Iv ... MYCOTAXON 123

MY COTAXON

VOLUME ONE HUNDRED TWENTY-THREE — TABLE OF CONTENTS

COVER SECTION

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SULOTTISSIOM PT OCEUIT ES Beles he cans, We 4 pee ater +A ide Sh poe ht OL eine Ay Bed Ones *

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RESEARCH ARTICLES

Scleroderma suthepense, a new ectomycorrhizal fungus from Thailand

JATURONG KUMLA, NAKARIN SUWANNARACH,

BOONSOM BuUSSABAN & SAISAMORN LUMYONG

Lichenochora tertia: the third species of the genus growing on

Xanthoria elegans JAVIER Etayo, ADAM FLAKUS,

MARTIN KUKWA & PAMELA RODRIGUEZ FLAKUS

Inonotus tramisetifer (Agaricomycetes), a new species from India

HARPREET Kaur, AVNEET P. SINGH & G.S. DHINGRA

Three new species of Hymenoscyphus from tropical China

Huan-D1 ZHENG & WEN-YING ZHUANG

Gloeophyllum protractum is synonymous with G. mexicanum

JiRki Kourt, JOSEF VLASAK JR. & JOSEF VLASAK

Pseudofusicoccum adansoniae isolated as an endophyte from

Jatropha podarica: new record for India

ROHIT SHARMA, GIRISH KULKARNI & YOGESH S. SHOUCHE

Pestalotiopsis species associated with Camellia sinensis (tea)

SAJEEWA S.N. MAHARACHCHIKUMBURA, EKACHAI CHUKEATIROTE,

LIANG-DONG Guo, PEDRO W. Crous, Eric H.C. MCKENZIE & KEVIN D. HYDE

Critical revision of myxomycetes in the Buenos Aires BAFC herbarium

—I GABRIEL MORENO, AURELIO CASTILLO & JORGE R. DESCHAMPS

New records of Melampsora species on willows in China

PENG ZHAO, CHENG-MING TIAN, YI-JIAN YAO, ZHEN-SHI Hou,

Qi WANG, YUICHI YAMAOKA & MAKOTO KAKISHIMA

Eight new combinations and a replacement name in the genus Hygrocybe

ALAN E. BESSETTE, ARLEEN R. BESSETTE,

WILLIAM C. Roopy & WALTER E. STURGEON

Tuber subglobosum and T. wenchuanense — two new species with

spino-reticulate ascospores Li FAN, JIN-ZHONG CAo & CHENG-LIN Hou

JANUARY-MARCH 2013... V

Cordochaete (Agaricomycetes), a new corticioid genus from India

S.K. SANYAL, SAMITA, G.S. DHINGRA & AVNEET P. SINGH 103

New or rare fungi from eastern Amazonia. I.

Circinoconiopsis amazonica gen. et sp. nov. ANTONIO HERNANDEZ-GUTIERREZ 107

Rediscovery of Pseudocolus garciae in southern Brazil

MARCELO A. SULZBACHER, VAGNER G. CorTEZ & IuRI G. BasE1a 113

Coccomyces pinicola sp. nov. on Pinus armandii from China

RoncG-Hua LEI, KUAN- YUE SUN, QIN ZHANG & CHENG-LIN Hou 121

Sporormiella octomegaspora, a new hairy species with eight-celled

ascospores from Spain FRANCESCO DOVERI & SABRINA SARROCCO 129

Neolinocarpon attaleae sp. nov. on Attalea funifera (Arecaceae)

from Brazil Napja SANTOS ViITORIA, Marta A.Q. CAVALCANTI,

CRISTIANE DUARTE DOS SANTOS, JADERGUDSON PEREIRA & JOSE LUIZ BEZERRA 141

Species of Puccinia (Basidiomycota; Uredinales) new to Indian subcontinent

(Northern Areas of Pakistan) M. SaBa & A.N. KHatip 147

New records of lichens from Chile Xin Yu WANG, IRIs PEREIRA,

SOON-OK OH, LI SONG WANG & JAE-SEOUN Hur 157

Endocarpon maritimum sp. nov. (lichenized Ascomycota) from

the maritime region of South Korea YOGESH JOSHI & JAE-SEOUN Hur 163

Two striking Inocybe species from Yunnan Province, China

YU-GUANG FAN & ToLGor Bau 169

Stipitate hydnums of the southern United States 1:

Phellodon mississippiensis sp. nov. RICHARD BaIrRD,

Lisa E. WALLACE, & GERALD BAKER 183

Two new species of Lignosus (Polyporaceae) from Malaysia -

L. tigris and L. cameronensis CHON-SENG TAN, Szu-TING NG & Jr TAN 193

Pionnotes, a synonym of Dacrymyces rather than Fusarium

KeITH A. SEIFERT 205

A new species of Stropharia from Western Ghats, India

GUNASEKARAN SENTHILARASU & SANJAY K. SINGH 213

Two new species of Endophragmiella from Spain

MARGARITA HERNANDEZ-RESTREPO,

JULIO MENA-PORTALES, JOSEP GUARRO & JOSEPA GENE 221

Sporisorium linderi, a new record for Asia

MUHAMMAD F1Az, ABDUL NASIR KHALID & HABIB AHMAD 229

New combinations, Scheffersomyces amazonensis and S. ergatensis

HECTOR URBINA & MEREDITH BLACKWELL 233

Additional information on Lecanora loekoesii Hal-YING WANG,

AN-Na Ge, HonG-ME! Li & ZUN-TIAN ZHAO 235

vI ... MYCOTAXON 123

Three new species and one new record of Lobothallia from China

XING-RAN Kou, SHU-XIA LI & QING REN 241

A new halotolerant species of Alternaria

from Qinghai-Tibet Plateau, China FaNnG WanG, AN-Na LI,

Donc-ME! Dal, XIAO-XUE XU & Duo-CHuaN Li 251

Additions to the smut fungi of Pakistan. 2

CVETOMIR M. DENCHEV, MUHAMMAD FIAZ,

TEODOR T. DENCHEV, HABIB AHMAD & ABDUL NASIR KHALID 255

Yunchangia, a new genus of smut fungi (Ustilaginaceae)

from China Lin Guo & Brao Xu 261

Peltaster fructicola, a newly recorded species from China

associated with sooty blotch and flyspeck CHEN CHEN, Liu Gao,

MEIYAN Qu, XIAOYONG WEI, WENHUAN LI,

RONG ZHANG, GUANGYU SUN & MARK L. GLEASON 265

A new greenish gilled species of Marasmius (Agaricales)

from Hainan Island, China Y1-Hua YANG, CHuN-YiNG DENG & Tat-Hut Li 271

A new species and new record of Chloridium from the

Qinghai-Tibet Plateau Area, China YugE-MING Wu & TIAN-Yu ZHANG 277

A new species of Heliocephala from Vietnam

VaDIM A. MEL’NIK, RAFAEL FE. CASTANEDA-RUIZ & MILAGRO GRANADOS 281

Lecanora gansuensis sp. nov. (subfusca group) from China

Ler LU, QraNG REN, DalI-FENG JIANG, HAI-YING WANG & ZUN-TIAN ZHAO 285

A new species of Gautieria from China ToLGor Bau & Yu Liu 289

Typification of Tuber formosanum

(Tuberaceae, Pezizales, Ascomycota) from Taiwan, China

PENG Q1A0, PEI-Gui Liu, HUNG Tao Hu & YUN WanG 293

Three new Inocephalus species with cuboid basidiospores

from New South Wales and Queensland, Australia

Davi L. LARGENT, SARAH E. BERGEMANN, SANDRA E. ABELL-DavISs,

Kerri L. KLUTING & GRIFFIN A. CuMMinGs 301

New Lecanora, Lecidea, Melaspilea, Placynthium, and Verrucaria

records for Turkey and Asia ALI ASLAN & KENAN Yazict 321

Ophiodothella angustissima comb. nov., a new name for

Acerviclypeatus poriformans and O. vaccinii

RICHARD T. HANLIN & Maria C. GONZALEZ 327

The genus Wrightoporia in Korea YEONGSEON JANG, SUNG WOOK LEE,

YOUNG WOON LIM, JIN SUNG LEE, TsUTOMU HATTORI & JAE-JIN Kim 335

Three new cercosporoid fungi from the Brazilian Atlantic forest

ANDRE L. FIRMINO, DANILO B. PINHO & OLINTO L. PEREIRA 343

JANUARY-MARCH 2013... VII

A new species of Entocybe (Entolomataceae, Agaricomycetes)

from Québec, Canada TIMOTHY J. BARONI & YVES LAMOUREUX 353

A new Species of Miriquidica from China

Xin ZHAO, Lu-Lu ZHANG & ZUN-TIAN ZHAO 363

Lenzitopsis oxycedri (Thelephoraceae, Basidiomycota)

newly recorded for the Balkan Peninsula

MITKO KARADELEV, KATERINA RUSEVSKA & OLIVER AVRAMOVSKI 369

Cortinarius of California: eight new species in subg. Telamonia

DIMITAR BOJANTCHEV 375

Acaulospora endographis (Glomeromycetes), a new fungus

with a complex spore wall Bruno T. GoTo,

CAMILLA M.R. PEREIRA, CAMILA P. NoBRE, NATALIA P. ZATORRE,

FERNANDA COVACEVICH, RICARDO L.L. BERBARA & LEONOR C. Mata 403

Basidiospore ultrastructure of some Dacrymycetales from Mexico

SIGFRIDO SIERRA, JOAQUIN CIFUENTES,

Oca M. ECHEVERRIA-MARTINEZ & SANDRA CASTRO-SANTIUSTE 409

Studies of two Corner types (Marasmius nigroimplicatus and

M. subrigidichorda) and new Gymnopus combinations

ZDENKO TKALCEC & ARMIN MESiI¢ 419

An emendation of Fusticeps and two new species from

the Brazilian Amazon Forest

JOSIANE SANTANA MONTEIRO & LUIS FERNANDO PASCHOLATI GUSMAO 431

Two Ropalospora lichens new to mainland China

Linc Hu, Hal-YING WANG’, JING LIu & ZUN-TIAN ZHAO 439

South Florida microfungi: a new species of Ellisembia (hyphomycetes)

with new records from the U.S.A. GreGorIO DELGADo 445

Aureoboletus zangii (Boletaceae), a new species from China

XIA0-FEI SHI & PEI-Gui Liu 451

Molecular characterization of Colletotrichum species causing

soybean anthracnose in Argentina ARACELI M. Ramos,

Luis FRANCO TADIC, ISABEL CINTO, MARCELO CARMONA & MARCELA GALLY 457

A new species of Cantharomyces (Laboulbeniales, Ascomycota)

from the Netherlands D. HAELEWATERS & A. DE KESEL 467

Lyophyllum rhombisporum sp. nov. from China X1a0-QiNG WANG,

DE-QUN ZHOU, YONG-CHANG ZHAO,

X1A0-LEI ZHANG, LIN LI & SHU-HonG Lr 473

New records of corticolous lichens from Vietnam SANTOSH JOSHI,

Tut THuy NGUYEN, NGUYEN ANH DZzuUNG,

UDENI JAYALAL, SOON-OK OH & JAE-SEOUN Hur 479

vill ... MYCOTAXON 123

NEW MYCOBIOTAS ONLINE (abstracts)

Gasteroid mycobiota (Basidiomycota) from Polylepis australis

woodlands of central Argentina

Maria L. HERNANDEZ-CAFFOT, GERARDO ROBLEDO & LAURA S. DOMINGUEZ 491

Checklist of the Argentinean Agaricales 6. Paxillaceae,

Gomphidiaceae, Boletaceae and Russulaceae N. NIVEIRO & E. ALBERTO 491

New lichen records from Armenia ARSEN GASPARYAN & HarRIE J. M. SrpmMAN 491

Corticioid fungi from arid and semiarid zones of the Canary Islands (Spain)

ESPERANZA BELTRAN-TEJERA, J. LAURA RODRIGUEZ-ARMAS,

M. TERESA TELLERIA, MARGARITA DUENAS, IRENEIA MELO,

M. JONATHAN DiAz-ARMAS, ISABEL SALCEDO & JOSE CARDOSO 492

The lichen biota of Igdir province (Turkey)

KENAN YAZICI, ANDRE APTROOT & ALI ASLAN 492

NOMENCLATURE

Nomenclatural novelties proposed in volume 123 493

PUBLICATION DATE FOR VOLUME ONE HUNDRED TWENTY-TWO

MYCOTAXON for OCTOBER—DECEMBER, VOLUME 122 (I-x1I + 1-494)

was issued on March 4, 2013

ERRATA FROM VOLUME 122

p-81, line 4 FOR: (No. U0836604) READ: (No. U1202262)

p.416, line 4 in Fic. FOR: Cantharellus atratus (AB445115)

READ: Craterellus atratus (AB445115)

JANUARY-MARCH 2013... IX

REVIEWERS — VOLUME ONE HUNDRED TWENTY-THREE

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.

N.S. Afshan

Artur Alves

Vladimir Antonin

André Aptroot

Timothy J. Baroni

Ann Bell

Shannon M. Berch

David Boertmann

Uwe Braun

Irwin M. Brodo

Rafael F. Castafieda Ruiz

Chee-Jen Chen

Vagner G. Cortez

Yu-Cheng Dai

Cony Decock

Cvetomir M. Denchev

Hasan Htiseyin Dogan

Eduardo Furrazola

Walter Gams

Genevieve Gates

Matteo Gelardi

Admir Jose Giachini

Paolo Giordani

Tom Grafenhan

Cecile Gueidan

Lin Guo

Shouyu Guo

Ying-Lan Guo

Gaston Guzman

Ian R. Hall

Nils Hallenberg

Roy E. Halling

Tsuyoshi Hosoya

Fabien C.C. Hountondji

Sabine Huhndorf

T.A.M. Jagadeesh Ram

Makoto Kakishima

Bryce Kendrick

Roland Kirschner

Kerry Knudsen

S.Y. Kondratyuk

Heikki Kotiranta

Cletus P. Kurtzman

David L. Largent

De-Wei Li

Tai-Hui Li

Ying-Ren Lin

Marcos Lizarraga Escobar

Laszl6 Lékés

Sandra Lupo

Eric H.C. McKenzie

James Mehl

Vadim A. Melnik

D.W. Minter

P. Brandon Matheny

Helmut Mayrhofer

D.W. Mitchell

Karen K. Nakasone

Pere Navarro-Rosinés

Nhu H. Nguyen

Machiel E. Noordeloos

Lorelei L. Norvell

Eduardo Nouhra

Jadson J.S. Oliveira

Yuko Ota

Clark Ovrebo

Khwanruan Papong

Omar Paino Perdomo

Ming Pei

Shaun R. Pennycook

Ronald H. Petersen

Olinto Liparini Pereira

Donald H. Pfister

Michele D. Piercey-

Normore

Christian Printzen

Scott A. Redhead

Scott C. Redlin

Erlei Melo Reis

M.J. Richardson

Amy Y. Rossman

Leif Ryvarden

Sergi Santamaria

Mark Seaward

B.M. Sharma

Roger Shivas

Rosa Mara Borges da

Silveira

H.J.M. Sipman

Matthew E. Smith

Dartanha J. Soares

Viacheslav Spirin

Marc Stadler

Steven L. Stephenson

Larissa Trierveiler Pereira

Steven A. Trudell

D.K. Upreti

Jukka Vauras

Else C. Vellinga

Yei-Zeng Wang

Yong Wang

Yun Wang

Zhu L. Yang

Ming Ye

Zhongyi Zhang

Rui-Lin Zhao

Wen-Ying Zhuang

X ... MYCOTAXON 123

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JANUARY-MARCH 2013... XI

FROM THE EDITOR-IN-CHIEF

FINALLY! — Despite our valiant efforts to bring Mycotaxon 123 to publication

in April as promised, medical science prevented our completing this volume as

rapidly as we had hoped. Our apologies to a very patient 203 authors (representing

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patient subscribers.

Within the pages of the delayed 2013 January-March volume are three new genera

(Circinoconiopsis from Brazil, Cordochaete from India, Yunchangia from China) and 57

species new to science representing Acaulospora, Fusticeps, Neolinocarpon, Passalora,

and Pseudocercospora from Brazil; Alternaria, Aureoboletus, Chloridium, Coccomyces,

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Miriquidica, and Tuber from China; Cantharomyces from the Netherlands; Cortinarius,

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Peru; Lignosus from Malaysia; and Pestalotiopsis and Scleroderma from ‘Thailand.

In addition to range extensions and/or new hosts for previously named taxa, we also

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24 July 2013

MY COTAXON

http://dx.doi.org/10.5248/123.1

Volume 123, pp. 1-7 January-March 2013

Scleroderma suthepense, a new ectomycorrhizal fungus

from Thailand

JATURONG KUMLA, NAKARIN SUWANNARACH,

BOONSOM BUSSABAN & SAISAMORN LUMYONG*

Department of Biology, Faculty of Science, Chiang Mai University,

Chiang Mai, 50200, Thailand

* CORRESPONDENCE TO: saisamorn.|@cmu.ac.th

ABSTRACT — A new species of earthball, described herein as Scleroderma suthepense, was

found under Prunus cerasoides in Doi Suthep-Pui National Park, Thailand. Morphological

characteristics indicate it belongs to section Scleroderma. Molecular analysis showed

differences from previously described Scleroderma species. A description, illustration, and

comparisons with related taxa are provided.

KEY worpDs — gasteromycete, phylogenetic analysis, taxonomy

Introduction

Ectomycorrhizal fungi have symbiotic associations with many tree species in

forests around the world and over 7000 species have been described (Taylor &

Alexander 2005). Scleroderma (Boletales; Binder & Hibbett 2006) has worldwide

distribution in temperate and tropical regions and forms ectomycorrhizas with

a broad range of woody plants including members of families Caesalpiniaceae,

Dipterocarpaceae, Fagaceae, Myrtaceae, Phyllanthaceae, and _ Pinaceae

(Munyanziza & Kuyper 1995; Sims et al. 1997; Sanon et al. 2009). There are

131 Scleroderma names in Index Fungorum (http://www.indexfungorum.

org/Names/Names.asp). In Thailand, only ten species of Scleroderma have

been reported, S. areolatum, S. bovista, S. cepa, S. citrinum, S. dictyosporum,

S. flavidum, S. lycoperdoides, S. polyrhizum, S. sinnamariense, and S. verrucosum

(Chandrasrikul et al. 2011). Scleroderma species have been used to increase the

growth of tree seedling, both in nurseries and in the field (Chen et al. 2006).

Several species of Scleroderma have been proposed based on morphological

characteristics of their basidiomes and basidiospores (Guzman 1970; Sims et

al. 1995; Guzman et al. 2004). Recent studies have also used molecular analysis

to analyze inter- and intraspecific variation in Scleroderma species (Phosri et al.

2009; Nouhra et al. 2012).

2 ... Kumla & al.

During an investigation of ectomycorrhizal fungi associated with tree species

in northern Thailand, we found an interesting species of Scleroderma under

Prunus cerasoides Buch.-Ham. ex D. Don (Rosaceae), which we describe as a new

species in the present paper. To confirm their taxonomic status the phylogenetic

relationship among 25 isolates of Scleroderma were also determined by analysis

of complete sequences of the internal transcribed spacer (ITS) regions of the

RNA gene.

Materials & methods

Morphology studies

Basidiomes of Scleroderma were collected from Medicinal Plant Garden (18°48'20"N

98°54'52"E, elevation 1072 m), Doi Suthep-Pui National Park, Chiang Mai Province,

Thailand in May 2012. Basidiomes were wrapped in aluminum foil or kept in plastic

specimen boxes until transportbacktothelaboratory where notes on macromorphological

features and photographs were taken within 24 h. Macromorphological data were

derived from fresh specimens, whereas micromorphological data were derived from

dried specimens mounted in 95% ethanol followed by distilled water, 3% KOH or

Melzer’s reagent. Size data of anatomical features are based on at least 50 measurements

of each structure. The specimens were dried at 40-45 °C and deposited at the Research

Laboratory for Excellence in Sustainable Development of Biological Resources, Faculty

of Science, Chiang Mai University, Thailand (SDBR-CMU).

Molecular studies

Genomic DNA of a dried specimen was extracted according to a CTAB method

(Kumla et al. 2012). The ITS regions of the RNA gene were amplified by polymerase

chain reaction (PCR) with primers ITS4 and ITS5 under the following thermal

conditions: 94 °C for 2 min; 35 cycles of 95 °C for 30 s, 50 °C for 30 s, 72 °C for 1 min, and

72 °C for 10 min. The PCR products were then checked on 1% agarose gels stained with

ethidium bromide and visualized under UV light. PCR products were purified by using

PCR clean up Gel extraction NucleoSpin’ Extract I] Purification Kit (Macherey-Nagel,

Germany, Catalog no. 740 609.50) following the manufacturer's protocol. The purified

PCR products were directly sequenced. Sequencing reactions were preformed and the

sequences were automatically determined in the genetic analyzer (1°' Base, Malaysia)

using the PCR primers mentioned above. Sequences were used to query GenBank via

BLAST (http://blast.ddbj.nig.ac.jp/top-e.html). For the phylogenetic analysis, a multiple

alignment subroutine in Clustal X (Thompson et al. 1997) and a maximum-parsimony

analysis using the PAUP beta 10 software version 4.0 (Swofford 2002) were carried out.

Results

Taxonomic description

Scleroderma suthepense Kumla, Suwannarach & Lumyong, sp. nov. PLATE 1

MycoBank MB800878

Differs from Scleroderma meridionale by its longer basidiospore spine length.

Scleroderma suthepense sp. nov. (Thailand) ... 3

PiaTE 1. Scleroderma suthepense. A, B: Basidiomes. C: Hypha with a clamp connection (arrowed).

D: Basidiospores as observed under a compound microscope. E: Basidiospore as observed with a

scanning electron microscope. Scale bars: A, B= 1 cm; C-E=5 um.

TypE— Thailand, Chiang Mai Province, Muang District, Doi Suthep-Pui National

Park, Medicinal Plant Garden, 18°48'20"N 98°54'52"E, elevation 1072 m, rainforest

dominated by Prunus cerasoides, Cinchona pubescens, and other trees, on sandy loam,

15 May 2012, Jaturong Kumla & Nakarin Suwannarach (Holotype, SDBR-CMU55-SC2;

GenBank sequence, JX205215).

ErymMoLoGcy—suthepense, referring to Doi Suthep-Pui National Park, where the new

species was found.

Basidiomes globose or subglobose 1.1-3.5 cm diameter, 1.0-3.9 cm height,

rhizomorphs well developed, white to yellow, 0.5-1.2 cm height. Peridium

0.5-1.0 mm thick when fresh, leathery, surface partially smooth with scattered,

small and thin scales, greyish yellow to greyish brown, consisting of two layers.

The outer layer consists of cylindrical, thick-walled, yellowish brown hypha up

to 8.0 um diameter, with scattered clamp connections, turning to reddish brown

with KOH. The inner layer consists of cylindrical, thick-walled, hyaline hyphae

up to 6.0 um diameter, with clamp connections. Gleba when mature dark

greyish brown to back and pulverulent. Basidia not observed. Basidiospores

globose to subglobose, strongly reticulate with spines, 8.0-13.0 um diameter

(n = 50) including ornamentation (PLATE 1D,E), spine 1.0-2.5 um in length,

dark yellowish brown in water or KOH and not changing in Melzer’s reagent.

4 ...Kumla & al.

ECOLOGY & DISTRIBUTION — Terrestrial on sandy loam, under Prunus

cerasoides in dipterocarp forest; known only from Thailand.

100; EU784414 Scleroderma citrinum

15 GQ166907 Scleroderma citrinum

too, EU819441 Scleroderma michiganense

FM213347 Scleroderma michiganense

72 JX205215 Scleroderma suthepense

EU718121 Scleroderma meridionale

60} GQ267487 Scleroderma bovista

100|' AB211267 Scleroderma bovista

EU819517 Scleroderma bovista

100; HQ688789 Scleroderma patagonicum

100

Clade 1

a HQ688788 Scleroderma patagonicum

FM2 13336 Scleroderma septentrionale

100) FJ840449 Scleroderma dictyosporum

FJ840443 Scleroderma dictyosporum

89 100} FM213354 Scleroderma cepa

85 EU783312 Scleroderma cepa

100 EU718117 Scleroderma laeve

97| 99 EU819518 Scleroderma areolatum

EU819438 Scleroderma areolatum

78 FJ840461 Scleroderma verrucosum

100| FM213349 Scleroderma polyrhizum

95} | FM213350 Scleroderma polyrhizum

EU718118 Scleroderma bermudense

woo) HQ687222 Scleroderma sinnamariense

100

Clade 2

FM213364 Scleroderma sinnamariense

Clade 3

AM947113 Pisolithus albus

AF374704 Pisolithus microcarpus

PLATE 2. A maximum parsimonious tree inferred from a heuristic search of ITS 1, 5.8S ribosomal

RNA gene, and ITS 2 alignments of 27 sequences. Pisolithus albus and P. microcarpus were used

to root the tree. Branches with bootstrap values >50% are shown above each branch and the bar

represents 10 substitutions per nucleotide position.

Molecular analysis

The 679-bp ITS sequence of S. suthepense (JX205215) was deposited in

GenBank and analyzed phylogenetically with 24 other Scleroderma sequences

and the outgroup (Pisolithus albus and P. microcarpus) obtained from the

GenBank database. Heuristic searches produced a tree length of 803 steps,

CI = 0.685, RI = 0.823, RC = 0.563 and HI = 0.315. One maximum-parsimony

tree is shown in PLATE 2. A phylogenetic dendrogram separates Scleroderma

fungi into three clades. Clade 1 contains species with reticulate spores (sect.

Scleroderma), while clades 2 and 3 include species with spiny or subreticulate

spores. Scleroderma suthepense stands within clade 1 together with S. bovista,

Scleroderma suthepense sp. nov. (Thailand) ... 5

S. citrinum, S. dictyosporum, S. meridionale, S. michiganense, S. patagonicum,

and S. septentrionale and forms a sister taxon with S. meridionale (72%

bootstrap support). Clade 2 includes six species (S. areolatum, S. bermudense,

S. cepa, S. laeve, S. polyrhizum, S. verrucosum). Clade 3 comprises a single species,

S. sinnamariense.

Discussion

Based on basidiospore morphology, Scleroderma species are separated into

three sections; sect. Aculeatispora (spiny spore), sect. Sclerangium (subreticulate

spore), and sect. Scleroderma (reticulate spore) (Guzman 1970; Sims et al. 1995,

1997). However, it is difficult to distinguish Scleroderma species based only on

morphology because basidiome size and shape change depending on the soil

and environment (Sanon et al. 2009). Scleroderma suthepense, described as a new

species in the present study, was found beneath Prunus cerasoides and is placed

in sect. Scleroderma based on its reticulate basidiospores and hyphal clamp

connections. Its basidiospores are similar in size to those of S. dictyosporum,

S. meridionale, and S. septentrionale (TABLE 1). Our ITS sequence analysis

clearly separates S. suthepense from the other reticulate spored Scleroderma

species in sect. Scleroderma, although S. suthepense appears closely related to

S. meridionale, which has a shorter basidiospore spine length (0.5-1.5 um) and

is reported only in North America (Sims et al. 1995; Guzman & Ovrebo 2000).

Our analysis separates the reticulate spored clade (clade 1) and the spiny or

subreticulate spored clades (clades 2 and 3) with 89% bootstrap support (PLATE

2). This result was similar to previous molecular phylogenetic studies, which

separated Scleroderma species into two moderately to strongly supported clades

(Phosri et al. 2009; Nouhra et al. 2012). Further studies are required to confirm

an ectomycorrhizal synthesis between S. suthepense and various host plants.

TABLE 1. Basidiospore sizes of Scleroderma suthepense and other species in

Scleroderma sect. Scleroderma.

Sauce ua) DIAMETER REFERENCES

S. bovista 11-16 Sims et al. (1995), Cortez et al. (2011)

S. citrinum 11-16 Sims et al. (1995), Cortez et al. (2011)

S. dictyosporum 9-14 Guzman (1970), Cortez et al. (2011)

S. hypogaeum 17-20 Sims et al. (1995)

S. meridionale 8-17 Sims et al. (1995), Guzman & Ovrebo (2000)

S. michiganense 14-20 Sims et al. (1995)

S. minutisporum 4-7 Alfredo et al. (2012)

S. patagonicum 19-28 Nouhra et al. (2012)

S. septentrionale 8-15 Jeppson & Piatek (2005)

S. suthepense 8-13 Present study

6 ... Kumla & al.

Such knowledge is important for selection and management of ectomycorrhizal

fungi for greenhouse and in field inoculation programs in Thailand.

Acknowledgements

This work was supported by grants from Thailand Research Fund for The Royal

Golden Jubilee Ph.D. Program (PHD/0309/2550) and Research-Team Promotion Grant

RTA5580007 and Graduate School of Chiang Mai University. We are grateful to Dr. Eric

H.C McKenzie and Dr. Steven L. Stephenson for presubmission reviews.

Literature cited

Alfredo DS, Leite AG, Braga-Neto R, Cortez VC, Baseia IG. 2012. Scleroderma minutisporum, a

new earthball from the Amazon rainforest. Mycosphere 3: 294-299.

http://dx.doi.org/10.5943/mycosphere/3/3/4

Binder M, Hibbett DS. 2006. Molecular systematics and biological diversification of Boletales.

Mycologia 98: 971-981. http://dx.doi.org/10.3852/mycologia.98.6.971

Chandrasrikul A, Suwanarit P, Sangwanit U, Lumyong S, Payapanon A, Sanoamuang N, Pukahuta

C, Petcharat V, Sardsud U, Duengkae K, Klinhom U, Thongkantha S, Thongklam S. 2011.

Mushroom (basidiomycetes) in Thailand. Office of Natural Resources and Environmental

Policy and Planning, Bangkok.

Chen YL, Dell B, Malajczuk N. 2006. Effect of Scleroderma spore density and age on mycorrhiza

formation and growth of containerized Eucalyptus globulus and E. urophylla seedlings. New

Forest 31: 453-467. http://dx.doi.org/10.1007/s11056-005-0880-1

Cortez VG, Baseia IG, Silveira RMB. 2011. Gasteroid mycobiota of Rio Grande do Sul, Brazil:

Boletales. JYFR 2: 44-52.

Guzman G. 1970. Monografia del género Scleroderma. Darwiniana 16: 233-407.

Guzman G, Ovrebo CL. 2000. New observation on sclerodermataceous fungi. Mycologia 92:

174-179.

Guzman G, Ramirez-Guillém F, Miller OK, Lodge DJ. 2004. Scleroderma stellatum versus

Scleroderma bermudense: the status of Scleroderma echinatum and the first record of Veligaster

nitidum from the Virgin Islands. Mycologia 96: 1370-1379.

Jeppson M, Piatek M. 2005. Scleroderma septentrionale (Fungi, Basidiomycetes) first records from

central Europe. Pol Bot J 50: 15-17.

Kumla J, Bussaban B, Suwannarach N, Lumyong S, Danell E. 2012. Basidiome formation of an

edible wild, putatively ectomycorrhizal fungus, Phlebopus portentosus without host plant.

Mycologia 104: 597-603. http://dx.doi.org/10.3852/11-074

Munyanziza E, Kuyper TW. 1995. Ectomycorrhizal synthesis on seedlings of Afzelia quanzensis

Welw. using various types of inoculum. Mycorrhiza 5: 283-287.

Nouhra ER, Caffot MLH, Pastor N, Crespo EM. 2012. The species of Scleroderma from

Argentina, including a new species from the Nothofagus forest. Mycologia 104: 488-495.

http://dx.doi.org/10.3852/11-082

Phosri C, Martin MP, Watling R, Jeppson M, Sihanonth P. 2009. Molecular phylogeny and re-

assessment of some Scleroderma spp. (gasteromycetes). An Jard Bot Madrid 166: 83-91.

http://dx.doi.org/10.3989/ajbm.2199

Sanon KB, Ba AM, Delaruelle C, Duponnois R, Matin EF 2009. Morphological and

molecular analyses in Scleroderma species associated with some caesalpinioid legumes,

Dipterocarpaceae and Phyllanthaceae trees in southern Burkina Faso. Mycorrhiza 19: 571-584.

http://dx.doi.org/10.1007/s00572-009-0272-z

Scleroderma suthepense sp. nov. (Thailand) ... 7

Sims KP, Watling R, Jeffries P. 1995. A revised key to the genus Scleroderma. Mycotaxon 56:

403-420.

Sims K, Watling R, De LA, Cruz R, Jeffries P. 1997. Ectomycorrhizal fungi of the Philippines: a

preliminary survey and notes on the geographic biodiversity of the Sclerodermatales. Biodivers

Conserv 6: 45-58.

Swofford DL. 2002. PAUP”: phylogenetic analysis using parsimony (*and other methods). Version

4.0 beta 10. Sunderland, Massachusetts.

Taylor AFS, Alexander I. 2005. The ectomycorrhizal symbiosis: life in the real world. Mycologist

19: 102-111. http://dx.doi.org/10.1017/S0269915X05003034

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.

Nucl Acid Res 24: 4876-4882.

MY COTAXON

http://dx.doi.org/10.5248/123.9

Volume 123, pp. 9-13 January-March 2013

Lichenochora tertia (Phyllachorales): the third species

of the genus growing on Xanthoria elegans

JAVIER ETAYo’, ADAM FLAKUS’, MARTIN KuKwA?

& PAMELA RODRIGUEZ FLAKUS*°

"Navarro Villoslada 16, 3° dcha., E-31003 Pamplona, Navarra, Spain

*Laboratory of Lichenology, W. Szafer Institute of Botany, Polish Academy of Sciences,

Lubicz 46, PL-31-512 Krakow, Poland

*Department of Plant Taxonomy and Nature Conservation, University of Gdansk,

Wita Stwosza 59, PL-80-308 Gdansk, Poland

‘Department of Botany and Molecular Evolution, Senckenberg Forschungsinstitut und

Naturmuseum, Senckenberganlage 25, D-60325 Frankfurt am Main, Germany

°Herbario Nacional de Bolivia, Instituto de Ecologia, Universidad Mayor de San Andrés,

Calle 27, Cota Cota, Casilla 10077, La Paz, Bolivia

CORRESPONDENCE TO: ‘jetayosa@educacion.navarra.es, *a.flakus@botany.

pl, *dokmak@ug.edu.pl, ‘pamela.rodriguez@senckenberg.de

AsBstTRAcT — Lichenochora tertia is a new lichenicolous fungus described from Peruvian

Andes. The species is characterized by small perithecia deeply immersed in the thallus of

Xanthoria elegans and small, narrowly ellipsoidal ascospores with subtle torus along the

septum; the fungus does not induce gall formation.

Key worps — Neotropics, South America, biodiversity, taxonomy

Introduction

During the study of the lichenicolous biota of Bolivia (Flakus & Kukwa

2012a,b, Kukwa et al. 2012) we examined an infected specimen of Xanthoria

elegans collected by the fourth author in Peru, which was found to host an

undescribed Lichenochora. ‘The lichenicolous biota of Peru is still very poorly

studied, and the only paper fully dedicated to this subject was published by

Etayo (2010). Therefore, it is not surprising that a new taxon was discovered in

a biologically rich Andean ecosystem.

The lichenicolous genus Lichenochora Hafellner (Phyllachorales) comprises

34 species worldwide, of which ten parasitize members of the family

Teloschistaceae. Two of them are known to grow on Xanthoria elegans (Link)

Th. Fr. (Triebel et al. 1991, Navarro-Rosinés & Etayo 2001, Etayo & Navarro-

Rosinés 2008, Hafellner et al. 2008, Lawrey & Diederich 2011).

10 ... Etayo & al.

Material & methods

The morphology and the anatomy were examined by using a NIKON SMZ800 and

a NIKON ECLIPSE 80i (DIC) microscopes. The anatomy was studied in water, KOH

solution (K) and, for the ascus structure, in Lugol’s iodine solution without (I) or with

pre-treatment with KOH (K/I). The measurements were made in tap water. Ascospores

measurements are presented as: arithmetic mean - standard deviation, arithmetic

mean, and arithmetic mean + standard deviation, flanked by the minimal and maximal

measurements in parentheses, and the length/breadth ratio (1/b) is presented in the

same way, followed by the number of measurements (n). Values in italics (e.g., -3.2-)

are arithmetic means. The holotype is conserved in Herbario Nacional de Bolivia,

Universidad Mayor de San Andrés, La Paz (LPB).

Results

Lichenochora tertia Etayo, Flakus & Rodr. Flakus, sp. nov. PLATE 1

MycoBank MB 800989

Differs from Lichenochora sinapispermae by its smaller perithecia, deeply immersed in

the host thallus and not forming galls, and by its more narrowly ellipsoidal ascospores.

Type — Peru: Dep. Cusco, Prov. Canchis, Cordillera de Vilcanota, cerca del Nevado

Japu Punta, vegetacion altoandina con suelo muy pedregoso con rocas de gran tamafio y

poca cobertura de plantas superiores, 5320 m, 13°45'41.4"S 71°04'58.8" W, en Xanthoria

elegans en piedra silicea, 13 March 2008, P. Rodriguez Flakus 498b (Holotype, LPB).

EryMoLoGy — fertius (Lat.), referring to this being the third species of the genus

Lichenochora found on Xanthoria elegans.

Fungus parasymbiotic, not inducing gall formation. Mycetium hyaline,

inconspicuous, observed only at the base of the perithecia. ASCOMATA

perithecia dispersed or aggregated in small groups, 120-190 um in diam. and

up to 200 um high, pyriform, strongly rough near apical part, immersed in 2/3

with only the upper part erumpent outside of host thallus; forming papillae.

ExcIpLe dark reddish-brown (K+ olive-brown), 20-40 um wide, formed by

several layers of polygonal cells, usually with many oil guttules in internal cells,

of dark brown thick-walled cells, 8-15 um in diam., with some cells protruding

to the outside giving it a verrucose appearance; innermost excipular layer of

pale brown to hyaline thin walled cells, 6-15 x 2-10 um. PAPILLAE of elongate

cells formed mainly in the upper part of perithecium (near ostiole), usually

grouped and forming wide appendages up to 20-35 um long. HyMENIUM

inspersed by oil droplets, 1-8 um in diam., large droplets (5-8 um in diam.)

abundant and obscuring the hymenium. PARAPHYSES rare in mature ascomata

and obscured by oil droplets, 3-8 um wide, with thin-walled cells. Asc

(2-)6-8-spored, 60-90 x 11-15 um, clavate, functionally unitunicate, K/I-,

with stalk up to 5 um long, thin-walled, not thickened in the apex. ASCOSPORES

narrowly ellipsoidal, sometimes slightly curved, with obtuse to slightly tapering

ends, smooth-walled, (0—)1l-septate, colourless, without perispore, formed

Lichenochora tertia sp. nov. (Peru) ... 11

Pate 1. Lichenochora tertia (holotype). A, B: habit of perithecia with rugose surface of Xanthoria

elegans; C: section through two perithecia immersed in host thallus and forming wide erumpent

appendages in upper part; D: section of perithecium showing structure of excipulum; E, F: asci with

ascospores inside; G: ascospores. Scale bars: A, B = 200 um; C = 50 um; D = 25 um; E-G = 10 um.

12 ... Etayo & al.

subtle torus along the septum, and usually with one large oil guttule in each

cell (sometimes splitting up into several small ones), walls c. 0.5 um thick,

(12-)15-16.5-18(-20) x (4-)4.9-5.3-5.7(-6.5) um, I/b = (2.2-)2.8-3.2-3.6

(-4.5) (n = 120).

DISTRIBUTION AND HABITAT — ‘The new species is known only from the

type locality in Peruvian Andes, where it was found on epilithic Xanthoria

elegans in high montane conditions.

ComMENTs — Lichenochora tertia is characterized by its small perithecia, small,

narrowly ellipsoidal ascospores with a subtle torus along the septa, and not

gall-forming. The most similar species is L. sinapispermae Etayo & Nav.-Ros.

due to its ascospore size and the presence of the torus, but it has comparatively

wider ascospores, (5.5—)6-6.9-8(-8.5) um, which are broadly ellipsoidal, with

I/b ratio (1.6-)1.9-2.2-2.5(-2.8), comparatively bigger perithecia, 200-250 um

in diam. (230-300 um high), and also shorter papillae, 8-13 um long (Navarro-

Rosinés & Etayo 2001). Moreover, the perithecia are at first half-immersed, but

soon almost sessile on the thallus of Caloplaca sinapisperma (Lam.) Maheu &

A. Gillet.

According to Hafellner et al. (2008) and Etayo & Navarro-Rosinés (2008)

two Lichenochora species are known to inhabit Xanthoria elegans, L. elegantis

Hafellner and L. xanthoriae Triebel & Rambold. Lichenochora elegantis has longer

ascospores, 28-33 um long, 4-spored asci and much bigger perithecia, 300-400

um in diam. (Hafellner et al. 2008), whereas in L. xanthoriae ascospores are

considerably wider, 8-9(-9.5) um wide, and the species induces the formation

of small cecidia (Triebel et al. 1991).

Acknowledgments

We are greatly indebted to Professor Pere Navarro-Rosinés (Barcelona) and Dr Kerry

Knudsen (Riverside) for reviewing the manuscript and Dr Mikhail P. Zhurbenko (St.-

Petersburg) for valuable comments on the text, as well as to Rosa I. Meneses Q., the

Director of Herbario Nacional de Bolivia, Universidad Mayor de San Andrés, La Paz

for generous cooperation. This research received support from the NCBiR in Poland

under the LIDER Programme for the years 2010-13 (no. 92/L-1/09) and the W. Szafer

Institute of Botany of the Polish Academy of Sciences through the program supporting

research of young scientists.

