, F = 10 um.

from the pileus context; numerous long, dextrinoid hairs present, these with

a basal clamp and 400-600 x 4-5 um, mostly hyaline not greenish in KOH;

context not dextrinoid. LAMELLAR TRAMA subparallel to interwoven; hyphae

hyaline sometimes lightly encrusted, clamped, 4-8 um wide; context not

dextrinoid. STIPE TISsUE parallel, smooth, hyaline, hyphae 4-8 um wide;

dextrinoid to weakly dextrinoid; stipe hairs numerous, covering the stipe apex

and base, dextrinoid to weakly dextrinoid, short and moderately thick-walled,

resembling setae, 52-85 x 5-10 um. BASIDIOSPORES 7.5-(8.7)-11.0 x 4-(4.6)-

6.0 um, Q = 1.8-2.1, amygdaliform, thin-walled, smooth, hyaline, inamyloid.

BASIDIA 26-30 x 6-7 um, clavate, typically four-spored. BASIDIOLES 22-25 x

4-5 um, clavate. PLEUROCYSTIDIA none. CHEILOCYSTIDIA 20-22 x 5-8 um,

numerous, hyaline, thin-walled, typically with several irregular apical, finger-

like appendages. CLamps present. RHIZOMORPHS absent.

HABITAT AND DISTRIBUTION occurring on fallen twigs and other woody

material in littoral forest.

ADDITIONAL SPECIMENS EXAMINED—AMERICAN SAMOA. TuTUILA, near Vaitogi,

on woody debris in wooded area near shoreline, 11 May 2009, leg. B.R. Kropp, BK11-

May-2009-11 (UTC253809); National Park of American Samoa, Tutuila Unit, on woody

debris in littoral forest along trail to Pola Island, 12 May 2009, leg. B.R. Kropp, BK12-

502 ... Kropp & Albee-Scott

May-2009-12 (UTC253824); BK12-May-2009-16 (UTC253828); BK12-May-2009-18

(UTC253830); BK12-May-2009-17 (UTC253829). Ta’u, National Park of American

Samoa, Tau Unit, on woody debris in littoral forest along trail to Siu point, 15 May

2009, leg. B.R. Kropp, BK15-May-2009-10a (UTC255959).

Acknowledgements

The National Park of American Samoa and the Department of Marine and Wildlife

Resources in American Samoa are thanked for facilitating the necessary collecting

permits and for providing helpful advice about collecting within the national park. Dr.

M.C. Aime and Dr. T.J. Baroni are gratefully acknowledged for their peer reviews of this

article.

Literature cited

Aime MC, Phillips-Mora W. 2005. The causal agent of witches’ broom and frosty pod of cacao

(chocolate, Theobroma cacao) form a new lineage of Marasmiaceae. Mycologia 97: 1012-1022.

http://doi.dx.org/10.3852/mycologia.97.51012

Brooks E. 2004. Wood decay fungi: pests and diseases of American Samoa. Land Grant Technical

Report No. 11, American Samoa Community College. 2 p.

Brooks FE. 2006. List of Plant Diseases in American Samoa. Land Grant Technical Report No. 44,

American Samoa Community College. 59 p.

Brown JKM, Hovmeller MS. 2002. Aerial dispersal of pathogens on the global and continental

scales and its impact on plant disease. Science 297: 537-541.

http://doi.dx.org/10.1126/science.1072678

Donald KM, Kennedy M, Spencer HG. 2005. Cladogenesis as the result of long-distance

rafting events in South Pacific topshells (Gastropoda, Trochidae). Evolution 59: 1701-1711.

http://doi.dx.org/10.1554/04-553.1

Evans HC, Stalpers JA, Samson RA, Benny GL. 1978. On the taxonomy of Monilia roreri, an

important pathogen of Theobroma cacao in South America. Canadian Journal of Botany 56:

2528-2532. http://doi.dx.org/10.1139/b78-305

Foighil D, Marshall BA, Hilbish TJ, Pino M. 1999. Trans-Pacific range extension by rafting

is inferred for the Flat Oyster Ostrea chilensis. The Biological Bulletin 196: 122-126.

http://doi.dx.org/10.2307/1542557

Halling R. 1993. Two new Crinipellinae (Tricholomataceae: Marasmiae) from South America.