Literature cited

Etayo J. 2010. Hongos liquenicolas de Pert. Homenaje a Rolf Santesson. Bulletin de la Société

Linnéenne de Provence 61: 83-128.

Etayo J, Navarro-Rosinés P. 2008. Una combinacion y tres especies nuevas de Lichenochora

(Phyllachorales, Ascomicetes liquenicolas), y notas adicionales para el género. Revista Catalana

de Micologia 30: 27-44.

Flakus A, Kukwa M. 2012a. New records of lichenicolous fungi from Bolivia. Opuscula

Philolichenum 11: 36-48.

Lichenochora tertia sp. nov. (Peru) ... 13

Flakus A, Kukwa M. 2012b. New species of lichenicolous fungi from Bolivia. Lichenologist 44(4):

469-477. http://dx.doi.org/10.1017/S0024282912000059

Hafellner J, Herzog G, Mayrhofer H. 2008. Zur Diversitat von lichenisierten und lichenicolen

Pilzen in den Ennstaler Alpen (Osterreigh: Steiermark, Oberésterreich). Mitteilungen des

Naturwissenschaftlichen Vereins fiir Steiermark 137: 131-204.

Kukwa M, Etayo J, Flakus A. 2012. Plectocarpon stereocaulicola: a new lichenicolous fungus

(Ascomycota: Roccellaceae) from Bolivia. Lichenologist 44(4): 479-482.

http://dx.doi.org/10.1017/S0024282912000151

Lawrey JD, Diederich P. 2011. Lichenicolous fungi - worldwide checklist, including isolated

cultures and sequences available. URL: http://www.lichenicolous.net [accessed 7.04.2012]

Navarro-Rosinés P, Etayo J. 2001. Lichenochora epinashii sp. nov. y L. sinapispermae sp. nov.

(Phyllachorales, Ascomicetes liquenicolas). Cryptogamie, Mycologie 22(2): 147-158.

Triebel D, Rambold G, Nash III TH. 1991. On lichenicolous fungi from continental North America.

Mycotaxon 42: 363-396.

MY COTAXON

http://dx.doi.org/10.5248/123.15

Volume 123, pp. 15-17 January-March 2013

Inonotus tramisetifer (Agaricomycetes),

a new species from India

HARPREET KAurR, AVNEET P. SINGH* & G.S. DHINGRA

Department of Botany, Punjabi University, Patiala 147 002, India

*CORRESPONDENCE TO: avneetbot@gmail.com

ABSTRACT - A new species, Inonotus tramisetifer (Hymenochaetaceae), is described in

association with Quercus incana from the state of Himachal Pradesh in India.

Key worps - Narkanda, tramal setal hyphae

While conducting fungal forays in Narkanda area of district Shimla, Himachal

Pradesh (India), Harpreet Kaur and G.S. Dhingra collected a poroid fungus

associated with tree of Quercus incana. After comparison of macroscopic and

microscopic features (Sharma 1995, Ryvarden 2004, 2005) it has been identified

as a new species of Inonotus close to I. cuticularis (Bull.) P. Karst., I. farlowii

(Lloyd) Gilb., and I. munzii (Lloyd) Gilb., having branched pilear setal hyphae

along with monomitic hyphal system and yellowish brown basidiospores.

Inonotus tramisetifer Har. Kaur, Avneet P. Singh & Dhingra, sp. nov. FIGS 1-10

MycoBAank 800985

Differs from Inonotus cuticularis, I. farlowii, and I. munzii by its tramal setal hyphae.

Tye: India, Himachal Pradesh: Narkanda, on way to Rampur, on living tree of Quercus

incana Bartram (Fagaceae), 13 August 2011, Harpreet 4746 (PUN, holotype).

Erymo.oey: The epithet refers to the presence of tramal setae.

Basidiocarps annual, pileate, poroid, effused-reflexed to dimidiate to applanate,

solitary to imbricate, up to 4 cm wide, 3 cm in radius, and 1.5 cm thick near

base, with brownish orange exuding droplets; abhymenial surface smooth

to velutinate, azonate, yellowish white to light orange to brown when fresh

becoming glabrous, light orange to bark brown on drying; hymenial surface

poroid, grayish yellow when fresh becoming dark blonde to clay brown on

drying; pores round to angular, 3-5 per mm; margins obtuse, yellowish white

when fresh becoming dark brown on drying, sterile <1 cm; pore tubes <1 mm

16 ... Kaur, Singh & Dhingra

PLATE 1. Inonotus tramisetifer (holotype). Fics 1-4: Basidiocarps. 1. Association with host. 2. Basidio-

carp exuding droplets. 3. Hymenial surface. 4. Abhymenial surface. Fics 5-10: Microscopic

structures. 5. Basidiospores. 6. Basidia. 7. Pilear setal hyphae. 8. Tramal setal hyphae. 9. Generative

hyphae. 10. Section.

Inonotus tramisetifer sp. nov. (India) ... 17

long, entire, brownish orange; dissepiments thin to thick, entire; context <1 cm

thick, firm-fibrous, brownish orange, upper tomentum delimited by a darker

compact layer that is formed by setigerous elements. Hyphal system monomitic;

generative hyphae <7.1 um wide, branched, septate, without clamps, thin- to

thick-walled, pale yellowish to yellowish brown. Setal hyphae of two types 1)

pilear setal hyphae <1350 x 11 um, abundant, monopodial to dichotomously

branched, branches with curved tips, thick-walled, dark brown; 2) tramal setal

hyphae <1300 x 6 um, abundant, monopodial to dichotomously branched,

branches with curved tips, extending into the hymenium <19 um and giving

the misleading appearance of hooked hymenial setae, thick-walled, brown

to dark brown. Hymenial setae absent. Basidia 13-16.5 x 4-4.7 um, clavate,

4-sterigmate, without basal clamp; sterigmata up to 2.4 um long. Basidiospores

5.9-6.5 x 3.5-4.7 um, broadly ellipsoidal, thick-walled, with oil droplets,

yellowish brown, inamyloid, acyanophilous.

REMARKS— Inonotus tramisetifer is similar to I. cuticularis, I. farlowii, and

I. munzii, but I. cuticularis and I. farlowii differ by their hymenial setae and lack

of tramal setal hyphae, and I. munzii differs by its duplex context and lack of

tramal setal hyphae (Ryvarden 2004, 2005).

Acknowledgements

The authors thank Head, Department of Botany, Punjabi University, Patiala, for

providing research facilities; Prof. Leif Ryvarden (Biological Institute, University of

Oslo, Norway) 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

Ryvarden L. 2004. Neotropical polypores Part 1 Introduction, Ganodermataceae & Hymenochaetaceae.

Fungiflora, Oslo, Norway. 227 p.

Ryvarden L. 2005. The genus Inonotus a synopsis. Fungiflora, Oslo, Norway. 147 p.

Sharma JR. 1995. Hymenochaetaceae of India. Botanical survey of India. Calcutta, India. 219 p.

MY COTAXON

http://dx.doi.org/10.5248/123.19

Volume 123, pp. 19-29 January-March 2013

Three new species of Hymenoscyphus from tropical China

HuaANn-D1 ZHENG & WEN- YING ZHUANG*

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

* CORRESPONDENCE TO: zhuangwy@im.ac.cn

ABSTRACT — Three new species of Hymenoscyphus collected from tropical regions of China

are described and illustrated. Hymenoscyphus magnicellulosus possesses whitish sessile

apothecia with large-celled textura prismatica at flank and isodiametric cells at the base, and

inequilateral ellipsoid ascospores 12.5-19(-22) x 4.5-6.7 um. Hymenoscyphus uniseptatus

has white stipitate apothecia, rectangular ectal excipular cells, broadly clavate asci, and

broadly fusoid 1-septate ascospores 16.7—20.5 x 5-6.5 um. Hymenoscyphus yunnanicus is

characterized by cream to light yellow stipitate apothecia, narrow ectal excipular hyphae, and

multiguttulate fusoid ascospores 16.5-19.5 x 5-5.6 um. Comparisons were made among the

new species and their morphologically similar fungi.

Key worps —Helotiales, morphology, taxonomy

Introduction

Hymenoscyphus Gray (Helotiaceae) is characterized by discoid and stipitate

to sessile apothecia, a white to yellowish hymenium surface, an ectal excipulum

of textura prismatica or sometimes mixed with textura angularis, a medullary

excipulum of textura intricata to textura porrecta, ascospores that are fusoid,

ellipsoid to scutuloid, and growth on plant materials (Gray 1821; Dennis 1956;

Sharma 1991; Lizon 1992; Zhang 2002). The main characters used to distinguish

species are color, shape and size of apothecia, structure of ectal excipulum,

size and apical apparatus of asci, shape, size and guttulation of ascospores,

and sometimes substrate specificity (Phillips 1887; Verkley 1993; White 1943;

Dennis 1956; Dumont 1981a,b; Lizon 1992).

Regional studies of Hymenoscyphus were carried out in Europe (Phillips

1887; Velenovsky 1934; Dennis 1956, 1962, 1968; Lizon 1992; Milekhin &

Prokhorov 2008), America (Seaver 1951; Dennis 1954, 1958a, 1960; Dumont

& Carpenter 1982; Gamundi 2003), Asia (Thind & Singh 1970, 1972; Thind

& Sharma 1980; Sharma 1991; Zhang 2002; Han & Shin 2008), Oceania

20 ... Zheng & Zhuang

(Dennis 1958b, 1961), and Africa (Dennis 1984; Descals et al. 1984; Fisher &

Spooner 1987). More than 500 names have been connected to Hymenoscyphus

(http://www.indexfungorum.org/Names/Names.asp), and approximately 155

species are currently accepted (Kirk et al. 2008). The genus needs a monographic

treatment.

Twenty-six species are known from China (Teng 1934, 1963; Tai 1979;

Zang 1983; Korf & Zhuang 1985; Zhuang & Korf 1989; Bi et al. 1990; Zhuang

1995, 1998a,b, 1999a,b, 2001, 2003a,b; Zhuang & Wang 1998a,b; Yu et al.

2000; Wang & Pei 2001; Zhang 2002; Zhang & Zhuang 2002a,b, 2004). The

species previously described from China were mainly based on materials from

temperate areas (Zhang 2002; Zhang & Zhuang 2002a,b, 2004), and very few

species were known from tropical China.

During our examinations of the Hymenoscyphus collections from tropical

Hainan and Yunnan provinces, we reported six species (Zheng & Zhuang

2011), of which five occur also in temperate China. As a continuation of the

work, we describe and name three new species in this paper: H. magnicellulosus,

H. uniseptatus and H. yunnanicus. Distinctions among the new species and

their related fungi are discussed.

Materials & methods

Specimens examined were collected from tropical areas of Hainan and Yunnan

provinces in 1999 and 2000. Field notes were taken from apothecia in fresh condition.

Dried apothecia were rehydrated with distilled water and sectioned at a thickness of

10-20 um with a freezing microtome (YD-1508A, Yidi Medical Instrument Co., Jinhua,

China). Measurements were taken from longitudinal sections and from squash mounts

in lacto-phenol cotton blue solution using an Olympus BH-2 microscope (Tokyo,

Japan). Iodine reactions of ascus apparatus were tested in Melzer’s reagent with 3%

KOH aqueous solution pretreatments. Features of apical ring were recorded according

to Baral (1987). Ascospore guttulation was examined in 3% KOH aqueous solution.

Photographs were taken using a Canon G5 digital camera (Tokyo, Japan) connected

to a Zeiss Axioskop 2 Plus microscope (G6ttingen, Germany) and a Zeiss Stemi 2000C

(G6ttingen, Germany) stereomicroscope. Line drawings were made using an Opton

microscope (West Germany) equipped with a drawing tube. Specimens examined are

deposited in the Mycological Herbarium, Institute of Microbiology, Chinese Academy

of Sciences (HMAS).

Taxonomy

Hymenoscyphus magnicellulosus H.D. Zheng & W.Y. Zhuang, sp. nov.

MycoBank MBs564248 Fics 1A, 2, 5A—C

Differs from Hymenoscyphus tamaricis by its much smaller whitish sessile apothecia,

narrower medullary hyphae, and shorter and wider asci.

Type: China, Yunnan: Pingbian, Daweishan, alt. 1900 m, on rotten bark, 4 Nov 1999,

W.Y. Zhuang & Z.H. Yu 3254 (Holotype, HMAS 188555).

Hymenoscyphus spp. nov. (China) ... 21

FIGURE 1 Gross morphology of dried apothecia on natural substrates. A. Hymenoscyphus

magnicellulosus (holotype); B. H. uniseptatus (holotype); C. H. yunnanicus (holotype). Scale bars

= 0.5 mm.

Erymo.ocy: The specific epithet refers to the large cells in ectal excipulum.

Apothecia turbinate, subsessile to sessile, scattered or 2—5 in a cluster, margin

smooth, 0.6-2 mm in diam.; hymenium surface flat to convex, drying slightly to

deeply concave, whitish when fresh, drying pale tan; receptacle surface smooth,

paler than hymenium. Outer covering layer absent. Ectal excipulum 20-55 um

thick, of textura porrecta at margin, hyphae 3-5 um wide; becoming textura

prismatica at flank, cells 8-30 x 4-11 um; and of textura angularis towards the

base, cells up to 25 x 11 um, or 5-15 um in diam. if isodiametric, cells hyaline,

thin- to slightly thick-walled. Medullary excipulum of textura intricata, 30-250

um thick, hyphae hyaline, 1.5-4 um wide, thin-walled. Subhymenium not

distinguishable. Hymenium 95-150 um thick. Asci8-spored, 88-129 x 9-12 um,

cylindric-clavate, gradually narrowed into a short-medium stalk, base attenuate,

arising from croziers, apex round to broadly papilliform, apical thickening 2-3

um thick, J+ in Melzer’s reagent with or without KOH pretreatment; apical ring

weakly to moderately bluing as two lines about 1-1.5 um long, even in width or

slightly thinner upward, of the Hymenoscyphus-type. Ascospores 12.5-19(-22)

x 4.5-6.7 um, obliquely uniseriate to irregularly biseriate, hyaline, ellipsoid,

slightly flattened on one side, occasionally 1-septate, with a dark stained area in

cotton blue, containing 1-2 large oil drops and several small ones in cotton blue

and KOH solution; guttules ellipsoid, rounded to irregular in shape, individuals

up to 10 x 5 um. Paraphyses filiform, slightly swelling and 2-3 um at the apex,

1-2 um at lower part, equal to or slightly exceeding the asci.

ADDITIONAL SPECIMENS EXAMINED: China, Hainan: Changjiang, Bawangling, alt. 1150

m, on rotten wood of broad-leaved tree, 7 Dec 2000, W.Y. Zhuang, Z.H. Yu & Y.H. Zhang

3669 (HMAS 188556); Lingshui, Diaoluoshan, alt. 1080 m, on rotten wood of broad-

leaved tree, 14 Dec 2000, W-Y. Zhuang, Z.H. Yu & Y.H. Zhang 3866 (HMAS 188557);

Tongza, Wuzhishan, alt. 1600 m, on rotten wood of broad-leaved tree, 16 Dec 2000, W.Y.

Zhuang, Z.H. Yu & Y.H. Zhang 3876 (HMAS 188558).

Notes: All examined H. magnicellulosus specimens share very similar

macroscopic and microscopic characteristics.

22 ... Zheng & Zhuang

FIGURE 2 Hymenoscyphus magnicellulosus (holotype). A. Structure of apothecium; B. Excipular

structure and hymenium at margin; C. Excipular structure at flank and near base; D-J. Ascospore

(E. 1-septate ascospore); K. Ascospores in asci; L-O. Asci. Scale bars: A = 100 um, B, C = 20 um,

D-O = 10 um.

Hymenoscyphus tamaricis R. Galan, Baral & A. Ortega resembles H. magni-

cellulosus in lignicolous habit, ectal excipulum structure, asci arising from

croziers, and guttulate and occasionally 1-septate ascospores of similar size and

shape, but differs in stipitate, pale to light yellowish-ochraceous apothecia which

are much larger [(1—)1.5-7(-9) mm in diam.], broader medullary hyphae (3-9

um wide), longer and narrower asci (125-140 x 8.2-8.5 um), and restricted

only on Tamarix host (Galan & Baral 1997).

A species with similarly sized white apothecia and isodiametric cells in ectal

excipulum is H. immutabilis (Fuckel) Dennis, which can be distinguished from

H. magnicellulosus by short-stipitate apothecia, smaller and fusoid ascospores

(10-13 x 4—4.5 um) biseriate in smaller asci (80-100 x 8-9 um), and mainly

foliicolous habit (White 1943; Dennis 1956; Dumont 1981b).

Hymenoscyphus fagineus (Pers.) Dennis has whitish and sessile to subsessile

apothecia similar to those of H. magnicellulosus. It is distinct in ectal excipulum

composed of short rectangular cells lying at a high angle to the surface, smaller

asci (65-80 x 8-9 um) J+ and with pore strongly blue in Melzer’s reagent,

smaller ascospores (8-16 x 4-5 um), and occurring only on fallen Fagus

sylvatica pericarps (Dennis 1956; Lizon 1992).

Hymenoscyphus spp. nov. (China) ... 23

FIGURE 3 Hymenoscyphus uniseptatus (holotype). A. Structure of apothecium; B. Excipular structure

and hymenium; C-H. Asci; I-M. Ascospore. Scale bars: A = 100 um, B = 20 um, C-M = 10 um.

Hymenoscyphus uniseptatus H.D. Zheng & W.-Y. Zhuang, sp. nov.

MycoBAnk MB564250 Fics 1B, 3, 5D-F

Differs from Hymenoscyphus jinggangensis by its smaller apothecia, larger asci, and

larger 1-septate multiguttulate ascospores.

Type: China, Yunnan: Malipo, Nanwenhe, alt. 900 m, on herbaceous stems and rotten

wood of broad-leaved tree, 9 Nov 1999, W.Y. Zhuang & Z.H. Yu 3356 (Holotype, HMAS

188559).

ErymMo.oey: The specific epithet refers to the uniseptate ascospores.

Apothecia stipitate, scattered or 2-3 in clusters, round, margin smooth, 0.7-1

mm in diam., stipe 0.3-0.8 mm long; hymenium surface flat to discoid, white

when fresh, becoming dull orange when dry; receptacle surface smooth,

concolorous with hymenium. Outer covering layer absent. Ectal excipulum

of textura prismatica, 15-30 um thick, cells hyaline, slightly thick-walled,

24 ... Zheng & Zhuang

rectangular to short rectangular, 10-22 x 7-15 um. Medullary excipulum with

two layers, outer layer of textura porrecta, 5-20 um thick, inner layer of textura

intricata, 15-30 um thick, hyphae hyaline, 2-3 um wide. Subhymenium not

distinguishable. Hymenium 90-105 um thick. Asci 8-spored, 80-95 x 8.5-14

um, broadly clavate, base attenuate, arising from simple septa, apex round to

conical, apical thickening 3-6 um thick, J+ in Melzer’s reagent with or without

KOH pretreatment; apical ring weakly bluing as two lines about 1.5-3 um long,

equal to or becoming thinner upward, of the Hymenoscyphus-type. Ascospores

16.7-20.5 x 5-6.5 um, biseriate to irregularly uniseriate, subfusoid, hyaline,

1-septate, containing 3-6 large oil drops and many small ones in cotton blue

and KOH solution; guttules round or irregular in shape, individual ones up to

5 x 4um. Paraphyses filiform, equal to or slightly exceeding the asci, 1.5-2 um

wide.

Notes: The type specimen of H. uniseptatus contains apothecia growing on

both herbaceous stems and rotten wood. The macroscopic and microscopic

characters of apothecia on different substrates are identical. In view of this, we

do not regard H. uniseptatus as host-specific.

Hymenoscyphus jinggangensis Yan H. Zhang & W.Y. Zhuang, originally

described from temperate China, is most similar to H. uniseptatus in stipitate

white apothecia, size of ectal excipulum cells, septate ascospores, and lignicolous

habit. However, H. jinggangensis differs in larger apothecia (1-2.8 mm in diam.),

thin-walled ectal excipulum cells, thicker medullary excipulum (up to 235 um

thick), smaller asci (65-83 x 7-8 um), and smaller (9-17 x 3-5 um) eguttulate

ascospores that are 1-3(—4)-septate instead of uniseptate (Zhang 2002; Zhang

& Zhuang 2002a).

Among the known Hymenoscyphus species, H. musicola (Dennis) Dennis

resembles H. uniseptatus in stipitate, whitish apothecia, ectal excipulum of

large-celled textura prismatica, 1-septate ascospores of similar size, shape and

guttulation. It can be easily separated from H. uniseptatus by convex apothecial

surface, longer asci [(90-—)100-110 x 9-12 um] arising from croziers, and

occurrence on Musa sapientum (Dennis 1958a; Dumont 198 1a).

Hymenoscyphus varicosporoides Tubaki is also similar in stipitate, white

apothecia, and 1-septate ascospores, but differs in narrower ectal excipulum

hyphae (Tubaki 1966: Pl. II fig. 2), much narrower asci (70-85 x 7-9 um),

smaller ascospores [(10—)12—15(-19) x 4-5(-7) um] with round ends, and

growth on rotting twigs in a semi-aquatic environment (Tubaki 1966).

Hymenoscyphus yunnanicus H.D. Zheng & W-Y. Zhuang, sp. nov. Fics 1C, 4, 5G-I

MycoBank MB564251

Differs from Hymenoscyphus crataegi by its larger apothecia, smaller ectal excipular

cells, larger asci, and larger ascospores.

Hymenoscyphus spp. nov. (China) ... 25

FiGuRE 4 Hymenoscyphus yunnanicus (holotype). A. Structure of apothecium; B. Excipular structure

and hymenium; C. Excipular structure between flank and stipe; D-G. Asci; H—P. Ascospore.

Scale bars: A = 100 um, B—C = 20 um, D-P = 10 um.

Type: China, Yunnan: Xichou, Xiaoqiaogou, alt. 1400 m, on rotten leaves of deciduous

tree, 11 Nov 1999, W.Y. Zhuang & Z.H. Yu 3424 (Holotype, HMAS 188560); W.Y.

Zhuang & Z.H. Yu 3430 (Isotype, HMAS 188561).

Erymo ocy: The specific epithet refers to the type locality of the fungus.

Apothecia stipitate, scattered, flat to discoid, 0.6—1.5 mm in diam., stipe 0.5—0.8

mm long; hymenium surface cream to light yellow when fresh, drying brownish

orange; receptacle surface smooth, paler than hymenium. Outer covering layer

of 1-2 cell layers, hyphae 1-2 um wide. Ectal excipulum of textura prismatica,

15-30 um thick, cells hyaline, 6-12 x 3-5 um. Medullary excipulum with two

layers, outer layer of textura porrecta, 6-40 um thick, inner layer of textura

intricata, 40-110 um thick, hyphae hyaline, 2-3 um wide. Subhymenium not

distinguishable (holotype) or 15-30 um thick (isotype). Hymenium 95-110

um thick. Asci (75—)87-120 x 7-11 um, clavate, 8-spored, some with a very

short stalk, base attenuate, arising from simple septa, apex round to somewhat

truncate, apical thickening 1-2 um thick, J+ in Melzer’s reagent with or without

KOH pretreatment; apical ring strongly bluing as two lines about 1-1.5 um

long and slightly thicker upward. Ascospores 16.5-19.5 x 5-5.6 um, obliquely

uniseriate to irregularly biseriate, hyaline, fusoid or rounded at anterior end,

26 ... Zheng & Zhuang

AR A) 0808

\ eeeee

ANS \s84008

FIGURE 5 Ascospores and asci of Hymenoscyphus spp. (holotypes). A-—C. H. magnicellulosus;

D-F. H. uniseptatus; G-I. H. yunnanicus. A, D, G. Apices of asci in Melzer’s reagent; B, E, H. Base of

the asci showing croziers or simple septa; C, F, I. Ascospores in KOH. Scale bars = 10 um.

slightly inequilateral, with a dark stained area in cotton blue, containing 1-3

large oil drops or multiguttulate in cotton blue and KOH solution; guttules

rounded or irregular in shape, large ones 2-4 um in diam. or in width.

Paraphyses filiform, equal to or slightly exceeding the asci, 1.5-2 um wide.

Notes: The two specimens examined are basically the same in morphology,

although the subhymenium, not distinguishable in the holotype, is well

developed in the isotype, which we regard as infraspecific variation.

Among the known Hymenoscyphus species possessing stipitate apothecia

and occurring on leaves, H. yunnanicus is most similar to H. phyllogenus

(Rehm) Kuntze in excipular structure. The latter differs in white and much

smaller (0.3-0.5 mm in diam.) apothecia, shorter (65-75 x 8-11 um) asci, and

ascospores that are smaller (11-16 x 4-6 um), eguttulate and ellipsoid (Dennis

1956; White 1943).

Another foliicolous Hymenoscyphus species that possesses stipitate

apothecia and fusoid multiguttulate ascospores is H. crataegi Baral & R. Galan

from Spain on leaves of Crataegus monogyna. It differs from H. yunnanicus by

Hymenoscyphus spp. nov. (China) ... 27

smaller apothecia (0.4—0.6 mm in diam.), slightly gelatinized ectal excipular

tissue composed of much larger cells (12-43 x 10-16 um), smaller asci (60-80

x 6-7.5 um), and smaller ascospores [14—17.5 x (3—)3.5—4(-4.5) um] (Baral et

al. 2006).

Acknowledgments

We thank Dr. Y.-Z. Wang and Dr. T. Hosoya for critically review of the manuscript

and invaluable suggestions, Dr. S.R. Pennycook and Dr. L.L. Norvell for detailed

corrections and valuable editorial help, Mr. H.-O. Baral for consultation, and Ms. Xia

Song for making sections of some materials for this study. This work was supported by

the National Natural Science Foundation of China (no. 31093440).

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MY COTAXON

http://dx.doi.org/10.5248/123.31

Volume 123, pp. 31-37 January-March 2013

Gloeophyllum protractum is synonymous with G. mexicanum

JiRf Kout’’, JOSEF VLASAK JR. & JOSEF VLASAK?

‘University of West Bohemia, Faculty of Education, Department of Biology, Geosciences and

Environmental Education, Klatovskd 51, CZ-306 19 Pilsen, Czech Republic

? Biol. Centre of the Academy of Sciences of the Czech Republic,

Branisovskdé 31, CZ-370 05 Ceské Budéjovice, Czech Republic

*CORRESPONDENCE TO: martial@seznam.cz

ABSTRACT — Gloeophyllum protractum, a boreal polypore, is synonymized with the

subtropical G. mexicanum. This taxonomic opinion is supported by sequencing of ITS and

nuLSU regions. Morphology and ecology of this species are discussed.

Key worps — Basidiomycota, Gloeophyllales, Gloeophyllaceae, molecular taxonomy

Introduction

The species of Gloeophyllum P. Karst. comprise a distinct group among

polypores distinguished by a brown rot, tough dark brown context, and

very variable (poroid, daedaleoid, or lamellate) hymenophore (Gilbertson &

Ryvarden 1986). The six species (G. abietinum (Bull.) P. Karst., G. carbonarium

(Berk. & M.A. Curtis) Ryvarden, G. odoratum (Wulfen) Imazeki, G. protractum

(Fr.) Imazeki, G. sepiarium (Wulfen) P. Karst., and G. trabeum (Pers.) Murrill)

known from Europe (Ryvarden & Gilbertson 1993, Niemela 2005) are

differentiated based on some critical morphological features and ecology,

because the hyphae, hymenium, and spores are very similar in all species. In

North America, these same taxa occur together with G. mexicanum (Mont.)

Ryvarden (mostly on pines in subtropical regions and with G. striatum (Sw.)

Murrill (a hardwood-specific, tropical species). Of the eight species found in

East Asia (Dai 2012), seven are also found in Europe and North America and

one (G. imponens (Ces.) Teng) is a tropical species growing on angiosperm

wood (Dai et al. 2011, Dai 2012). All European species represent very common

wood-inhabiting fungi except for G. protractum, which is very rare in Europe,

limited to inner Scandinavia (Ryvarden & Gilbertson 1993) and some recently

described localities in the Mediterranean region (Bernicchia et al. 2007, Tura et

32 ... Kout, Vlasak Jr. & Vlasak

al. 2010). In contrast, in the USA G. protractum is described as widespread from

Florida to Alaska (Gilbertson & Ryvarden 1986) and is occasionally recorded

also in East Asia (Dai et al. 2004, Dai 2012).

The unique phylogenetic position of the Gloeophyllum clade, separated

from other polypores, was extensively studied by Garcia-Sandoval et al.

(2011). Although many Gloeophyllum spp. sequences have been published,

G. protractum is represented in GenBank by only one sequence, which is,

however, identical with G. odoratum sequences. To clarify this issue, we sequenced

variable rRNA regions of typical specimens of G. odoratum, G. protractum, and

G. mexicanum.

Materials & methods

Specimens of Gloeophyllum were collected by the authors in Europe and USA or

obtained from University of Helsinki herbarium, Finland (H). Our collected specimens

are deposited in private herbarium of J. Vlasak (JV); some duplicates were also deposited

in Prague Museum Herbarium (PRM).

SPECIMENS EXAMINED: Gloeophyllum carbonarium: USA, FLoripa, Everglades NP,

pine, 29.VIII.2010, leg. & det. J. Vlasak (JV1008/67; GenBank JX266860, JX266862);

Mahagony Hammock, pine, 21.XII.2003, leg. J. Vlasak Jr., det. J. Viasak (JV0312/21.10-J;

GenBank JX266861, JX266863).

Gloeophyllum mexicanum: USA, FLoripa, Everglades NP, pine, 29.VIII.2010, leg.

& det. J. Vlasak (JV1008/67; GenBank JX266857); Big Cypress Nat. Preserve, Collier-

Seminole State Park, 25.XII.2003, leg. J. Vlasak Jr., det. J. Viasak (JV0312/25.14-J); Ocala,

Ocala Nat. Forest, pine, 19.XII.2002, leg. J. Vlasak Jr., det. J. Vlasak (JV0212/35-J, PRM

860472; GenBank JX266856, JX266864).

Gloeophyllum odoratum: CZECH REPUBLIC, Hluboka nad Vltavou, Borek, spruce

stump, 2.X1.2004, leg. & det. J. Vlasak (JV0411/2; GenBank JX266858, JX266859).

Gloeophyllum protractum: USA, CALIFORNIA, Sierra Nevada, Tioga Pass, lodgepole

pine, 18.VIHI.2001, leg. & det. J. Vlasak (JV0108/100, PRM 860473; and JV 0108/101,

PRM 860474; GenBank JX266850, JX266865 [sequences of both specimens identical]);

OREGON, Crater Lake, pine, 15.1X.2007, leg. J. Vlasak Jr., det. J. Vlasak (JV 0709/135-

J; GenBank JX266851, JX266852). FINLAND, INaRIN Lappt, Inari, Lemmenjoki

National Park, fallen charred trunk of Pinus sylvestris, 1.[X.1974, leg. & det. T. Niemela

(H 6012949; GenBank JX266853, JX266854); fallen decorticated trunk in dry site,

3.1X.1980, leg. & det. T. Niemela (H 6012948; GenBank JX266855).

DNA methods

DNA was isolated and sequenced as described by Vlasak & Kout (2011). The

evolutionary history was inferred by using the Maximum Likelihood method; the

analyses were conducted in MEGA 5 (Tamura et al. 2011).

Results & discussion

We sequenced rRNA ITS region of several Gloeophyllum mexicanum and

G. protractum specimens and one typical G. odoratum specimen. To root

the phylogeny we used our sequences of G. carbonarium (considered closely

related to G. odoratum by Garcia-Sandoval et al. 2011). Other G. carbonarium,

Gloeophyllum protractum, a synonym of G. mexicanum ... 33

JV0709/135-J hapl2 USA G. protractum

H 6012948 Finland G. protractum

JV0212/35-J USA G. mexicanum

H 6012949-hapI1 Finland G. protractum

JV1008/67A USA G. mexicanum

8 | HM536088 G. mexicanum

H 6012949-hapl2 Finland G. protractum

JV0709/135-J hapl1 USA G. protractum

JV0108/101 USA G. protractum

56! JV0108/100 USA G. protractum

JV0411/2 hapl1 G. odoratum

JV0411/2 hapl2 G. odoratum

100 | HM536090 G. protractum USDA strain H-80

HM536089 G. odoratum

FJ627257 China G. carbonarium

io | ;JV1008/67 USA G. carbonarium

35! J¥0312/21.10-J USA G. carbonarium

G. mexicanum 34

G. odoratum

G. carbonarium

——— +3 10 changes

Fic. 1: Phylogenetic relationships of Gloeophyllum species inferred with ITS rRNA sequences.

Sequenced specimens were determined according to their morphology; other sequences were

retrieved with their names from GenBank. Gloeophyllum carbonarium was used to root the tree.

Topology from maximum likelihood analysis. Support values along branches from maximum

likelihood bootstrap. Branch lengths are drawn proportional to genetic distances (bar indicates

10 changes per ITS1-5.8S-ITS2 region). Some of the specimens show two haplotypes differing by

short insertions/deletions. GenBank numbers indicate sequences retrieved from GenBank; other

GenBank numbers in Materials & method.

G. mexicanum, and G. odoratum sequences were retrieved from GenBank.

Alignment and simple analysis showed that the G. mexicanum sequences are

identical with G. protractum sequences (Fie. 1). In particular, the G. protractum

and G. mexicanum ITS sequences differ on average in only two positions,

which corresponds to differences between individual G. mexicanum specimens

or between different haplotypes in one collection. On the other hand, in

G. odoratum approximately 10% of the positions differ from G. mexicanum/

protractum sequences. The same result — identity of G. mexicanum with

G. protractum and a pronounced divergence in G. odoratum — was obtained

using nuLSU sequence analysis (not shown). The ITS2 region in one of two

European G. protractum specimens had a unique insertion (4b) not present in

the other specimen from the same locality. Accordingly, identity of European

and American G. mexicanum/protractum specimens was also confirmed.

The name Gloeophyllum mexicanum [originally proposed in 1843

from Mexico as Lenzites mexicana Mont. (Montagne 1843)] has priority

34 ... Kout, Vlasak Jr. & Vlasdk

tee

nny

Fic. 2: Gloeophyllum mexicanum JV0212/35-J, Florida, typical, photo in collection.

over G. protractum [originally described in 1851 from northern Europe as

Trametes protracta Fr. (Fries 1851)]. Gilbertson & Ryvarden (1986) separate

G. mexicanum (Fic. 2) from G. protractum based primarily on the daedaleoid

to lamellate hymenophore (which they describe as poroid to radially elongated

in G. protractum). Nevertheless, in our collections both from California (Fics 3,

4) and Finland, one specimen had a more or less poroid hymenophore while the

other from the same locality showed a daedaleoid to lamellate structure. We infer

that this feature is not diagnostic. In other respects — especially in the glabrous

to semi-glossy pileal surface unique in the genus as well as microscopically

— the two specimens appear identical. Particularly, G. mexicanum is described

as pileate polypore with sessile and broadly attached fruitbodies elongated

along the substrate, 3-10 x 1-3 cm, <1 cm thick, mostly applanate, more rarely

triquetrous, with upper surface at first finely tomentose but soon glabrous with

a poroid to daedaleoid hymenophore, occasionally with few lamellae 8-12 per

cm, trimitic hyphal system, and basidiospores 9-12 x 3.5-4 um; G. protractum

is described as a pileate polypore with sessile fruitbodies distinctly elongated

along the substrate, 10 x 4 cm, 5-15 mm thick, often slightly triquetrous in

section, with a glabrous to semi-glossy pileal surface when juvenile, pores

entire to weakly elongated radially, 1-2 per mm, trimitic hyphal system, and

basidiospores 8.5-11(-12) x 3-4(-4.5) um (Gilbertson & Ryvarden 1986). The

main difference is their ecology, with G. mexicanum regarded as a subtropical

or tropical American species and G. protractum as boreal or alpine in Europe

where it was described. In the USA, we also found the typically thick poroid

G. protractum only in the Sierra Nevada at 3000 ma.s.L, not at lower elevations.

But we believe that as it descends the fungus becomes more applanate and

lamellate so that it forms a “typical” G. mexicanum fructification near the

Gloeophyllum protractum, a synonym of G. mexicanum ... 35

Fic. 3: Gloeophyllum mexicanum JV0108/101, Sierra Nevada, Tioga Pass,

determined originally as G. protractum, photo in situ.

Fic. 4: Gloeophyllum mexicanum JV0108/100, Sierra Nevada, Tioga Pass,

determined originally as G. protractum, photo in collection.

36 ... Kout, Vlasak Jr. & Vlasak

southern USA border. Intermediate forms may be sometimes determined as

G. mexicanum, sometimes as G. protractum, and this is the reason, in our

opinion, why so many G. protractum finds are reported from lowland and

southern USA (Gilbertson & Ryvarden 1986).

From the above descriptions, inspection of our own specimens, and molecular

data we infer that the crucial features of G. mexicanum and G. protractum are

in total agreement. We show that different ecology and associated differences

in morphology need not result in species differentiation as now understood

(Taylor 2000). Similar results were recently obtained with very variable

American specimens of Fuscoporia viticola (Schwein.) Murrill analyzed by

molecular methods (Vlasak et al., unpublished).

There is something odd also with two other Gloeophyllum species recorded

from the USA: G. odoratum and G. abietinum. These extremely common

European fungi are reported from only rather limited areas in the western USA

(Gilbertson & Ryvarden 1986) in a region known for its phytogeographical

uniqueness. We regard the distribution maps of G. odoratum as unreliable

because G. protractum has probably been misidentified as G. odoratum in the

USA. We were not able to find either species when we visited the putative areas

of G. odoratum and G. abietinum occurrence in the USA.

Acknowledgments

Prof. Yu-Cheng Dai and Dr. Viacheslav Spirin have kindly acted as presubmission

reviewers and their help is acknowledged. This research was funded with institutional

support RVO: 60077344.

Literature cited

Bernicchia A, Savino E, Gorjén SP. 2007. Aphyllophoraceous wood-fungi on Pinus spp. in Italy.

Mycotaxon 101: 5-8.

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, Wei YL, Wang Z. 2004. Wood-inhabiting fungi in southern China 2. Polypores from

Sichuan Province. Ann. Bot. Fenn. 41: 319-329.

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. Fenn. 48: 219-231.

Fries EM. 1851. Nya Svamparter. Ofvers. K. Vetensk.-Akad. Forh. 8: 42-54.

Garcia-Sandoval R, Wang Z, Binder M, Hibbet DS. 2011. Molecular phylogenetics of the

Gloeophyllales and relative ages of clades of Agaricomycotina producing a brown rot. Mycologia

103: 510-524. http://dx.doi.org/10.3852/10-209

Gilbertson RL, Ryvarden L. 1986. North American polypores, Vol. 1. Fungiflora. Oslo. 1-433.

Montagne JPFC. 1843. Quatri¢me centurie de plantes cellulaires exotiques nouvelles, Décades VIII,

IX et X. Ann. Sci. Nat., Bot., 2e sér., 20: 352-379.

Niemela T. 2005. Polypores, lignicolous fungi. Noorrlinia 13: 1-320.

Ryvarden L, Gilbertson RL. 1993. European polypores 1. Synopsis Fungorum 6: 1-387.

Gloeophyllum protractum, a synonym of G. mexicanum ... 37

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 28: 2731-2739.

http://dx.doi.org/10.1093/molbev/msr121

Taylor JW, Jacobson DJ, Kroken S, Kasuga T, Geiser DM, Hibbett DS, Fisher MC. 2000. Phylogenetic

species recognition and species concepts in fungi. Fungal Genetics and Biology 31: 21-32.

http://dx.doi.org/10.1006/fgbi.2000.1228

Tura D, Zmitrovich IV, Wasser SP, Nevo E. 2010. Checklist of Hymenomycetes (Aphyllophorales

s.l.) and Heterobasidiomycetes in Israel. Mycobiology 38: 256-273. http://dx.doi.org/10.4489/

MYCO.2010.38.4.256

Vlasak J, Kout J. 2011. Tropical Trametes lactinea is widely distributed in the eastern USA.

Mycotaxon 115: 271-279. http://dx.doi.org/10.5248/115.271

MY COTAXON

http://dx.doi.org/10.5248/123.39

Volume 123, pp. 39-45 January-March 2013

Pseudofusicoccum adansoniae isolated as an endophyte

from Jatropha podagrica: new record for India

ROHIT SHARMA’, GIRISH KULKARNI’? & YOGESH S. SHOUCHE!

Microbial Culture Collection, National Centre for Cell Science &

?Molecular Biology Unit, National Centre for Cell Science

NCCS Complex, Ganeshkhind, Pune- 411 007 (Maharashtra), India

* CORRESPONDENCE TO: rohit@nccs.res.in

ABSTRACT — Pseudofusicoccum adansoniae (Botryosphaeriaceae), representing a new generic

record for India, is described and illustrated. The species, isolated as an endophyte from

healthy leaf midrib and fruit of Jatropha podagrica, is characterized by large conidiomata and

hyaline ellipsoid conidia with round apices. Its teleomorph is unknown. Comparison of the

internal transcribed spacer 1 and 2 loci and 5.88 rDNA sequences showed 100% sequence

similarity with that of CBS 122055, the ex-type strain of P. adansoniae.

Key worps — anamorph, Botryosphaeriaceae, ITS, phylogeny, taxonomy

Introduction

Pseudofusicoccum Mohali et al. (Botryosphaeriaceae) is a relatively new

genus typified by Pseudofusicoccum stromaticum (Mohali et al.) Mohali et al.

(Crous et al. 2006). Recent morphological studies and ITS1-5.8S-ITS2 sequence

analyses indicate that Pseudofusicoccum species occur in Venezuela and Brazil

(P. stromaticum), Western Australia (P adansoniae Pavlic et al., P ardesiacum

Pavlic et al., P kimberleyense Pavlic et al.), and South Africa (P olivaceum

J.W.M. Mehl & B. Slippers, P. violaceum J.W.M. Mehl & B. Slippers) (Mohali et

al. 2006, 2007; Pavlic et al. 2008; Mehl et al. 2011; Marques et al. 2012) (TABLE

1). Although some species have been isolated from dead wood, most have

been isolated from healthy plant tissues where these fungi occur seemingly as

endophytes. Recently, during an investigation of endophytic fungal diversity

of Jatropha podagrica Hook. (Euphorbiaceae), two isolates were recovered with

the morphological characteristics of Pseudofusicoccum adansoniae. The plant

is considered toxic (Ojewole & Odebiyi 1980) and used for tanning leather

and the production of a red dye (Bassam 2010). A literature survey indicated

AO ... Sharma, Kulkarni & Shouche

that Pseudofusicoccum has not previously reported from India (Bilgrami et al.

1991, Jamaluddin et al. 2004), and P adansoniae has been reported only from

Australia. The present communication describes and illustrates these isolates in

light of the molecular sequence data from ITS rDNA loci.

Materials & methods

The fungus was isolated from healthy leaf midrib and fruit of the perennial plant

Jatropha podagrica (Buddha belly or Gout plant) from Pune (Pimpri), Maharashtra,

India (18°37'07.04"N 73°48'13.43"E). Plant parts were washed in running tap water,

sterile distilled water (3 washes) and cut into fragments (1-2 cm). Pieces of the plant

were dipped and washed with 75% ethanol (5 min, twice) and 100% ethanol (30 sec),

placed on Potato Dextrose agar (PDA) (Hi-Media, India) and Czapek Dox agar (CDA)

(Hi-Media, India) media, and incubated at room temperatures (RT; c. 28-30 °C). After

4-5 days, mycelial growth was observed on the CDA Petri dish. The cultures were

purified on 2% Malt Extract agar (MEA) plates and strains designated as MMI00062

(from leaf midrib) and MMI00064 (from fruit). Unstained microscopic observations

were made using lactophenol (Hi-Media, India) mounts and observed under a Nikon

YS100 microscope (Nikon, Japan). Measurements of morpho-taxonomic characters

were recorded and compared with type descriptions of known species (Mohali et al.

2006, Pavlic et al. 2008, Mehl et al. 2011). Rayner’s colour chart (1970) was used as a

reference for culture colours.

Scanning Electron Microscopy (SEM) of air-dried pycnidia was done by coating with

platinum in Jeol sputter coater (JFC 1600) and examining on a Jeol-JSM 6360A SEM at

10 kV. The culture MMI00064 has been deposited in the Microbial Culture Collection

(WDCM-930), Pune, India as MCC 1020.

For sequencing of the ITS loci, DNA was extracted from pure cultures grown in

Petri dishes using the QIAamp® DNA Mini Kit (Qiagen, Inc., Valencia CA). DNA

concentration was estimated using a Nanodrop ND-1000 machine (Thermo scientific,

USA). The genomic DNA was amplified using ITS1 and ITS4 primers (White et al. 1990)

in a PE 9700 thermocycler (PE, Applied Biosystems, Singapore) with the following

conditions: initial denaturation at 94 °C for 2 min, 35 cycles of denaturation at 94 °C for

1 min, annealing at 55 °C for 1 min, extension at 72 °C for 1 min, and a final extension

step at 72 °C for 10 min. The positive amplicons were purified using a PCR Cleanup Kit

(Qiagen, Inc., Valencia CA) and purified PCR products were sequenced (both strands)

on an ABI 3730 xl DNA analyzer using the Big Dye terminator kit (Applied Biosystems,

Inc., Foster City, CA). An NCBI BLASTn search was conducted for sequence similarity

(Zhang et al. 2000).

For phylogenetic relationships with Pseudofusicoccum adansoniae MCC 1020,

11 related sequences were retrieved from GenBank database. Neoscytalidium

novaehollandiae CBS 122071 (EF585540.1) was used as the outgroup and sequence

alignment carried out using MUSCLE (Edgar 2004). Maximum parsimony (MP)

analysis and phylogenetic tree construction was conducted using MEGA 5.0 (Tamura

et al. 2011).

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Pseudofusicoccum kimberleyense CBS122058 (EU144057.1)

Pseudofusicoccum kimberleyense CBS 122061 (EU144059.1)

Pseudofusicoccum violaceum CBS 124936 (FJ888474.1)

2 'Pseudofusicoccum violaceum CMW22671 (F J888472.1)

Pseudofusicoccum ardesiacum CBS 122062 (EU144060.1)

52 | Pseudofusicoccum ardesiacum CBS 122063 (EU144061.1)

Pseudofusicoccum olivaceum CBS 124939 (F J888459.1)

98 ' Pseudofusicoccum olivaceum CMW22639 (FJ888463.1)

Pseudofusicoccum adansoniae MCC 1020 (JQ585586)

93 'Pseudofusicoccum adansoniae CBS 122055 (EF585523.1)

Pseudofusicoccum stromaticum CBS117449 (DQ436935.1)

90 ' Pseudofusicoccum stromaticum CMW13426 (EF 118041.1)

Neoscytalidium novaehollandiae CBS122071 (EF 585540. 1)

PLATE 1. Maximum parsimony tree (by complete deletion of gaps) based on ITS sequences (ITS1-

5.8S-ITS2) depicting the position of P adansoniae MCC 1020 within Pseudofusicoccum and its relation

with other closely related phylospecies. Bootstrap values (1000 replicates) shown above branches.

Results

The cultures isolated were identified as Pseudofusicoccum adansoniae

based on colony size and colour, conidial size, and conidiophore morphology

as described by Pavlic et al. (2008). When the ITS sequence data of six

Pseudofusicoccum species procured from GenBank were aligned with our

isolate (JQ585586) sequence, 44 ambiguous base pairs bp were removed

from the total 568 (bp) prior to analysis. Of the 524 bp used to reconstruct

the phylogeny, 457 were conserved, 64 were variable, 16 were parsimony-

informative, and 48 were singletons. Maximum parsimony analysis produced

eleven equally parsimonious trees with a tree length of 68 (CI = 0.941176,

RI = 0.857143, and RCI = 0.806723 for all sites). One such most parsimonious

tree (PLATE 1) exhibits three major clades: Clade 1 (P kimberleyense,

P. violaceum, P. ardesiacum), Clade 2 (P. stromaticum, PR. adansoniae MCC

1020, P adansoniae CBS 122055), and Clade 3 (consisting of P. olivaceum). The

Maximum parsimony (MP) tree showed P adansoniae MCC 1020 forming

a separate clade along with P adansoniae CBS 122055, but forming a sister

branch to P stromaticum. Moreover, the strain MCC 1020 and P. adansoniae

CBS 122055 showed 100% similarity within 507nt of the ITS region when

compared in MEGA 5.0 (Tamura et al. 2011).

Taxonomy

Pseudofusicoccum adansoniae Pavlic, T.I. Burgess & M.J. Wingf., Mycologia 100:

855. 2008. PLATE 2

Teleomorph: Unknown.

Colonies initially form a moderately dense mycelial mat. Initially mycelium

pale olivaceous grey (21''""’d) to olivaceous grey (21''""’b) from the middle of

Pseudofusicoccum adansoniae new to India ... 43

S188 P2B6n0m-88

PLATE 2. Pseudofusicoccum adansoniae. Fic. 1. Conidiomata detail showing mycelial hairs covering

the surface (SEM). Fic. 2. Conidia. Scale bar = 25um.

vey very

the colony. At maturity, grey olivaceous (21''i) to olivaceous black (27'"’m)

and after 3-5 d becoming dark greenish-black (33'""""k) with age, covering the

90 mm diam Petri dish after 4 d in the dark on 2% MEA medium. Colonies

slightly irregular, occasionally radially striated with zones or irregular circles.

Aerial mycelium slightly fluffy, becoming dense and cottony with age, turning

pale olivaceous grey (21'’"’d). Radiations are evident on front and reverse of

colony. Conidiomata 1-2 mm, some 5 mm in diam, readily formed from the

middle of colony within 7-10 d, covering the entire surface of colony and deeply

immersed in the medium (seen as a round black structures on the reverse side

of Petri dishes 14-16 d after incubation), eustromatic, multilocular, immersed.

Hyphae brown, branched. Conidiogenous cells holoblastic, smooth, cylindrical,

hyaline, the first conidium produced holoblastically and subsequent conidia

enteroblastically. Conidia ellipsoid, occasionally slightly bent or irregularly

shaped, 9.7-25.8 x 6.5-9.7 um (av. 16.5 x 7.3 um), apices rounded, smooth

with fine granular content, hyaline, thin-walled, unicellular, surrounded by a

mucous layer.

DISTRIBUTION AND HABITAT: Western Australia (from infected Adansonia gregorii and

Mangifera indica, and from asymptomatic Acacia synchronicia, A. gregorii, Eucalyptus

sp., and Ficus opposita); Maharashtra, India (endophytic in Jatropha podagrica).

MATERIAL EXAMINED: INDIA, MAHARASHTRA STATE, Pune, Pimpri, 18°37'07.04"N

73°48'13.43"E, endophytic in leaf midrib of Jatropha podagrica, 25.IV.2011, Rohit

Sharma, culture MMI00062; endophytic in fruit of J. podagrica, 25.IV.2011, Rohit

Sharma, culture MMI00064 (MCC 1020; GenBank JQ585586).

Discussion

Pseudofusicoccum adansoniae MCC 1020 is characterized by multilocular

hairy conidiomata with several pycnidia whose inner margin is covered

with hyaline mycelia. The fungus produces several conidiomata (10-15) in

44 ... Sharma, Kulkarni & Shouche

culture covered by mycelial hairs (Fic. 1). It contains several hyaline, ellipsoid

conidia (Fic. 2) formed on hyaline conidiophores covered in a mucous layer.

However, the conidia of P adansoniae MCC 1020 average 16.5 x 7.3 um

(TABLE 1) and are narrower and broader compared to the average 22.5 x 5.2

um of P adansoniae CBS 122055 (Pavlic et al. 2008). TaBLE 1 clearly shows the

similarity of a combination of characters (e.g., colony colour, growth characters,

conidiogenous cell and conidial morphologies) for MCC 1020 and the other

analyzed P adansoniae strain. Phylogenetic analysis suggests that P adansoniae

is only distantly related to all other Pseudofusicoccum species. ITS1-5.8S-ITS2

sequence analyses place P. adansoniae MCC 1020 and P. adansoniae CBS 122055

together in a distinct clade forming a sister branch to P. stromaticum. The

maximum parsimony analysis and resulting tree (PLATE 1) clearly indicates that

apart from the P adansoniae and P. stromaticum branches, Pseudofusicoccum

is divided into two major groups, one containing P. violaceum, P. ardesiacum,

and P. kimberleyense and the other containing P. olivaceum. Neighbour Joining

analysis also generates similar tree topologies.

Most species (including Pseudofusicoccum) of the Botryosphaeriaceae are

endophytic, and some can be latent pathogens (Crous et al. 2006, Slippers &

Wingfield 2007). Pseudofusicoccum adansoniae CBS 122055 was originally

described from Derby, Western Australia, and isolated from infected branches

of Adansonia gregorii F. Muell. (=Adansonia gibbosa Cunn. et al.) (Malvaceae)

and healthy branches of Acacia synchronicia Maslin (Leguminosae), Eucalyptus

sp., and Ficus opposita Miq. (Moraceae) (Pavlic et al. 2008). The fungus has also

been isolated as an endophyte from Adansonia gregorii (Sakaladis et al. 201 1a)

and from diseased stems of Mangifera indica L. (Anacardiaceae) from Western

Australia (Sakaladis et al. 2011b). Hence, detection of P adansoniae MCC 1020

from healthy leaf midrib and fruit of Jatropha podagrica is not unusual, as it

has been isolated from asymptomatic branches of other trees. As it represents

a first species record outside Australia and the first Pseudofusicoccum record

from India, it is reported as new to the Indian fungal flora. Reports of

Pseudofusicoccum from other parts of world — especially as endophytes —

should be expected.

Acknowledgments

The authors thank DBT, New Delhi, India for funding, Director (NCCS, India) for

laboratory facilities and Dr. S.V. Shinde (University of Pune, India) for SEM. Mr. J.W.M.

Mehl (University of Pretoria, Pretoria) and Dr. Artur Alves (Universidade de Aveiro,

Portugal) are also acknowledged for suggestions and pre-review of the manuscript. JM

was kind enough to provide the colour chart for description. Authors also thank Dr.

Shaun Pennycook (Landcare Research Private, New Zealand) for nomenclature review

and suggestions.

Pseudofusicoccum adansoniae new to India... 45

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MY COTAXON

http://dx.doi.org/10.5248/123.47

Volume 123, pp. 47-61 January-March 2013

Pestalotiopsis species associated with Camellia sinensis (tea)

SAJEEWA S.N. MAHARACHCHIKUMBURA’”?, EKACHAI CHUKEATIROTE’”,

L1ANG-DonG Guo’, PEDRO W. Crous*, ERIC H.C. MCKENZIE?

& Kevin D. Hype?”

‘Institute of Excellence in Fungal Research, Mae Fah Luang University,

Chiang Rai 57100, Thailand

School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand

*Key Laboratory of Systematic Mycology & Lichenology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing 100190, PR. China

‘CBS-KNAW Fungal Biodiversity Centre, PO. Box 85167, 3508, AD, Utrecht, The Netherlands

‘Landcare Research, Private Bag 92170, Auckland, New Zealand

* CORRESPONDENCE TO: kdhyde3@gmail.com

ABSTRACT — We describe a new species Pestalotiopsis furcata isolated from Camellia sinensis

(tea), which is distinguished morphologically by its relatively large conidia (29-39 x 8.5-10.5

um) and 5-9 apical appendages, some of which are branched and lack basal appendages. A

phylogenetic tree based on a combination of ITS, B-tubulin and TEF1 clearly distinguishes

P. furcata from other species in the genus. We examined syntype material of P theae, which we

designate as lectotype, and a fresh collection from Thailand, which we designate as epitype.

Key worps — Amphisphaeriaceae, grey blight, phylogeny, systematics, tea disease

Introduction

Camellia sinensis (L.) Kuntze (Theaceae) is widely planted in the tropics

and subtropics and probably originated at the point of confluence of the lands

of northeast India, north Burma, southwest China and Tibet (Wight 1959;

Mondal et al. 2004). Commonly known as tea, it is prepared as a beverage with a

cooling, slightly bitter, astringent flavor and is the most widely consumed liquid

in the world after water (Mondal et al. 2004). Several fungi are known to cause

diseases of foliage, stems and roots of C. sinensis. Brown blight (Colletotrichum

camelliae Massee), leaf blotch (Colletotrichum sp.), grey blight (Pestalotiopsis

longiseta (Speg.) K. Dai & Tak. Kobay. and P. theae, blister blight (Exobasidium

vexans Massee), twig die-back and stem canker (Macrophoma theicola Petch),

and horse hair blight (Marasmius crinis-equi F. Muell. ex Kalchbr.) are common

fungal diseases affecting tea plantations (Gadd 1949; Hainsworth 1952; Chen

et al. 1982).

A8 ... Maharachchikumbura & al.

The genus Pestalotiopsis contains several species responsible for plant

diseases. Pestalotiopsis species have been isolated as both endophytes and

pathogens in Camellia sinensis (Wei et al. 2007; Joshi et al. 2009). Pestalotiopsis

is an anamorphic genus in Amphisphaeriaceae (Barr 1975, 1990; Kang et al.

1998, 1999; Maharachchikumbura et al. 2011, 2012). In southern India grey

blight disease of tea caused by Pestalotiopsis spp. resulted in 17% production

loss (Joshi et al. 2009) and 10-20% yield loss in Japan (Horikawa 1986). Five

Pestalotiopsis species have been recorded from tea (Agnihothrudu 1964),

although P. longiseta and P. theae are considered to be the major species causing

grey blight (Joshi et al. 2009).

The taxonomic status of species in Pestalotiopsis is unresolved, as many

species are generally not host-specific, conidial characters vary, and species

limits overlap. The species concepts adopted by Steyaert (1949) and Guba

(1961) are problematic (Maharachchikumbura et al. 2011). Epitypification

of important species using molecular data and analyses is needed to resolve

the species and to establish their important diagnostic characters, as has been

done for several other plant pathogenic genera such as Colletotrichum (Hyde

et al. 2009), Phomopsis (Udayanga et al. 2011), Cochliobolus (Manamgoda et

al. 2011), and Phyllosticta (Wikee et al. 2011). Re-examination of type material

and establishment of epitypes from fresh collections relying on sequence

analyses from living cultures and full descriptions are needed to resolve species

in Pestalotiopsis and other important pathogenic genera (Hyde et al. 2010; Cai

et al. 2011; Koko et al. 2011).

In order to re-evaluate Pestalotiopsis species associated with tea (C. sinensis),

we examined syntype material of P theae, which we here describe, illustrate,

and designate as lectotype. We also epitypify this economically important

taxon with a fresh collection from tea in Thailand and fully describe the new

collection. Sequence data from the epitype allows us to distinguish P. theae

from other Pestalotiopsis collections from tea. We also describe P. furcata, a

new Pestalotiopsis species from tea in Thailand, which is differentiated by

morphological and molecular differences.

Materials and methods

Study site and plant material

Diseased leaves of tea were collected from Chiang Mai Province, northern Thailand,

near the Mushroom Research Centre, at an elevation of 900 mat 19°17.123'N 98°44.009’E.

The region has a mean annual temperature of 24°C and mean annual precipitation of

2121 mm (http://www.tmd.go.th/en/). Leaf samples were placed in clean polythene bags

and symptoms were recorded.

Isolation and identification of pathogen

A single conidium culture technique was used to obtain pure fungal colonies

according to Chomnunti et al. (2011). The colonies were transferred to 2% potato-

Pestalotiopsis furcata sp. nov. (Thailand) ... 49

dextrose agar (PDA) medium and incubated at room temperature (25°C). Sporulation

was induced on sterilized carnation leaves that were aseptically placed on the surface

of PDA with growing mycelium. The morphology of fungal colonies was recorded

following the method of Hu et al. (2007). Fungal mycelium and spores were observed

under the light microscope and photographed. Microtome sections were made with a

Leica CM1850 freezing microtome. All the microscopic measurements were measured

with Tarosoft image framework (v. 0.9.0.7) and 30 conidial measurements were taken

for each isolates.

All other cultures were obtained from China General Microbiological Culture

Collection (CGMCC) and a syntype herbarium specimen of P. theae (BPI 406804) was

borrowed from BPI and studied. Methods of examination, photography, and isolation

followed Boonmee et al. (2011).

DNA extraction and PCR condition

Total genomic DNA was extracted from fresh cultures using a modified protocol

of Doyle & Doyle (1987) and Lee & Taylor (1990). Fresh fungal mycelia (500 mg)

was scraped from the margin of a PDA plate incubated at 25°C for 7 to 10 days and

transferred into a 1.5 mL centrifuge tube with 100 uL of preheated (60°C) 2x CTAB

extraction buffer (2% (w/v) CTAB, 100 mM Tris-HCl, 1.4 M NaCl, 20 mM EDTA, pH

8.0), and 200 mg sterilized quartz sand. Mycelia were ground using a glass pestle for 5

min and an extra 500 uL 2x CTAB preheated (60°C) was added and incubated in a 65°C

water bath for 30 min with occasional shaking. 500 uL of phenol:chloroform (1:1) was

added to each tube and shaken thoroughly to form an emulsion. The mixture was spun

at 11900 g for 15 min at 25°C in a microcentrifuge and the supernatant phase decanted

into a fresh 1.5 mL tube. Supernatant containing DNA was re-extracted with phenol:

chloroform (1:1) at 4°C until no interface was visible. 50 uL of 5M KOAc was added

into the supernatant followed by 400 uL of isopropanol and inverted gently to mix. The

genomic DNA was precipitated at 9200 g for 2 min at 4°C in a microcentrifuge. The

DNA pellet was washed with 70% ethanol twice and dried using SpeedVac® (AES 1010;

Savant, Holbrook, NY, USA) until dry. The DNA pellet was then resuspended in 100 uL

TE buffer (10 mM Tris-HCl, 1 mM EDTA).

PCR amplification

The ITS and 5.8S region of rDNA molecule was amplified using primer pairs ITS4

(5'-TCCTCCGCTTATTGATATGC-3') and ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3')

(White et al. 1990), B-tubulin gene region was amplified with primer pairs BT2A (5'-

GGTAACCAAATCGGTGCTGCTTTC-3') and BT2B (5'-ACCCTCAGTGTAGTGACCCTTGGC-3’)

(Glass & Donaldson 1995; O’Donnell & Cigelnik 1997) and TEF1 was amplified using

the primer pairs EF1-526F (5'-GTCGTYGTYATYGGHCAYGT-3’) and EF1-1567R (5’-

ACHGTRCCRATACCACCRATCTT-3’) (Rehner 2001).

PCR was performed with the 25 uL reaction system consisting of 19.75 uL of double

distilled water, 2.5 uL of 10x Taq buffer with MgCL, 0.5 uL of dNTP (10 mM each),

0.5 wL of each primer (10 uM), 0.25 uL Taq DNA polymerase (5 U/uL), 1.0 uL of DNA

template. The thermal cycling program was as follows:

For ITS an initial denaturing step of 95°C for 3 min, followed by 35 amplification

cycles of 95°C for 30 s, 52°C for 45 s, and 72°C for 90 s, and a final extension step of 72°C

50 ... Maharachchikumbura & al.

TABLE 1. Pestalotiopsis isolates (and Seiridium outgroup) considered in the

phylogenetic study

ISOLATE GENBANK ACCESSION NUMBER

ITS B-tubulin TEF1

P. cf. algeriensis SD077 JQ683718 JQ683702 JQ683734

P. cf. disseminata SD034 JQ683716 JQ683700 JQ683732

P. cf. menezesiana SG064 JQ683719 JQ683703 JQ683735

SD072 JQ683713 JQ683697 JQ683729

P. cf. microspora SD056 JQ683722 JQ683706 JQ683738

P. cf. versicolor SG100 JQ683712 JQ683696 JQ683728

SD047 JQ683715 JQ683699 JQ683731

SDO91 JQ683714 JQ683698 JQ683730

sD040 JQ683717 JQ683701 JQ683733

P. cf. virgatula SD004 JQ683723 JQ683707 JQ683739

P. furcata (ex-holotype) MFLUCC12-0054 JQ683724 JQ683708 JQ683740

P. hainanensis (ex-type) = GQ869902 — —

P jesteri (ex-type) _ AF377282 = _

P. kunmingensis (ex-type) — AY373376 — —

P. pallidotheae (ex-type) — AB482220 _ —

P. theae (ex-epitype) MFLUCC12-0055 JQ683727 JQ683711 JQ683743

SCol1l JQ683726 JQ683710 JQ683742

Pestalotiopsis sp. SsD012 JQ683720 JQ683704 JQ683736

SD072 JQ683713 JQ683697 JQ683729

Seiridium sp. SD096 JQ683725 JQ683709 JQ683741

for 10 min. For B-tubulin PCR conditions were an initial step of 3 min at 95°C, 35 cycles

of 1 min at 94°C, 50 s at 55°C, and 1 min at 72°C, followed by 10 min at 72°C. For TEF1,

an initial step of 5 min at 94°C, 10 cycles of 30 s at 94°C, 55 s at 63°C or 66°C (decreasing

1°C per cycle), 90 s at 72°C, plus 36 cycles of 30 s at 94°C, 55 s at 53°C or 56°C, 90 s

at 72°C, followed by 7 min at 72°C. The PCR products were verified by staining with

Goldview (Guangzhou Geneshun Biotech, China) on 1% agarose electrophoresis gels.

Phylogenetic analysis

DNAStar, SeqMan was used to obtain consensus sequences from sequences generated

from B-tubulin and TEF1, forward and reverse primers. Combination sequence data

obtained from three gene regions were aligned using CLUSTALX (v. 1.83) (Thompson

et al. 1997). The sequences were manually adjusted using BioEdit (Hall 1999), to allow

maximum alignment and maximum sequence similarity. A maximum parsimony

analysis (MP) was performed using PAUP (Phylogenetic Analysis Using Parsimony)

v. 4.0b10 (Swofford 2002). Ambiguously aligned regions were excluded and gaps were

treated as missing data. Trees were inferred using the heuristic search option with TBR

branch swapping and 1000 random sequence additions. Maxtrees were set up to 5000,

branches of zero length were collapsed and all multiple parsimonious trees were saved.

Tree length [TL], consistency index [CI], retention index [RI], rescaled consistency

index [RC], homoplasy index [HI], and log likelihood [-In L] (HKY model) were

calculated for trees generated under different optimality criteria. The robustness of the

Pestalotiopsis furcata sp. nov. (Thailand) ... 51

most parsimonious trees was evaluated by 100 bootstrap replications resulting from

maximum parsimony analysis, each with 10 replicates of random stepwise addition of

taxa (Felsenstein 1985). The Kishino-Hasegawa tests (Kishino & Hasegawa 1989) were

performed in order to determine whether the trees inferred under different optimality

criteria were significantly different. Trees were viewed in Treeview (Page 1996).

The same procedure was used for the ITS gene for the 16 Pestalotiopsis isolates and

the type sequences of P hainanensis, P. jesteri, P. kunmingensis, and P. pallidotheae were

downloaded from GenBank.

Results

A phylogenetic tree was constructed using combined ITS, 6-tubulin

and TEF1 sequences of 16 isolates of Pestalotiopsis, with Seiridium sp. as the

outgroup (TaBLE 1). The aligned data matrix comprised 2019 characters of

which 1490 were constant, 199 were variable and parsimony-uninformative,

and 330 were parsimony-informative. The Kishino-Hasegawa (KH) test

showed that the two trees generated from parsimonious analysis did not differ

significantly (length = 768 steps, CI = 0.848, RI = 0.905, HI = 0.152, RC =

0.767). The aligned data matrix of ITS sequences consisted of 528 characters of

which 395 were constant, 62 were variable and parsimony-uninformative, and

71 were parsimony-informative. Kishino-Hasegawa (KH) test showed length =

178 steps, CI = 0.826, RI = 0.913, HI = 0.174, RC = 0.754).

Taxonomy

Pestalotiopsis theae (Sawada) Steyaert, Bull. Jard. bot. Etat Brux. 19(3): 327 (1949)

FIGS 1, 2

= Pestalotia theae Sawada, Spec. Report Agric. Exp. Station

Formosa 11: 113 (1915), as “Pestalozzia”

Type: Taiwan, Taipei [Taihokuché, Rigyokutsu (Tanaka 1917)], on living leaves of

Camellia sinensis, 13 July 1908, coll. Y. Fujikiro, det. K. Sawada (Lectotype designated

here, BPI 406804). Thailand, Chiang Mai Prov., Mae Taeng Distr., Ban Pha Deng,

Mushroom Research Centre, 19°17.123'N 98°44.009’E, 900 m, rainforest, on living

leaves of Camellia sinensis, January 20, 2010, S.S.N. Maharachchikumbura St200110

(Epitype designated here, MFLU 12-0116; ex-epitype culture, MFLUCC 12-0055 =

CPC 20281; GenBank, JQ683727, JQ683711, JQ683743).

LECTOTYPE

LEAF SPOTS initially brown on leaves of tea, becoming 1 cm in diam.,

and grey with brown margins when mature, or covering up to half of the

leaf; dotted with acervuli. ACERVULI initially subepidermal, later erumpent,

finally exposed. CONIDIOPHORES in clusters, simple, short, filiform, fugacious.

Conip1a fusiform, slightly constricted at septa, 4-septate, 24-28 x 6.6-8.3 um

(mean = 26.5 x 7.4 um); basal cell obconic, hyaline, thin and smooth-walled,

4.3-5.6 um long (mean = 5 um); 3 median cells, with thick verruculose walls,

52 ... Maharachchikumbura & al.

dark brown, septa and periclinal walls darker than the rest of the cell, together

15-20 um long (mean = 18.5 um) second cell from base 6.5-7 um (mean =

6.8 um); third cell 4.8-5.5 um (mean = 5.3 um; fourth cell 5.5-6.0 um (mean

= 5.7 um); apical cell hyaline, conic to cylindrical 4.2-5.8 um long (mean =

UNITED STATES DEPARTMENT OF AGRICULTURE

Bureau oF PLant InpustRy

MYCOLOGICAL COLLECTIONS =

Pestalozzia theae Sawada “hy

on Thea sinensis

Taipeh, Taiwan

July 13, 1908

Coll. Y. Fujikiro :

~US04068040 0

Fic. 1. Pestalotiopsis theae (lectotype, BPI 406804). A. Lectotype herbarium material. B,C. Leaf

blight symptoms on leaf of Camellia sinensis. D,E. Conidia. F. Conidia (drawing from Steyaert

1943) G. Conidia (drawing from Guba 1961). Scale bars: D-G= 20 um.

Pestalotiopsis furcata sp. nov. (Thailand) ... 53

4.9 um); apical appendages tubular, 3-4, arising from the upper portion of the

apical cell, 23-33 um long (mean = 27 um), slightly swollen at the apex; basal

appendages, filiform, 5-9 um, sometimes knobbed.

Fic. 2. Pestalotiopsis theae (epitype, MFLU 12-0116) A. Conidia in culture. B. Conidiogenous

cells. C,D. Conidia. E,F, Colony in culture (E. from above; F. from below). Scale bars: A,B= 20 um,

C,D=15 um.

EPITYPE

CONIDIOPHORES growing in clusters, simple, short, filiform, fugacious,

smooth, thin-walled, hyaline, 4-8 x 1-2 um (mean = 6 x 1.5 um). CONIDIA

fusiform to ellipsoid, straight to slightly curved, 4-septate 22.5-28 x 6.7-8.2

um (mean = 25.5 x 7.6 um), basal cell conic or obconic, hyaline, thin and

smooth walled, 3.9-5.3 um long (mean = 4.55 um), with 3 median cells, thick

verruculose walls, constricted at the septa, concolorous, dark brown, septa and

periclinal walls darker than the rest of the cell, together 14.5-18.5 um long

54 ... Maharachchikumbura & al.

(mean = 16.7 um) (second cell from base 5-—7.2 um (mean = 6.3 um); third cell

4.8-6 um (mean = 5.4 um); fourth cell 5-6.8 um (mean = 5.7 um)); apical cell

hyaline, cylindrical 4.2-5.9 um long (mean = 5.2 um); 3-4 apical appendages,

tubular, arising from the upper portion of the apical cell, 22.5-31 um long

(mean = 26.5 um), slightly swollen at the apex; basal appendages, filiform,

4—7 um.

Colonies growing relatively fast on PDA, reaching 7 cm after 5 days at 25°C,

fimbriate, whitish, dense, aerial mycelium on surface, fruiting bodies black;

reverse of the culture yellowish white.

The syntypes of P theae were recorded from diseased leaves of Camellia

sinensis growing in Taiwan. The specimen from BPI corresponds with one of

the collections listed in the translated protologue, and therefore constitutes a

syntype specimen (Tanaka 1917, as “Taihokucho, Rigyokutsu, July 13, 1908,

Y. Fujikuro”). Since no ex-type culture is available and the lectotype is in poor

condition, an epitype with a living culture is designated from a sample collected

in Thailand.

Pestalotiopsis furcata Maharachch. & K.D. Hyde, sp. nov. FIG. 3

MycoBank MB564563

Differs from other Pestalotiopsis species with large conidia by its wider conidia with 5-9

apical, branched appendages.

Type: Thailand, Chiang Mai Prov., Mae Taeng Distr., Ban Pha Deng, Mushroom

Research Centre, 19°17.123'N 98°44.009’E, elevation 900 m, rainforest, on living leaves

of Camellia sinensis, 20 January 2010, S.S.N. Maharachchikumbura $200110 (Holotype,

MFLU 12-0112; ex-holotype culture MFLUCC 12-0054 = CPC 20280; GenBank,

JQ683724, JQ683708, JQ683740).

Erymo.ocy: The specific epithet refers to the branching nature of the apical

appendages.

Associated with grey blight on leaves of Camellia sinensis, small, rounded,

yellow-green spots on the leaves become brown to grey, with concentric rings

bearing black, scattered conidiomata (Fic. 3). Conidiomata acervuli scattered

or gregarious, rarely confluent, subepidermal in origin, erumpent when mature,

round to oval in outline, conical to oval in longitudinal section, 180-300 um

wide, 70-160 um high, unilocular, glabrous; wall tissue (stroma and parietal

cells) only a few cells thick (14-22 um), forming a textura angularis, cell walls

thick, outermost layer hyaline, inner layers pale brown to brown, encrusted.

Conidiophores reduced to conidiogenous cells lining the inner wall of the

conidiomatal cavity. Conidiogenous cells discrete, lageniform, smooth, thin-

walled, hyaline, with 2-3 proliferations. Conidia fusoid to ellipsoid, straight to

slightly curved, 4-septate, 29-39 x 8.5-10.5 um (mean = 35.5 x 9.7 um), basal

cell obconic, hyaline or slightly olivaceous, thin- and smooth-walled, 4.9-6.4

Pestalotiopsis furcata sp. nov. (Thailand) ... 55

Fic. 3. Pestalotiopsis furcata (holotype, MFLU 12-0112). A. Blight on leaf of Camellia sinensis.

B. Conidiomata, split irregularly. C. Section of conidiomata. D. Conidiophores/conidiogenous

cells. E-H. Conidia with branched appendages. I,J. Colony on PDA (I from above, J from below).

Scale bars: C= 50 um, D-H= 20 um.

um long (mean = 5.8 um), with 3 median cells, doliiform to subcylindrical,

with thick verruculose walls, constricted at the septa, concolorous, olivaceous,

septa and periclinal walls darker than the rest of the cell, wall rugose, together

20.7-25 um long (mean = 23.4 um) (second cell from base 7-9 um (mean = 7.9

um); third cell 7.5-9.1 um (mean = 8.2 um); fourth cell 7.2-9.2 um (mean =

8.0 um); apical cell hyaline, conic to cylindrical 6.3-8.44 um long (mean = 7.48

um); 5-9 tubular apical appendages, some appendages branched, arising from

the upper portion of the apical cell, 20-35 um long (mean = 27.7 um), unequal;

basal appendages absent.

Colonies on PDA reaching 7 cm after 7 days at 25°C, edge entire, whitish,

with dense, aerial mycelium on surface, fruiting bodies black, gregarious;

reverse of culture white.

56 ... Maharachchikumbura & al.

HABITAT/DISTRIBUTION: Known to inhabit living leaves of Camellia sinensis,

Thailand.

ADDITIONAL MATERIAL EXAMINED: THAILAND, CHIANG Mat Prov., MAE TAENG

Distr., Ban Pha Deng, Mushroom Research Centre, 19°17.123'N 98°44.009’E, elevation

900 m, rainforest, on living leaves of Camellia sinensis, 10 July 2010, S.C. Karunarathna

$100710 (MFLU12-0113); 9 September 2011, S.S.N. Maharachchikumbura S110911

(MFLU12-0114); 9 December 2011, S.S.N. Maharachchikumbura $91211 (MFLU12-

0115).

TABLE 2. Synopsis of Pestalotiopsis furcata and related species.

P. furcata P. nattrassii * P leucopogonis® P macrospora* _ P. hainanensi“

Conidia size 29-39 x 27-33 X 27-32 Xx 30-40 x 19-22 x

(um) 8.5-10.5 8-9 7.5-9.5 7-9 5-6

Median Concolorous, | Concolorous, | Concolorous, Concolorous, Cascoloreas)

: ; Brown to

cells olivaceous brown brown olivaceous ;

olivaceous

Apical appendages

—number 5-9 1-4 7-11 3-5 1-3

-length (um) 20-35 25-44 12-19 15-22 1-10

-branching Branched No No Branched No

—position Apex Apex 3 rows (top, Apex Apex

middle, and

bottom)

Basal Lacking Lacking or Lacking or Present Lacking

appendages short present

*= from Guba (1961); > = from Nag Raj (1993); * = from Liu (2007)

Notes: Pestalotiopsis furcata is both morphologically (TaBLe 2) and

phylogenetically (Fics. 4-5) distinct. Its conidia (29-39 x 8.5-10.5 um)

are larger than those of other Pestalotiopsis species as well as wider than

such morphologically similar species as P hainanensis (19-22 x 5-6 um),

P. leucopogonis (27-32 x 7.5-9.5 um), PB macrospora (30-40 x 7-9 um),

and P. nattrassii (27-33 x 8-9 um). Its most characteristic feature is its 5-9

apical, branched appendages. Although P. leucopogonis also has 7-11 apical

appendages, they arise from three levels attached to the apical cell (apex, middle,

and base) and at 12-19 um long are shorter than those of P. furcata (20-35 um).

Pestalotiopsis macrospora has shorter (15-22 um) branched apical appendages.

Pestalotiopsis furcata also lacks basal appendages, which are otherwise present

in most Pestalotiopsis species.

Discussion

In the combined dataset Pestalotiopsis species cluster in two main clades

with high (100%) bootstrap support (Fic. 5). These two clades can be

Pestalotiopsis furcata sp. nov. (Thailand) ... 57

differentiated by the color of the median cells, confirming earlier reports

(Jeewon et al. 2003; Liu et al. 2010; Maharachchikumbura et al. 2011, 2012).

In the multigene analysis (Fic. 5), clade A (P. theae) was separate from clade

B with high bootstrap support. When additional ITS sequences downloaded

from GenBank were analyzed (results not shown), putatively named P. theae

strains clustered in both clades, indicating that many have misapplied names.

The median conidial cells of clade B strains are olivaceous, while clade A strains

have brown median conidial cells similar to the type (Fic. 5). We blasted the

SG064 P. cf. menezesiana

SD047 P. cf. versicolor

$D004 P. cf. virgatula

100

SD091 P. cf. versicolor

SD072 P. cf. menezesiana

SD040 P. cf. versicolor

88 SG100 P. cf. versicolor

P. theae (epitype)

87 Clade A

SC011 P. theae

SD012 Pestalotiopsis sp.

Clade B

SD073 Pestalotiopsis sp.

100 AY373376 P. kunmingensis

AB482220 P. pallidotheae

P. furcata

GQ869902 P. hainanensis

‘ SD034 P. cf. disseminata

SD056 P. cf. microspora

99 51

SD077 P. cf. algeriensis

AF377282 P. jesteri

SDO096 Seiridium sp.

-1 changes

Fic. 4. Maximum parsimony phylogram generated from ITS gene region analysis of species of

Pestalotiopsis recorded from tea and other available Pestalotiopsis spp. type sequences. Data were

analyzed with random addition sequence, unweighted parsimony and treating gaps as missing

data. Seiridium sp. is placed as outgroup, and all ex-type sequences are set in bold font.

58 ... Maharachchikumbura & al.

$SG100 P. cf. versicolor

$D047 P. cf. versicolor

SD091 P. cf. versicolor

$D040 P. cf. versicolor

SG064 P. cf. menezesiana

100

SD072 P. cf. menezesiana

$D004 P. cf. virgatula

P. theae (epitype)

100

Clade A

SC011 P. theae

100

$D012 Pestalotiopsis sp.

100 Clade B

SD073 Pestalotiopsis sp.

SD034 P. cf. disseminata

SD056 P. cf. microspora

$D077 P. cf. algeriensis

P. furcata

SDO096 Seiridium sp.

10 changes

Fic. 5. Maximum parsimony phylogram generated from combine three genes (ITS, B-tubulin and

TEF1) analysis of species of Pestalotiopsis recorded from tea. Data were analyzed with random

addition sequence, unweighted parsimony and treating gaps as missing data. Seiridium sp. is placed

as outgroup, and all ex-type sequences are set in bold font.

isolates from tea plants at the Mushroom Research Centre with ITS sequences in

GenBank and there was one similar species. The type strain of P. hainanensis is

similar to the ITS sequence of the Thailand tea isolate, although morphological

characters differentiate these species. In the phylogram (Fic. 5) the Thailand tea

species forms a monophyletic subclade and is distinguished as the new species,

P. furcata. Our analyses confirm the observations by Hu et. al. (2007) and Liu

et al. (2010) that a combined multigene dataset better resolves the taxonomy of

Pestalotiopsis. Hu et al. (2007) showed that the ITS gene is less informative than

the B-tubulin gene in differentiating Pestalotiopsis species. Liu et al. (2010),

however, disagreed and indicated that proper analysis and alignment of the ITS

Pestalotiopsis furcata sp. nov. (Thailand) ... 59

region can usefully group Pestalotiopsis species according to different types of

conidial pigmentation, which can be used as a key character for the phylogeny

of the genus. We constructed phylogenetic trees separately for each gene region

(results not shown for 6-tubulin and TEF1) with ITS < §-tubulin < TEF1 in their

degree of species resolution, but best resolution results were with all three genes

combined.