Mycotaxon 47: 379-385.

Hennings P. 1902. Fungi camerunensis novi. III. Botanische Jahrbiicher fiir Systematik 30: 39-57.

Kerekes JF, Desjardin DE. 2009. A monograph of the genera Crinipellis and Moniliophthora from

Southeast Asia including a molecular phylogeny of the nrITS region. Fungal Diversity 37:

101-152.

Kropp BR, Albee-Scott S. 2010. Inocybe tauensis, a new species from the Samoan Archipelago

with biogeographic evidence for a Paleotropical origin. Fungal Biology 114: 790-796.

http://doi.dx.org/10.1016/j.funbio.2010.07.005

Kropp BR, Hansen D, Flint KM, Thomson SV. 1996. Artificial inoculation and colonization of

Dyer’s Woad (Isatis tinctoria) by the systemic fungus Puccinia thlaspeos. Phytopathology 86:

891-896. http://doi.dx.org/10.1094/Phyto-86-891

McDougall I. 1985. Age and evolution of the volcanoes of Tutuila, AS. Pacific Science 39:

311-320.

Moniliophthora aurantiaca sp. nov. (Samoa) ... 503

Maddison DR. 1994. Phylogenetic methods for inferring the evolutionary history and processes

of change in discretely valued characters. Annual Review of Entomology 39: 267-292.

http://doi.dx.org/10.1146/annurev.ento.39. 1.267

Moncalvo J-M, Lutzoni FM, Rehner SA, Johnson J, Vilgalys R. 2000. Phylogenetic relationships of

agaric fungi based on nuclear large subunit ribosomal DNA sequences. Systematic Biology. 49:

278-305. http://doi.dx.org/10.1093/sysbio/49.2.278

Munsell Color. 2000. Munsell soil color charts. Gretag Macbeth, New Windsor, NY.

Rodriguez F, Oliver JL, Marin A, Medina JR. 1990. The general stochastic model of nucleotide

substitution. Journal of Theoretical Biology 142: 485-501.

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. http://doi.dx.org/10.1093/bioinformatics/btg180

Singer R. 1989. New taxa and new combinations of Agaricales (Diagnoses fungorum novorum

Agaricalium IV). Fieldiana, Botany New Series 21: 1-133.

http://doi.dx.org/10.5962/bhI.title.2537

Takahashi H. 2000. Three new species of Crinipellis found in Iriomote Island, southwestern Japan,

and central Honshu, Japan. Mycoscience 41: 171-182. http://doi.dx.org/10.1007/BF02464328

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The ClustalX windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 24: 4876-4882. http://doi.dx.org/10.1093/nar/25.24.4876

Van Balgooy MMJ, Hovenkamp PH, Van Welzen PC. 1996. Phytogeography of the Pacific — floristic

and historical distribution patterns in plants. 191-213, in: A Keast, SE Miller (eds). The origin

and evolution of Pacific island biotas, New Guinea to Eastern Polynesia: patterns and processes.

SPB Academic Publishing, Amsterdam.

Wanscher JH, Kornerup A. 1991. Farver i farver: bestemmelse af farver. Politikens Forlag,

Copenhagen. 248 p.

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR protocols: a guide to

methods and applications. Academic Press, Inc., New York.