Most DNA sequences deposited for Pestalotiopsis species in GenBank are

problematic (Maharachchikumbura et al. 2011), as they are likely to have

been wrongly named. There are very few ex-type sequences and therefore the

resolution of species and their authentic naming are difficult and confused. Re-

examination of type materials and establishment of epitypes with sequenced

living cultures are needed to advance understanding of species in this genus.

Acknowledgments

Drs. Roger Shivas (Biosecurity Queensland, Brisbane, Australia) and Dr. Yong

Wang (Guizhou University, Guizhou, P.R. China) are thanked for critical review of the

manuscript. We also thank the King Saud University; Key Lab of Systematic Mycology

and Lichenology, Institute of Microbiology, Chinese Academy of Sciences, Beijing,

China; Mushroom Research Foundation, Chiang Mai, Thailand, the National Research

Council of Thailand (grant for Pestalotiopsis No: 55201020008) and Mae Fah Luang

University (grant for Pestalotiopsis No: 55101020004); for supporting this research.

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MYCOTAXON

http://dx.doi.org/10.5248/123.63

Volume 123, pp. 63-79 January-March 2013

Critical revision of myxomycetes in the

Buenos Aires BAFC herbarium — 1

GABRIEL MORENO *, AURELIO CASTILLO ? & JORGE R. DESCHAMPS ”

' Dpto. de Biologia Vegetal, Facultad de Biologia, Universidad de Alcala,

28871Alcald de Henares, Madrid, Spain

? Universidad de Belgrano, Facultad de Ciencias Agrarias,

Federico Lacroze 1955, Cap. Fed. 1426. Buenos Aires, Argentina

* CORRESPONDENCE TO: gabriel.moreno@uah.es

ABSTRACT — ‘The first analytical results of 32 myxomycetes specimens kept at the BAFC

herbarium are presented. The following species stand out: Craterium obovatum, Diderma

meyerae, Diderma stellulum, Didymium ovoideum, Stemonitis pallida, S. webberi, Trichia

decipiens var. olivacea, and T. subfusca. Type material descriptions of Stemonitis pallida and

S. webberi are also provided. Optical microscope (LM) and scanning electron microscope

(SEM) micrographs illustrate the most representative characteristics of species that are rare

or not often mentioned in the literature.

Key worps — Protozoa, myxobiota, slime moulds, Argentina, USA, taxonomy

Introduction

The BAFC herbarium currently at the Biological Sciences Department of

the School of Exact and Natural Sciences, Buenos Aires University, was created

in the 1960s from Rolf Singer’s 1960-66 collections and other materials. The

herbarium later added donations from Irma Gamundi, Jorge E. Wright, Carlos

Goémez, Emilio del Busto, and Maria E. Ranalli (among others), and Jorge W.

Wright arranged for shipping mycological materials from the old Ministry of

Agriculture and the Instituto Forestal Nacional to BAFC. During the 1970s,

Jorge Deschamps, Alicia Godeas, Daniel Cabral, Silvia Lopez, Monica Adler,

Andrea Romero and others continued with this expansion. During the last

decade following the death of Jorge E. Wright, the fungal collections in the

herbarium have not increased appreciably. The herbarium currently curates

approximately 28,000 mycological specimens, including 476 myxomycetes.

In this paper, we present critical revisions of some rare or taxonomically

significant myxomycetes kept in the BAFC herbarium. Some of this material

originates from other countries.

64 ... Moreno, Castillo & Deschamps

Materials & methods

The material studied originated from Argentina (Buenos Aires, Santa Fé, Misiones,

Tucuman, and Tierra del Fuego provinces), USA (New York, Maryland, and California

states), and Brazil (Rio Grande do Sul state). The collections are preserved in the BAFC

herbarium (School of Exact and Natural Sciences, University City, II Pavilion, 4th. floor,

University of Buenos Aires, Argentina).

Slide mounts in Hoyer’s medium of each specimen are preserved in the Plant

Biology herbarium of the University of Alcala, Madrid (AH, Alcala de Henares). Spore

measurements were made using an oil immersion objective and include such surface

structures as warts and spines.

Scanning electron microscopy (SEM) micrographs were taken at the University of

Alcala de Henares using a Zeiss DSM-950 instrument. Sporocarps were rehydrated in

concentrated ammonium hydroxide (28-30%) for 30 minutes, dehydrated in aqueous

ethanol (70%) for 30 minutes, fixed for 2 hours in pure ethylene glycol dimethyl ether

(= 1,2-dimethoxymethane) and finally immersed in pure acetone for at least 2 hours

followed by critical point drying and sputtering with gold-palladium. This technique

allows the use of very little material (part of a single sporocarp or even no more than a

few spores).

The terminology used for the spore-producing stages follows Dérfelt & Marx (1990)

and Lado & Pando (1997). Spore wall ornamentation as seen by SEM is described

according to the terminology proposed by Rammeloo (1974, 1975) and abbreviations

for author citations follow Kirk & Ansell (1992). The nomenclature used follows Lado

(2001, 2005-11).

Taxonomy

Arcyria globosa Schwein., Schriften Naturf. Ges. Leipzig 1: 64 (1822) Fics. 1-4

SPECIMEN EXAMINED: USA: NEw York, Centreport, on crown debris of Castanea sp.,

VIII-1891, leg. O.F Cook (BAFC 30398, ex “BPI 3405” [= BPI 805828]).

ComMENTS — This species is characterized by its isolated or gregarious, stipitate

sporocarps (0.3-0.7 x 0.5-1.5 mm), globose, white to greyish white in colour, a

fugacious peridium with a wide calyculus and concolorous stipe of 0.2-0.8 mm

length. Capillitium elastic, formed by 3.5-4.5 um diam. filaments with warts

which are usually arranged in more or less marked spirals. Spores 7.5-8.5 um

in diam., hyaline, with dispersed warts. Under SEM, spore ornamentation is

seen to comprise dispersed groups of irregular verrucae. The remaining spore

wall contains a surface covered by tiny bacula, which cannot be seen under the

light microscope (LM). The capillitial filaments contain warts that are joined in

short crests with a helicoidal arrangement.

Badhamia affinis Rostaf., Sluzowce monogr.: 143 (1874) Figs. 5-7

SPECIMEN EXAMINED: ARGENTINA: BUENOS AIRES, Banfield, on semi-rotten bark

of Manihot grahamii Hook., 5-IV-1973, leg. J. Deschamps (BAFC 22822, as Badhamia

cinerascens).

Myxomycetes of BAFC (Argentina) ... 65

Fics. 1-10 Arcyria globosa BAFC 30398 (1-4): 1-3. Capillitium. 4. Spore (SEM). Badhamia

affinis BAFC 22822 (5-7, SEM): Spores. Badhamia versicolor BAFC 22854 (8-10. SEM):

8, 9. Packets of spores. 10. Spore. Bars: 1, 2, 4-7, 10 = 2 um; 3, 8,9=5 um.

ComMMENTS — ‘This species is characterized by its globose to subglobose

sporocarps which are laterally squashed (0.5-1 x 0.4—1 mm) and isolated or

in groups of 2-5, sometimes forming short substipitate plasmodiocarps which

are dark brown at the insertion with the substrate. The peridium is whitish

and warted by the accumulation of calcium carbonate. Dehiscence is apical

and irregular. The capillitium is a three-dimensional net of calcium carbonate

tubules, which join in the centre of the sporotheca to form a whitish strongly

marked pseudocolumella. Spores are 14-15(-16) um in diam., globose to

subglobose, dark violet, with abundant dense warts. Regularly distributed

bacula form the spore ornamentation (SEM).

The presence of a marked white pseudocolumella at the centre of the

sporotheca and the large spore size are two important diagnostic characters for

B. affinis. See Moreno & Oltra (2010) for more information, including LM and

SEM photographs, can be found in.

Badhamia cinerascens G.W. Martin, which is rarely cited, is a rare and critical

species from Colombia. It differs from B. affinis by its lack of a pseudocolumella

and thin peridium.

66 ... Moreno, Castillo & Deschamps

Badhamia versicolor Lister, J. Bot. 39: 81 (1901) Fics. 8-10.

SPECIMEN EXAMINED: ARGENTINA, TIERRA DEL FUEGO, Puerto Harberton, on bark

of possibly Nothofagus pumilio (Poepp. & Endl.) Krasser “lenga’, 2-II-1973, leg. J.E.

Wright & E. del Busto 2177 (BAFC 22854).

ComMENTs — ‘This material was previously cited by Deschamps (1976). The

specimen lacks calcium carbonate in the peridium, although it has some

dispersed granules. Its clustered spores form thick hollow balls of 30-40 spores

in each cluster. Individual spores are ovoid to triangular with obtuse extremes

of 11-13 x 10-11 um diam. and are more strongly warted on the outer exposed

surface. Irregularly distributed bacula form the spore ornamentation (SEM).

This species is known only from Tierra del Fuego, Argentina (Arambarri

1975; Deschamps 1976) and Chubut in Patagonia (Wrigley de Basanta et al.

2010).

Craterium obovatum Peck, Bull. Buffalo Soc. Nat. Sci. 1: 64 (1873) Figs. 11-13

= Badhamia rubiginosa (Rostaf.) Rostaf., Sluzowce monogr. suppl.: 5 (1876)

SPECIMEN EXAMINED: USA, NEW YoRK, SUFFOLK Co., Centerport, VIII-1891, leg. O.F

Cook 1366, as B. rubiginosa (BAFC 28684).

ComMENTS — ‘This species is characterized by its stipitate sporocarps, which

are isolated, gregarious to caespitose and form dense fructifications, 1-2 mm

total height. Sporothecae are obovoid, 0.8-1 x 0.4-0.6 mm, brownish-purple

and with a thin peridium. Dehiscence is apical and operculate. Stipe cylindrical,

0.5-1 mm in length and usually equal or shorter than the sporotheca.

Pseudocolumella is not very well formed. The capillitium is formed by very

abundant whitish nodules. Hypothallus dark brown, membranous. Spores are

12-15 um in diam., globose to slightly polyhedric, dark violet, with tightly

arranged warts that sometimes form crests. Dense bacula with warty coralloid

apices form the spore ornamentation (SEM).

Excellent photographs of the sporocarps of this species can be found in

Poulain et al. (2011). This typical and strange spore ornamentation (SEM) has

been observed in specimens from Baja California and Sonora (Mexico) by

Lizarraga et al. (1999, 2007).

Diderma meyerae H. Singer, G. Moreno, Ilana & A. Sanchez, in Moreno et al.,

Cryptog. Mycol. 24: 53 (2003) Figs. 14-16

SPECIMEN EXAMINED: USA, CALIFORNIA, SHASTA Co.,Sunflower Flat, Lassen Volcanic

National Park, elevation 6000 feet, on conifer wood, 7-II-1965, leg. D.T. Kowalsky.

(BAFC [ex Bridge Cooke Herbarium 34923] as Diderma niveum).

ComMENTSs — ‘The whitish to straw-colored fructifications are arranged in

dense groups. Sporothecae are 1.5-3 mm diam., globose to subglobose, and

sometimes compacted together. Dehiscence is apical and irregular. Peridium

Myxomycetes of BAFC (Argentina) ... 67

Fics. 11-16. Craterium obovatum (BAFC 28684): 11, 12. Spores (SEM). 13. Spore ornamentation.

Diderma meyerae (BAFC [ex Bridge Cooke Herbarium 34923]): 14, 15. Spores (SEM). 16. Spore

ornamentation. Bars: 11-12, 14-15 = 2 um; 13, 16=1 um.

double, the external layer crustose and calcareous, and internal layer transparent

to slightly iridescent. Pseudocolumella central, of variable morphology,

globose to subglobose or conical with dark straw to ferruginous cream colours.

Capillitium with dark filaments (1-2 um width), lighter and bifurcate at the

tips. Spores dark in mass, (7-)8-9 um in diam., globose to subglobose, light

violet (LM), with thick warts to short crests. Verrucae, some joining to form

short crests, comprise the ornamentation (SEM).

Diderma meyerae is easily recognised under the LM from its scarcely

projecting, non-spinose ornamentation formed by warts that join to form short

crests. Moreno & al. (2003) described D. meyerae from European nivicolous

material (Austria and France). This species is often confused with D. niveum.

The European material has larger spores (10-13(-15) um diam.), but the spore

ornamentation and other characteristics are similar to those of the American

specimen.

Diderma stellulum M.L. Farr, Int. J. Mycol. Lichenol. 3: 208 (1988) Figs. 17-22

SPECIMEN EXAMINED: BRAZIL, R10 GRANDE DO SUL, Porto Alegre, Morro Santana,

31-VIII-1984, leg. C. Rodriguez (BAFC 32313, as D. stellula).

COMMENTS — Sporocarps isolated to gregarious, stipitate, dark brown,

reminiscent of the colour of Lepidoderma trevelyanii (Grev.) Poulain & Mar.

68 ... Moreno, Castillo & Deschamps

Fics. 17-22. Diderma stellulum (BAFC 32313: 17. Capillitium. 18-21. Spores (SEM). 22. Spore

ornamentation. Bars: 17 = 10 um; 18-21 = 2 um; 22=1 um.

Mey, 1.5-2 mm width when opened. Stipe approximately 1 mm height, 0.2

mm wide, dark brown and widened at its base. Peridium with three layers,

the outer layer cartilaginous of a brown colour, the middle layer is whitish,

thick with calcareous granules, the inner layer is attached to the middle layer

which is membranous and iridescent. Dehiscence stellate, with recurved

rays. Pseudocolumella not very prominent, 0.4-0.6 mm, flat, rough, whitish

to straw-coloured. Hypothallus membranous. Capillitium, dark, reticulate,

formed by thin filaments of 0.5-1 um diam. and with wider nodules of 5-40

um diam. Spores 12-14 um diam., globose, dark brown and strongly warted.

These warts are joined forming crests or a marked subreticulum; SEM shows

that the spore ornamentation is formed by bacula joined in long sinuous crests,

giving a somewhat subreticulate appearance.

Farr (1988), who described D. stellulum from a single specimen from Brazil,

did not indicate where the species type is deposited. The BAFC specimen comes

from the same type locality and was collected by the same person. Although the

collection date is two years before the one indicated by Farr as “comm. 1986”,

it is possible that this specimen is part of the type, although Farr described a

slightly larger spore (14-16 um).

Diderma stellulum is macroscopically similar to Lepidoderma trevelyanii,

which is differentiated by its peridial crystalline scales and smaller (11-12 um)

warted spores.

Myxomycetes of BAFC (Argentina) ... 69

Diderma asteroides (Lister & G. Lister) G. Lister has different sporocarps

and smaller (11-12 um) warted spores.

Diderma antarcticum (Speg.) Sturgis differs in its sessile sporothecae,

circumscissile dehiscence, globose columella, rugged, double peridium,

capillitium filaments bearing small spines and spinose spores (Arambarri 1973;

Wrigley de Basanta et al. 2010).

Diderma gracile Aramb. is a similar species which differs in its long stipe

which is approximately twice the size of the sporotheca, irregular dehiscence

forming petaloid non-recurved lobes, a pulvinate strongly marked columella

and thin capillitial nodules (Arambarri 1973; Wrigley de Basanta et al. 2010).

Diderma subasteroides M.L. Farr, which is also stipitate, differs due to the

irregular dehiscence, capillitium without marked nodules, and warted spores

(Farr 1971). By SEM, the African specimens (Malawi and Rwanda) from Buyck

(1983) have large warts that form a subreticulum similar to the specimen

studied in this paper.

Didymium ovoideum Nann.-Bremek., Acta Bot. Neerl. 7: 780 (1958) Fics. 23-26

SPECIMEN EXAMINED: USA, MARYLAND, Beltsville, on dead leaves, twigs, 7.VII-1967,

leg. M.L. Farr 4072 (BAFC 26535).

ComMMENTs — ‘This species is characterized by its large sporocarps, 1-1.2

mm total height and its stipitate and gregarious habit. Sporothecae 0.6-0.8 x

0.4-0.5 mm, subglobose to ovoid. Peridium thin, fragile with irregular

dehiscence, whitish and pulverulent due to the accumulation of calcium

carbonate crystals. Stipe approximately 1 mm long, cylindrical, reddish and

longitudinally grooved. Pseudocolumella ovoid, yellow, covered with calcium

carbonate crystals. The capillitium is branched and anastomosed and formed

of hyaline to pale yellowish filaments approximately 1 um diam. Spores 7-8.5

um in diam., globose, dark to light violet, covered by small warts. Short, dense

verrucae joined into low crests compose the spore ornamentation (SEM).

This species is known only in the Northern Hemisphere (www.discoverlife.

org) and Central America (Costa Rica; Lado & Wrigley de Basanta 2008).

Physarum compressum Alb. & Schwein., Consp. fung. lusat.: 97 (1805)

SPECIMEN EXAMINED: ARGENTINA, MIsIONngEs, El Pesado, on remains of Bromeliaceae,

V-1964, leg. C.E. Gomez (BAFC 22130, as P. polycephalum).

ComMMENTS — ‘This species is characterized by its laterally compressed,

whitish sporothecae with a black to whitish stipe, abundant short and rounded

capillitial nodules, whitish in colour and globose spores of 10-12 um diam. and

warted ornamentation. In Argentina, according to (Deschamps 1976; Crespo &

Lugo 2003), this species was recorded in the provinces of Buenos Aires, Jujuy,

Misiones, and Tucuman.

70 ... Moreno, Castillo & Deschamps

Fics. 23-29. Didymium ovoideum. BAFC 26535 (24-26): 23. Sporocarp and columella (SEM).

24. Calcium carbonate crystals. 25. Spore (SEM). 26. Spore ornamentation. Reticularia jurana

BAFC 22846 (27-29): 27. Pseudocapillitium. 28. Spore (SEM). 29. Spore ornamentation reticulate

on one hemisphere and smooth on the other. Bars: 23 = 200 um; 24 = 5 um; 25, 28 = 2 um;

26, 29 = 1 um; 27 = 10 um.

Physarum polycephalum Schwein., Schriften Naturf. Ges. Leipzig 1: 63 (1822)

SPECIMENS EXAMINED: ARGENTINA, BuENOos AIRES, Tigre, 12-X-1968, leg. E.

Barbetti (BAFC 22238); Belén de Escobar, El Cazador, on a rotting trunk of Salicaceae,

10-V-1969, leg. J. Deschamps & E. del Busto (BAFC 22232): As. Llavallol, Inst. Fit. Santa

Catalina, on dry leaves and bark, III-1972, leg. A. Fortuny (BAFC 22610).

CoMMENTS — A widely distributed species, mostly found in the American

continent (www.discoverlife.org). One cited collection has yellowish

sporothecae, but two others are whitish as described in Martin & Alexopoulos

(1969).

Reticularia jurana Meyl., Bull. Soc. Vaud. Sci. Nat. 44: 297 (1908) Figs. 27-29

SPECIMEN EXAMINED: ARGENTINA, BUENOS ArRES, La Plata, Los Hornos, on

living Myoporum laetum G. Forst., 6-1-1972, leg. J. Deschamps (BAFC 22846, as R.

intermedia).

ComMENTSs — ‘The studied specimen is fragmented but can be seen to develop

from a dark reddish, pulvinate aethalium with a brilliant and very fragile

peridium; abundant pseudocapillitium, delicate and non-rigid, formed by

filaments that are flattened to form a three-dimensional net. Spores, dark in

mass, 6-7 um diam., reticulate on one hemisphere and smooth on the other.

The SEM reveals that the spore ornamentation of one hemisphere is formed

by a reticulum of 8-12 angular to hexagonal meshes with solid, unperforated

muri; the other hemisphere is smooth or with a faint ornamentation.

Myxomycetes of BAFC (Argentina) ... 71

Reticularia jurana has been recently cited from Patagonia (Argentina) in

Nothofagus woods (Wrigley de Basanta et al. 2010). In Argentina it has been

confused with Reticularia intermedia Nann.-Bremek., which is distinguished

by a pseudocapillitium formed by branched filaments that do not form a three-

dimensional net.

Stemonitis axifera (Bull.) T. Macbr., N. Amer. Slime-moulds: 120 (1899) | Fics. 30-35

= Stemonitis smithii T. Macbr., Bull. Iowa Univ. Lab. Nat.Hist. 2: 381 (1893)

SPECIMENS EXAMINED: ARGENTINA, BUENOS AIRES, Llavallol, Santa Catalina, in logs,

2-II-1970, leg. L. Frias & G. Rovetta (BAFC 22306, as S. flavogenita); 2-III-1970, leg. J.

Deschamps (BAFC 22310, as S. flavogenita).

COMMENTS — Sporocarps stipitate, 3.5-6 mm total height. Sporothecae dark to

light reddish. Stipe cylindrical, dark reddish, 0.8-1.5(-2) mm length. Columella

reaching to the apex. Capillitium with very open and wide meshes, more

than 35 um diam. Spores hyaline to slightly dark yellowish, (6-)7-8 um diam.,

globose, almost smooth to lightly warted; (SEM) ornamentation of irregularly

distributed verrucae to short bacula and a spore surface covered by a complete

reticulum only visible in the SEM.

In the BAFC 22306 specimen, the columella apex has a small expansion

resembling that of Stemonitis flavogenita but the spore characteristics confirm

that it is S. axifera. Some collections of other Stemonitis species such as S. fusca

Fics. 30-35. Stemonitis axifera. BAFC 22310 (30-32): 30. Columella and capillitium. 31 Spore

(SEM). 32. Spore ornamentation. BAFC 22306 (33-35): 33. Capillitium. 34 Spore (SEM). 35. Spore

ornamentation. Bars: 30, 33 = 20 um; 31, 34 = 2 um; 32, 35 = 0.5 um.

72 ... Moreno, Castillo & Deschamps

occasionally exhibit funnel-shaped expansions at the apex of the columella.

The spore and capillitium characteristics need to be studied carefully to avoid

misidentification.

Stemonitis axifera is a cosmopolitan species (Martin & Alexopoulos 1969).

Moreno & al. (2004) synonymized Stemonitis smithii with S. axifera, noting that

although the spore dimensions varied, the spore ornamentation was the same.

Stemonitis flavogenita E. Jahn, Verh. Bot. Vereins Prov. Brandenburg 45: 165 (1904)

Fics. 36-42

SPECIMENS EXAMINED: ARGENTINA, BUENOS AIRES, Castelar, INTA, I-1970, leg. E.

del Busto & J. Deschamps (BAFC 22298): Llavallol, Santa Catalina, 6-IV-1970, leg. J.

Deschamps (BAFC 22291).

CoMMENTS — The specimens studied are characteristic for their stipitate

aggregated sporocarps, 15-18(-20) mm total height. Sporothecae dark brown,

cylindrical, 9-12 x 0.4-0.5 mm. Stalk cylindrical, dark reddish, 4-5 mm total

height. Capillitium formed by filaments with short spines, forming an peridial

net with meshes of 15-55 um diam. and an internal net of very wide meshes.

Columella widened into an apical funnel up to 120 um diam. Spores globose,

7-8 um diam., dark violet in mass, very light violet by LM, slightly warted. The

spore ornamentation is formed by regularly distributed bacula (SEM).

Stemonitis flavogenita is very similar to S. splendens in spore ornamentation,

but S. splendens lacks a columella funnel. This is a very widely distributed species,

already recorded in Argentina (Deschamps 1976; Crespo & Lugo 2003).

6 oe 37 380 oe

Fics. 36-42. Stemonitis flavogenita. BAFC 22291 (36-38): 36. Capillitium and columella.

37, 38. Spores (SEM). BAFC 22298 (39-42): 39. Capillitium. 40. Apex of columella. 41. Apex of

columella (detail). 42. Spore (SEM). Bars: 36, 40 = 50 um; 37, 42 = 2 um; 38 = 1 um; 39, 41 = 20 um.

Myxomycetes of BAFC (Argentina) ... 73

Stemonitis fusca Roth, Bot. Mag. (Rémer & Usteri) 1(2): 26 (1787)

SPECIMEN EXAMINED: ARGENTINA, BUENOs ArREs, Llavallol, Santa Catalina, Instituto

Fitotécnico, 5-VI-1970, leg. J.E. Deschamps (BAFC 22296, as S. virginiensis).

ComMMENTS — ‘The spore ornamentation is reticulate with 8-10 meshes per

hemisphere. This ornamentation and the larger spore size distinguishes

Stemonitis fusca from Stemonitis virginiensis Rex. More information about

these species can be found in Castillo et al. (1997).

Stemonitis pallida Wingate, in Macbride, N. Amer. Slime-moulds: 123 (1899)

Figs. 43-48

SPECIMENS EXAMINED: ARGENTINA, Buenos Arrgs, Llavallol, Santa Catalina,

leg. C. Naranjo, 26-VI-1964, (BAFC 22309); La Plata, Punta Lara, Arroyo Las Cafias,

on log remains, 10-II-1971, leg. del Busto & J. Deschamps (BAFC 22289). USA:

PENNSYLVANIA, DELAWARE Co., on rotten wood, coll. Hugo Bilgram, Ellis & Everhart,

North American Fungi Second series 3498 (BPI 802957, isotype).

ComMENTsS — ‘This specimen is characterized by its aggregated and stipitate

sporocarps of 4-6 mm total height. Stalk 1-2 mm long, less than half the height

of the sporocarp. Sporotheca cylindrical, dark brown, 0.4-0.5 x 0.3-0.4 mm.

Capillitium an internal net with wide expansions (10-15 um) and large meshes.

Peridial net irregular, formed by hyaline to very pale small meshes, 7-20 um

diam. with short spines which make it distinct from otherwise similar species.

‘ : uke eatin alll

Fics. 43-46 Stemonitis pallida. BAFC 22309 (43-46): 43. Capillitium (detail). 44, 45. Spores

(SEM). 46. Spore ornamentation. BPI 802957 (Isotype): 47, 48. Spores (SEM). Bars: 43 = 20 um;

44, 45 = 2 um; 46-48 = 1 um.

74 ... Moreno, Castillo & Deschamps

Spores globose, 7-8 um diam., pale lilaceous by LM, with abundant and regularly

distributed warts. The ornamentation is formed by dense, long, narrow bacula

(SEM), similar to that of Wingate’s type material from Pennsylvania (USA).

Stemonitis pallida is a species close to S. splendens, which produces taller and

darker sporocarps, a uniformly coloured capillitium, and a larger mesh size in

the peripheral net. Stemonitis pallida is a widely distributed species, already

cited from Argentina (Deschamps 1972; Crespo & Lugo 2003).

Stemonitis splendens Rostaf., Sluzowce monogr.: 195 (1874)

SPECIMENS EXAMINED: ARGENTINA, BuENos Arrgs, Lavallol, Santa Catalina, on

rotten wood, 3-XI-1968, leg. J. Deschamps (BAFC 22196, as S. webberi); II-1970, leg.

J. Deschamps (BAFC 22294. as S. pallida); possibly on bark of Ulmus procera Salisb.,

2-II-1970, leg. J. Deschamps (BAFC 22290, as S. carolinensis); San Clemente del Tuyu,

on fallen log possibly of Populus sp., 24-II-1971, leg. G. Pousadela (BAFC 22285, as

S. flavogenita). USA, VirGINIA, Radnor Heights, on Pinus virginiana, 20-V1-1935, leg.

C.L. Shear (BAFC 22562).

COMMENTS — Stemonitis splendens is characterized by its grouped stipitate

dark red sporocarps, 6-17(-20) mm total height. Sporothecae cylindrical,

(8-)10-15 x 0.4-0.6 mm. Stalk dark red, cylindrical, hollow, 3-7 mm total

height. Capillitium with branches more or less perpendicular to the columella,

externally forming a net of wide meshes 50-80 um diam. Spores globose,

6.5-8(-9) um in diam., with regularly distributed warts. The SEM shows a spore

ornamentation formed by abundant and evenly distributed bacula.

Stemonitis splendens is a species of variable size and morphology (Martin

& Alexopoulos 1969). Stemonitis flavogenita, which displays a similar size

and spore ornamentation under the SEM, is distinguished by a columellar

apex lacking a membrane expanded into a cup-shape and smaller (15-55 um)

capillitial meshes. Stemonitis pallida can be differentiated by its shorter total

height (4-6 mm) and a peridial net of hyaline to very pale meshes, less than

7-20 um diam. and with short spines. SEM reveals spores with longer, denser

bacula.

Stemonitis webberi Rex, Proc. Acad. Nat. Sci. Philadelphia 43: 390 (1891) Fics. 49-57

SPECIMENS EXAMINED: ARGENTINA: BUENOs AIRES, Lavallol, Santa Catalina, on

rotten wood, 9-V-1970, leg. L. Frias & G. Rovetta (BAFC 22297, as S. carolinensis);

bark of Ulmus procera, 22-X-1972, leg. J. Deschamps & G. Rovetta (BAFC 22862, as S.

splendens). USA: NEBRASKA, Lincoln, 1988, leg. H.J. Webber 15 (PH, type of Stemonitis

webberi).

ComMENTs — ‘The Argentinean material is characterized by its sporocarps

forming groups of 15-20 mm total height. Sporothecae cylindrical, dark

brown, 9-12 mm high and 0.5-0.8 mm wide. Stalk dark red, 4-8 mm in length.

Columella reaches to the apex where it gets thinner and becomes sinuous.

Myxomycetes of BAFC (Argentina) ... 75

Fics. 49-57 Stemonitis webberi. BAFC 22297 (49-54): 49. Capillitium and columella. 50. Capillitium.

51, 52. Spores by SEM. 53, 54. Spore ornamentation. Type in PH: 55-57. 55. Spore (SEM). 56, 57.

Spore ornamentation. Bars: 49 = 50 um; 50 = 20 um; 51, 55 = 2 um; 52 = 1 um; 53, 54, 56, 57 = 0.5 um.

Capillitium is formed by an internal net of wide and angular meshes with

an external net of wide meshes of more than 25 um diam., formed by thin

filaments, approximately 1-2 um diam. and without spines. Spores globose to

subglobose, (6.5-)7-8 um in diam., dark violet, homogeneously warted. SEM

reveals that the ornamentation is formed by regularly distributed thick bacula

and that the spore surface is covered by a complete reticulum.

Stemonitis webberi type description: Sporocarps in groups; dark brown.

Sporotheca cylindrical, 4-7 mm height. Stalk reddish of 3-4 mm height.

Capillitium formed by an internal net of large wide meshes, membranous

expansions in the anastomoses and a peripheral net slightly formed. Spores

globose, 7-7.5 um diam., light grey with a paler outline and evenly distributed

spines. By SEM the spores have an ornamentation formed by regularly scattered

bacula distributed over the entire spore surface, which appears to have a slightly

reticulate ornamentation.

76 ... Moreno, Castillo & Deschamps

Stemonitis webberi, which shares with S. splendens a similar capillitium

morphology and spore size, can be differentiated only by SEM due to its specific

spore ornamentation.

Stemonitis axifera also has a similar reticulum on the spore surface

that is observable only by SEM and which cannot be seen by LM. It can be

distinguished by its capillitium with different meshes, different spore size, and

an ornamentation with more scarce, irregularly distributed warts.

(

Fics. 58-61 Trichia subfusca (BAFC 22180): 58. Capillitial elater. 59. Free end of elater. 60. Spore

(SEM). 61. Spore ornamentation. Bars: 58, 59 = 5 um; 60 = 2 um; 61 = 0.5 um.

Trichia subfusca Rex, Proc. Acad. Nat. Sci. Philadelphia 42: 192 (1890) Frias. 58-61

SPECIMENS EXAMINED: ARGENTINA, BUENOS AIRES, Punta Lara, Boca Cerrada, on

rotten wood, 10-5-1969, leg. J. Deschamps (BAFC 22180, as T. pusilla). USA, WEST

VIRGINIA, MONONGALIA Co., Cooper’s Rock State Park, Morgantown, 500 m altitude,

on white-rotted logs of Tsuga canadensis, 16-1X-2000, leg. M. Schnittler & D.W. Mitchell

(DWM 6171).

COMMENTS — Sporocarps scattered and stipitate, 1.1-1.5 mm total height,

stalk fibrous, darker than the sporotheca, 2-3 mm total height. Sporothecae

globose to subpyriform, (0.4-0.5 mm diam.), dark brown with red shades.

Peridium with apical irregular dehiscence, without dehiscence lines marked

as in Trichia botrytis (J.F. Gmel.) Pers. Capillitium formed by yellow elaters of

3-4 um diam., with three smooth spirals and free ends approximately 25 um

long. Spores pale yellow, 9-11 um diam., regularly warted by LM. By SEM, the

spores have relatively large capitate pila that tend to coalesce with adjacent pila

(Rammeloo 1974).

Our material has a longer stalk than found in descriptions of T’ subfusca by

other authors as well as smaller spores (cf. 11-15 um; Martin & Alexopoulos

1969). The American specimen of T’ subfusca (DWM 6171), however, resembles

our specimen in spore size and other dimensions.

Trichia decipiens var. olivacea (Meyl.) Meyl., Bull. Soc. Vaud. Sci. Nat. 55: 244

(1924)

SPECIMENS EXAMINED: ARGENTINA, TIERRA DEL FUEGO, Valle Carbajal, in moss and

hepatics on wood, 20-II-1961, leg. I. Gamundi (BAFC 22187, as Trichia pusilla); on the

Myxomycetes of BAFC (Argentina) ... 77

road to Martial glacier, on rotten wood, 16-I-1964, leg. I. Gamundi (BAFC 22193, as

Trichia pusilla).

ComMENTs — ‘This variety is characterized by its abruptly stipitate sporocarps,

more or less operculate dehiscence, and spores with low crests.

Tubifera ferruginosa (Batsch) J.F Gmel., Syst. nat. 2: 1472 (1792)

SPECIMEN EXAMINED: ARGENTINA, ENTRE R1os, Departamento Victoria, Molino

Doll, on bark of Populus nigra var. italica Koehne, 1-1-1972, leg. J. Deschamps (BAFC

22552, as T. casparyi).

ComMMENTS — ‘This cosmopolitan species forms large pseudoaethalia with a

well-developed hypothallus, usually over log debris. It is characterized by its

reticulate spores (6-8 um diam.) with uniformly sized meshes.

BAFC 22552 was misdetermined as Tubifera casparyi (Rostaf.) T. Macbr., a

species with a central pseudocolumella, a structure absent from T. ferruginosa

and BAFC 22552.

Tubifera microsperma (Berk. & M.A. Curtis) G.W. Martin, Mycologia 39(4): 461

(1947)

SPECIMENS EXAMINED: ARGENTINA, TucuMAN, Chicligasta, Alpachiri, 1-XII-1946,

leg. A.Garolera (BAFC 30276 [ex LIL C-7550]). SANTA FE, Venado Tuerto, on fallen log,

23-II-1970, leg. D. Murray (BAFC 22253).

ComMENnTs — This species is easily recognized by its fructifications that usually

form a stipitate pseudoaethalium, by small spores (4-5.5 um diam.) and an

inner verrucose peridium by LM and which, by SEM, appears as a surface with

small raised tubules reminiscent of octopus suckers (Moreno et al. 2004).

Results

In this first contribution to the myxomycetes of BAFC, we concluded from

our study of 32 specimens (26 from Argentina, 5 from the USA, 1 from Brazil)

that:

(1) The 32 revised specimens represent 20 taxa (19 species and 1 variety) and

resulted in redetermination of the following collections (TABLE 1):

(2) Trichia decipiens var. olivacea is added to the Argentinean myxomycete

catalogue.

(3) Diderma meyerae is added to the USA and North American myxomycete

catalogues.

(4) Badhamia cinerascens, Reticularia intermedia, S. virginiensis, and Tubifera

casparyi are deleted from the Argentinean myxomycete catalogue.

(5) Type materials of Stemonitis pallida and S. webberi are revised and

described.

78 ... Moreno, Castillo & Deschamps

TABLE 1. BAFC collections redetermined as a result of the current study

PREVIOUS INCORRECT LABEL CORRECT NAME

__Badhamia cinerascens BAFC 228220 Badhamia affinis oo ccccsssssssasssse

__Diderma niveum BAFC [ex Bridge Cooke Herbarium 34923] = Diderma meyerae cesses

Physarum polycephalum BAFC 22130 ceeeae Physarum compressum ae

__Reticularia intermedia BAFC 22846 oc csesaesaste Reticularia jurana cc ccssssssssssen

___ Stemonitis carolinensis BAFC 22290 Stemonitis splendens

__ Stemonitis carolinensis BAFC 22297 cee Stemonitis webberi

___ Stemonitis flavogenita BAPC 22285 cece Hemonitis splendens

___ Stemonitis flavogenita BAFC 22306 and BAFC 22310 0 Stemonitis axifera. ae

___ Stemonitis pallida BAFC 22294 ee SHemonitis splendens

__ Stemonitis splendens BAFC 22862 ceeeeenstemonitis webberi

___ Stemonitis virginiensis BAFC 22296 Stemonnitis fusca cae

__ Stemonitis webberi BAFC 22196 cesaesnesseenestemonitis splendens

_Prichia pusilla BAPC 22180 cc csssesnstaesansennenne Trichia subfUsca oo cccasususnsenae

___prichia pusilla BAFC 22187 and BAFC 22193 Trichia decipiens var. olivacea

Tubifera casparyi BAFC 22552 Tubifera ferruginosa

Acknowledgements

We wish to express our gratitude to Mr. A. Priego and Mr. J.A. Pérez of the Electron

Microscopy Service of the University of Alcala de Henares for their invaluable help with

the SEM. We also thank Luis Monje and Angel Pueblas of the Department of Drawing

and Scientific Photography at the Alcala University for his help in the digital preparation

of the photographs, to Dr. J. Rejos, curator of the AH herbarium for his assistance

with the specimens examined in the present study and to Maria L. Deschamps for the

translation and to D.W. Mitchell and M. Lizarraga for revision of the manuscript and

adding a number of useful comments. We would also like to highlight the help provided

for this work by the technicians Susana Pereira and Laura del Busto and to the curator

of the BAFC herbarium Andrea I. Romero.

Literature cited

Arambarri AM. 1973. Myxomycetes de Tierra del Fuego I. Especies nuevas y criticas del género

Diderma (Didymiaceae). Bol. Soc. Argent. Bot. 15: 175-182.

Arambarri AM. 1975. Division Myxophyta. Clase Myxomycetes. Flora Criptogamica de Tierra del

Fuego II: 1-107. FECIC. Buenos Aires.

Castillo A, Moreno G, Illana C, Lago J. 1997. A critical study of some Stemonitales. Mycol. Res. 101:

1329-1340. http://dx.doi.org/10.1017/S0953756297004103

Crespo E. Lugo MA. 2003. Catalogue of the Myxomycetes from Argentina. Mycotaxon 87: 91-102.

Deschamps JR. 1972. El género Stemonitis (Myxomycetidae) en Argentina. Bol. Soc. Argent. Bot.

14: 139-153.

Deschamps JR. 1976. Los Myxomycetes de la Argentina. Catalogo critico, distribucion y clave de las

especies. Physis 35(91): 319-339.

Dorfelt H, Marx G. 1990. Zur terminologie der sporenbildenden stadien der Myxomyceten. Beitr.

Kenntn. Pilze Mitteleurop. 6: 5-14.

Myxomycetes of BAFC (Argentina) ... 79

Farr ML. 1988. Notes on Mycetozoa. V. Corrections, redisposition, and new taxa. Int. J. Mycol. Lich.

3(2-3): 199-213.

Kirk PM, Ansell AE. 1992. Authors of fungal names. A list of authors of scientific names of fungi,

with recommended standard forms of their names, including abbreviations. Index of fungi

Supplement. CABI.

Lado C. 2001. Nomenmyx a nomenclatural taxabase of Myxomycetes. Cuadernos de trabajo de

Flora Micoldgica Ibérica 16: 1-221.

Lado C. 2005-11. An on line nomenclatural information system of Eumycetozoa.

http://www.nomen.eumycetozoa.com (accessed 7-VI-2011).

Lado C, Pando F 1997. Flora Mycologica Iberica. Vol. 2. Myxomycetes, I. Ceratiomyxales,

Echinosteliales, Liceales, Trichiales. Real Jardin Botanico Madrid, J. Cramer. 323 p.

Lado, C, Wrigley de Basanta D. 2008. A review of neotropical myxomycetes (1828-2008). Anales

del Jardin Botanico de Madrid. 65(2): 211-254.

Lizarraga M, Illana C, Moreno G. 1999. SEM studies of the Myxomycetes from the Peninsula of Baja

California (Mexico), I. Arcyria to Fuligo. Ann. Bot. Fenn. 35: 287-306.

Lizarraga M, Moreno G, Esqueda M, Coronado M. 2007. Myxomycetes from Sonora, México. III:

National Forest reserve and wildlife refuge Ajos-Bavispe. Mycotaxon 99: 291-301.

Martin GW, Alexopoulos CJ. 1969. The Myxomycetes. Iowa, University of lowa Press. 560 p.

Moreno G, Oltra M. 2010. Notas sobre los géneros Badhamia, Badhamiopsis y Craterium

(Myxomycetes) en Esparia. Bol. Soc. Micol. Madrid 34: 161-197.

Moreno G, Singer H, Illana C, Sanchez A. 2003. SEM-studies on nivicolous Myxomycetes. The

Diderma niveum complex in Europe. Crytogamie Mycologie 24: 39-58.

Moreno G, Singer H, Sanchez A, Illana C. 2004. A critical study of some Stemonitales of North

American herbaria and comparison with European nivicolous collections. Bol. Soc. Micol.

Madrid 28: 21-41.

Poulain M, Meyer M, Bozonnet J. 2011. Les myxomycétes tomes 1-2. Fédération mycologique et

botanique Dauphiné-Savoie, Sevrier, France.

Rammeloo J. 1974. Structure of the epispore in the Trichiaceae (Trichiales, Myxomycetes) as seen

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in subantarctic forest of Patagonia and Tierra del Fuego, Argentina. Nova Hedwigia 90: 45-79.

http://dx.doi.org/10.1127/0029-5035/2010/0090-0045

MY COTAXON

http://dx.doi.org/10.5248/123.81

Volume 123, pp. 81-89 January-March 2013

New records of Melampsora species on willows in China

PENG ZHAO’, CHENG-MING TIAN’, YI-JIAN YAO?, ZHEN-SHI Hov%,

Qi WANG, YUICHI YAMAOKA° & MAKOTO KAKISHIMA” ©

‘Graduate School of Life and Environmental Sciences, University of Tsukuba,

Ibaraki, 305-8572, Japan

College of Forestry, Beijing Forestry University, Beijing, 100083, China

°State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing, 100101, China

‘College of Agronomy, Inner Mongolia Agricultural University,

Hohhot, Inner Mongolia, 010018, China

°Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,

Jilin Agricultural University, Changchun, Jilin Province, 130118, China

°Faculty of Life and Environmental Sciences, University of Tsukuba, Ibaraki, 305-8572, Japan

*CORRESPONDENCE TO: kakishimal6@poem.ocn.ne.jp

ABSTRACT — Three rust species, Melampsora chelidonii-pierotii, M. humilis, and M. kiusiana,

are recorded for the first time in China based on morphological features and rDNA ITS1-

5.8S-ITS2 sequence information. New hosts are also reported: Salix raddeana for M. chelidonii-

pierotii; S. caprea and S. siuzevii for M. humilis; and S. triandra and S. starkeana for M. kiusiana.

The morphology, host range, and distribution of these species in China are provided.

Key worps — Melampsoraceae, Salicaceae, taxonomy

Introduction

The genus Melampsora Castagne (Melampsoraceae, Pucciniales) contains

about 90 species (Kirk et al. 2008). The species are characterized by formation

of crust-like telia comprising sessile, laterally adherent single-celled teliospores

under the host epidermis (Pei et al. 2005). Melampsora species cause serious

damage to cultivated willows, which are a significant resource for renewable

biomass production (Royle et al. 1992). In China, 25 Melampsora species have

been recorded on willows (Farr & Rossman 2011), of which 24 species were

reported on ~72 cultivars representing 18 sections in Salix, while one species,

M. kamikotica, was reported on Chosenia (Liu 2005, Tai 1979, Wang et al. 1980,

Zhuang 1986, 1989, 1994, Zhuang & Wang 2006, Zhuang & Wei 2002, 2003).

However, the taxonomic identities of these reported species need reevaluation

due to taxonomic confusion and morphological similarity among species.

82 ... Zhao & al.

Melampsora species on willows deposited in several herbaria in China

were investigated by morphological examinations and rDNA ITS1-5.8S-ITS2

sequence analyses. Three species — M. chelidonii-pierotii, M. humilis, and

M. kiusiana — were represent new records for China. Their morphology, host

plants, and geographical distribution in China are described in the present

paper.

Materials & methods

Herbarium specimens used for morphological examinations were borrowed

from the Mycological Herbarium of Institute of Microbiology, Chinese Academy of

Sciences (HMAS); Inner Mongolia Agricultural University (HNMAP); the Mycological

Herbarium of College of Forestry, Northwest A & F University, China (HMNWFC);

Systematic Mycology and Microbiology Laboratory, Agricultural Research Service,

USDA, USA (BPI); and the Mycological Herbarium of the Graduate School of Life and

Environmental Sciences, University of Tsukuba, Tsukuba, Japan (TSH).

The collections were examined morphologically using light microscope (LM) and

scanning electron microscopy (SEM). For LM examination, the urediniospores or

thin sections of uredinia and telia were mounted in a drop of lactophenol solution on

a microscopic slide. Germ pores of urediniospores were observed by the method of

Liang (2006). For SEM, urediniospores or sori obtained from specimens were attached

to specimen holders by double-sided adhesive tape and coated with platinum-palladium

with an E-1030 Ion Sputter Coater (Hitachi, Tokyo, Japan). They were examined using

an S-4200 SEM (Hitachi, Tokyo, Japan) operating at 15 kV.

DNA was extracted from specimens by the methods of Tian et al. (2004) and

fragments of rDNA ITS1-5.8S-ITS2 were amplified by polymerase chain reaction (PCR)

techniques. Amplified PCR products were directly sequenced, and obtained sequences

were compared with data deposited in GenBank using BLAST to calculate similarity

values.

Taxonomy

Melampsora chelidonii-pierotii Tak. Matsumoto, Bot. Mag. (Tokyo) 40: 46, 1926

FIG:.1

SPERMOGONIA and AECIA not seen in Chinese specimens. UREDINIA mostly

hypophyllous, 0.4-0.8 mm; Urepiniospores obovoid, ellipsoid or broadly

ellipsoid, 16-26 x 11-18 um, wall 1.5-2.6 um thick, echinulate, germ pores

3-6, bizonate or tending to be scattered; PARAPHYSES mostly pear- or lollipop-

shaped and interspersed throughout uredinia, evenly thickened at apex or

slightly thickened at apex, up to 7 um. TELIA amphigenous or epiphyllous,

0.2-0.4 mm, subcuticular; TEL1IosporEs 12-41 x 5-14 um, wall 1.6 um thick,

not thickened at apex.

SPECIMENS EXAMINED: CHINA, SHAANXI PROVINCE, Shang Bai Yun, Tai Bai

Shan, II, III on Salix chaenomeloides Kimura, 4 September 1985, Cheng-Ming

Tian (HMNWEC13); JILIN PROVINCE, Antu county, II, III on S. raddeana

Melampsora spp. (China) ... 83

FiGuRE 1. Melampsora chelidonii-pierotii (HMNWFC13). Morphology — A: Uredinium observed by

SEM. B: Obovoid or ellipsoid urediniospores observed by LM. C: Paraphyses with evenly thickened

apex or slightly thickened apex. D: Scattered germ pores (arrows). E: Echinulate urediniospores

observed by SEM. F: Subcuticular telium (e = epidermal cells of the host). Scale bars: A = 100 um;

B, D = 20 um; C = 10 um; E= 15 um.

Laksch. ex Nasarow, 16 July 1960, Yu-Chuan Yang, (BPI1109442); ANHUI

PROVINCE, Qingyang County, IJ, III on Salix sp., 9 November 1932, Cheo S.Y.

(BP1023209, BPI023210).

84 ... Zhao & al.

ComMENTs: Matsumoto (1926) described M. chelidonii-pierotii based on a

rust fungus on S. pierotii from Honshu, Japan. This species is characterized by

subcuticular telia and mostly obovoid or ellipsoid urediniospores. Based on

the morphology, one Chinese specimen on S. chaenomeloides from Shaanxi

Province, which had previously been identified as M. coleosporioides (Cao

& Li 1999), was redetermined to M. chelidonii-pierotii, as were three other

specimens from China labeled as M. larici-epitea (one on S. raddeana and two

on an unidentified Salix sp.). These specimens possessed subcuticular telia, and

urediniospore dimensions agreed well with the descriptions of M. chelidonii-

pierotii (Hiratsuka & Kaneko 1982). Two rDNA ITS1-5.8S-ITS2 sequences

obtained from specimens BPI1109442 and BPI023210 also supported the

morphological identification. A BLAST search of the GenBank database showed

a 100% similarity of these sequences with M. chelidonii-pierotii strain E-1g

(AB646769.1, deposited by Yamaoka et al. 2010). The teliospores of the Chinese

specimens were smaller (12-41 x 5-14 um) than in the Japanese specimens

(18-60 x 14-23 um, Hiratsuka & Kaneko 1982). However, the teliospore

position separated these specimens from the morphologically similar species,

M. coleosporioides.

Melampsora chelidonii-pierotii has been reported on Salix pierotii and

S. chaenomeloides from Japan and Russia (Hiratsuka & Kaneko 1982; Azbukina

1974). Here we report S. raddeana as a new host. Although this rust species

was identified on willows from Shaanxi, Jilin, and Anhui Provinces in China,

a wide distribution has been expected throughout China based on the wide

distribution of S. chaenomeloides in the northeast, northwest, and central regions

the country (Zheng 1983). Japanese mycologists (Matsumoto 1926, Kaneko &

Hiratsuka 1981) reported its aecial stage on Chelidonium majus vat. asiaticum

and Corydalis incisa and demonstrated experimentally the heteroecious life

cycle between S. chaenomeloides and Corydalis incisa. However, host alternation

of M. chelidonii-pierotii in China is still unknown.

Melampsora humilis Dietel, Bot. Jahrb. 32: 50, 1902 Fic. 2

SPERMOGONIA and AECIA not seen in Chinese specimens. UREDINIA mainly

hypophyllous, 0.1-0.4 mm; Urepinrospores globoid or broadly ellipsoid,

14-22 x 11-19 um, wall 1.5-2.8 um thick, echinulate, germ pores 3-7, scattered;

PARAPHYSES mostly pear- or lollipop-shaped and peripheral or interspersed

throughout uredinia, evenly thickened or slightly thickened at apex, up to 4.6

um. TELIA mainly hypophyllous, 0.1-0.4 mm, subepidermal; TELIOsPpoORES

11-36 x 5-14 um, wall sometimes slightly thickened at apex, 1.5-3.2 um.

SPECIMENS EXAMINED: CHINA, HEBEI PROVINCE, Xiao Wu Tai Shan, IJ, III on S.

caprea L., 22 September 1990, Jian-Yun Zhuang (HMAS67392); INNER MONGOLIA,

Genhe City, II, III on S. siuzevii Seemen, 19 August 1993, Zhen-Shi Hou (HNMAP1594.

HNMAP3197).

Melampsora spp. (China) ... 85

FiGurRE 2. Melampsora humilis (HNMAP1594). Morphology — A: Uredinium with peripheral

paraphyses. B: Globoid or broadly ellipsoid urediniospores observed by LM. C: Echinulate

urediniospore observed by SEM. D: Scattered germ pores (arrows). E: Paraphyses slightly

thickened at apex. F: Subepidermal telium with teliospores slightly thickened at apex. Scale bars:

A = 50 um; B = 20 um; C = 6 um; D, E = 15um; F = 25 um.

86 ... Zhao & al.

ComMENTs: Dietel (1902) proposed M. humilis based on a rust fungus on

S. koriyanagi in Japan. Morphologically, this species is closely related to

M. epitea but differs from the latter by its shorter teliospores (Hiratsuka &

Kaneko 1982). A Chinese rust specimen on S. caprea, collected from Hebei

Province and previously identified as M. larici-capraearum (Tai 1979), was

redetermined to M. humilis. Two rust specimens on S. siuzevii from Inner

Mongolia, previously identified as M. repentis in China (Liu 2005), also represents

M. humilis. Although the urediniospore dimensions and shape in these Chinese

specimens agree with M. epitea as reported in Japan (Hiratsuka & Kaneko 1982),

the teliospores are shorter (<36 um) than in M. epitea (<50 um). Sequence

comparison also supported separation of the rust from M. epitea. The rDNA

ITS1-5.8S-ITS2 sequences obtained from two specimens, HMAS67392 and

HNMAP1594, showed a 99% similarity with sequences from M. humilis (TSH-

R10561 and TSH-R10220) from S. integra in Japan (unpublished). The data

were distinct from sequences of two formae specials of M. epitea (AY444777.2

and AY444778.2; 98% similarity) deposited by Pei et al. (2005).

Two willow hosts, S. integra and S. koriyanagi, were reported as hosts of

M. humilis from Sakhalin, Taiwan, and Japan (Hiratsuka & Kaneko 1982). Here

we report M. humilis for the first time for mainland China and S. caprea and S.

siuzevii as new hosts. The aecial host and life cycle information of M. humilis

have not been reported yet, and host alternation is also unknown for China.

Melampsora kiusiana Hirats. f., Bot. Mag. (Tokyo) 57: 281, 1943 Fig. 3

SPERMOGONIA and AECIA not seen in Chinese specimens. UREDINIA

hypophyllous, 0.1-0.4 mm; Urepinrospores globoid or broadly ellipsoid,

9-18 x 6-14 um, wall 1.5-2.1 um thick, echinulate, germ pore scattered or

tending to bizonate, 2-5; PARAPHYsIS pear- or lollipop-shaped and interspersed

throughout uredinia, wall thickened at apex, up to 8 um. TELIA amphigenous,

0.1-0.4 mm, subepidermal; TELIosporEs 14-56 x 5-12 um, wall 1 um thick,

slightly thickened at apex.

SPECIMENS EXAMINED: CHINA, INNER MONGOLIA, Gen He City, H, III on S. triandra

L., 23 August 2005, Zhen-Shi Hou (HNMAP3161, HNMAP3181); Orogen Qi, II, III on

S. starkeana Willd., 23 August 1995, Zhen-Shi Hou (HNMAP1972).

ComMENTs: Hiratsuka (1943), who described M. kiusiana on S. subopposita

in Kyushu, Japan, characterized it with small urediniospores and slender

teliospores. We redetermined two rust specimens on S. triandra from Inner

Mongolia, previously reported as M. repentis, as M. kiusiana. A rust specimen

on S. starkeana, identified as M. lapponum (Liu 2005), also resembled

M. kiusiana. Morphological examinations revealed that the rusts specimens

from S. triandra and S. starkeana had smaller urediniospores (9-18 x 6-14 um)

than other morphologically similar species, M. epitea (13-23 x 11-18 um) and

Melampsora spp. (China) ... 87

FiGurE 3. Melampsora kiusiana (HNMAP3181). Morphology — A: Uredinium with abundant

paraphyses. B: Globoid or broadly ellipsoid urediniospores observed by LM. C: Echinulate

urediniospores observed by SEM. D: Scattered germ pores (arrows). E: Paraphyses with evenly

thickened or slightly thickened apex. F: Subepidermal telium with slender teliospores. Scale bars:

A = 150 um; B = 10 um; C, E= 20 um; D = 5 um; F = 25 um.

88 ... Zhao & al.

M. humilis (16-20 x 13-18 um) (Hiratsuka & Kaneko 1982). The urediniospores

of these specimens were also smaller than M. repentis (13-17 x 12-14 um) and

M. lapponum (20-21 x 15-16 um) (Klebahn 1914). The teliospores were slightly

larger than those of M. epitea (25-50 x 8-14 um), M. humilis (13-33 x 6.5-15

um), M. repentis (16-48 x 7-14 um), and M. lapponum (30-50 x 6-12 um)

(Hiratsuka & Kaneko 1982, Klebahn 1914). Rusts on S. triandra and S. starkeana

agreed with the morphological description of M. kiusiana (Hiratsuka & Kaneko

1982). Sequences from these specimens also differed from M. epitea and M.

humilis. The rDNA ITS1-5.8S-ITS2 sequences obtained from HNMAP3161,

HNMAP3181, and HNMAP1972 were somewhat distinct from M. epitea f.

laricis-epitea (AY444778.2; 98% similarity) deposited by Pei et al. (2005). The

sequences were also distinct from M. humilis (98% similarity) obtained from

S. caprea and S. siuzevii in China. Morphological and molecular comparisons

confirmed these Chinese specimens as M. kiusiana, and this represents the first

report of M. kiusiana from Inner Mongolia in China. In Japan, S. subopposita

was reported as a host of this species (Hiratsuka 1943), and the two willow

species, S. triandra and S. starkeana, are new hosts of M. kiusiana. An aecial

host of this species has not yet been reported for M. kiusiana, and life cycle

information is also unknown for China.

Acknowledgments

We are grateful to Dr. Ming-Hao Pei (Centre for Bioenergy and Climate Change,

Plant & Invertebrate Ecology Division, Rothamsted Research, Harpenden, Hertfordshire,

UK) and Dr. Eric H.C. McKenzie (Herbarium PDD, Landcare Research, Auckland, New

Zealand) for serving as pre-submission reviewers and for their valuable comments and

suggestions. Dr. Zhi-Min Cao (Forestry College, Northwest A & F University, Yangling,

Shaanxi, China) is thanked for kindly providing some important herbarium specimens

in this study.

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Society of Japan 22: 219-230.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi,

10th edn. CABI, Wallingford, UK.

Melampsora spp. (China) ... 89

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Chelidonium majus L. and Corydalis incisa Pers. Botanical Magazine (Tokyo) 40: 43-47.

Pei MH, Bayon C, Ruiz C. 2005. Phylogenetic relationships in some Melampsora rusts on

Salicaceae assessed using rDNA sequence information. Mycological Research 109: 401-409.

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45-54. http://dx.doi.org/10.1016/0961-9534(92)90087-7

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Microbiologica Sinica 20: 16-28.

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MY COTAXON

http://dx.doi.org/10.5248/123.91

Volume 123, pp. 91-93 January-March 2013

Eight new combinations and a replacement name

in the genus Hygrocybe

ALAN E. BESSETTE*, ARLEEN R. BESSETTE,

WILLIAM C. RooDY & WALTER E. STURGEON

* CORRESPONDENCE TO: alanb1@tds.net

ABSTRACT — Eight invalidly published combinations and one nomen novum in the genus

Hygrocybe are validated in this paper.

Key worps — Hygrophorus, valid publication

In WAxcAP MUSHROOMS OF EASTERN NoRTH AMERICA (Bessette et al. 2012),

eleven Hygrophorus names were invalidly transferred into the genus Hygrocybe

because they lacked full and direct basionym references. ‘Three of these eleven

taxa (H. murina, H. conica var. atrosanguinea and H. pratensis var. robusta)

had previously been validly combined by Malloch (2010). The remaining

eight combinations are validated here. According to current interpretations of

Hygrocybe by Bon (1976), Pegler (1983), Singer (1986), Young & Wood (1997),

Young (2005), and Boertmann (2010), all these taxa should be transferred

based on both their macroscopic characters and micromorphology.

It should, however, be noted that the genus Hygrocybe currently is under

revision based on molecular information, and that significant changes in the

taxonomy are expected (Lodge et al. 2006, Boertmann 2010).

Each of the taxa for which we propose a new combination has slender

basidia, parallel to interwoven hymenophoral trama, and a pileipellis that is a

cutis. Several of these taxa have been cited as Hygrocybe species in mushroom

club newsletters and species lists as well as in the literature (e.g., Arora 1986,

Barron 1999) without a valid combination (Boertmann 2002). In accordance

with the rules of the International Code of Nomenclature for algae, fungi, and

plants (McNeill et al. 2012), we propose the following eight new combinations

and one avowed substitute (nomen novum):

92 ... Bessette & al.

Hygrocybe americana Bessette, A.R. Bessette, Roody & W.E. Sturgeon, nom. nov.

MycoBank MB801106

= Hygrophorus acutus A.H. Sm. & Hesler, Lloydia 5: 57. 1942.

non Hygrocybe acuta F.H. Moller 1945

Hygrocybe atro-olivacea (A.H. Sm. & Hesler) Bessette, A.R. Bessette,

Roody & W.E. Sturgeon, comb. nov.

MycoBank MB801107

= Hygrophorus atro-olivaceus A.H. Sm. & Hesler, Sydowia 8: 326. 1954.

Hygrocybe auratocephala (Ellis) Bessette, A.R. Bessette, Roody & W.E. Sturgeon,

comb. nov.

MycoBank MB801108

= Agaricus auratocephalus Ellis, Bulletin of the Torrey Botanical Club 6: 75. 1876.

Hygrocybe basidiosa (Peck) Bessette, A.R. Bessette, Roody & WE. Sturgeon,

comb. nov.

MycoBank MB801109

= Clitocybe basidiosa Peck, Bulletin of the New York State

Museum of Natural History 1(2): 5. 1887.

Hygrocybe marginata var. concolor (A.H. Sm.) Bessette, A.R. Bessette,

Roody & W.E. Sturgeon, comb. nov.

MycoBank MB801110

= Hygrophorus marginatus var. concolor A.H. Sm., Papers of the

Michigan Academy of Science, Arts and Letters 38: 59. 1953.

Hygrocybe marginata var. olivacea (A.H. Sm. & Hesler) Bessette,

A.R. Bessette, Roody & W.E. Sturgeon, comb. nov.

MycoBank MB801111

= Hygrophorus marginatus var. olivaceus A.H. Sm. & Hesler, Lloydia 5: 40. 1942.

Hygrocybe recurvata (Peck) Bessette, A.R. Bessette, Roody & W.E. Sturgeon,

comb. nov.

MycoBank MB801112

= Hygrophorus recurvatus Peck, New York State Museum Bulletin 157: 28. 1912.

Hygrocybe subaustralis (A.H. Sm. & Hesler) Bessette, AR. Bessette, Roody &

W.E. Sturgeon, comb. nov.

MycoBank MB801113

= Hygrophorus subaustralis A.H. Sm. & Hesler, Lloydia 5: 46. 1942.

Hygrocybe tahquamenonensis (A.H. Sm. & Hesler) Bessette, A.R. Bessette,

Roody & W.E. Sturgeon, comb. nov.

MycoBAnkK MB801114

= Hygrophorus tahquamenonensis A.H. Sm. & Hesler, Sydowia 8: 331. 1954.

Hygrocybe comb. nov. and nom. nov. ... 93

Acknowledgments

We wish to thank David Boertmann for his recommendation to validate these

names and for providing transfer data information. We also wish to thank both David

Boertmann and Dr. Steve Trudell for reviewing this manuscript and offering valuable

suggestions to improve it.

Literature cited

Arora D. 1986. Mushrooms demystified, 2" ed. Ten Speed Press, Berkeley, California. 959 p.

Barron GL. 1999. Mushrooms of northeast North America. Lone Pine Publishing, Vancouver, BC.

336 p.

Bessette AE, Roody WC, Sturgeon WE, Bessette AR. 2012. Waxcap mushrooms of eastern North

America. Syracuse University Press, Syracuse, New York. 179 p.

Boertmann D. 2002. Index Hygrocybearum. A catalogue to names and potential names in Tribus

Hygrocybeae Kihner (Tricholomatales, Fungi). Bibliotheca Mycologica 192. 168 p.

Boertmann D. 2010. The genus Hygrocybe, 2°‘ revised edition. -Fungi of Northern Europe 1.

Danish Mycological Society, Copenhagen. 200 p.

Bon M. 1976. Clé monographique des Hygrophoraceae Roze. Doc. Myc. 7(25): 1-24.

Lodge DJ, Matheny P, Cantrell SA, Moncalvo J-M, Vilgalys R, Redhead S. 2006. Delineating the

Hygrophoraceae: character myths vs. gene trees. Poster at the Joint Meeting of the American

Phytopathological Society, The Canadian Phytopathological Society and the Mycological

Society of America, Québec, Canada, 29 July - 2 August 2006.

Malloch D. 2010. Assessment of species diversity in the Atlantic Maritime Ecozone. NRC Research

Press. Ottawa, Canada. 758 p.

McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W. Hawksworth DL, Herendeen PS, Knapp

S, Marhold K. Prado J, Prud’ homme van Reine WF, Smith GF, Wiersema JH, Turland NJ. 2012.

International Code of Nomenclature for algae, fungi, and plants (Melbourne Code ), adopted

by the Eighteenth International Botanical Congress, Melbourne, Australia, July 2011. Regnum

Vegetabile 154. 240 p. http://www.iapt-taxon.org/nomen/main.php

Pegler DN. 1983. Agaric flora of the Lesser Antilles, Kew Bull. Add. Ser. 9. HMSO, London.

http://dx.doi.org/10.2307/4107846

Singer R, 1986. Agaricales in Modern Taxonomy. Koeltz Scientific Books, Koenigstein. 981 p. + 81

pl.

Young AM, 2005. Fungi of Australia. Hygrophoraceae. CSIRO Publishing, Melbourne. Young AM,

Wood AE. 1997. Studies on the Hygrophoraceae (Fungi, Homobasidiomycetes, Agaricales) of

Australia. Australian Systematic Botany 10: 911-1030. http://dx.doi.org/10.1071/SB96005

MY COTAXON

http://dx.doi.org/10.5248/123.95

Volume 123, pp. 95-101 January-March 2013

Tuber subglobosum and T. wenchuanense —

two new species with spino-reticulate ascospores

Li FAN’, JIN-ZHONG CAO? & CHENG-LIN Hou?

" 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 Tuber species from China with spino-reticulate ascospores are

described and illustrated. Tuber subglobosum is characterized by its subglobose ascospores

with distinct spino-reticulate ornamentation, and T. wenchuanense is recognized by the very

irregular spino-reticulate ornamentation on the ascospore surface, with the spines connected

to each other at the base and forming U-shaped ridges at the apex. ITS sequence analyses

support both new species.

KEY worps — Ascomycota, Tuberaceae, taxonomy

Introduction

Arguably the most useful feature for distinguishing Tuber species is the type

of ascospore ornamentation. Some European species such as T. aestivum Vittad.

and T: magnatum Picco have a more or less regular reticulum (mesh) covering

the spore surface (Riousset et al. 2001). The other species group, which includes

T. melanosporum Vittad. and T: rufum Vittad., has ascospores covered by spines.

In contrast, many Chinese species have spino-reticulate ornamentation on the

ascospore surface where a regular arrangement of spines is linked by U-shaped

ridges. ‘This was first described by Liu (1985) for T! taiyuanense B. Liu and

again by Wang (1990) for T: liaotongense Y. Wang from Northeast China. More

recently, other species with spino-reticulate ascospore ornamentation that have

been described include T: huidongense Y. Wang, T: microspermum L. Fan & J.Z.

Cao, T. microspiculatum L. Fan & Yu Li, T: polyspermum L. Fan & C.L. Hou,

T. sinoalbidum L. Fan & J.Z. Cao, and T: umbilicatum Juan Chen & P.G. Liu

(Chen et al. 2005, Fan et al. 2012a,b; Wang & He 2002).

Curiously, so far only one European species, Tuber malacodermum E. Fischer,

and two North American species, T. spinoreticulatum Uecker & Burds. and

96 ... Fan, Cao & Hou

T: lyonii Butters, have been described with spino-reticulate ascospores (Chen

et al. 2005). In this paper two additional species of Tuber with spino-reticulate

ascospores are described from China.

Materials & methods

Morphological studies

The specimens of one Tuber species (collected in Sichuan Province) were already

dried and deposited in HMAS (Herbarium Mycologicum Academiae Sinicae, Institute

of Microbiology, Chinese Academy of Sciences). The specimens of the other new

species were collected fresh from Sichuan and Yunnan Provinces and deposited in

BJTC (Herbarium Biology Department, Capital Normal University). Macroscopic

characteristics are described from both fresh and rehydrated specimens. Microscopic

characters were described from razor-blade sections mounted in 3% KOH, Melzer’s

reagent, or 0.1% (w/v) cotton blue in lactic acid. For scanning electron microscopy

(SEM), spores were scraped from the dried gleba, placed onto double-sided tape,

mounted directly on an SEM stub, coated with gold-palladium, and then examined and

photographed with a HITACHI S-4800 SEM.

Molecular methods

Herbarium samples were crushed by shaking for 3 min at 30 Hz (Mixer Mill MM

301, Retsch, Haan, Germany) ina 1.5 ml tube together with one 3 mm diameter tungsten

carbide ball. Total genomic DNA was then extracted using the PeqLab E.Z.N.A. Fungal

DNA kit following the manufacturer's protocol. The ITS region was amplified with

PCR using the primers ITS1/ITS4 (White et al. 1990). PCR was performed in 50 ul

reactions containing DNA template 2 ul, primer (10 uM) 2 ul each, 2x Master Mix

TABLE 1. Tuber specimens and sequence numbers used in molecular studies.

SPECIES VOUCHER SPECIMEN ITS ITS1 ITS2

T. aestivum C. TaeW028I1-E157 (UPS) AJ888118 a =

C. TaeW064S-W140 (UPS) AJ888062 - -

T. huidongense Thui-pzh08 DQ486032 ~ -

Thui-pzho9 DQ486031 - -

T. liaotongense IFS Y. Wang 88059 - DQ478672 DQ478634

IFS Y. Wang 88061 - DQ478671 DQ478635

T. lyonii GB 112 EU394704 - -

MOHARCN5S5Cfb EU268568 - -

T. rufum Picco Truf-eu01 DQ329375 - -

Rl AF106892 - -

Morphotype 5 AY940646 - -

1506 AY112894 - -

T. spinoreticulatum RH158 GQ221454 - -

U188 FJ809884 - -

T. subglobosum BJTC FAN153 JX267043 - -

T. taiyuanense T42_HMAS 75888 GU979033 a -

HMAS 60234 - DQ478664 DQ478650

T. wenchuanense HMAS 60239 JX267044 - s

Tuber spp. nov. (China) ... 97

(Tiangen Biotech (Beijing) Co. Ltd.) 25 ul. PCR reactions were run as follows: an initial

denaturation at 95 °C for 3 min, followed by 30 cycles at 95 °C for 2 min, 55 °C for 25 s,

72 °C for 2 min, and a final extension at 72 °C for 10 min. The PCR products were sent

to Invitrogen Biotechnology Co. Ltd. (Beijing, China) for purifying, sequencing, and

editing. The other sequence data of ITS rDNA included in this study were downloaded

from GenBank. GenBank numbers are shown in TABLE 1.

Phylogenetic analyses

DNA sequences were aligned with Clustal X (Thompson et al. 1997). The alignment

was manually adjusted with Se-Al v.2.03a (Rambaut 2000). The aligned dataset was

analyzed with maximum parsimony (MP) using PAUP*4.0b10 (Swofford 2002).

MP analysis was conducted using heuristic searches with 1000 replicates of random-

addition sequence, tree bisection reconnection (TBR) branch swapping algorithm. All

characters were equally weighted and unordered. Gaps were treated as missing data to

minimize homology assumptions. A bootstrap (BP) analysis was performed with 1000

replicates, each with 10 random taxon addition sequences. TBR branch swapping was

employed. The Bayesian analysis was performed with MrBayes 3.1.2 (Huelsenbeck et al.

2001; Ronquist & Huelsenbeck 2003) with two sets of four chains (one cold and three

heated) and the stoprule option in effect, halting the analyses at an average standard

deviation of split frequencies of 0.01. The sample frequency was set to 100, and the first

25% trees were removed as burn-in. Bayesian posterior probabilities (PP) were obtained

from the 50% majority rule consensus of the remaining trees. Tuber aestivum was used

as outgroup.

400 Tuber liaotongense IFS Y.Wang88059

1.00

85 Tuber liaotongense IFS Y.Wang88061

Tuber subglobosum JX267043

1o9f Tuber huidongense DQ486032

1.00 Tuber huidongense DQ486031

— Tuber taiyuanense GU979033

1.00

100

1.00|_ Tuber taiyunaense HMAS60234

1.00 4000 Tuber lyonii EU394704

1.00l_ Tyber lyonii EU268568

0.99

Tuber wenchuanense JX267044

4100 Tuber rufum AY 112894

af 1.00|_ FS Tuber rufum AY940646

0.98

s00f Tuber rufum DQ329375

1.001 Tuber rufum AF106892

100] Tuber spinoreticulatum GQ221454

1.00|

Tuber spinoreticulatum FJ809884

'- Tuber aestivum AJ888118

__ Tuber aestivum AJ888062

Fic.1. Phylogeny derived from maximum parsimony analysis of the ITS rDNA sequences of selected

Tuber species with spino-reticulate ascospores, using T. aestivum as outgroup. Bootstrap values of

more than 70% from 1000 replications are shown above the respective branches. Bayesian posterior

probabilities were estimated and clades with PP>0.95 (95%) are marked under the branches.

98 ... Fan, Cao & Hou

Results

Molecular phylogenetics

The maximum parsimony analysis of ITS sequences produced one most

parsimonious tree (Fic. 1) with a length (TL) = 708 steps, consistency index

(CI) = 0.7345, retention index (RI) = 0.7968, homoplasy index (HI) = 0.2655,

and rescaled consistency index (RC) = 0.5852.

The ITS sequence analysis (Fic. 1) revealed that the sequence of the new

species T: subglobosum grouped in a clade with the T! liaotongense sequences

with moderate support (BP = 85, PP = 1.00). The T: wenchuanense sequence

was placed as a separate clade in Tuber.

Taxonomy

Tuber subglobosum L. Fan & C.L. Hou, sp. nov. Fic. 2

MycoBank MB 800576

Differs from other Tuber species by the combination of subglobose ascospores and

spino-reticulate spore ornamentation.

Type: China. Sichuan Province, Huili County, in the soil under mixed woods, 20 Dec.

2010, Jin-Zhong Cao116 (Holotype, BJTC FAN153).

ErymMo oey: subglobosum (Lat.), referring to the subglobose ascospores.

AscoMATA subglobose, 1-2 cm in diam., firm, solid, surface very minutely

flattened verrucose, glabrous, brown at maturity. Odor slight, not pungent.

PeripIuM 200-300 tum thick, two layers; outer layer 100-150 um,

pseudoparenchymatous, composed of small subangular or subglobose cells

mostly 7.5-15 um in diam., with thickened and yellowish-brown walls, darker

towards the outer surface; inner layer 100-150 um, composed of intricately

interwoven hyphae, hyaline, thin-walled, branched, septate, 2.5-5 um in diam.

GLEBA grey brown to brown at maturity, marbled with numerous, narrow,

branched, white to white-yellow veins continuous with inner peridium. Asc1

subglobose, ellipsoid, clavate, or irregular, hyaline, thick-walled, mostly 2-4-

spored, rarely 1-spored, with a short or a very long stalk, 75-100 x 45-70

um excluding stalk. Ascospores mostly subglobose, a few broadly ellipsoid,

dark brown at maturity, 20-32.5 x 20-27.5 um excluding ornamentation;

ornamentation distinctly spino-reticulate, with spines 3-6 um high, straight or

bent at apex, the meshes generally 4-6 across the ascospore width.

ADDITIONAL SPECIMEN EXAMINED: CHINA. YUNNAN PROVINCE, Huize County, in the

soil under mixed woodland, 2 Nov. 2011, Shao-Ping Li 007 (BJTC FAN 168).

Comments — Tuber subglobosum is very similar to T: liaotongense, an endemic

species from Northern China. Tuber liaotongense also has subglobose to broad

elliptic ascospores with spino-reticulate ornamentations, but its spines are

much lower on the reticulum, making the ornamentation look like a typical

reticulum (Cao et al. 2011). This alone is sufficient to distinguish the new

Tuber spp. nov. (China) ... 99

Fic.2. Tuber subglobosum (BJTC FANI153, holotype) a. Ascomata. b, c. Asci and ascospores

observed under light microscope. d. Ascospore observed under SEM.

species. The phylogenetic analysis (Fic. 1) also grouped T. subglobosum in a

clade with T. liaotongense but with only moderate support (BS = 85, PP = 1.00).

This indicates that the two species are closely related but clearly distinct.

Other Tuber species with spino-reticulate ascospores differ from T: subglobosum

by their more or less regular ellipsoid ascospores.

Tuber wenchuanense L. Fan & J.Z. Cao sp. nov. FIG.3

MycoBank MB 800577

Differs from the other Tuber species by its complete, deep and irregular spino-reticulate

ascospore ornamentation with the spines connected to each other at both the bases and

the tips with U-shaped ridges.

Type: China. Sichuan Province, Wenchuan County, in soil under Larix mastersiana

Rehder & E.H. Wilson, 22 Sept. 1989, Bin-Cheng Zhang 618 (HMAS 60239, Holotype);

Bin-Cheng Zhang 622 (Isotype, HMAS 602414).

ETryMOLoGy: wenchuanense (Lat.), referring to the type locality of the taxon.

AscoMaTa globose or subglobose, 0.5-1 cm in diam., grey brown when

fresh, slightly pitted and whitish at the base, surface smooth. Odor not know

100 ... Fan, Cao & Hou

Fic.3. Tuber we

under light microscope. c, d. Ascospore observed under light microscope. e. Ascospore observed

under SEM.

when fresh. PERIDIUM 200-250 um thick, two layers; outer layer 50-100 um,

pseudoparenchymatous, composed of subglobose or subangular cells 5-15 um

in diam., with slightly thickened walls, yellowish-brown towards the surface;

inner layer 150-200 um, composed of intricately interwoven hyphae, hyaline,

thin-walled, branched, septate, 2.5-5 um diam. GLEBA white, pale white or

grey brown at maturity, marbled with numerous, narrow, branched, white

veins continuous with inner peridium. Ascr subglobose, ellipsoid or irregular,

hyaline, thin-walled, 65-95 x 50-70 um, sessile, 1-5-spored. ASCOSPORES

ellipsoid or broadly ellipsoid, yellow brown, covered with spines at first, then

adjacent spines being joined by U-shaped ridges forming a complete irregular

spino-reticulum, 25-45 x 17.5-30 um excluding the ornamentation of 3-4 um

in height, occasionally on some ascospores the tips of spines remain isolated

when the spines reaching up to 5-7.5 um in height and reflexed at the tips,

meshes numerous and various, typically 8-14 across the ascospore width.

COMMENTS — ‘The type specimen of T: wenchuanense in HMAS had been

labeled as T. lyonii, a species originally from North America which, according

to Trappe et al. (1996), is a species with ascospores with isolated spines or spines

with bases partially connected by low ridges that form a complete or incomplete

reticulum. However, our molecular analysis showed that the Chinese material

Tuber spp. nov. (China) ... 101

did not group in the same clade as T. lyonii, and instead belonged to a clade of

its own (Fic. 1). Because of this and the unique ornamentation of its ascospores,

we treat the Chinese material as a new species.

Acknowledgments

We are grateful to Prof. Wen-Ying Zhuang and Dr. Ian R. Hall for serving as the

pre-submission reviewers. The study was supported by the NSFC (No. 30870008), the

Beijing Natural Science Foundation (No. 5072006) and the Key Program of Beijing

Education Commission (KZ201110028036).

Literature cited

Cao JZ, Chi WD, Fan L Li Y. 2011. Notes on the neotype of Tuber taiyuanense. Mycotaxon 116:

7-11. http://dx.doi.org/10.5248/116.7

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, Hou CL, Cao JZ. 2012a [“2011”]. Tuber sinoalbidum and T. polyspermum — new species

from China. Mycotaxon118: 403-418. 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. Mycotaxon119: 391-395. http://dx.doi.org/10.5248/119.391

Huelsenbeck JP, Ronquist F, Nielsen R. Bollback JP. 2001. Bayesian inference of phylogeny and its

impact on evolutionary biology. Science 294: 2310-2314.

http://dx.doi.org/10.1126/science.1065889

Liu B. 1985. New species and new records of hypogeous fungi from China (I). Acta Mycologica

Sinica 4(2): 84-89.

Rambaut A. 2000. Estimating the rate of molecular evolution: incorporating non-contemporaneous

sequences into maximum likelihood phylogenies. Bioinformatics 16: 395-399.

doi:10.1093/bioinformatics/16.4.395

Riousset L, Riousset G, Chevalier G, Bardet MC. 2001. Truffes d'Europe et de Chine. Institut

National de la Recherche Agronomique, Paris. 181 p.

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

Swofford DL. 2002. PAUP*, phylogenetic analysis using parsimony (*and other methods), version

4. Sunderland, MA, USA, Sinauer Associates.

Trappe JM, Jumpponen AM, Cazares E. 1996. NATS truffle and truffle-like fungi 5: Tuber lyonii

(= T. texense), with a key to the spiny-spored Tuber species groups. Mycotaxon 60: 365-372.

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 Res. 24: 4876-4882. doi:10.1093/nar/25.24.4876

Wang Y. 1990. First report of study on Tuber species from China. 45-50. in: Atti del II Congresso

Internazionale sul Tartufo, Spoleto, Nov. 24-27, 1988.

Wang Y, He XY. 2002. Tuber huidongense sp. nov. from China. Mycotaxon 83: 191-194.

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/10.5248/123.103

Volume 123, pp. 103-106 January-March 2013

Cordochaete (Agaricomycetes), a new corticioid genus from India

S.K. SANYAL, SAMITA, G.S. DHINGRA* & AVNEET P. SINGH

Department of Botany, Punjabi University, Patiala 147 002, India

*CORRESPONDENCE TO: dhingragurpaul@gmail.com

ABSTRACT - A new monotypic corticioid genus, Cordochaete, is described with the new

species C. cystidiata, from Haridwar in Uttarakhand, India.

Key worps - hyphal cordons, Chandighat, angiospermous wood

While conducting a fungal foray in Chandighat area in district Haridwar of

Uttarakhand, India, S.K. Sanyal collected a specimen on decaying angiospermous

wood in a mixed forest. The specimen was compared macroscopically and

microscopically with similar genera in Corticiaceae (Rattan 1977, Eriksson et

al. 1981, Burdsall 1985, Dhingra 1987, Nakasone & Sytsma 1993, Greslebin

et al. 2004, Martinez &Nakasone 2005, Zmitrovich & Ezhov 2011) but

could not be assigned to any already known, and is described here as a new

genus. Morphological traits show similarities with the genera Phlebiopsis and

Rhizochaete.

Cordochaete Sanyal, Samita, Dhingra & Avneet P. Singh, gen. nov.

MycoBAank 801039

Differs from Phlebiopsis in having hyphal cordons and is distinguished from Rhizochaete

by the absence of a red or violet KOH reaction in both the hymenium and hyphal

cordons.