MYCOTAXON

http://dx.doi.org/10.5248/119.505

Volume 120, pp. 505-506 April-June 2012

Regional annotated mycobiotas new to the Mycotaxon website

ABASTRACT — Five new species distribution lists are added to the MycoTaxon ‘web-

list’ page covering Coprinaceae & Strophariaceae and Bolbitiaceae & Crepidotaceae in

Argentina (both by Niveiro & Albert6); macromycetes from Collestrada forest ecosystems

in Perugia, Italy (by Angelina & al.); lichens from Pollino National Park, Italy (by

Puntillo & al.); and saprobes on Alnus alnobetula in the Swiss alps (by Senn-Irlet & al.).

This brings to 94 the free access mycobiotas now available on the MycoTaxon website

<http://www.mycotaxon.com/resources/weblists.htm]>.

SOUTH AMERICA

Argentina

Niveiro, N.; Alberté, E. Checklist of the Argentinean Agaricales 2. Coprinaceae and

Strophariaceae. 38 p.

Asstract — A checklist of species belonging to the families Coprinaceae and

Strophariaceae was made for Argentina. The list includes all species published till year

2011. Twenty-one genera and 251 species were recorded, 121 species from the family

Coprinaceae and 130 from Strophariaceae.

Niveiro N. & E. Alberté. Checklist of the Argentine Agaricales 3. Bolbitiaceae and

Crepidotaceae. 28 p.

Asstract — A checklist of Agaricales species belonging to Bolbitiaceae and

Crepidotaceae has been made for Argentina. The list included all species published

until 2011. Fourteen genera and 187 species are recorded, 94 species from the

Bolbitiaceae and 93 from Crepidotaceae.

EUROPE

Italy

Angelini, Paola, Giancarlo Bistocchi, Andrea Arcangeli & Roberto Venanzoni.

Preliminary checklist of the macromycetes from Collestrada forest ecosystems

in Perugia (Italy). 14 p.

ABSTRACT — A preliminary taxonomic list of the macromycetes growing in forest

ecosystems in Perugia (Italy) is presented based on mycological research carried

506 ... New regional mycobiotas online

out in the most widespread local plant communities from the forest of Collestrada:

Quercus spp. woodlands, Carpinus betulus L. woodland and plantations with Pinus

pinea e/o Pinus pinaster. In the period from Jan. 2011 to Dec. 2011 133 taxa belonging

to 70 genera were recorded. For each taxa the following items were reported: Latin

name, author, WGS-84 Global Position System (GPS) coordinates, coordinate grids

from a Google Earth Collestrada image, date of the survey and habitat. This work

contributes to the Umbrian regional checklist, which will eventually be integrated

with the Italian national checklist.

Puntillo, D., M. Puntillo, G. Potenza & S. Fascetti. New, rare, and noteworthy lichens

in the Pollino National Park (Basilicata, southern Italy). 13 pp.

Asstract — 42 lichens new to the Basilicata Region were found in the

territory of the Pollino National Park (southern Italy). There are two

very interesting groups of species: the first, the Caliciales, indicator of the

ecological continuity of the forest, are present on old trees (Abies alba Mill.).

The second one consists of foliicolous species that are present in a small

humid valley with subtropical microclimatic conditions. From the results

of this work, the number of taxa of the lichen flora of Basilicata rises to 558

species, with an increase of 9%.

Switzerland

Senn-Irlet, Beatrice, Rolf Miirner, Elia Martini, Nicolas Kiiffer, Romano de Marchi &

Guido Bieri. Saprobic fungi on wood and litter of Alnus alnobetula in the Swiss

Alps. 34 pp.

ABSTRACT — 246 species representing 73 genera and 90 species of ascomycetes,

basidiomycetes being represented with 44 genera of aphyllophoralean fungi with 77

species, 23 genera of agarics with 68 species and 8 genera of tremelloid fungi with

12 species growing on wood and litter of Alnus alnobetula in Switzerland are given.

Clitocybe and Mycena species dominate among the leaf litter inhabiting species. Fallen

branches have the highest species diversity. The host-specific Peniophora aurantiaca is

one of the most conspicuous and most frequent species.

MY COTAXON

http://dx.doi.org/10.5248/120.507

Volume 120, pp. 507-510 April-June 2012

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 120

Acrogenospora hainanensis Jian Ma & X.G. Zhang, p. 59

Annulatascus menglensis D.M. Hu, L. Cai & K.D. Hyde, p. 82

Aqualignicola vaginata D.M. Hu, L. Cai & K.D. Hyde, p. 84

Arcyria galericulata B. Zhang & Yu Li, p. 402

Basidiobolomycetes Humber, p. 484

Bipolaris salkadehensis Anmadpour & Heidarian, p. 302

Caeoma ahmadii Afshan, Niazi & Khalid, p. 241

Caeoma khanspurense Khalid, Afshan & Niazi, p. 242

Caeoma rosicola Afshan, Khalid & Niazi, p. 243

Cantharellus zangii X.F. Tian, P.G. Liu & Buyck, p. 100

Clavulicium hallenbergii Avneet P. Singh, J. Kaur & Dhingra, p. 353

Colletotrichum populi C.M. Tian & Zheng Li, p. 283

Conidiobolus sinensis Y. Nie, X.Y. Liu & B. Huang, p. 432

Coriolopsis psila (Lloyd) Ryvarden, p. 228

Crepidotus variabilis var. subsphaerosporus J.E. Lange 1940,

lectotypified, epitypified, p. 425

Entoloma aurantioquadratum C.K. Pradeep & K.B. Vrinda, p. 338

Entoloma crassum C.K. Pradeep & K.B. Vrinda, p. 335

Entoloma suaveolens C.K. Pradeep & K.B. Vrinda, p. 332

Entomophthoromycetes Humber, p. 486

Entomophthoromycota Humber, p. 481

Erysiphe javanica Meeboon & S. Takam., p. 191

Exobasidium ferrugineae Minnis, A.H. Kenn. & N.A. Goldberg, p. 455

Fusarium bactridioides Wollenw. 1934, lectotypified, p. 418

Glomus trufemii B.T. Goto, G.A. Silva & Oehl, p. 3

Gomphus crassipes (L.M. Dufour) Maire 1937, lectotypified, p. 394

Infundibulicybe lateritia (J. Favre) Vizzini & Contu, p. 368

Infundichalara minuta Koukol, p. 346

Inonotus multisetifer Abrahao & Gugliotta, p. 36

508 ... MycoOTAXON 120

Lactifluus subg. Piperati Verbeken, p. 449

Lactifluus sect. Allardii (Hesler & A.H. Sm.) De Crop, p. 449

Lactifluus sect. Aurantiifolii (Verbeken) Verbeken, p. 450

Lactifluus sect. Phlebonemi (R. Heim ex Verbeken) Verbeken, p. 446

Lactifluus sect. Piperati (Fr.) Verbeken, p. 449

Lactifluus sect. Polysphaerophori (Singer) Verbeken, p. 445

Lactifluus sect. Pseudogymnocarpi (Verbeken) Verbeken, p. 447

Lactifluus sect. Rubroviolascentini (Singer) Verbeken, p. 447

Lactifluus sect. Tomentosi (McNabb) Verbeken, p. 448

Lactifluus acicularis (Van de Putte & Verbeken) Van de Putte, p. 444

Lactifluus albocinctus (Verbeken) Verbeken, p. 445

Lactifluus allardii (Coker) De Crop, p. 450

Lactifluus angustus (R. Heim & Gooss.-Font.) Verbeken, p. 446

Lactifluus arsenei (R. Heim) Verbeken, p. 446

Lactifluus aurantiifolius (Verbeken) Verbeken, p. 450

Lactifluus austrovolemus (Hongo) Verbeken, p. 444

Lactifluus brunneoviolascens (Bon) Verbeken, p. 446

Lactifluus brunnescens (Verbeken) Verbeken, p. 445

Lactifluus caperatus (R. Heim & Gooss.-Font.) Verbeken, p. 450

Lactifluus caribaeus (Pegler) Verbeken, p. 446

Lactifluus carmineus (Verbeken & Walleyn) Verbeken, p. 447

Lactifluus clarkeae (Cleland) Verbeken, p. 448

Lactifluus cocosmus (Van de Putte & De Kesel) Van de Putte, p. 450

Lactifluus crocatus (Van de Putte & Verbeken) Van de Putte, p. 444

Lactifluus denigricans (Verbeken & Karhula) Verbeken, p. 447

Lactifluus distantifolius (Van de Putte, Stubbe & Verbeken) Van de Putte, p. 444

Lactifluus dwaliensis (K. Das, J.R. Sharma & Verbeken) K. Das, p. 449

Lactifluus flammans (Verbeken) Verbeken, p. 445

Lactifluus foetens (Verbeken & Van Rooij) Verbeken, p. 445

Lactifluus glaucescens (Crossl.) Verbeken, p. 449

Lactifluus goossensiae (Beeli) Verbeken, p. 445

Lactifluus gymnocarpoides (Verbeken) Verbeken, p. 447

Lactifluus gymnocarpus (R. Heim ex Singer) Verbeken, p. 445

Lactifluus kivuensis (Verbeken) Verbeken, p. 448

Lactifluus lamprocystidiatus (Verbeken & E. Horak) Verbeken, p. 444

Lactifluus leucophaeus (Verbeken & E. Horak) Verbeken, p. 449

Lactifluus longipilus (Van de Putte, H.T. Le & Verbeken) Van de Putte, p. 444

Lactifluus longisporus (Verbeken) Verbeken, p. 447

Lactifluus longivelutinus (X.H. Wang & Verbeken) X.H. Wang, p. 446

NOMENCLATURAL NOVELTIES & TYPIFICATIONS ...