TYPE SPECIES: Cordochaete cystidiata Sanyal et al.

Erymotocy: The generic name is based on the resemblance to Rhizochaete and the

presence of hyphal cordons.

Basidiocarp resupinate, adnate, effused; hymenial surface corneous as dry;

hyphal system monomitic; generative hyphae without clamps; basal zone

very thin, context conspicuous, of loosely interwoven hyphae; subhymenium

compact, hyphal cordons both in context and subhymenium; cystidia

thick-walled, encrusted; basidia clavate, 4-sterigmate, without basal clamp;

basidiospores ellipsoidal to subcylindrical, inamyloid, acyanophilous.

104 ... Sanyal & al.

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PLATE 1. Cordochaete cystidiata (holotype). Fics 1-2: Basidiocarps. 1. Fresh. 2. Dried.

Fics 3-7: Microscopic structures. 3. Basidiospores. 4. Basidia. 5. Generative hyphae. 6. Cystidia.

7. Section. Fics 8-9. Micrographs. 8. Encrusted cystidia and basidiospores. 9. Section.

Cordochaete cystidiata gen. & sp. nov. (India) ... 105

REMARKS— ‘This genus can be differentiated from Rhizochaete in lacking

a reddish or purplish change of the basidiome and hyphal cordons in KOH

solution and from Phlebiopsis in lacking very compact context.

Cordochaete cystidiata Sanyal, Samita, Dhingra & Avneet P. Singh, sp. nov.

MycoBank 801040 FIGs 1-9

Differs from Phlebiopsis gigantea by its loosely arranged context and presence of cystidia

only in the hymenium and from Rhizochaete radicata by its compact hymenium and

subhymenium.

Type: India, Uttarakhand: Haridwar, Chandighat, on decaying angiospermous wood, 20

August 2011, S.K. Sanyal 4745 (PUN, holotype).

Erymo.ocy: The epithet refers to the presence of encrusted cystidia.

Basidiocarp resupinate, adnate, effused, < 850 um thick in section; hymenial

surface reticulate, corneous as dry, pale orange to grayish orange when fresh,

orange gray to brownish gray on drying; margins thinning, fibrillose, paler

concolorous to whitish, to indeterminate. Hyphal system monomitic; generative

hyphae < 6.5 um wide; generative hyphae branched, septate, without clamps;

basal zone of a very few horizontal hyphae, followed by conspicuous zone of

loosely interwoven hyphae interspersed with hyphal cordons; subhymenial zone

mainly of compact vertical hyphae and hyphal cordons; hyphae in the cordons

thick-walled, without clamps. Cystidia 85-95 x 11-13.5 um, subfusiform to

fusiform, thick-walled, encrusted, projecting beyond the hymenial surface,

negative to sulphovanillin. Basidia 28-39 x 5-6.5 um, clavate, 4-sterigmate,

thin- to somewhat thick-walled, without basal clamp; sterigmata < 5 um

long. Basidiospores 6.5-8 x 3-3.5 um, ellipsoidal to subcylindrical, thin- to

somewhat thick-walled, inamyloid, acyanophilous.

REMARKS— Rhizochaete radicata differs in having basidiome that turns reddish

purple in KOH solution and subicular hyphae with brown granules that dissolve

in KOH. Phlebiopsis gigantea differs in having cystidia distributed throughout

the basidiome.

Acknowledgements

The authors thank Head, Department of Botany, Punjabi University, Patiala,

for providing research facilities; Prof. Nils Hallenberg (University of Gothenburg,

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

Burdsall HH. 1985. A contribution to the taxonomy of the genus Phanerochaete (Corticiaceae,

Aphyllophorales). Mycologia Mem. 10. 165 p.

Dhingra GS. 1987. The genus Phlebiopsis in the Eastern Himalayas. Nova Hedwigia 44: 221-227.

106 ... Sanyal & al.

Eriksson J, Hjortstam K, Ryvarden L. 1981. The Corticiaceae of North Europe -VI. Fungiflora, Oslo.

pp. 1051-1276.

Greslebin A, Nakasone KK, Rajchenberg M. 2004. Rhizochaete, new genus of phanerochaetoid

fungi. Mycologia 96: 260-271. http://dx.doi.org/10.2307/3762062

Martinez S, Nakasone KK. 2005. The genus Phanerochaete (Corticiaceae, Basidiomycotina) sensu

lato in Uruguay. Sydowia 57(1): 94-101.

Nakasone KK, Sytsma KJ. 1993. Biosystematic Studies on Phlebia acerina, P. rufa, and P. radiata in

North America. Mycologia 85(6): 996-1016. http://dx.doi.org/10.2307/3760683

Rattan SS. 1977. The resupinate Aphyllophorales of the North Western Himalayas. Bibliotheca

Mycologica 60. 427 p.

Zmitrovich IV, Ezhov ON. 2011. Ecology and plectology of Phlebia tremelloidea (Polyporales,

Agaricomycetes). Acta Mycologica 46(1): 19-25.

MY COTAXON

http://dx.doi.org/10.5248/123.107

Volume 123, pp. 107-111 January-March 2013

New or rare fungi from eastern Amazonia. 1.

Circinoconiopsis amazonica gen. and sp. nov.

ANTONIO HERNANDEZ GUTIERREZ

Laboratorio de Micologia, Instituto de Ciéncias Biologicas, Universidade Federal do Para,

Rua Augusto Corréa 01, Bairro do Guamd, Belém, PA, Brasil, CEP: 66071-110

* CORRESPONDENCE TO: anther@ufpa.br

ABSTRACT — Circinoconiopsis amazonica, a new fungal genus and species, is described and

illustrated from specimens collected during an excursion to the Scientific Station Ferreira

Penna at the Floresta Nacional de Caxiuana, eastern Amazonia, Para, Brazil.

Key worps — helicosporous microfungi, palm tree, systematics, tropical rainforest

Introduction

The helicosporous anamorphic fungi include a rather small group of fungi

recognized for their peculiar coiled conidia. These may be coiled in one, two,

or three planes. In some genera such as Candelabrum Beverw., Helicofilia

Matsush., and Spirosphaera Beverw., bizarrely shaped conidia form after a very

complicated coiling and branching process.

From the morphological and ecological point of view, the helicosporous

microfungi are fascinating fungi that are mostly saprobes associated with plant

litter, rotten wood, and decaying twigs in moist places or around water (Zhao

et al. 2007).

In an impressive paper on helicosporous microfungi from China, Zhao et

al. (2007) published a checklist including the fifty-four known genera of this

group of fungi. Intensive studies of mycobiota, mainly in tropical regions, have

led to the discovery of new taxa among these captivating fungi.

The dematiaceous anamorphic fungus that is the subject of this paper was

collected on decaying material of Oenocarpus sp., a kind of palm tree. Its conidia

closely resemble those of the monotypic genus Circinoconis Boedijn. Boedijn

erected Circinoconis in 1942 based on specimens associated with Flagellaria

indica L. (Flagellariaceae) in Sumatra, Imperata arundinacea Cirillo (Poaceae)

in Malaysia, and Plectocomia elongata Mart. ex Blume (Arecaceae) in Java. For

108 ... Hernandez-Gutiérrez

many years no new Circinoconis records were published in any part of the world.

During palynological studies of peat samples from Tripura, India, Prasad &

Ramesh (1984) found dispersed conidia, which were radiocarbon dated at

3340 + 140 Y.B.P. Comparing the fossil conidia with those of C. paradoxa, the

authors concluded that “... spores dealt presently are not known to be borne

by any other fungus, except Circinoconis: Circinoconis was found again, 67

years after the first record, on Calamus (Arecaceae) in Thailand by Pinnoi et al.

(2009). The present fungus is sufficiently different to be proposed here as a new

genus, Circinoconiopsis.

Materials & methods

During an excursion in October 2003 to the Scientific Station Ferreira Pena at

the Floresta Nacional de Caxiuana, eastern Amazonia, Para, Brazil, (1°42.5'-2°15'S

51°15'-51°56'W) samples of decaying parts of palm trees were collected. Conidiophores

and conidia were extracted from the substratum using a fine sharpen needle under

the dissecting microscope and mounted in lactic acid in glycerol (Kirk et al. 2008).

Microscopic observations were made under clear light compound microscope.

Herbarium acronyms follow the Index Herbariorum (Holmgren & Holmgren 2012).

Taxonomy

Circinoconiopsis A. Hern.-Gut., gen. nov.

INDEXFUNGORUM IF 550073

Differs from Circinoconis in lacking trichotomous apical branching of conidiophores

and in having conidiogenous cells that produce only one conidium each.

TYPE SPECIES: Circinoconiopsis amazonica A. Hern.-Gut.

ErymMo.oey: Latin, circinoconi- referring to the hyphomycete genus Circinoconis =

resemblance; Greek, -opsis, (Oyic).

Anamorphic. Conidiophores macronematous, mononematous, erect, rarely

branched. Conidiogenous cells monoblastic, integrated, determinate. Conidial

secession schizolytic. Conidia solitary, coiled in one or two planes, rostrate,

dry.

Circinoconiopsis amazonica A. Hern.-Gut. sp. nov. Fic. 1

INDEXFUNGORUM IF 550072

Differs from Circinoconis paradoxa in lacking trichotomous apical branching of

conidiophores, and in having conidiogenous cells that produce only one conidium

each.

Type: Brazil, Para, Melgaco, Caxiuana, Scientific Station Ferreira Penna, on decaying

leaves of Oenocarpus sp. (Arecaceae), 2. X. 2003, A. Hernandez. (Holotype, MG-

203121).

EryMo_oey: the specific epithet refers to the Amazon forest, the place of collection.

ANAMORPHIC fungus. CONIDIOPHORES in natural substratum effuse, hairy,

originated from mycelium immersed in the substratum; macronematous,

110 ... Hernandez-Gutiérrez

mononematous, mainly unbranched, erect, 9-14-septate, smooth, brown

to dark brown 114-273(-165.8) x 6-9(-7) um. CONIDIOGENOUS CELLS

monoblastic, slightly lageniform to subulate, determinate 37-40 x 1.25-2 um.

Conidiogenous loci complanate, apical. CoNIDIUM SECEssION schizolytic.

Conia helicoid, 1-1%-coils, solitary, 84—150(-335) um high, 28-39 um wide

in the coiled part, individual filaments 14-16 um in the broadest part, 9-18

(— 34)-septate, rostrum 26-100(- 253) um high, brown, paler towards the apex

up to almost sub-hyaline, smooth, sometimes verrucose at basal part, dry.

Teleomorph unknown.

ComMENts: The main differences between this new genus and Circinoconis lie

in conidiophore morphology and pattern of conidiogenesis.

Circinoconis paradoxa Boedijn, the type and only species of that genus,

bears conidiophores that are always trichotomously apically branched. The two

lateral branches are fertile and light brown, while the central one is sterile and

subhyaline. Fertile branches have integrated conidiogenous cells with percurrent

proliferations (Ellis 1971). Conidia are racquet-shaped with a twisted dark

brown basal part and a short to more or less rostrate paler apex. Conidiophores

of Circinoconiopsis are, in comparison, mostly simple or rarely two-branched,

but in that case the branching is neither apical nor trichotomous.

From the original description of Circinoconis the pattern of conidiogenesis

is not clear. It was described thus: “... the development of the conidia starts with

a more or less horizontally directed, nearly hyaline thread ... which forms one

coil and at the point of contact the cell walls are attached to each other, giving

rise to a closed ring” (Boedijn 1942). Nothing was said about the conidiogenous

cells or the mode of origin of conidia (for example, whether holoblastic or

enteroblastic), nor about secession. Ellis (1971) provided more information,

although it is not clear whether Ellis examined the type material (the only

specimen in the collection where Ellis worked, IMI 45731a, was collected on

Imperata arundinacea from Malaysia, but information for that number, added

to the IMI accessions book on 31 May 1951, does not include date of collection

or any indication that it might be part of the type). He described conidiogenous

cells as “.... monoblastic, integrated, terminating branches, percurrent,

cylindrical” Although conidiogenous cells are illustrated by Ellis as annellidic,

as a result of successive conidial production, the term annellidic was not used.

Conidia, according to Ellis (1971), are circinate and rostrate; his illustration

differs from that of Boedijn regarding conidial morphology and shows conidia

that are not ringed. In Circinoconiopsis, by comparison, conidiogenous cells are

determinate, acutely pointed, and sub-lageniform in shape.

Conidiogenesis in this new genus follows the holoblastic pattern, and no new

conidia are produced after the conidium detaches. The conidiophore branching

pattern differ absolutely in the two genera: in Circinoconis branches are always

Circinoconiopsis amazonica gen. et sp. nov (Brazil) ... L111

present in apical position and are associated with conidial production, while

in Circinoconiopsis branches are not always present and, when they occur, are

neither apical nor related to the conidiogenesis process.

Regarding conidial shape, although in both genera conidia are circinate and

rostrate, in Circinoconis only one coil is completed and a laterally protruding

cell is observed just before the beginning of the beaked part, while in

Circinoconiopsis, conidia are one to one and half coiled and the beaked part

is narrower, ending in a longer and thinner rostrum than in Circinoconis. The

conidial coil (40-60 um) is wider in Circinoconis than in Circinoconiopsis.

These remarkable differences between these very close genera are, in the

opinion of the author, enough to justify the establishment of a new genus.

Acknowledgments

The author is grateful to Dr. Rafael Felipe Castafieda Ruiz and Dr. David W. Minter

for their critical review of the manuscript and suggestions for revision. The author also

wants to express his gratitude to the CNPq/PNOPG for financial support and to Museu

Paraense Emilio Goeldi for the laboratory facilities.

Literature cited

Boedijn KB. 1942. Circinoconis, a new genus of Dematiaceae (Fungi Imperfecti). 150th Anniv. Vol.

R. Bot. Gard. Calcutta: 209-211.

Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute. Kew, Surrey.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10 ed. CAB International, Wallingford.

Holmgren PK, Holmgren NH. 2012. [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/

Pinnoi A, Phongpaichit S, Hyde KG, Jones EBG. 2009. Biodiversity of fungi on Calamus (Palmae)

in Thailand. Cryptogamie, Mycologie 30(2): 181-190.

Prasad MNV, Ramesh NR. 1984. First record of Circinoconis conidia from Holocene formation of

Tripura, India. Current Science 53: 38-39.

Zhao GZ, Liu XZ, Wu WP. 2007. Helicosporous hyphomycetes from China. Fungal Diversity

26: 313-524.

MY COTAXON

http://dx.doi.org/10.5248/123.113

Volume 123, pp. 113-119 January-March 2013

Rediscovery of Pseudocolus garciae in southern Brazil

MARCELO A. SULZBACHER”? , VAGNER G. CORTEZ? & IURI G. BASEIA?

"Universidade Federal de Pernambuco, Programa de Pés-graduagao em Biologia de Fungos,

Departamento de Micologia, Recife, PE, Brazil

? Universidade Federal do Parand, Campus Palotina, Palotina, PR, Brazil

° Universidade Federal do Rio Grande do Norte, Departamento de Botanica, Ecologia e Zoologia,

Natal, RN, Brazil

* CORRESPONDENCE TO: marcelo_sulzbacher@yahoo.com.br

ABSTRACT —Pseudocolus garciae is described and illustrated based on fresh specimens

collected in southern Brazil. This is the third known report for the species since its discovery

in 1895. This clathraceous fungus is characterized by the white receptacle, which differs from

the pinkish to red receptacle of P. fusiformis. A detailed description accompanies photographs,

SEM images, and line drawings.

KEY worpDs — gasteromycetes, Phallales, subtropical fungi, taxonomy

Introduction

Pseudocolus Lloyd is characterized by a shortly stipitate receptacle with three

to four unbranched columns bearing the slimy gleba in their internal surface,

which are connected at the apex or seldom become free (Dring 1980). The

genus is a poorly known due to the scarcity of collections and is regarded as

one of the most difficult phalloid genera to treat satisfactorily at the species

level using morphological features (Dring 1980). Colus Cavalier & Séchier is

similar but differs in the receptacle, which is composed of columns forming

an apical lattice (Dring 1980). Pseudocolus (Clathraceae, Phallales; Hosaka et

al. 2006) currently comprises two species: P. fusiformis (E. Fisch.) Lloyd, with

a pinkish to red receptacle widely distributed in Australasia, Europe, and the

Americas (Wright 1960, Burk 1978, Dring 1980, Hemmes & Desjardin 2009,

Akata & Dogan 2011), and P. garciae, with a white receptacle and a restricted

distribution in the subtropical zone of southern Brazil (Moller 1895, Braun

1932, Rick 1961). The present paper reports the occurrence of P. garciae, a rare

and poorly known clathraceous fungus, almost 120 years after its discovery.

It follows recent observations on south Brazilian phalloid fungi (Cortez et al.

2011a, 2011b, 2011c, Trierveiler-Pereira et al. 2009).

114 ... Sulzbacher, Cortez & Baseia

Materials & methods

Specimens were gathered in a Eucalyptus plantation in Minas do Camaqua, located

in the municipality of Cacapava do Sul, southeast region of the state of Rio Grande do

Sul, southern Brazil (30°54'S, 53°25'W). The relict native vegetation in the area, typical

of the Campos Biome of southern South America, is composed of grasses and shrubs of

several families, such as Asteraceae, Cyperaceae, Leguminosae, and Poaceae (Overbeck et

al. 2007), but some areas contain exotic tree species (mostly Pinus and Eucalyptus).

Fresh basidiomata were collected and analyzed macro- and microscopically

in accordance to Miller & Miller (1988). Color terminology follows Kornerup &

Wanscher (1978). Microscopic analysis of basidiomata comprised measurements of

20 basidiospores and hyphae (from the receptacle and rhizomorphs). For scanning

electron microscopy (SEM) study, dried material was mounted directly on aluminum

stubs and coated with a 5 nm thick layer of gold using a Balzers SCD 050 Sputter. Stubs

were examined in a Jeol JSM-6360 LV scanning electron microscope at the Universidade

Federal do Parana (UFPR), Centro de Microscopia Eletrénica (CME). Specimens are

deposited in UFRN-Fungi Herbarium (Holmgren & Holmgren 1998).

Taxonomy

Pseudocolus garciae (Méller) Lloyd, Mycol. Notes 28: 358. 1907. FIGs 1-9

= Colus garciae Moller, Brasil. Pilzb.: 146. 1895.

IMMATURE BASIDIOMATA (mycoeggs) 5-8 mm diam., white (1A1) to

yellowish white (4A2); globose to subglobose, opening by splitting from the

apex and forming three slightly irregular lobes. MATURE BASIDIOMATA 26-35

mm high when expanded. RECEPTACLE 9-13 x 3-4 mm, yellowish white (4A2)

to orange white (5A2) throughout; with a long-cylindrical, rugose and hollow

pseudostipe, splitting upwards into three tapered columns connected at their

tips. COLUMNS with one large tube occupying an entire half of the inner face and

a group of about three small tubes, forming the outer portion, which carry a pair

of flanges running longitudinally down the arm. VoLvaA membranous, 5-7 mm

diam., white (7A1), with numerous white and branched (7A1) rhizomorphs

projecting from the base. GLEBa olive (1F7), produced over the inner surface of

the columns. Opor fetid, resembling decaying fish. BAsIDIOSPORES 3-4.5 (-5)

x 1-1.5 um, ellipsoid to bacilliform, smooth, thin-walled, hyaline in 3% KOH;

surface smooth with SEM. RECEPTACLE composed of pseudoparenchymatous

hyphae, 23.5-37 x 13.5-20 um, globose to subglobose, thin-walled. CLamp

CONNECTIONS present. Basip1A not observed. RHIZOMORPHS constituted by

hyaline, thin-walled hyphae, 3-12 um diam., smooth or encrusted by numerous

angular crystals 1.5-5 um diam., slowly dissolving in 3% KOH. Other hyphae

present smooth thick walls, up to 1.5 um diam., with brown contents, 2-3 um

diam.; terminal hyphae rounded.

Hasirat: Saprophytic on litter and soil, in Eucalyptus plantation.

DISTRIBUTION: only known from southern Brazil, states of Santa Catarina

(Moller 1895) and Rio Grande do Sul (Braun 1932, Rick 1961).

Pseudocolus garciae in Brazil ... 115

Fics 1-3. Pseudocolus garciae. 1. Fresh mature basidioma. 2. Mycoegg in cross-section.

3. Immature and mature specimens showing abundant rhizomorphs. Scale bar: 10 mm.

EXAMINED SPECIMENS: BRAZIL. R10 GRANDE DO SUL, Cacapava do Sul, Minas do

Camaqua, 03.XII.2009, leg. M.A. Sulzbacher 209 (UFRN-Fungi 1522).

Comments: Moller (1895) originally described P garciae (as a Colus species)

from the Atlantic rainforest of Southern Brazil. The holotype was collected near

116 ... Sulzbacher, Cortez & Baseia

Fics 4-5. Pseudocolus garciae SEM photographs.

4. Basidiospores. 5. Rhizomorph surface covered by crystals.

Blumenau, state of Santa Catarina. Lloyd (1907) later transferred the species

to the recently proposed genus Pseudocolus, but others have also considered it

a Lysurus (Hennings 1902) or Anthurus (Cunningham 1931). The receptacle

tubular structure of Pseudocolus is a good character to distinguish the genus

within the anthuroid fungi (Dring 1980).

Pseudocolus garciae in Brazil ... 117

Fics 6-9. Pseudocolus garciae. 6. Rhizomorphs: external hyphae bearing crystals.

7. Rhizomorphs: internal hyphae. 8. Basidiospores. 9. Pseudoparenchymatic hyphae of the receptacle.

Since the original description, little information on this species has been

available. In fact, the only known specimen except for the type collection was

gathered in 1905 by J. Rick, reported later by Braun (1932) and Rick (1961).

It is important to note that Dring (1980) translated his description from the

118 ... Sulzbacher, Cortez & Baseia

original protologue by Méller (1895), and both Lloyd (1907) and Braun (1932)

reproduced the holotype photos. Thus, the present study reports the third

collection for the species.

Our specimens fit in all aspects the descriptions provided by Mdller (1895)

and Braun (1932). There is no doubt the fungus is rare, since it is known only

from the region in which the type collection was originally described. The new

collection’s habitat, however, differs from the original. Examination of new

fresh material allowed observation of both immature and mature specimens

(Fics 1-3) and provided new morphological data not described elsewhere:

long and abundant white rhizomorphs present at the base of the mycoeggs

(Fic 3), as well as the hyphal structure forming them (Fics 5-7).

SEM analysis revealed that hyphae in the rhizomorphs are covered by

numerous prismatic to irregular shaped crystals (Fics 5-6) and confirmed

that the basidiospore surface is, in fact, smooth (Fic 4). Pseudocolus comprises

two species, both occurring in southern Brazil: P fusiformis and P. garciae. In

P. fusiformis, the receptacle is pinkish to red, the stipe is hollow and divided into

3 or 4 vertical columns tapering upwards, and the basidiospores are elliptical,

3-4 x 1-2 um (Dring 1980). The preserved material of P garciae (Brazil. Rio

Grande do Sul: Sao Leopoldo, 1905, leg. J. Rick, PACA 12816, as Colus garciae) is

in poor condition and therefore taxonomically inconclusive. However, judging

from the descriptive notes by Braun (1932) and Rick (1961), it fits well with

Moller’s fungus. The present finding expands the morphological knowledge of

P. garciae from Brazilian subtropics. Additional phylogenetic studies are needed

to clarify the systematics of the Neotropical Phallales.

Acknowledgments

We thank Dr. Admir J. Giachini (Universidade Federal de Santa Catarina, Brazil)

and Dr. Eduardo R. Nouhra (Universidad Nacional de Cérdoba, Argentina) for critical

review of the manuscript, Dr. Maria S. Marchioreto (Herbarium PACA), CME/UFPR

for facilities during SEM analysis and CNPq (Brazil) for financial support to research.

Literature cited

Akata I, Dogan HH. 2011. Pseudocolus fusiformis, an uncommon stinkhorn new to Turkish

mycobiota. Mycotaxon 115: 259-262. http://dx.doi.org/10.5248/115.259

Braun B. 1932. Estudo sobre as Phalloideas Riograndenses. Relatério do Gindsio Anchieta (Porto

Alegre) 1932: 5-28 + 8 pl.

Burk WR. 1978. Pseudocolus fusiformis: synonymy and distributional records. Mycologia 70:

900-905.

Cortez VG, Baseia IG, Silveira RMB. 2011la. Gasteroid mycobiota of Rio Grande do Sul State,

Brazil: Lysuraceae (Basidiomycota). Acta Sci., Biol. Sci. 33: 87-92. http://dx.doi.org/10.4025/

actascibiolsci.v33il.6726

Cortez VG, Baseia IG, Silveira RMB. 2011b. Two noteworthy Phallus from southern Brazil.

Mycoscience 52: 436-438. http://dx.doi.org/10.1007/s10267-011-0124-5

Pseudocolus garciae in Brazil ... 119

Cortez VG, Sulzbacher MA, Baseia IG, Antoniolli ZI, Silveira RMB. 2011c. New records of

Hysterangium (Basidiomycota) from a Eucalyptus plantation in southern Brazil. R. Bras. Bioci.

9:.220"223.

Cunningham GH. 1931. The Gasteromycetes of Australasia XI. The Phallales, Part II. Proc. Linn.

Soc. New South Wales 56: 182-200.

Dring DM. 1980. Contributions towards a rational arrangement of the Clathraceae. Kew Bull. 35:

1-96.

Hemmes DE, Desjardin DE. 2009. Stinkhorns of the Hawaiian Islands. Fungi 2: 8-10.

Hennings P. 1902. Eine neue norddeutsche Phalloidee (Anthurus borealis Burt var. n. klitzingii

P. Hennings). Hedw. Beibl. 41: 169-174.

Holmgren PK, Holmgren NH. 1998. Index Herbariorum: a global directory of public herbaria and

associated staff. Available at: http://sweetgum.nybg.org/ih/. Accessed: 16 May 2011.

Hosaka K, Bates ST, Beever RE, Castellano MA, Colgan III 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

Kornerup A, Wanscher JE. 1978. Methuen handbook of colour. 3" ed. London: Eyre Methuen.

Lloyd CG. 1907. Concerning the Phalloids. Mycol. Notes 28: 349-364.

Miller Jr. OK, Miller HH. 1988. Gasteromycetes: morphology and developmental features. Mad

River Press, Eureka.

Moller A. 1895. Brasilische Pilzblumen. Bot. Mitth. Tropen 7: 1-152.

Overbeck GE, Miiller SC, Fidelis A, Pfadenhauer J, Pillar VD, Blanco CC, Boldrini II, Both R,

Forneck ED. 2007. Brazil’s neglected biome: the South Brazilian Campos. Persp. Pl. Ecol., Evol.

Syst. 9: 101-116. http://dx.doi.org/10.1016/j.ppees.2007.07.005

Rick J. 1961. Basidiomycetes Eubasidii in Rio Grande do Sul - Brasilia 6. Iheringia, Sér. Bot.

9: 451-479.

Trierveiler-Pereira L, Loguercio-Leite C, Calonge FD, Baseia IG. 2009. An emendation of Phallus

glutinolens. Mycol. Progr. 8: 377-380. http://dx.doi.org/10.1007/s11557-009-0603-7

Wright JE. 1960. Notas sobre faloideas Sud y Centroamericanas. Lilloa 30: 339-359 + 3 pl.

MY COTAXON

http://dx.doi.org/10.5248/123.121

Volume 123, pp. 121-128 January-March 2013

Coccomyces pinicola sp. nov. on Pinus armandii from China

RONG-HuaA LEI, KUAN-YUE SUN, QIN ZHANG & CHENG-LIN Hou

College of Life Science, Capital Normal University

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

* CORRESPONDENCE TO: houchenglincn@yahoo.com

ABSTRACT —A new species of Coccomyces, C. pinicola on twigs of Pinus armandii, is described

from Yunnan Province, China. Coccomyces pinicola differs from other species on twigs of

Pinus by its much smaller and stalked ascomata. A phylogenetic analysis based on ITS rDNA

sequences indicates that C. pinicola is more closely related to Colpoma quercinum (but only

with weak support) than to any Coccomyces spp. included in the analysis.

Key worps —Ascomycota, Rhytismataceae, taxonomy, phylogeny

Introduction

Coccomyces De Not. is the second largest genus in Rhytismatales, with

over 100 species (Hawksworth 2008). Molecular analyses have shown that

Coccomyces is polyphyletic, and that the type species, C. tumidus (Fr.) De Not.,

is not closely related to most Coccomyces species but to Lophodermium eucalypti

(Rodway) P.R. Johnst. (Lantz et al. 2011). However, the current out-dated and

unsatisfactory systematics of Rhytismatales must be used until its phylogenetics

are revised.

There are seven Coccomyces species recorded on twigs of Pinus L. (Sherwood

1980, Lin et al. 1994, Hou & Piepenbring 2007). We recently collected another

Coccomyces-like specimen on twigs of Pinus armandii Franch. with a unique

combination of morphological characters that we propose as a new species.

Materials & methods

Morphological methods

Material with mature ascomata was selected for morphological observation.

External shape, size, color, mechanism of opening of ascomata and conidiomata, as well

as the characteristics of zone lines, were observed under the dissecting microscope. The

detailed methods of morphological studies followed Hou et al. (2009).

122 ... Lei & al.

TABLE 1. ITS rDNA sequences used in molecular studies

SPECIES (VOUCHER SPECIMEN) SEQUENCE

Coccomyces australis EF191240

C. dentatus DQ491499

C. guizhouensis (HOU439A) JX317677

C. guizhouensis (HOU439B) JX317678

C. leptideus GU138727

C. pinicola (HOU486A) JX317676

Colpoma quercinum U92306

Cudonia sichuanensis AF433147

Dothidea sambuci AY883094

Elytroderma deformans AF203469

Hypoderma rubi GU138750

Lirula macrospora HQ902159

Lophodermium agathidis AY100662

L. nitens AY100640

L. piceae AY775683

L. pinastri AY100649

Meloderma desmaczieresii AF426056

Meria laricis U92299

Pezicula carpinea AF141197

Potebniamyces pyri AY608642

Rhytisma acerinum GQ253100

R. salicinum AY465515

Spathularia flavida AF433154

Soleella chinensis GU138755

Terriera minor AY100664

Therrya fuckelii JF793672

T. pini JF793676

Tryblidiopsis pinastri JF268769

T. pinastri JF793678

DNA extraction, PCR and sequencing

Total genomic DNA was extracted from ascomata following the protocol of Hou

et al. (2009). Three new sequences for DNA regions nuclear ribosomal DNA large

subunit (nrLSU), internal transcriber spacer (ITS) and mitochondrial DNA small

subunit (mtSSU) were obtained for Coccomyces pinicola (HOU486A) and Coccomyces

guizhouensis Y.R. Lin & B.F Hu (HOU439A and 439B). The LROR/LR5 primers was

used for nrLSU, and the ITS1-f/ITS4 primers for ITS (Vilgalys & Hester 1990, White et

al. 1990); and the mrSSU1/mrSSU3R primers were used for mtSSU (Zoller et al. 1999).

The PCR products were purified, sequenced and edited by Invitrogen Biotechnology Co.

Ltd. (Beijing, China). The other ITS sequences included in this study were downloaded

from GenBank (TABLE 1).

Phylogenetic analyses

We inserted mtSSU and nrLSU DNA sequences obtained from our samples into the

matrix by Lantz et al. (2011) in order to determine the initial phylogenetic position.

Coccomyces pinicola sp. nov. (China) ... 123

The preliminary phylogeny grouped both Coccomyces pinicola and C. guizhouensis were

placed in the core clade, with C. pinicola closely related to Colpoma quercinum (Pers.)

Wallr. and C. guizhouensis close to C. strobi J. Reid & Cain and Therrya sp., although

both clades had very low support (not shown). We next analyzed ITS rDNA sequences

from our samples and related rhytismatalean species recorded on conifer twigs recently

deposited in GenBank (Solheim et al. 2012) for additional preliminary information. We

aligned the ITS rDNA dataset with Clustal X (Thompson et al. 1997) and then manually

corrected by eye in Se-Al v.2.03a (Rambaut 2000). Ambiguously aligned regions were

excluded from further analyses. The sequence data were first prepared and analyzed with

maximum parsimony using PAUP* 4.0b10 (Swofford 1998). Dothidea sambuci, Pezicula

carpinea, Meria laricis and Potebniamyces pyri were chosen as outgroup based on Lantz

et al. (2011). The analysis was conducted using heuristic searches with 1000 replicates

of random-addition sequence, tree bisection reconnection (TBR) branch swapping

and no MAXTREE limit. All characters were equally weighted and unordered. Gaps

were treated as missing data to minimize homology assumptions. A bootstrap analysis

was performed with 1000 replicates, random additions, MAXTREES set to 1000, and

TBR branch swapping. For the Bayesian analysis MrModeltest 3.7 with the Akaike

information criterion (AIC) was used to choose the substitution model for the separate

dataset (Nylander 2004). The model GRT + I + G was chosen for the ITS sequences. The

Bayesian analysis was performed with MrBayes 3.1.2 (Huelsenbeck et al. 2001, Ronquist

& Huelsenbeck 2003) with two sets of four chains (one cold and three heated) and the

stoprule option in effect, halting the analyses at an average standard deviation of split

frequencies of 0.01. The sample frequency was set to 100, and the first 25% of trees were

removed as burn-in. Bayesian posterior probabilities (PP) were obtained from the 50%

majority consensus of the remaining trees.

Results

Molecular phylogenetics

The ITS rDNA matrix included 29 taxa. The ITS rDNA sequence alignment

comprised 540 characters, with 233 phylogenetically informative positions. The

maximum parsimony analysis produced one most parsimonious tree (Fic. 1)

with a length (TL) = 1210 steps, consistency index (CI) = 0.4802, retention

index (RI) = 0.4844, homoplasy index (HI) = 0.5198 and rescaled consistency

index (RC) = 0.2326. Phylogenetic analysis supported the Rhytismatales

(excluding Meria laricis and Potebniamyces pyri) as a clade (BP=87). This main

clade was further divided into three poorly supported clades. Coccomyces

pinicola clustered weakly with Colpoma quercinum, and they were sister to a

moderately supported clade composed of bark-inhabiting species, Coccomyces

guizhouensis, Therrya fuckelii (Rehm) Kujala, Therrya pini (Alb. & Schwein.)

Hohn., and Tryblidiopsis pinastri (Pers.) P. Karst. (see Fic. 1.). The remaining

clades in the phylogenetic tree were similar to a previous study (Lantz et al.

2012).

124 ... Lei & al.

Tryblidiopsis pinastri JF793678

— Tryblidiopsis pinastri JF268769

we Therrya fuckelii JF793672

87 Therrya pini JF793676

ab 36 Coccomyces guizhouensis JX317677

1.00

Coccomyces guizhouensis JX317678

Coccomyces pinicola JX317676

Colpoma quercinum U92306

Terriera minor AY 100664

$4 Lophodermium piceae AY775683

0.99)

Lirula macrospora HQ902159

He Spathularia flavida AF433154

1.00

Cudonia sichuanensis AF433147

ae Coccomyces leptideus GU138727

1.00

Coccomyces dentatus DQ491499

Meloderma desmazieresii AF426056

92 Soleella chinensis GU138755

70 Elytroderma deformans AF203469

Bs Lophodermium pinastri AY 100649

87 1.00

Lophodermium nitens AY 100640

Coccomyces australis EF 191240

Hypoderma rubi GU138750

Rhytisma salicinum AY 465515

Lophodermium agathidis AY100662

Rhytisma acerinum GQ253100

Pezicula carpinea AF141197

97 Meria laricis U92299

Potebniamyces pyri AY608642

Dothidea sambuci AY 883094

Fic. 1. Phylogenetic hypothesis derived from maximum parsimony analysis of the ITS rDNA

sequences of rhytismatalean species and other related species, using Dothidea sambuci, Meria

laricis, Pezicula carpinea and Potebniamyces pyri as outgroup. Bootstrap values of more than 70%

from 1000 replications are shown above the respective branches. Bayesian posterior probabilities

(PP) were estimated and clades with PP>0.95 are marked under the branches.

Coccomyces pinicola sp. nov. (China) ... 125

Ours

Cory

ZS X©S

Fics 2-4. Coccomyces pinicola. 2. A twig bearing ascomata. 3. Ascoma in median vertical section.

4. Paraphyses, asci with ascospores and discharged ascospores.

Coccomyces pinicola R.H. Lei & C.L. Hou, sp. nov. Fics 2-9

MycoBAnk 800963

Differs from Coccomyces guizhouensis by its smaller, stalked, black ascomata.

Type-China, Yunnan Province, Lijiang, Tiejiashan, alt. ca. 2000 m, on dead and

dying twigs of Pinus armandii. 11 July 2007, C.L. Hou 486A (Holotype, BJTC 201202;

GenBank, JX317676); C.L. Hou 486B (Isotype, BJTC 201212).

EryMoLoGy-pinicola, referring to the host genus Pinus.

Ascomata erumpent from the bark, stalked, circular, or slightly irregular,

200-400 um diam., 350-500 um high, single, occasional coalescing, with a black

and crimpled surface, opening by irregular splits to expose a slightly yellow

hymenium. Lips absent. In median vertical section, covering stroma 70-100

um thick near the center of ascomata, and thinner towards the edge, consisting

of an outer layer of 3 rows of dark brown, thick-walled angular cells and an

inner layer of 2-4 rows of hyaline, thin-walled angular cells. Basal stroma well

developed, <100 um thick, consisting of short, hyaline hyphae. Subhymenium

126 ... Lei & al.

under a dissecting microscope. 7. Ascoma in median vertical section. 8. Paraphyses asci, and

ascospores. 9. Discharged ascospores.

40-50 um thick, consisting of small, hyaline cells. Paraphyses 80-130 x 1 um

filiform, not branched, often circinate at the apex. Asci ripening sequentially,

thin-walled, cylindrical-clavate, 60-110 x 7-11 um, with a 30-40 um long stalk,

truncate at the apex, without circumapical thickening, J—, 8-spored. Ascospores

25-30 x 2-3 um, long-fusiform, acute at both ends, hyaline, aseptate, with a

thin or indistinguishable gelatinous sheath.

Conidiomata and zone lines not observed.

DISTRIBUTION. Coccomyces pinicola is known only from the type locality,

Yunnan, China.

Discussion

There are seven Coccomyces species recorded on twigs of Pinus, namely

C. cembrae Rehm, C. guizhouensis, C. irretitus Sherwood, C. lijiangensis C.L. Hou

& M. Piepenbr., C. papillatus Sherwood, C. parvulus Sherwood, and C. strobi.

Coccomyces pinicola is distinguished from these other species on Pinus twigs by

its much smaller ascomata with a conspicuous stalk. The asci and ascospores

of C. pinicola are similar to those of C. guizhouensis, a species that also occurs

on Pinus armandii, but C. guizhouensis has much larger, discoid ascomata

<1.5 mm diam (Lin et al. 1994). Furthermore, host tissue tightly covers the

C. guizhouensis ascomatal surface, resulting in a grey to dark grey ascomatal

Coccomyces pinicola sp. nov. (China) ... 127

surface in contrast to C. pinicola where the host tissue on the covering layer is

detached and the black ascoma is exposed. The other Coccomyces species on

Pinus are easily distinguished by their more or less filiform ascospores. Asci

and ascospores of C. pinicola also resemble Therrya abieticola C. L. Hou &

M. Piepenbr. on Abies. However, the T. abieticola ascomata are inconspicuously

discoid in surface view and the paraphyses are typical of Therrya, i.e. filiform

paraphyses with apical knobs.

The combined nrLSU and mtSSU DNA sequence analyses by Lantz et al.

(2011) indicated that a Therrya sp. and Coccomyces strobi, both occurring on

Pinus twigs, formed a weak clade, which weakly groups with an adjacent clade

containing C. crystalligerus Sherwood and Colpoma quercinum. Tryblidiopsis

pinastri on Picea twigs weakly groups with Cudoniaceae species (Lantz et al.

2011). In the present ITS rDNA sequence analysis, Tryblidiopsis pinastri is

not closely related to the monoclade of Cudonia sichuanensis and Spathularia

flavida, but to a strongly supported clade composed of most bark-inhabiting

species on conifers, Coccomyces guizhouensis, Therrya fuckelii, and Therrya

pini. Interestingly, the new species Coccomyces pinicola is distally related to

C. guizhouensis but weakly sister to Colpoma quercinum, which occurs on the

bark of Quercus and has larger elliptic ascomata opening by a longitudinal split

and filiform ascospores. The results of the phylogenetic analysis was inconsistent

with the morphological characters and it cannot be explained clearly now.

Our molecular analysis suggests that bark-inhabiting species on conifers

(except C. pinicola) are genetically highly related, although they differ in many

aspects, such as ascoma shape and opening patterns and ascospore characters.

Lantz et al. (2011) considered that ascospore characters and ascoma shapes

are limited for generic delimitation and suggested that Therrya, Hypohelion,

Coccomyces (excluding the type species C. tumidus), Colpoma, and Duplicariella

be recombined in the future. Our current analysis indicates that Tryblidiopsis

might also belong in this group.

Acknowledgments

We are grateful to Prof Ying-Ren Lin and Prof. Ming Ye for serving as pre-submission

reviewers, to Joanne E. Taylor for her assistance with English language, and to Dr. PR.