Lactifluus luteolus (Peck) Verbeken, p. 446

Lactifluus luteopus (Verbeken) Verbeken, p. 447

Lactifluus medusae (Verbeken) Verbeken, p. 447

Lactifluus nonpiscis (Verbeken) Verbeken, p. 446

Lactifluus novoguineensis (Henn.) Verbeken, p. 449

Lactifluus olivescens (Verbeken & E. Horak) Verbeken, p. 449

Lactifluus paleus (Verbeken & E. Horak) Verbeken, p. 449

Lactifluus pallidilamellatus (Montoya & Bandala) Van de Putte, p. 444

Lactifluus phlebonemus (R. Heim & Gooss.-Font.) Verbeken, p. 446

Lactifluus pinguis (Van de Putte & Verbeken) Van de Putte, p. 445

Lactifluus pisciodorus (R. Heim) Verbeken, p. 446

Lactifluus pseudogymnocarpus (Verbeken) Verbeken, p. 447

Lactifluus pseudoluteopus (X.H. Wang & Verbeken) X.H. Wang, p. 448

Lactifluus pseudovolemus (R. Heim) Verbeken, p. 448

Lactifluus pumilus (Verbeken) Verbeken, p. 447

Lactifluus putidus (Pegler) Verbeken, p. 446

Lactifluus roseophyllus (R. Heim) De Crop, p. 449

Lactifluus rubiginosus (Verbeken) Verbeken, p. 6

Lactifluus rubrobrunnescens (Verbeken, E. Horak & Desjardin) Verbeken, p. 446

Lactifluus rubroviolascens (R. Heim) Verbeken, p. 447

Lactifluus rugatus (Kiihner & Romagn.) Verbeken, p. 448

Lactifluus subclarkeae (Grgur.) Verbeken, p. 450

Lactifluus subpiperatus (Hongo) Verbeken, p. 449

Lactifluus tanzanicus (Karhula & Verbeken) Verbeken, p. 445

Lactifluus tenuicystidiatus (X.H. Wang & Verbeken) X.H. Wang, p. 447

Lactifluus veraecrucis (Singer) Verbeken, p. 445

Lactifluus vitellinus (Van de Putte & Verbeken) Van de Putte, p. 445

Lactifluus volemoides (Karhula) Verbeken, p. 448

Lactifluus xerampelinus (Karhula & Verbeken) Verbeken, p. 448

Leptogium subjuressianum Marcelli & Kitaura, p. 218

Leptogium subjuressianum var. caparoense Kitaura & Marcelli, p. 220

Manoharachariella indica Rajeshkumar & S.K. Singh, p. 44

Moniliophthora aurantiaca Kropp & Albee-Scott, p. 499

Nectria zangii Z.Q. Zeng & W.Y. Zhuang, p. 69

Neozygitales Humber, p. 486

Neozygitomycetes Humber, p. 485

Octaviania violascens Choeyklin, Boonprat. & Somrith., p. 150

Omphalina pyxidata var. cystidiata M. Curti, Contu & Vizzini, p. 364

Paradoxa sinensis L. Fan & J.Z. Cao, p. 473

509

510... MYCOTAXON 120

Paralepista abdita (Dé6rfelt) Vizzini, p. 262

Paralepista ameliae (Arcang.) Vizzini, p. 262

Paralepista biformis (Peck) Vizzini, p. 262

Paralepista femoralis (H.E. Bigelow) Vizzini, p. 262

Paralepista flaccida (Sowerby) Vizzini, p. 262

Paralepista flaccida var. fibrillosa (Malencgon) Vizzini, p. 262

Paralepista gilva (Pers.) Vizzini, p. 262

Paralepista lentiginosa (Fr.) Vizzini, p. 262

Paralepista maculosa (Sacc.) Vizzini, p. 262

Paralepista pseudoparilis (Enderle & Contu) Vizzini, p. 263

Paralepista shafferi (H.E. Bigelow) Vizzini, p. 263

Paralepista splendens (Pers.) Vizzini, p. 263

Paralepistopsis Vizzini, p. 257

Paralepistopsis acromelalga (Ichimura) Vizzini, p. 261

Paralepistopsis amoenolens (Malencon) Vizzini, p. 257

Passalora aseptata R. Singh, Chaurasia, K. Shukla & Upadhyaya, p. 462

Phaeocollybia nigripes Wartchow & V. Coimbra, p. 172

Phlyctis communis Chitale & Makhija, p. 76

Pisolithus capsulifer (Sowerby) Watling, Phosri & M.P. Martin

(also epitypified, lectotypified), p. 202

Pisolithus orientalis Watling, Phosri & M.P. Martin, p. 205

Plectania seaveri M. Carbone, Agnello & LaGreca, p. 318

Pluteus crenulatus Justo, Battistin & Angelini, p. 14

Pluteus stenotrichus Justo, Battistin & Angelini, p. 17

Postia subplacenta B.K. Cui, p. 232

Pseudocercospora danaicola (Vienn.-Bourg.) Pirnia & Zare

(also neotypified), p. 164

Rossbeevera yunnanensis Orihara & M.E. Sm., p. 141

Russula jilinensis G.J. Li & H.A. Wen, p. 51

Septobasidium capparis S.Z. Chen & L. Guo, p. 269

Septobasidium dacrydii S.Z. Chen & L. Guo, p. 274

Septobasidium reevesiae S.Z. Chen & L. Guo, p. 272

Sympodioplanus yunnanensis G.Z. Yang & Z.F. Yu, p. 289

Terriera simplex Y.R. Lin, X.M. Gao & C.T. Zheng, p. 210

Tuber microsphaerosporum L. Fan & Yu Li, p. 471

Tubifera applanata Leontyev & Fefelov, p. 248

Vararia longicystidiata Samita, Sanyal, Dhingra & Avneet P. Singh, p. 357

Wrightoporia nigrolimitata Jia J. Chen, p. 296