Johnston for giving valuable suggestions for the phylogenetic position of the new

species. The study was supported by the National Natural Science Foundation of China

(No. 30870014, 31170019 and 31270065) and PHR (KZ201110028036).

Literature cited

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Yunnan Province, China. Mycotaxon. 102: 165-170.

Hou CL, Li L, Piepenbring M. 2009. Lophodermium pini-mugonis sp. nov. on needles of Pinus

mugo from the Alps based on morphological and molecular data. Mycol. Prog. 8: 39-33.

http://dx.doi.org/10.1007/s11557-008-0575-z.

128 ... Lei & al.

Huelsenbeck JP, Ronquist F, Nielsen R, Bollback JP. 2001. Bayesian inference of phylogeny and its

impact on evolutionary biology. Science. 294: 2310-2314.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s Dictionary of the Fungi,

10th edn. CAB International, Wallingford, 771p.

Lantz H, Johnston PR, Park D, Minter DW. 2011. Molecular phylogeny reveals a core clade of

Rhytismatales. Mycologia 103: 57-74. http://dx.doi.org/10.3852/10-060.

Lin YR, Liu HY, Tang YP, Hu BF. 1994. Two new species of the genus of Coccomyces from China.

Acta Mycol. Sin. 13: 8-12. (in Chinese)

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574.

Sherwood MA. 1980. Taxonomic studies in Phacidiales: the genus Coccomyces (Rhytismataceae).

Occ. Pap. Farlow Herb. Crypt. Bot. 15: 1-120.

Solheim H, Brynthe T. Hietala AM. 2013. Characterization of the ascomycetes Therrya fuckelii

and T. pini fruiting on Scots pine branches in Nordic countries. Mycol. Prog. 12: 37-44.

http://dx.doi.org/10.1007/s11557-012-0813-2.

Swofford DL (1998) PAUP*: phylogenetic analysis using parsimony (and other methods). Sinauerm

Sunderland, MA.

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Nucleic. Acids. Res. 25: 4876-4882.

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several species of Cryptococcus. J. Bacteriol. 172: 4238-4246.

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

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small subunit ribosomal DNA of lichen-forming ascomycetes. Lichenologist 31: 511-516.

MY COTAXON

http://dx.doi.org/10.5248/123.129

Volume 123, pp. 129-140 January-March 2013

Sporormiella octomegaspora, a new hairy species

with eight-celled ascospores from Spain

FRANCESCO DOVERI* & SABRINA SARROCCO

Department of Agriculture, Food and Environment, University of Pisa,

80 via del Borghetto, 56124 Pisa, Italy

* CORRESPONDENCE TO: f.doveri@sysnet.it

Axsstract — An ascolocular ascomycete with semi-immersed, hairy and pyriform

pseudothecia, abundant pseudoparaphyses, fissitunicate 8-spored asci, and dark, very

large 8-celled ascospores has been isolated from deer dung in Andalusia (Spain). Based on

morphological features, a new species is erected and accommodated in Sporormiella, which

the authors regard as a genus independent of Preussia. The new species is discussed and

placed in a key, and a previous worldwide key to Sporormiella species with 8-celled spores

is updated.

Key worps — coprophily, phylogeny, Pleosporales, relationships, Sporormiaceae

Introduction

Sporormiella Ellis & Everh. (Sporormiaceae Munk, Pleosporales) is

characterised by ascoloculate, aperiphysate, ostiolate pseudothecia, fissitunicate,

elongated, 8-spored asci with a scarcely developed apical apparatus, dark

colored, transversely septate, 4- to poly-celled ascospores with germ-slits and

usually with a gelatinous envelopment, and preferable growth on dung (Ellis &

Everhart 1892, Ahmed & Cain 1972, Barr 2000).

Preussia Fuckel, in the same family, has morphological features so similar to

Sporormiella that its independence has been questioned. We refer to previous

works on this subject (Doveri 2004, 2005, 2007, 2011; Doveri & Coué 2008) to

explain why the senior author regards Sporormiella as distinct from Preussia

(Cain 1961; Ahmed & Cain 1972; Barrasa & Checa 1991; Lumbsch & Huhndorf

2007, 2010; Kirk et al. 2008). That view has been strengthened by an extensive

phylogenetic study on Sporormiaceae (Kruys & Wedin 2009), which does not

definitely resolve the question of synonymy.

Another view (von Arx 1973, Guarro et al. 1997a, Chang & Wang 2009) that

regards Sporormiella as a later synonym of Preussia has led to the recombination

of several Sporormiella species in Preussia (von Arx 1973; Valldosera & Guarro

130 ... Doveri & Sarrocco

1990; Guarro et al. 1997a,b; Abdullah et al. 1999; Arenal et al. 2004, 2005, 2007;

Chang & Wang 2009, Kruys & Wedin 2009) or even the description of new

Sporormiella species in Preussia sensu lato (Arenal et al. 2005, 2007; Chang &

Wang 2009, Asgari & Zare 2010).

Sporormiella encompasses, at the present, more than 80 species, mostly with

glabrous pseudothecia and 4-celled ascospores, a minority (12) with hairs,

and/or more than 4-celled ascospores, very few with ascospores consisting of a

variable number of cells (2-10).

A recent collection from Andalusia (Spain) of a hairy species with

consistently 8-celled ascospores has been submitted to our observation. Its

morphological features are unique, thus allowing us to recognise it as a new

Sporormiella species.

The aim of our study is to define the new species and establish its relationships

with other hairy Sporormiella spp. and a group of Sporormiella with 8-celled

ascospores.

Materials & methods

Dried and fresh specimens of Sporormiella octomegaspora were sent us from Spain,

the fresh material still on small pieces of red-deer (Cervus elaphus) dung. Fresh material

(five specimens) was utilised for macro- and microscopic studies and for attempting,

unsuccessfully, isolation in axenic culture. The specimens were picked from the dung

with a sterile needle, washed in distilled water, and placed in a drop of water on a

microscope slide. Microscopic examinations were carried out on specimens mounted

in water, Melzer’s reagent, and methyl blue. Spore size was measured in water and

calculated on 50 ascospores discharged from mature asci in 4 ascomata. The collection

has been preserved as dried material and slides (MCVE). Herbarium abbreviation

follows Holmgren & Holmgren (2012).

Sporormiella spp. that have not been recombined in that genus, are indicated by

presenting the current binomials with the genus enclosed in quotation marks (e.g.,

“Sporormia’ carpinea, “Preussia” variispora).

Taxonomy

Sporormiella octomegaspora Doveri & Sarrocco, sp. nov. PL. 1-3

MycoBAank 803276

Differs from all other Sporormiella species by its larger 8-celled ascospores.

Type: Spain, Andalusia, Huelva, Dofiana National Park (approx. 37°N 6.5°W), on red

deer dung, 28.10.2008, leg. A. Suarez (Holotype, MCVE 27409)

EryMo_oey: From the Greek octo = eight, megas = large, spora = spore, referring to the

large, 8-celled ascospores

MACROCHARACTERS — PSEUDOTHECIA 720-900 x 400-500 um, ostiolate,

pyriform, semi-membranous, dark brown to blackish, rough, tomentose owing

to a dense net of hyphoid hairs spread all over the ascoma. Neck differentiated,

200-250 x 180-200 um, cylindrical, somewhat darker, subcoriaceous, with the

Sporormiella octomegaspora sp. nov. (Spain) ... 131

\. TT

PLATE 1. Sporormiella octomegaspora (holotype): A = ascoma on dung; B = hairs at the neck apex;

C = detail of exoperidium at the neck; D-F = peridial hairs; G = centrum; H = detail of exoperidium;

I = Pseudoparaphyses. Scale bars: A = 300 um; B, E-F = 20 um; C = 80 um; D, H = 40 um; G = 150

um; H = 25 um.

132 ... Doveri & Sarrocco

same net of hairs at its base and shorter, more isolated hairs at the apex, hardly

observable at low magnification (10 x).

MICROCHARACTERS — PERIDIUM pseudoparenchymatous, two-layered:

endostratum of pale brown to almost hyaline, thin-walled, polygonal cells,

13-18 x 12-15 um, exostratum a textura angularis of dark brown, thick-

walled, polygonal cells, 6-10 x 5-8 um, with intervals of cylindrical cells in

rows (textura prismatica), a textura globulosa-angularis of blunt angular

to roundish and somewhat darker cells at the neck, 5-9 um diam., papillate

around the ostiole. Some carbonaceous deposits sprinkled all over, but more

abundant on the neck; PERIDIAL HAIRS hyphoid, very long, wavy, strongly

intertwined, fairly thick-walled, quite dark brown, septate, often branched,

sometimes anastomosed and somewhat encrusted, blunt at the tips, 2-3.5 um

diam.; NECK HAIRS hyphoid at the base, similar in size and shape to peridial

hairs, polymorphous, paler, stiffer and shorter at the apex, 15-30 um long,

usually with a rounded, sometimes clavate tip; PPEUDOPARAPHYSES mixed with

the asci and exceeding them, cylindric-filiform, sometimes submoniliform,

3-7 um diam., often branched, septate, usually slightly tapering upwards,

somewhat constricted at the septa, containing abundant hyaline vacuoles; Asc

325-450 x 47-72 um, fissitunicate, inamyloid, 8-spored, cylindric-clavate to

clavate, lacking an apical apparatus, roundish or dome-shaped or even slightly

flattened at the apex, usually quite abruptly narrowing below in a short stalk,

12-30 um long; ascospores (148-)152-175(-180) x 16-18(-20) um, usually

three to five bundled in the upper portion of the ascus, the others placed at

various levels below, and one usually the lowest, hyaline in the early stages,

pale brown later, finally dark brown, cylindric-subfusiform, individually

surrounded by a broad gelatinous envelope, smooth, thick-walled, constantly

8-celled, deeply and transversely septate. Cells easily separable at any level, each

with a sigmoidal, parallel germ slit. End cells conical, with blunt tips, narrower

and usually longer than the middle ones, but often not longer than the second

and the seventh, third to sixth cells from the upper end sub-cubical, almost as

long as wide, and the third usually the broadest, sometimes slightly wider than

long, the second and seventh cell cylindrical, usually longer than wide, all cells

with fairly acute angles.

EcoLoeGy & DisTRIBUTION—About fifteen scattered specimens on red-deer

(Cervus elaphus) dung in the field in October. To date only known from the

type locality.

Discussion

Sporormiella octomegaspora is characterised by large and densely hairy

ascomata, with hairs widespread both on the pseudothecial venter and neck,

large and short-stalked asci, and very large, constantly 8-celled, subfusiform

ascospores with sigmoidal germ slits and a particular arrangement inside

Sporormiella octomegaspora sp. nov. (Spain) ... 133

PLATE 2. Sporormiella octomegaspora (holotype): A-B = upper cells of ascospores at the ascus apex;

C-E = upper and middle parts of asci with ascospores; F = ascus base; G = detail of fissitunicate

(arrows) asci; H = ascus bases with ascospores. Scale bars: A-B = 20 um; C-H = 50 um; I= 150 um.

134 ... Doveri & Sarrocco

PLATE 3. Sporormiella octomegaspora (holotype): A-B = ascospores; C-E = detail of ascospores;

F-G = spore cells. Scale bars: A—B, F = 35 um; C = 18 um; D-E = 15 um; G = 30 um.

Sporormiella octomegaspora sp. nov. (Spain) ... 135

the asci. The whole of these features allows us to separate S. octomegaspora

from species with hairy pseudothecia and/or 8-celled ascospores and easily

distinguish it from all other Sporormiella spp.

In their basic monograph on Sporormiella, Ahmed & Cain (1972) reported

3 hairy species with 4-celled ascospores and 18 species with constantly 8-celled

ascospores, including the coprophilous “Sporormia” octoloculata Fabre and

“Sporormia” pulchra E.C. Hansen (both cited as doubtful or not examined by

them) and the non-coprophilous, non-examined “Sporormia” carpinea Fautrey,

“Sporormia” gigaspora Fuckel, S. subticinensis (Mouton) Dugan & R.G. Roberts,

and S. ticinensis (Pirotta) Doveri (Doveri 2004). They also reported 2 species

with inconsistently 8-celled ascospores, S. commutata (Niessl) S.I. Ahmed

& Cain (7-9-celled), and “Sporormia” variabilis G. Winter (5-8-celled). All

Sporormiella species described by Ahmed & Cain (1972) — with the exception

of S. insignis (Niessl) S.I. Ahmed & Cain and S. splendens (Cain) S.I. Ahmed

& Cain — whether with constantly or inconstantly 8-celled ascospores, differ

from S. octomegaspora due to ascospores that are much shorter than 100 um.

Sporormiella splendens has, like S. octomegaspora, short-stalked asci, but can

be easily distinguished by its glabrous ascomata, smaller pseudothecia and asci,

and somewhat shorter but notably narrower cylindrical ascospores (9-13 um

diam., Cain 1934, Ahmed & Cain 1972, Furuya & Udagawa 1972, Leenurm

1998, Treigiené 2004), with middle cells longer than broad and an oblique to

diagonal germ slit.

Although it has short-stalked asci, and ascospores with cubical middle cells

and an arrangement inside the asci similar to S. octomegaspora, S. insignis

differs in having glabrous and much smaller pseudothecia, smaller asci (Winter

1887, Bayer 1924, Prokhorov & Armenskaya 2003), and somewhat narrower

and notably shorter (100-122 x 14-15 um; Niessl 1878, Ahmed & Cain 1972)

ascospores, with middle cells equal in size to each other (so the spore shape is

cylindrical rather than cylindric-fusiform) and diagonal germ slits.

Bell (2005), who collected the species in Australia on Eastern Grey kangaroo

dung and which she described as S. insignis, straddles S. insignis and S. splendens,

“having the ascospore width of the former and the ascospore length of the latter,”

but in our opinion drawings and measurements reported in her work match

those of S. octomegaspora. Although the Australian material was unfortunately

inadequate for a herbarium sample (Bell, pers. comm.), our examination of a

slide from Australia strengthens our original opinion, as Bell’s material has only

slightly wider spores on average.

Doveri (2004) mentioned and briefly described or discussed (sometimes

as Preussia) all new Sporormiella spp. published after Ahmed & Cain's (1972)

monograph, some of which are characterised, like S. octomegaspora, by hairy

pseudothecia and/or 8-celled ascospores. Among them, the hairy species,

136 ... Doveri & Sarrocco

S. lasiocarpa Lorenzo and “Preussia” aquilirostrata Guarro et al. (Guarro et

al. 1997b), have 4-celled and much smaller ascospores [respectively 62-76 x

10-12 um (Lorenzo 1994), 34-44 x 8.8-11.5 um (Guarro et al. 1997b)] than

S. octomegaspora; on the contrary, the species with 8-celled ascospores —

S. oblongiclavata M.E. Barr & Malloch, S. tomilinii O.V. Korol., and “Preussia”

variispora Abdullah et al. — do not have hairy pseudothecia, and “Preussia”

variispora has both 7- and 8-celled ascospores, while all have much

smaller ascospores (Abdullah et al. 1999, Barr 2000, Korolyova 2000) than

S. octomegaspora.

Besides establishing new species of Sporormiella, Doveri (2005, 2007),

Doveri & Coué (2008), and Chang & Wang (2009) updated the list of new

Sporormiella spp. (sometimes as Preussia) published since 2004. None of them

has consistently 8-celled ascospores.

The Welt & Heine (2007) worldwide key to Sporormiella spp. with 8-celled

ascospores is here updated with the addition of S. octomegaspora and other

species not previously included:

Worldwide key to Sporormiella species with 8-celled ascospores

[N.B.: “Sporormia octoloculata” and “Sporormia” carpinea have been omitted from the

key, as the original descriptions are meagre]

la. Ascospores 5-10-celled (never consistently 8-celled) ...............00. eee eeee 2

LbrAscospores-conisistently Saceled s fs-scick dessvcied feds seven tie peben hes deboa tbss- dougie Sob ephickeg % A

2a. Ascospores 5-8-celled, 62-75 x 14-19 um (Winter 1874), cylindrical, with middle

cells usually broader thanlong ....................... “Sporormia” variabilis

2b: Ascospores:/—9-_celled citlerentant chape Asai 5 tant eM se tall Soke wre cla) 3

3a. Ascospores (7—)8-9(-10)-celled, 45-53 x 8-10 um, fusiform with middle cells

wider than long, and the third cell usually the broadest ....... Sporormiella sp.

(described by Richardson 2004b)

Sb Ascospores longer eylindrigs clavate: s ot.ns Fuk.) ees net eee aR See ak 4

4a. Ascospores 50-60 x 8-10.5 um (Ahmed & Cain 1972), usually 9-celled, sometimes

7-8-celled, with the middle cells broader than long, and the third cell from the

Lio PEC PHEDIOAGEST a So ents eso Mutts Mandily Bement Maguey tea oct S. commutata

Ab. 9-celled ascospores rare or absent ......... 0... cece eect eee eee teens 5

5a. Ascospores 65-75 x 7-12 um, 7-8-celled, with middle cells as long as broad, and

the third cell from the upper end the largest ............. “Preussia” variispora

5b. Ascospores mostly 8-celled, sometimes 7-9-celled ........... cece eee eee 6

6a. Ascospores 70-90 x 11-13 um, with the third or fourth cell from the upper end

hives citi Ree he Rie Raita Siete ot Ak Seal ete See RR Sporormiella sp.1

(described by Richardson 2004a)

6b. Ascospores 58—72 x 8—9.5 um, cylindrical, with isodiametric cells, to cylindric-

clavate, with the upper cells shorter than wide .............. Sporormiella sp. 2

(described by Richardson 2004a)

Sporormiella octomegaspora sp. nov. (Spain) ... 137

7atlb): Ascogpores: 1.00) it lone..y F.e nn ch e een cp new cg oe SES eee ay 8

CD ASCOSPOLES <0 WOM ONG: AML tM A UM UM AIM AlN AA dlle OM cages oR ae 10

8a. Ascomata hairy, 720-900 x 400-500 um. Ascospores 152-175 x 16-18 um,

cylindric-subfusiform with third to sixth cells from the upper end almost

as long as wide, and the third usually the broadest. Germ slits sigmoid and

Pratl ey Se Se Fk Sten epee tester ent Wem dete cent eae esse S. octomegaspora

8b. Ascomata glabrous, smaller. Ascospores narrower, cylindrical. Germ slits oblique

{ROMO F TELE) 1 Peak edd abet peed dee Ae a Air rE A is Eon is rt Ale le alo Mis Boe oe Bele 9

9a. Ascomata 550-600 x 350-400 um. Ascospores 135-165 x 9.5-12.5 um (Cain

1934), with middle cells longer than broad, cylindrical ............ S. splendens

9b. Ascomata subglobose, 250-330 um diam. Ascospores 100-122 x 14-15 um

(Ahmed & Cain 1972), with middle cells almost equal in size, cubical

truth ven Pamite Maas Mevatts Mena ha Wena te Mevaty Mona tita Meva te teann oe arans oe; S. insignis

10a. (7b) Ascospores variable in shape, cylindrical to cylindric-fusoid or slightly

cylindric-clavate, 50-58 x 6-8 um, with middle cells equal in width, or the

fourth and/or the fifth, or even some other cell, the broadest ... Sporormiella sp.

(described by Cribb 1994)

10b; Ascospores-quite:conistant:in-shape sci 4.5 scba dre peta tre acea de aces deep tee dg eecdreg Om 11

1la. Ascospores cylindrical, with middle cells almost equal in width ............. 12

11b. Ascospores (sub)clavate, with one broader cell .............. 0... e ee eee 16

12a: AScospotesanore-than.6U MimMelOne Cte Kan Ie, lees nett gta tnt asa ltadbat 9 13

12b:-Ascospores:less:than-60 (timdone: 2. 2.7.26 oo. Ene ne eine 5 ole cee Sle ee SG 14

13a. Ascospores 79-95 x 14-16 um (Ahmed & Cain 1972). Middle cells broader

than long and with an obliquely transverse germ slit ............. S. platymera

13b. Ascospores 72 x 9 um (Fuckel 1871), with middle cells as long as wide

obs Sori ck eae tah kL dah LP ee ck LR 2 aR “Sporormia” gigaspora

(S. subticinensis with ascospores 62—75 x 9-12 um (von Arx & Storm 1967), is hardly

separable from “Sporormia” gigaspora, although Mouton (1897) noted that it differs in

having clavate rather than oblong asci and much smaller ascomata)

14a (12b). Ascospores 47-57 x 12-14 um, with middle cells usually broader than

1S hse a PARE AS EAR rE Re CAR UTE ARS tr Air) Mame A “Sporormia” pulchra

14b. Ascospores narrower, with middle cells almost as long as wide .............. 15

15a. Ascospores 48-58 x 6—7 um with diagonal germ slits ................ S. bipartis

15b. Ascospores 40—44 x 8 um (Pirotta 1878). Germ slits not described .. S. ticinensis

16a (11b). Ascospores 30-48 x 4—7 um, with the second cell from the upper end the

broadest, and with almost parallel germ slits. Non-coprophilous

Divey aN 5 Eek sesh + Mag t > Tggh a etlgats atta slag ses ate ask S. oblongiclavata

16b. Third or fourth cell the broadest. Germ slits oblique to diagonal

(not described in S. tomilinii). Coprophilous species ..................00- 17

17a. Third cell the broadest. Cells easily separable to non-separable .............. 18

17b. Fourth cell the broadest. Cells hardly separable .....................00004. 23

138 ... Doveri & Sarrocco

18a. Ascospores 38—43.5 x 8-9 um (Doveri 2008) (40 x 5-6 um, Auerswald 1868;

40-48 x 7-8 um, Ahmed & Cain 1972). Cells easily separable from each

other, and the end ones conic- ovoid. Asci gently narrowing below in a long

Stalks tts va PA siconrey Seton Uh ovate, Morar El ons LM cea cll ora iM ona esther et trons S. octomera

bSbeAscosnoreslarger Lyf See ay A ee A ee ee ee Ey oo ye 19

19a--Asci abruptly-contractéed inva very short stipe... eee ee es need 20

19b. Asci gently narrowed in a stipe. Ascospores with (sub)conical end cells ...... 21

20a. Ascospores 48.5—63 x 12.5-14.5 um (Doveri 2008) (48-58 x 12-14 um,

Ahmed & Cain 1972). Spore cells hardly separable, end cells hemispheric

aitey etdanty Meads Meet, Upumantly apehy Bava. hy Meee ten Dy UMN, At ne nla e onl S. octonalis

20b. Ascospores 52-57 x 8-9 um (Ahmed & Cain 1972), with cells easily separable,

CTIACCELS SUDCOMICANE: drs. Besant PRlmarts Miche Berna Mende d Bande Benrey t S. schadospora

21a (19b). Ascospores 71-90 x 12-16 um (Bommer & Rousseau 1886) (65-80 x

12-15 um, Ahmed & Cain 1972). Asci short-stipitate ................ S. affinis

ZU: ASCOSPOKES-SHOTLET sg Foe ag Oy et te, ee ae On Oe ER, ARE WW Ra kona ee! 22

22a. Ascospores 52.5-60 x 9.5—10.5 um (Doveri 2008) (50-59 x 10-11.5 um,

Ahmed & Cain 1972). Cells not separable. Asci long-stipitate ....S. corynespora

22b. Ascospores 55-70 x 13-15 um. Cells easily separable. Asci short-stipitate

ER PASAT PEN 0 ERs PORTE 3 PRENATAL AEN ORE IEE S. tomilinii

23a (17b). Ascospores 49-58 x 9-10 um (Ahmed & Cain 1972). Asci abruptly short-

SEDILALC: 9.0 Poe ROe Ae PRS BRS Bee! Tle! UENO! TAM Oae Lee RL UE S. ontariensis

23D; ASCOsporecsmmaller™ fa. psd hay ddiba-e gd. weery d-neer gd neera di neerg di parte sd peeue snore 24

24a. Ascospores 40-51.5 x 7.5-8.5 um (Doveri 2008) (40-49 x 8-9 um, Ahmed &

Cain 1972) Ascrabruptly-short-stipitate ss. «..7 Fo... reas hPa 2 S. pascua

24b. Ascospores 34-42 x 7 um (Doveri 2008) (32-36 x 5.5-6.5 um, Ahmed & Cain

1972) -ASel pently-short=stipitate v1.9 penta dclt por Pugh pat eae eat d ap foe S. minipascua

Acknowledgements

The authors wish to thank A. Bell and M.J. Richardson and the MycoTAxon editors

for critical revision of the manuscript, A. Suarez and E. Rubio Dominguez for providing

us with the material subject of our study.

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MY COTAXON

http://dx.doi.org/10.5248/123.141

Volume 123, pp. 141-145 January-March 2013

Neolinocarpon attaleae sp. nov.

on Attalea funifera (Arecaceae) from Brazil

NapjA SANTOS VITORIA*”, MARIA A.Q. CAVALCANTI,

CRISTIANE DUARTE DOS SANTOS?, JADERGUDSON PEREIRA?

& JOSE LUIZ BEZERRA**

"Departamento de Micologia, Universidade Federal de Pernambuco,

Av. Prof. Nelson Chaves, s/n°, 50670-901, Recife, PE, Brazil

*Departamento de Educagao, Universidade do Estado da Bahia,

Rua do Gangorra, n° 503, 48.608-240, Bairro Alves de Souza, Paulo Afonso, BA, Brazil

*Departamento de Ciéncias Agrdrias e Ambientais, Universidade Estadual de Santa Cruz,

Rodovia Ilhéus-Itabuna, Km 16, 45662-900, Ilhéus, BA, Brazil

‘Centro de Ciéncias, Agrdrias e Biologicas, Universidade Federal do Recéncavo da Bahia,

Rua Rui Barbosa, 710, 44.380-00, Cruz das Almas, BA, Brazil

* CORRESPONDENCE TO: nadjasv@hotmail.com

ABSTRACT — Neolinocarpon attaleae is described from the palm Attalea funifera collected in

Brazil. This species is illustrated and compared with similar species. A key to all species of

Neolinocarpon is provided.

Key worps — Ascomycota, biodiversity, palm fungi, taxonomy, Brazilian mycota

Introduction

Hyde (1992) introduced the genus Neolinocarpon K.D. Hyde to accommodate

N. globosicarpum K.D. Hyde, a Linocarpon-like specimen, with deeply immersed

ascomata formed beneath a slightly raised or flattened clypeus. Since then,

seven new species and a new combination have been added to Neolinocarpon:

N. australiense K.D. Hyde et al., N. calami K.D. Hyde et al., N. enshiense K.D.

Hyde et al., N. eutypoides (Penz. & Sacc.) K.D. Hyde et al., N. inconspicuum

K.D. Hyde et al., N. nonappendiculatum K.D. Hyde et al., N. nypicola K.D. Hyde

& Alias, and N. penniseti Bhilabutra & K.D. Hyde (Hyde et al. 1998, Hyde &

Alias 1999, Bhilabutra et al. 2006). All species of Neolinocarpon have so far been

collected on palms, except N. penniseti, a species from grass.

During palm microfungi surveys of Pernambuco and Bahia States

along the Brazilian Northeastern coast 81 ascomycetes were identified

142 ... Vitoria & al.

(Souza et al. 2008, Vitoria et al. 2008, 2010, 2011a,b, 2012). In this paper a

new species, Neolinocarpon attaleae from Attalea funifera Mart. ex Spreng.

(Arecaceae), is illustrated, described, and compared with similar species within

Neolinocarpon.

Materials & methods

Dead leaves of palms were collected in the municipality of Una, Bahia state.

The specimens collected were incorporated into CEPEC Herbarium (Mycological

Collection) in Itabuna, Bahia. Observations under the stereomicroscope were followed

by the study of squash preparations and vertical free hand sections of the ascomata.

Morphological features were described and measured using a Zeiss-Axiophot E

microscope and photographed were taken using a digital camera Cyber-shot 4.1 Mega

Pixels DSC-P73x, adjusted to the eyepieces of the microscope and the stereomicroscope.

All measurements were made in water preparations. Samples stained with lacto-glycerol

cotton blue, Melzer’s reagent and acid lactofuchsin were also prepared.

Neolinocarpon attaleae Vitoria & J.L. Bezerra, sp. nov. PLATES 1-2

MycoBANnk 800126

Differs from Neolinocarpon nonappendiculatum by its shorter, wider, slightly curved to

straight, clavate ascospores.

Type: Brazil. Bahia, Una, Estacao Experimental Lemos Maia-CEPLAC, 15°16'207"S

39°05'532"W, on dead leaf (rachis) of Attalea funifera, 21.VII.2009, Nadja Vitoria, 156

(Holotype, CEPEC 2320).

EryMo_oey: attaleae, in reference to the host genus, Attalea.

ASCOMATA deeply immersed in host tissue, externally visible as black, shiny,

raised and spherical bodies, due its conspicuous clypeate ostioles; in vertical

section 350-880 x 220-650 um (including ostiole), subglobose with a

blackened clypeus surrounding the periphysate ostiole. OSTIOLE ca 117.5-250

um diam, 170-250 um high, central. PERrpIUM up to 55 um wide, composed

of several layers of brown, elongated, flattened cells, sometimes comprising

two darker outer strata and one lighter inner stratum. PARAPHYSES up to 5

uum wide, numerous, hypha-like, septate, unbranched. Asci1 137.5-227.5 x

7.5-14(-15) um, 8-spored, cylindric-clavate, usually curved, sometimes straight,

long pedicellate, unitunicate, apically rounded, with a refractive, discoid, non-

amyloid subapical ring ca 1.5-2 x 1-2 um. Ascosporgs (52.5-)57.5-93(-105) x

3-4(-5) um, fasciculate, hyaline, guttulate, unicellular, smooth, slightly curved

to straight, filiform-cylindrical, filiform-fusoid to + clavate, apex rounded,

attenuated towards rounded base, with refractive septum-like bands, lacking

appendages.

ADDITIONAL SPECIMENS EXAMINED — BRAZIL. BAuntA: Una, Estacao Experimental

Lemos Maia-CEPLAC, 18.XII.2008, Nadja Vitoria, 154,155 (CEPEC 2318, 2319);

15.11.2010, Nadja Vitéria, 157, 158 (CEPEC 2321, 2322); 06.VII.2010, Nadja Vitoria,

159A GEPEG 2323).

Neolinocarpon attaleae sp. nov. (Brazil) ... 143

st me ——

PLATE 1. Neolinocarpon attaleae (from holotype): A. Appearance of the ascomata on the host

surface. B-C. Vertical section through an ascoma. D. Section through peridium.

ComMENTs — ‘The collections from Attalea funifera were identified as

Neolinocarpon according to Hyde (1992), but differ from other species of the

genus mainly by ascospore morphology. As in N. penniseti, N. inconspicuum,

and N. nonappendiculatum, N. attaleae lacks appendages at the apices of the

ascospores (Bhilabutra et al. 2006). Ascospores of N. attaleae have similar

dimensions to N. penniseti ((52.5-)57-64(-84) x 2.5-3 um; Bhilabutra et al.

2006) and N. inconspicuum (76-98 x 2-3 um; Hyde et al. 1998), but the new

species differs in ascoma size and shape, asci and subapical ring, and ascospore

morphology.

Key to species of Neolinocarpon

Las tFASCOSPOTES WILMA PPETIGACED srad, "rates Mar arees Naval Maracas Meracche orarsty wean actouema cea 2

ib Ascospontes Wil hOUtappendagesd 6604 hark Gi Teh ft Ph TR ee 8 oe he Sh 7

Zo, ASCOSPOkes GM er CNANGD: MIN tr e3 disse dhe ok dudypceS dap od dobaoced die 48d agen dasa ded 3

2b. Ascospores 42-64 x 2-3.5 um; mucilaginous pad at narrow truncated base

Su pghton ea dreaeg anes ta dipes gaint diner gd ery acherg ache tai tG N. enshiense

UTICA ai ea, creel tae Me tae bes hae WR ER SE meh eae Ol gence 4

SD RETEEGRIT A tae ist sebsd auch setod arch satsd qin sated anton cial entender fcbnivn tent clair ten SAR chin f IB choy Uh ey A 5

4a. Ascospores 70-119.5 x 2-3 um; ascomata solitary .............. N. globosicarpum

4b. Ascospores 92-117 x 2-3.8 um; ascomata in groups of 1-4............ N. nypicola

5a. Basal appendage keel-like; ascospores 81-107(-126) x 2.5-3.5 um ...N. australiense

5b. Appendage a crescent-shaped or mucilaginous pad .............. 0. eee eee eee 6

144 ... Vitoria & al.

ve *

12um

20 um

20 wm

20 um

PiaTE 2. Neolinocarpon attaleae (from holotype): A. Paraphyses and _ asci.

B. Ascus. C. Non-amyloid, discoid subapical ring (arrow). D-F. Ascospores.

Neolinocarpon attaleae sp. nov. (Brazil) ... 145

6a. Crescent-shaped mucilaginous pad at apex; ascospores 68-85 x

DSS SS LUT Pl Reel aM ieee NAME, 18s Brel AN, ba alls tevaatles acs tills fac saben foc oak esd N. calami

6b. Mucilaginous appendage at base; ascospores 73-95(-106) x

TESA 2 LM eee oe ee re esas its ilcctts Wiecdt wltedar tet tet N. eutypoides

7a. Ascomata inconspicuous; ascospores 76-98 x 2-3 uM........... N. inconspicuum

ADRASCONIABA -CONSDICUOUSHs , iunctary Wine its Miwebey ube waa gdaly hah ey hie Sethe teh Ble eed 8

8a. On grass (Poaceae); ascospores (52.5—)57-64(-84) x 2.5-3 um ....... N. penniseti

SPLON- paleriseCArecdecae) tas se nk catlena tatters tkesh Ns elesee oon tase ra a ary eit ore iebie Sugita 9

9a. Ascospores (52.5-)57.5-93(-105) x 3-4 um, filiform-cylindrical,

filiform fuse@id ost eelay diene 54) yh sary ld, rae cate ats Cooley N. attaleae

9b. Ascospores 114-138 x 2-2.5 um, filiform, curved.......... N. nonappendiculatum

Acknowledgments

The authors thank CAPES-MEC Brazil, CNPq-Brazil for scholarships and CEPLAC

for facilities and laboratories used to lead the research. Also sincere thanks to Drs.

Dartanha J. Soares and Olinto Liparini Pereira for the pre-submission review for our

manuscript.

Literature cited

Bhilabutra W, Lumyong S, Jeewon R, McKenzie EHC, Hyde KD. 2006. Neolinocarpon penniseti sp.

nov. on the grass Pennisetum purpureum (Poaceae). Cryptogamie Mycologie 27(4): 1-6.

Hyde KD. 1992. Fungi from decaying intertidal fronds of Nypa fruticans including three

new genera and four new species. Botanical Journal of the Linnean Society 110: 95-110.

http://dx.doi.org/10.1111/j.1095-8339.1992.tb00284.x

Hyde KD, Alias SA. 1999. Linocarpon angustatum sp. nov., and Neolinocarpon nypicola sp. nov.

from petioles of Nypa fruticans, and a list of fungi from aerial parts of this host. Mycoscience

40: 145-149.

Hyde KD, Taylor JE, Frohlich J. 1998. Fungi from palms XXXIV. The genus Neolinocarpon with five

new species and one new recombination. Fungal Diversity 1: 115-131.

Souza CAP, Vitoria NS, Bezerra JL, Luz EDMN, Inacio CA, Dianese JC. 2008. Camarotella

brasiliensis sp. nov. (Phyllachoraceae) on Syagrus schizophylla (Arecaceae) from Brazil.

Mycotaxon 103: 313-317.

Vitoria NS, Bezerra JL, Gramacho KP, Luz EDMN. 2008. Camaroetella torrendiella comb. nov. e

C. acrocomiae: agentes etioldgicos das lixas do coqueiro. Tropical Plant Pathology 33(4):

295-301. http://dx.doi.org/10.1590/S1982-56762008000400006

Vitoria NS, Bezerra JL, Gramacho KP. 2010. A simplified DNA extraction method for PCR

analysis of Camarotella spp. Brazilian Archives of Biology and Technology 53(2): 249-252.

http://dx.doi.org/doi:10.1590/S1516-89132010000200001

Vitoria NS, Cavalcanti MAQ, Hyde KD, Bezerra JL. 2011a. Arecomyces new to Brazil, including

A. attaleae sp. nov. Cryptogamie Mycologie 32(1): 103-108.

Vitoria NS, Cavalcanti MAQ, Luz EDMN, Bezerra JL. 2011b. Endocalyx melanoxanthus var.

melanoxanthus (Ascomycota): a new to Brazil and three new hosts. Mycotaxon 117: 109-113.

http://dx.doi.org/10.5248/117.109

Vitoria NS, Cavalcanti MAQ, Hyde KD, Bezerra JL. 2012. Brunneiapiospora brasiliensis

sp. nov. (Clypeosphaeriaceae) on palms from Brazil. Nova Hedwigia 94(1-2): 245-250.

http://dx.doi.org/10.1127/0029-5035/2012/0094-0245

MY COTAXON

http://dx.doi.org/10.5248/123.147

Volume 123, pp. 147-155 January-March 2013

New rust species and hosts from Northern Areas of Pakistan

M. SABA'* & A.N. KHALID’?

"Department of Botany, University of the Punjab

Quaid-e-Azam Campus, Lahore, 54590, Pakistan

* CORRESPONDENCE TO: rustflora@gmail.com

ABsTRACT — During a survey of rust fungi of Northern Areas of Pakistan, Puccinia aegopodii

and P. rhytismoides were collected as new to the Indian subcontinent. The uredinial state of

Chrysomyxa pyrolae is described for the first time from Pakistan on Pyrola rotundifolia subsp.

karakoramica, a new host. Puccinia swertiae is reported as a new record for Northern Areas,

and P. oniwaensis is recorded on Persicaria orientalis, a new host.

Key worps — Deosai Plains, Fairy Meadows, Uredinales

Introduction

Most parts of Northern Areas lie within the watersheds of the Himalaya,

Hindu Kush, and Karakoram mountain ranges. Climatic conditions vary widely

in the Northern Areas ranging from the monsoon influenced moist temperate

zone in the western Himalayas to the semi-arid cold desert in the Northern

Karakoram and Hindu Kush. The Northern Areas of Pakistan are well known

for their biodiversity. Although these areas contain about 3000 plant species

and are floristically rich, they have not been thoroughly surveyed for rust fungi

and only approximately 71 species have been reported to date (Ahmed 1956a,b;

Gjaerum & Iqbal 1969; Kakishima et al. 1993a,b; Kaneko 1993; Khalid et al.

1995; Khalid & Iqbal 1996, 1997a,b; Ahmed et al. 1997; Sultan et al. 2006a,b,

2008; Iqbal et al. 2008, 2009; Afshan & Khalid 2009; Afshan et al. 2009, 2010,

2011; Ishaq et al. 2011; Khalid & Saba 2011; Saba & Khalid 2011). During our

2010-2011 rust collection trips in the Northern Areas, we discovered Puccinia

aegopodii and P. rhytismoides, both new for the Indian subcontinent and which

we report with new hosts and other rust fungi new for these areas of Pakistan.

Materials & methods

Rust fungi were collected from Deosai Plains and Fairy Meadows, Northern Areas,

Pakistan. Healthy plants with inflorescences and fruits were collected for accurate

148 ... Saba & Khalid

identification. Host plants were identified by comparing them with specimens in the

herbarium of the Department of Botany, University of the Punjab, Lahore (LAH).

Free hand sections of infected portions of material and spores were mounted in

lactophenol. Semi-permanent slides were prepared by cementing cover slips with nail

lacquer (Dade & Gunnell 1969). Preparations were observed under a NIKON YS 100

microscope. Drawings of spores were made by using a Camera Lucida (Ernst Leitz,

Wetzlar, Germany). Spores were measured using an ocular micrometer (Zeiss, St

Albans, Hertfordshire, England). At least thirty spores were measured for each spore

stage. Measurements include the usual range and the arithmetic means; extremes are

given in parentheses.

Taxonomy

Chrysomyxa pyrolae (DC.) Rostr., Bot. Centralbl. 5: 127 (1881). Fic. 1

SPERMOGONIA, AECIA and TELIA not seen. UREDINIA scattered evenly on

abaxial surface of leaves, Caeoma-like with delicate, one-layered peridium,

orange-yellow, round, surrounded by ruptured peridium. UREDINIOSPORES

globose, obovoid, ellipsoid to polyhedral, hyaline to subhyaline, sometimes

with yellowish contents, produced in chains, (15-)18-22 x (18-)22-29 um

(mean 19.8 x 24.7 um); wall hyaline, 1-2 um thick, densely verrucose.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Fairy Meadows, on leaves of

Pyrola rotundifolia subsp. karakoramica (Kiisa) Y.J. Nasir (Ericaceae), with II stage, 23

Jul 2010, coll. & det. Malka Saba 12 (LAH).

ComMENTs: The aecial stage of Chrysomyxa pyrolae was reported on cones

of Picea smithiana (Wall.) Boiss. from Fairy Meadows by Kaneko (1993) and

Ahmad et al. (1997). Its uredinial stage from the same locality is newly recorded

for Pakistan, and Pyrola rotundifolia subsp. karakoramica is a new host for this

rust fungus.

Puccinia aegopodii (Schumach.) H. Mart., Prodr. Fl. Mosq., Ed. 2: 226 (1817). Fic. 2

SPERMOGONIA, AECIA and UREDINIA not seen. TELIA amphigenous,

predominantly hypophyllous, minute, rounded, aggregated, surrounded by

ruptured epidermis, pulverulent, black. TeLIospores ellipsoid or oblong,

cinnamon brown to dark brown, not or slightly constricted at septum, rounded

at both ends, sometimes attenuated below, (12-) 16-22 x (22-)26-35 um (mean

18.7 x 30.3 um); wall dark brown to chestnut brown, 1-2(-2.5) um, germ pores

apical, sometimes subapical or near septum in distal cell, adjacent to septum

in proximal cell, mostly with hyaline dome-shaped, up to 3 um high papilla;

pedicel hyaline, short, deciduous, 5-6 x 2-9 um.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Fairy Meadows, on leaves of

Aegopodium alpestre Ledeb. (Apiaceae), with III stage, 23 Jul 2010, coll. & det. Malka

Saba 13 (LAH). GERMANY, BADEN-WURTTEMBERG, Donautal, Inzigkofen, am

Strasschen zum Nickhof, kurz vor dem Nickhof, on leaves of Aegopodium podagraria L.,

16 May 2010, leg. O. Holdenrieder and R. Berndt (ZT Myc 3027).

Chrysomyxa & Puccinia spp. new to Pakistan ... 149

A 18kU X33@ SO ms

“ERE eUOP

B 10K ex2/S6G_ 161m CRL UOP

eet ee ,

Fic. 1. Chrysomyxa pyrolae (SEM). (A) Uredinium with urediniospores and one-layered peridium

(square). (B) Urediniospores with densely verrucose ornamentation.

150 ... Saba & Khalid

Fic. 2. Puccinia aegopodii. Lucida drawings showing position of germ pores on teliospores.

Scale bar =15 um.

ComMENTs: Farr & Rossman 2011 have reported P aegopodii from China and

Mongolia on Aegopodium alpestre. Our report is an addition to the mycobiota

of the Indian subcontinent.

Puccinia oniwaensis Hirats. f. & S. Sato, Trans. Mycol. Soc. Japan 3: 56 (1962). Fic. 3

SPERMOGONIA and AECIA not seen. UREDINIA hypophyllous, pulverulent,

brown, intermixed with telia. UREDINIosPpoREs subglobose, ellipsoid to

obovoid, pale brown to brown, 15-20 x 22-27 um; wall 1.2-2 um, minutely

echinulate, germ pores 2, supraequatorial. Telia hypophyllous, sometimes

aggregated, pulverulent, black. TEL1osporgs ellipsoid to oblong, rounded at

both ends, chestnut brown, slightly or not constricted at septa, 20-22 x 27-34

um; wall chestnut brown, verrucose, more dense in upper cell; 2-3 um; germ

pore apical in upper cell and near pedicel in lower cell; apex 3-4.5 um, brown.

One and three celled teliospores rarely present.

Chrysomyxa & Puccinia spp. new to Pakistan ... 151

B—_

Fic. 3. Puccinia oniwaensis (Lucida drawings).

(A) Urediniospores showing minutely echinulate ornamentation.

(B) Teliospores with densely verrucose ornamentation. Scale bar =10 um.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Deosai Plains, 3300 m a.s.1., on

leaves of Persicaria orientalis (L.) Spach (Polygonaceae), with II + III stages, 10 Sep 2011,

coll. Abdul Nasir Khalid, det. Malka Saba 15 (LAH).

COMMENTS: Puccinia oniwaensis has been reported on Polygonum sibiricum

Laxm. from Astore, Rama Lake (Baltistan, Northern Areas) by Khalid et al.

(1995). Persicaria orientalis is a new host and P. oniwaensis is a new report for

Deosai Plains.

Puccinia rhytismoides Johanson, Bot. Not.: 173 (1886). Fic. 4

SPERMOGONIA, AECIA and UREDINIA not seen. TELIA hypophyllous,

compact, subepidermal, loculate, surrounded by cylindrical paraphyses, 60-80

x 100-200 um. TELIosPoregs clavate, ellipsoid or oblong, 10-14 x 26-58 um

(mean 11.8 x 40.3 um), pale brown or brown at apex, cinnamon brown at base,

constricted at septum, lower cell longer than apical cell, attenuated at base;

152 ... Saba & Khalid

Fic. 4. Puccinia rhytismoides. Lucida drawings showing 1-3 celled teliospores.

Scale bar =15 um.

wall smooth, hyaline to pale brown, 0.5-1 um thick; apex conical, truncated

or rounded, 3-6 um thick; pedicel dark brown, persistent, 4-7 x 3-13 um.

Mesospores common, pale brown, clavate or oblong, 8-10 x 18-30 um; wall

up to 1 um thick, smooth; apex conical or rounded 4-6 um; pedicel short, dark

brown. Three-celled teliospores rare.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Deosai Plains, on leaves of

Thalictrum alpinum L. (Ranunculaceae), with III stage, 10 Sep 2011, coll. Abdul Nasir

Khalid, det. Malka Saba 16 (LAH). FINLAND, KiLpisjARVI, on leaves of T: alpinum, 12

Jul 1935, coll. & det. J.I. Liro (ZT Myc s. n.). NORWAY, TRONDELAG, near Kongsvold,

Doorefjeld, on leaves of T: alpinum, 1925, leg. A.G. Eliasson (ZT Myc s.n.).

ComMENTs: Puccinia rhytismoides, which represents a new record for the

Indian subcontinent, has previously been reported on Thalictrum alpinum from

Alaska, Finland, Iceland, Norway, Russia, and Sweden and on T! alpinum var.

stipitatum Y. Yabe and T: tuberiferum Maxim. in Japan (Farr & Rossman 2011).

However Hiratsuka et al. (1992) placed P. rhytismoides in synonymy under

P. ustalis Berk., which Ahmad et al. (1997) from Pakistan reported on Anemone

obtusiloba D. Don.

Chrysomyxa & Puccinia spp. new to Pakistan ... 153

Fic. 5. Puccinia swertiae (Lucida drawings).

(A) Peridial cells showing densely verrucose ornamentation.

(B) Aeciospores with minutely verrucose ornamentation. Scale bar =10 um.

Puccinia swertiae (Opiz) G. Winter, Rabenh. Krypt.-Fl., Ed. 2, 1(1): 205 (1881)

[“1884”]. Fig. 5

SPERMOGONIA and UREDINIA not seen. AECIA on abaxial side, cupulate,

yellowish, gregarious, 200-300 x 250-450 um. Peridial cells yellowish brown,

rhomboidal to oblong or irregular, 17.7-39 x 22-53 um (mean 35.3-37.9 um);

wall densely verrucose, 2.8-8.5 um. AEcIOsPORES obovoid or subglobose,

hyaline to brown, 19.6-25.2 x 20.5-27(-36) um (mean 21.8 x 24.4 um); wall

finely verrucose, 0.5-1.2 um. TELIA abaxial, black, pulverulent, covered by

ruptured epidermis, scattered, naked, 100-200 x 200-800 um. TELIOSPORES

obovoid or ellipsoid, rounded at both ends, not or hardly constricted at septum,

dark brown to chestnut brown, 25-30.4 x 31-39 um (mean 27 x 35 um); wall

verrucose, 1.8-3.5(-4) um; germ pores two, next to septum in both upper and

lower cells, rarely apical in upper cell; apex dark brown to chestnut brown,

not or slightly thickened, 1.2-2 um; pedicel short, hyaline, persistent, 4-9.4 x

2.6-8.5 um.

MATERIAL EXAMINED: PAKISTAN, NORTHERN AREAS, Deosai Plains, on leaves of

Swertia petiolata D. Don (Gentianaceae), with I + III stages, 10 Sep 2011, coll. Abdul

Nasir Khalid, det. Malka Saba 17 (LAH).

ComMENTSs: Puccinia swertiae has been reported on Swertia petiolata and

S. speciosa Wall. from Kalam (Swat, Khyber Pakhtunkhwa), Sargodha, and

Shogran (Kaghan Valley, Khyber Pakhtunkhwa) (Ahmad 1956a,b; Jorstad &

Iqbal 1967; Ono & Kakishima 1992; Ono 1992). Puccinia swertiae represents a

new record for Deosai Plains.

154 ... Saba & Khalid

Acknowledgments

We are highly obliged to the Pakistan Science Foundation, Islamabad, for funding

this research. We are sincerely thankful to Dr. Amy Rossman (Systematic Mycology

and Microbiology Laboratory, USDA-ARS, Beltsville) and Dr. Omar Paino Perdomo

(Dominican Society of Mycology, Santo Domingo, Dominican Republic) for acting

as presubmission reviewer. We are cordially thankful to Dr. Shaun R. Pennycook and

Dr. Lorelei L. Norvell for their critical reviews to improve the manuscript.

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, Khalid AN, Iqbal SH, Niazi AR, Sultan A. 2009. Puccinia subepidermalis sp. nov. and

new records of rust fungi from Fairy Meadows, Northern Pakistan. Mycotaxon 110: 173-182.

http://dx.doi.org/10.5248/110.173

Afshan NS, Khalid AN, Iqbal SH, Niazi AR, Sultan A. 2010. Puccinia anaphalidis-virgatae, a new

species, and a new variety of rust fungi from Fairy Meadows, Northern Pakistan. Mycotaxon

110: 173-182. http://dx.doi.org/10.5248/112.483

Afshan NS, Khalid AN, Niazi AR, Iqbal SH. 2011. New records of Uredinales from Fairy Meadows,

Pakistan. Mycotaxon 115: 203-213. http://dx.doi.org/10.5248/115.203

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 §, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Nabiza Printing Press, Karachi, Pakistan.

Dade HA, Gunnell J. 1969. Class work with fungi. Commonwealth Mycological Institute Kew.

Farr DF, Rossman AY. 2011. Fungal Databases, Systematic Mycology and Microbiology Laboratory,

ARS, USDA. http://nt.ars-grin.gov/fungaldatabases/

Gjaerum HB, Iqbal SH. 1969. Some rust fungi from West Pakistan. Nytt. Mag. Bot. 16: 221-223.

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y,

Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba Shuppankai, Ibaraki.

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, the Northern Areas of Pakistan. Mycotaxon 109: 1-7.

http://dx.doi.org/10.5248/109.1

Ishaq A, Afshan NS, Khalid AN. 2011. New records of poaceous rust from Pakistan. Mycotaxon

118: 237-244. http://dx.doi.org/10.5248/118.237

Jorstad I, Iqbal SH. 1967. Uredinales from West Pakistan. Nytt. Mag. Bot. 14: 31-38.

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.

Kaneko S. 1993. Parasitic fungi on woody plants from Pakistan. Cryptogamic Flora of Pakistan,

Vol. 2 (Eds. T. Nakaike and S. Malik), Nat. Sci. Mus., Tokyo, Japan. pp. 149-168.

Khalid AN, Iqbal SH. 1996. Puccinia eriocyclae sp. nov. on Eriocycla stewartii from Northern

Pakistan. Mycoscience 37: 437-438. http://dx.doi.org/10.1007/BF02461000

Khalid AN, Iqbal SH. 1997. Puccinia deosaiensis sp. nov. on Epipactis helleborine (L.) Crantz. Can.

J. Bot. 75(5): 864-866. http://dx.doi.org/10.1139/b97-095

Chrysomyxa & Puccinia spp. new to Pakistan ... 155

Khalid AN, Saba M. 2011. Puccinia cortusae (Basidiomycota; Uredinales) on Cortusa brotheri

(Primulaceae), new to southern Asia (Fairy Meadows, Pakistan). Mycotaxon 117: 317-320.

http://dx.doi.org/10.5248/117.317

Khalid AN, Iqbal SH, Masood A. 1995. New records of Uredinales from Pakistan. Sci. Int. (Lahore)

7(4): 531-532.

Ono Y. 1992. Uredinales collected in the Kaghan Valley, Pakistan. Cryptogamic flora of Pakistan

1: 217-240.

Ono Y, Kakishima M. 1992. Uredinales collected in the Swat Valley, Pakistan. Cryptogamic flora of

Pakistan 1: 197-216.

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MY COTAXON

http://dx.doi.org/10.5248/123.157

Volume 123, pp. 157-162 January-March 2013

New records of lichens from Chile

XIN Yu WANG*?, IRIS PEREIRA*, SOON-OK OH;?,

L1 SONG WANG?! & JAE-SEOUN Hur?

' Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany,

Chinese Academy of Sciences, Yunnan 650204, China

? Instituto Biologia Vegetal y Biotecnologia, Universidad de Talca, 2 Norte 685, Talca, Chile

° Korean Lichen Research Institute, Sunchon National University,

Sunchon 540-742, South Korea

* CORRESPONDENCE TO: jshur1@sunchon.ac.kr

ABSTRACT — After fieldwork in central Chile, five lichen species were established as new for

the country: Bryoria lanestris, Buellia disciformis, Haematomma sorediatum, Lecidea capensis,

and Punctelia rudecta. They were discovered in two national parks in the Maule Region

and the area surrounding the Termas de Chillan, Bio-Bio Region. Each species is briefly

described, with habit illustrations and comments on chemistry, habitat, and morphology.

These collections suggest a high potential for finding more unreported lichen species in

Chile’s national parks.

Key worps — lichenized fungi, South America, floristic survey, taxonomy

Introduction

In Chile, the climate varies from arid desert in the north, through humid

subtropical in Easter Island, to near polar in the south, extending across 38

degrees of latitude. The five species reported here were collected in two national

parks with a Mediterranean climate in the central part of Chile. About 1400

lichen species have been previously reported from Chile (Galloway & Quilhot

1998). Our new findings cover only a very small part of the country, but indicate

that as it has such a wide range of different climates, many more species may

still be discovered, and the total number of lichen species occurring in Chile is

likely to be far higher than currently reported.

Materials & methods

The 405 specimens sampled during the current collections are deposited in the

Korean Lichen Research Institute, Sunchon National University, South Korea (KoLRI).

158 ... Wang & al.

The specimens were examined by standard microscopical techniques, hand-

sectioned under a Nikon SMZ 645 dissecting microscope. Anatomical descriptions were

based on observations under a Nikon Eclipse E200 microscope. Secondary metabolites

were identified by TLC according to Elix et al. (1987), Orange et al. (2001), and White

& James (1985). Medullas were tested with IKI (10% aqueous potassium iodide) for

identification.

Results

Bryoria lanestris (Ach.) Brodo & D. Hawksw. FIG. 1A, B

Thallus fruticose to subpendent, attached to the substrate by one holdfast,

surface pale olive brown to pale brown, smooth, 0.2-0.4 mm thick, 1-5 cm

long, main branch usually isotomic, with few side branches, surface with white

soralia, growing from the fissures, without pseudocyphellae.

CuHemistry: Thallus P+ red or P-, medulla P-, soralia C-, K-, P+ red,

containing fumarprotocetraric acid (by TLC).

Hasirat: On the bark of Nothofagus sp. Mainly distributed in the northern

boreal zone (Brodo & Hawksworth 1977).

SPECIMEN EXAMINED: CHILE, MAuLe REGION, TALcA PROVINCE, National Reserve

“Altos de Lircay’, 35°36'08.2"S 71°02'56.7"W, alt. 1367 m a.s.L, on bark, 26 January 2012,

J.-S. Hur CL-120099-1 (KoLRI 014265).

REMARKS: The only Bryoria species previously reported in Chile is B.

chalybeiformis (Galloway & Quilhot 1998), which grows on rock or soil,

without soralia but with spinulose branches, its color is much darker, and it is

shiny. Bryoria simplicior, which might also be confused with B. lanestris, can be

distinguished by its roundish and greenish black soralia that are broader than

the branch and by its lack of compounds.

Buellia disciformis (Fr.) Mudd FIG. 1C, D

Thallus crustose, thin and rimose, immersed or superficial, surface pale

white to whitish green, prothallus present when meeting with other thalli,

black. Apothecia lecideine, disc black and plane, without pruina, with smooth

margin, 0.2-0.4 mm in diam., spores 8 per ascus, brown, 1-septate, 14-21 x

6-9 um.

CuHeEMiIstRy: Thallus K+ yellow, containing atranorin (by TLC).

Hasitat: On bark, mainly distributed in temperate areas (Bungartz et al.

2007)

SPECIMEN EXAMINED: CHILE, Maule REGION, CAUQUENES PROVINCE, National

Reserve “Los Ruiles’, 35°49'49.3"S 72°30'25.5"W, alt. 342 m a.s.l., on bark, 28 January

2012, J.-S. Hur CL-120240 (KoLRI 014407).

REMARKS: Buellia disciformis might be confused with some Lecidella species,

which also grow on bark and are morphologically quite similar, but it can be

distinguished by its brown septate spores and K+ yellow reaction (atranorin).

Lichens new to Chile ... 159

Fic. 1 Habit of lichens new to Chile. A-B. Bryoria lanestris.

C-D. Buellia disciformis. E-F. Haematomma sorediatum.

Buellia chloroleuca could be easily confused with B. disciformis, but its distinctly

C+ orange (xanthone) reaction readily distinguishes it.

Haematomma sorediatum R.W. Rogers FIG. 1E, F

Thallus crustose, white to greenish gray, granulose, soralia present, scattered

on the surface of the thallus or on the margin of apothecia, orbicular, white.

Apothecia lecanorine, up to 1 mm in diam., disc bright red, flat, epruinose,

margin persistent, crenulate, spores 8 per ascus, hyaline, 7-12 septate, straight

fusiform, 40-70 x 5-7 um.

160 ... Wang & al.

CuHEMIstTry: Thallus and soralia K+ yellow, C-, KC+ yellow, P+ pale yellow,

containing atranorin and placodialic acid (by TLC).

Hapsirat: On bark. It has cosmopolitan distribution (Brodo et al. 2008).

SPECIMEN EXAMINED: CHILE, MAuLE REGION, CAUQUENES PROVINCE, National

Reserve “Los Ruiles”, 35°50'02.4"S 72°'30'35.3"W, alt. 175 m a.s.l., 28 January 2012, J.-S.

Hur CL-120158 (KoLRI 014324).

REMARKS: The only Haematomma species from Chile can easily be separated

from other species by its roundish soralia on the apothecial surface and margin

and a red apothecial disc that is sometimes covered with pruina. Staiger & Kalb

(1995) treated H. neglectum as a synonym of H. sorediatum, but Brodo et al.

(2008) still had doubts about this. As the two species need more study, we use

the name H. sorediatum here in the strict sense.

Lecidea capensis Zahlbr. FIG. 2A, B

Thallus squamulose to areolate, reddish brown to brown, smooth, convex,

usually shiny, marginal part white, prothallus black, medulla I-. Apothecia

black, round and subimmersed, pruinose, 0.5-1.3 mm in diam., hymenium

40-60 um high, hyaline, epihymenium greenish black, spores ellipsoid, 10-14

x 3-4 um.

CuHEMIsTrY: Thallus and medulla all negative in spot reaction, containing

2’-O-methylperlatolic acid (by TLC).

Hasirat: On rock, known from coastal forest and South Africa and Australia

(Galloway 2007).

SPECIMEN EXAMINED: CHILE, Bio-Bio REGION, CHILLAN PROVINCE, Vicinity of

Termas de Chillan, 36°54'21.6"S 71°24'05.5"W, alt. 1980 m a.s.l., 3 February 2012, J.-S.

Hur CL-120360 (KoLRI 014529).

REMARKS: The species is characterized by saxicolous habit and a convex

shiny thallus that is areolate and pale brown to reddish brown. It is similar to

L. fuscoatrula, which differs in its I+ violet medulla reaction, smaller (<10 um)

ascospores, dark brown hymenium, and 2’-O-methylmicrophyllinic acid as the

main compound.

Punctelia rudecta (Ach.) Krog FIG. 2C, D

Thallus foliose, greenish gray, lobes plane and linear, 3-7 mm wide, covered

with white pseudocyphellae, irregular in shape, isidia mostly on the central part

of the thallus, simple or branched, sometimes lobule-like, brown-tipped, lower

surface pale brown, with light brown rhizines, simple or forked. Apothecia not

seen.

CuHEMistry: Thallus K+ yellow, C-, medulla K-, C+ red, containing

atranorin and lecanoric acid (by TLC).

Hasitat: On rock; common in North America, Europe, Australia, and Asia

(Egan & Aptroot, 2004)

Lichens new to Chile... 161

Fic. 2 Habit of lichens new to Chile. A~-B. Lecidea capensis. C-D. Punctelia rudecta.

SPECIMEN EXAMINED: CHILE, MAULE REGION, CAUQUENES PROVINCE, National

Reserve “Los Ruiles’, 35°50'02.4"S 72°30'35.3"W, alt. 175 m a.s.l., 28 January 2012, J.-S.

Hur CL-120142 (KoLRI 014308).

REMARKS: This is the only Punctelia species with isidia reported from Chile. It

is similar to P. borreri, which has soredia on the lobe margins instead of isidia.

As all other Punctelia species reported in Chile before have soralia, they can

easily be separated from P. rudecta. It is common and widespread in eastern

and southeastern North America (Brodo et al. 2001).

Acknowledgments

This work was supported by: Instituto Biologia Vegetal y Biotecnologia and Instituto

de Quimica de Recursos Naturales, Universidad de Talca, Chile; the National Research

Foundation of Korea (No. 2011-0030491); and the foundation of Key Laboratory of

Biodiversity and Biogeography, Kunming Institute of Botany, CAS (KLBB-201210)

and National Natural Science Foundation of China (No. 31170023, KSCX2—EW-Z-9,

Y3216111W1).

Literature cited

Brodo IM, Hawksworth DL. 1977. Alectoria and allied genera in North America. Opera Botanica

42: 1-164.

Brodo IM, Sharnoff D, Sharnoff S. 2001. Lichens of North America. New Haven: Yale University

Press: 795:p.

162 ... Wang & al.

Brodo IM, Culberson WL, Culberson CF. 2008. Haematomma (Lecanoraceae) in North

and Central America, including the West Indies. The Bryologist 111(3): 363-423.

http://dx.doi.org/10.1639/0007-2745(2008)111[363:HLINAC]2.0.CO;2

Bungartz F, Nordin A, Grube M. 2007: Buellia. 113-179, in: TH Nash III et al. (eds). Lichen Flora

of the Greater Sonoran Desert Region. Volume 3. Lichens Unlimited, Arizona State University,

Tempe..

Egan RS, Aptroot A. 2004. Punctelia. 431-436, in: TH Nash III et al. (eds). Lichen Flora of the

Greater Sonoran Desert Region, Vol. 2. Lichens Unlimited, Arizona State University, Tempe,

Arizona.

Elix JA, Johnston J, Parker JL. 1987. A catalogue of standard thin layer chromatographic data and

biosynthetic relationships for lichen substances. Canberra: Australian National University.

Galloway DJ. 2007. Lecidea Ach. 1803. 740-787, in: Flora of New Zealand. Lichens. Revised Second

Edition including Lichen-Forming and Lichenicolous Fungi. Vol. 1. Manaaki Whenua Press,

Lincoln, New Zealand.

Galloway DJ, Quilhot W. 1998. Checklist of Chilean lichen-forming and lichenicolous fungi.

Gayana Botanica 55(2): 111-185.

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

London: British Lichen Society.

Staiger B, Kalb K. 1995. Haematomma-Studien. I. Die Flechtengattung Haematomma. Bibliotheca

Lichenologica 59: 1-198.

White FJ, James PW. 1985. A revised guide to the microchemical techniques for the identification

of lichen substances. British Lichen Society Bulletin 57: 1-41.

MY COTAXON

http://dx.doi.org/10.5248/123.163

Volume 123, pp. 163-167 January-March 2013

Endocarpon maritima sp. nov. (lichenized Ascomycota)

from the maritime region of South Korea

YOGESH JOSHI! & JAE-SEOUN HurR”*

"Department of Botany, Kumaun University, S.S.J. Campus,

Almora 263601, Uttarakhand, India

*Korean Lichen Research Institute, Sunchon National University,

Sunchon 540-742, South Korea

*CORRESPONDENCE TO: jshur1@sunchon.ac.kr

ABSTRACT — A new lichenized fungus, Endocarpon maritima, was collected from the rocky

seashores of Bogil Island, South Korea. ‘The species is characterized by a squamulose to lobate

brown to olive-green thallus attached to the substratum by rhizohyphae, blackish hypothallus,

2-spored asci, spores 25-37.5(-50) x (10.5-)13-17(-20) um, and a maritime distribution.

Lichenized species of Aspicilia, Buellia, Caloplaca, Phaeophyscia, Physcia, and Pterygiopsis

occur in close association with E. maritima.

KEY worps — pyrenocarpous lichen, taxonomy, Verrucariaceae

Introduction

The cosmopolitan pyrenocarpous squamulose to subfoliose lichen genus

Endocarpon Hedw. is represented by ca. 50 species (Kirk et al. 2008) and

occupies a wide range of habitats, such as terrestrial, maritime, or fresh water,

depending on the species. The genus has been well worked out in India (Singh

& Upreti 1984), Australia (McCarthy 1991), New Zealand (Galloway 2007),

Japan (Harada 1993), Mexico (Breuss 2000), the Greater Sonoran Desert

(Breuss 2002), Costa Rica (Aptroot et al. 2008), and Great Britain (Orange

2008, Giavarini & Fox 2009). However, the genus is not well researched from

South Korea and so far is represented by only 4 species (Joshi et al. 2009, Moon

& Aptroot 2009).

The maritime Endocarpon species comprise important elements in the

intertidal and splash zones at the boundary of terrestrial biota, frequently

forming with Verrucaria a blackish zone. Extensive field surveys of the maritime

regions conducted by one author (YJ) during his stay in South Korea revealed

an Endocarpon species new to science, described here.

164 ... Joshi & Hur

Materials & methods

Studied specimens are deposited in KoLRI (Korean Lichen Research Institute,

Sunchon, South Korea) and CBM (Natural History Museum and Institute, Chiba,

Japan). Description and photographs of external morphology are based on air-dried

material observed under a dissecting stereomicroscope (Nikon SMZ 645). Sections

made by a razor blade under the stereomicroscope were mounted in lactophenol cotton

blue. Anatomical descriptions are based on these preparations studied by a compound

microscope (Nikon Eclipse E200). Measurements of ascospores were made at x400

magnification mounted in water; only free ascospores lying outside the asci were

measured. Secondary metabolites were identified by TLC as described by White & James

(1985) and Orange et al. (2001) using solvent system C. The mycological terminology

follows Kirk et al. (2008).

New species

Endocarpon maritima Y. Joshi & Hur, sp. nov. Fic. 1

MycoBank MB 801275

Similis Endocarpon ramulosum, E. neopallidulum et E. japonicum, sed differt in

maritimum habitio und hypothallus praesens.

Type: South Korea. Jeollanam-do: Bogil-myeon, Bogil Island, Tong-ri, near Tongri

Beach, 34°09'68.4"N 126°35'15.3"E, alt. ca 3 ma.s.l., on exposed rocks, 6 February 2010,

Y. Joshi, H. S. Jeon, M. H. Jeong 100197 (holotype, KoLRI; isotype CBM).

EryMoLocy — ‘The specific epithet refers to its occurrence in the maritime region of

South Korea.

THALLUS squamulose to lobate, brownish to olive-green, 120-130 um thick,

composed of loosely aggregated squamules, squamules usually lobate at

maturity; hypothallus +present, black, extending beyond the squamules or lobes.

SQUAMULES usually adnate or apparently ascending near the margin. LOBULES

0.2-0.6 x 0.2-0.4 mm, sparsely or repeatedly, irregularly branched, frequently

imbricate, discrete to +contiguous, margins entire, +recurved upwards at

margin; shape of lobules regular; apically rounded, +darker than rest of the

thallus. Upper surface brown to +pale brown with olive tinge, slightly glossy,

smooth, flat to slightly convex. Lower surface almost black around attaching

parts, with sparse rhizohyphae, lacking rhizines. Squamules attached to the

substratum at basal ends or central part. UPPER CORTEX paraplectenchymatous,

12.5-20 um thick, +pale brown in the uppermost part, hyaline in the remainder.

MEDULLA white, composed of spherical hyphal cells, 20-35 um thick. ALGAL

LAYER 50 um thick, photobiont solitary, in vertical rows in the upper part of

layer. LOwER CORTEX undifferentiated from the medulla, dark brown to almost

black, 25 um thick.

PERITHECIA numerous, 1-4 per squamule/lobule, black, laminal, immersed,

globose to somewhat depressed, generally a small black area around ostiole

is emerging like a prominent protuberance, ostioles like pale to dark dots or

Endocarpon maritima sp. nov. (South Korea) ... 165

>-=

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ifs TA

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FiGuRE 1: Endocarpon maritima (habit). Scale bar = 5 mm.

indistinct, perithecia pyriform in cross sections, (220-)230 x 250(-270) um.

ExcIPLe dark brown to almost black, 50 um thick. SUBHYMENIUM 25 um thick.

PERIPHYSES 20-50 um long. HYMENIUM 70-87.5 x 187.5-237.5 um, hymenial

algae spherical, 5-7.5 um across. Asci clavate, 75-100 x 20-25 um, 2-spored.

Ascosporss hyaline to very pale brown, muriform, ellipsoidal to bacilliform,

25-37.5(-50) x (10.5-)13-17(-20) um. Pycnip1a and conidia not seen.

CHEMISTRY — Spot test reactions: thallus K-, C-, P-. TLC: no lichen

substances detected.

ECOLOGY & DISTRIBUTION — The new taxon is thus far confined to the

maritime region of South Korea, where it grows luxuriantly over non-calcareous

(limestone) rocks in close association with Aspicilia caesiocinerea (Nyl. ex

Malbr.) Arnold, Buellia badia (Fr.) A. Massal., Caloplaca bogilana Y. Joshi

& Hur, C. cinnabarina (Ach.) Zahlbr., Phaeophyscia hirtuosa (Kremp.) Essl.,

Physcia stellaris (L.) Nyl., and Pterygiopsis affinis (A. Massal.) Henssen. This is a

second new species from Bogil Island (the other being Caloplaca bogilana) and

is so far known only from this island.

REMARKS — ‘The new taxon is characterized by a squamulose to lobate, brown

to olive-green thallus attached to substratum via rhizohyphae, +blackish

hypothallus, 2-spored asci, and maritime distribution. Endocarpon ramulosum

166 ... Joshi & Hur

H. Harada and E. neopallidulum H. Harada from Japan differ in lacking a

hypothallus and being strictly inland species growing between elevations of

200-900 m and 100-150 m, respectively. Endocarpon japonicum H. Harada,

another closely related species, differs in its different substrate specificity (growth

at the edge of streams on partially submerged rocks), absence of a hypothallus,

and its inland distribution between elevations of 50-700 m. In its lobate

form, E. petrolepideum (Nyl.) Nyl. would not be confused with E. maritima,

because its squamules are mono-lobed and generally scattered to adjacent

but not overlapping. Endocarpon nigromarginatum H. Harada resembles

E. maritima in having squamules with conspicuous black borders (i.e. margins)

but differs in having both rhizines and rhizohyphae on the lower surface and a

+pachydermatous upper cortex. The presence of a black hypothallus confuses

the new taxon with Endocarpon nigrozonatum Ajay Singh & Upreti, which

differs in having an inland distribution and strictly squamulose morphology.

Acknowledgments

This work was supported by a grant from the Korea National Research Resource

Center Program, and the Korean Forest Service Program (KNA 2012) through Korea

National Arboretum. One of the authors (YJ) is thankful to Head, Dept. of Botany,

Kumaun University, S.S.J. Campus, Almora for providing laboratory facilities to work.

The authors are thankful to Drs Laszlo L6k6és and T.A.M. Jagadeesh Ram for reviewing

the manuscript and providing valuable comments.

Literature cited

Aptroot A, Licking R, Sipman HJM, Umafia L, Chaves JL. 2008. Pyrenocarpous lichens with

bitunicate asci: a first assessment of the lichen biodiversity inventory of Costa Rica. Biblioth.

Lichenol. 97: 1-162.

Breuss O. 2000. Eine ungewohnliche Endocarpon-Art (Lichenisierte Ascomyceten, Verrucariales)

aus Mexiko. Osterr. Z. Pilzk. 9: 147-149.

Breuss O. 2002. Endocarpon. 181-187, in TH Nash III et al. (eds). Lichen flora of the Greater

Sonoran Desert Region. Vol. I. Lichens Unlimited Tempe, AZ.

Galloway DJ. 2007. Flora of New Zealand. Lichens, 2nd ed. Manaaki Whenua Press, Lincoln, NZ.

Giavarini V, Fox BW. 2009. Endocarpon. 385-386, in CW Smith et al. (eds). The lichens of Great

Britain and Ireland. British Lichen Society, Natural History Museum Publications, London.

Harada H. 1993. A taxonomic study of the lichen genus Endocarpon (Verrucariaceae) in Japan.

Nova Hedwigia 56: 335-353.

Joshi Y, Wang XY, Lee YM, Byun B-K, Koh YJ, Hur J-S. 2009. Notes on some new records of macro-

and micro-lichens from Korea. Mycobiology 37(3): 197-202.

http://dx.doi.org/10.4489/MYCO.2009.37.3.197

Kirk PM, Cannon PF, Minter DW, Stalpers JA. (eds.) 2008. Dictionary of the fungi, 10th edition.

CABI Publishing, Wallingford, UK.

McCarthy PM. 1991. The lichen genus Endocarpon Hedwig in Australia. Lichenologist 23: 27-52.

Moon KH, Aptroot A. 2009. Pyrenocarpous lichens in Korea. Biblioth. Lichenol. 99: 297-314.

Orange A. 2008. British pyrenocarpous lichens. [(viewed online on 07 July 2012)

http://www.thebls.org.uk/content/documents/British_Pyrenocarpous_Lichens.zip].

Endocarpon maritima sp. nov. (South Korea) ... 167

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society, London.

Singh A, Upreti DK. 1984. The lichen genus Endocarpon from India. Candollea 39: 539-548.

White FJ, James PW. 1985. A revised guide to microchemical techniques for the identification of

lichen substances. British Lichen Society Bulletin 57(supplement): 1-41.

MY COTAXON

http://dx.doi.org/10.5248/123.169

Volume 123, pp. 169-181 January-March 2013

Two striking Inocybe species from Yunnan Province, China

YU-GUANG FAN & TOLGOR BAuU*

Institute of Mycology, Jilin Agricultural University, Changchun 130118, PB. R. China

* CORRESPONDENCE TO: junwusuo@126.com

ABSTRACT —Two striking species, Inocybe caroticolor sp. nov. and I. olivaceonigra, are

documented in this paper. Inocybe caroticoloris characterized by its carrot-colored basidiomata,

entirely pruinose stipe, aromatic odor, and nodulose basidiospores. Inocybe olivaceonigra,

originally described from Papua New Guinea, is new to China. Illustrated descriptions of

the two species are provided based on the Chinese collections. DNA sequences of internal

transcribed spacer regions (ITS) of the Chinese materials were generated, analyzed, and

submitted to the GenBank.

Key worps — Agaricales, Inocybaceae, taxonomy

Introduction

Inocybe (Fr.) Fr. is a large agaric genus, which was established as a “tribe” of

Agaricus by Fries in 1821 and elevated to genus rank in 1863. Currently, there

are about 500 (Kirk et al. 2008) to 700 (Matheny et al. 2009) species of Inocybe

recognized worldwide. In the last few years, new species of Inocybe continue

to be described from around the world (Matheny & Bougher 2005, Kobayashi

2005, 2009; Jacobsson & Larsson 2009, Vauras & Kokkonen 2009, Kobayashi &

Onishi 2010, Kropp & Albee-Scott 2010, Kropp et al. 2010, Kokkonen & Vauras

2011, Vauras & Larsson 2012).

In East Asia, more than 140 species of Inocybe have been recorded in Japan

(Kobayashi 2002, 2005, 2009). This number is still increasing (Kobayashi &

Onishi 2010). At present, there are ninety-eight names of Inocybe reported

from China (Fan & Bau 2010). Unfortunately, many of the names recorded

in China lack detailed descriptions or information on voucher specimens.

Although a few studies on species diversity of Inocybe have been undertaken

in some provinces (Liu 1987, Zheng et al. 1985), rather few species have been

originally described as new from China (Wang 1973). Monographic studies on

species diversity of Inocybe in China are needed.

During taxonomic studies on Inocybaceae from China, we encountered two

impressive species easy to recognize by their conspicuous colors in the field.

170 ... Fan & Bau

Inocybe caroticolor is proposed as new based on comparative studies of available

collections. Another species is found to be conspecific with I. olivaceonigra after

examining the holotype. It is re-described based on Chinese collections. DNA

sequences of internal transcribed spacer regions (ITS) of the Chinese materials

were generated and analyzed.

Materials & methods

Materials were collected in Castanopsis forests with a subtropical highland climate

at 1900 m elevation in Yunnan Province (25°03'N 102°37'E and 25°08'N 102°44'E), and

examined specimens were deposited in the Herbarium of Mycology, Jilin Agricultural

University (HMJAU). Additional Chinese collections from KUN (with HKAS numbers)

and authentic specimens of I. lutea Kobayasi & Hongo from TNS and I. olivaceonigra

and “Astrosporina lutea” from Z+ZT were examined (Herbarium abbreviations follow

Thiers (2012)). Macroscopic characters were described based on field notes. Microscopic

features were made from slide preparations mounted in 5% KOH solution and Congo

Red. Measurements of microscopic features were made at 1000x on an Nikon 80i

microscope. Color notations follow Kornerup & Wanscher (1981). Measurement

methods follow Kobayashi (2002, 2009).

Genomic DNA was extracted from silica-dried materials with a modified CTAB

protocol (Doyle & Doyle 1987). ITS gene fragments were amplified with primers ITS1F

and ITS4B (Gardes & Bruns 1993). Sequencing was performed with an ABI 3730 DNA

analyzer and an ABI BigDye 3.1 terminator cycle sequencing kit (Shanghai Sangon

Biological Engineering Technology & Services Co. Ltd, Shanghai, China). Five new

sequences (JX025772-JX025776) have been deposited at GenBank. The new sequences

were used as queries of the NCBI nucleotide sequence database using the BLASTn

search algorithm.

Taxonomy

Inocybe caroticolor T. Bau & Y.G. Fan sp. nov. FIGS 1, 2

MycoBank: MB 564971

Differs from Inocybe lutea by its hymenial cystidia with thicker walls, its stipe covered

uniformly by caulocystidia, and its aromatic odor.

Type: China, Yunnan Province, Kunming, Heilongtan Park, under Quercus variabilis

Blume, 26 Aug 2010, Y.G. Fan 2010126 (Holotype, HMJAU 23271; GenBank

JX025774).

EtyMo.oay: Refers to the carrot color of the basidioma.

BASIDIOMA small. PrL—Eus 17-33 mm in diam, conical-convex when young,

then plano-convex, dry, with a prominent obtuse umbo, covered with appressed

squamules, radially arranged, fibrillose, rimulose to usually rimose at margin,

margin inrolled when young, occasionally recurved when mature, orange (5A7-

8) to apricot (4A7-8), squamules concolorous when young, then turning brown

(5C8) to reddish brown (6C8), background orange-apricot (4A8) to ochraceous

(3A6). LAMELLAE adnexed, moderately crowded, up to 3 mm in width, pale

Inocybe caroticolor sp. nov. (China) ... 171

Fic. 1. Inocybe caroticolor basidiomata (HMJAU 24614).

orange to apricot (4A7-8) when young, dirty apricot (4B7) to brown (5B7)

when mature, gill edge concolorous or paler, not smooth. Stripe 30-42 x 2-3

mm, solid, pale orange (5A6) to apricot (4A7), equal with subbulbous to non-

marginate bulbous base, occasionally base distinctly wider, entirely pruinose

on the stipe surface, densely pruinose at upper part, longitudinally striate

downwards, stipe base with whitish tomentum. CorTINA absent. CONTEXT

with conspicuous aromatic odor, fleshy in pileus, whitish to pale apricot (4A5),

1-3 mm in width, striate in stipe, pale apricot (4A5).

BASIDIOSPORES 6.3-9.3(-9.5) x (4.4-)4.7-6.0(-6.2) um, Q = (1.43-)1.47-

1.58(-1.61), weakly to prominently nodulose (with 7-9 nodules), yellowish

brown. Basip1a 24-37 x 5-9 um, with 4 sterigmata, occasionally with 1 or 2

sterigmata, clavate, narrower downwards, at times slightly enlarged at the base,

usually with yellowish pigments. PLEUROCYSTIDIA 48-63 x 12-16 um, fusiform

with obtuse apices, occasionally utriform, crystalliferous at apex, base obtuse

or tapering into pedicel, walls bright yellow, 2-3(-4) um thick, hyaline inside,

rarely with yellow pigments. CHEILOCYSTIDIA similar to pleurocystidia, more

variable, occasionally cylindric; PARACysTIDIA abundant among cheilocystidia,

clavate to obovoid, thin-walled, hyaline, mixed with basidia. CAULOCYSTIDIA

descending to stipe base, 48-79 x 9-14 um, abundant, similar to pleurocystidia,

but more variable in shape, usually with slender habit and thinner walls

(0.8-2.4 um); CAULOPARACYSTIDIA abundant, thin-walled, fusoid to lageniform,

at times apex tapered, mixed with filamentous hyphae. PILEIPELLIs a cutis,

172 ... Fan & Bau

composed of cylindric, smooth hyphae 2.0-7.0 um wide, regularly arranged,

with oil-yellow to dark-yellow intra-hyphal pigments in upper layer, almost

hyaline in lower layer. CLAMP CONNECTIONS present in all tissues, but not at

every septum.

HABITAT AND ECOLOGY: Singly in groups, common in roadsides near forests,

all known collections under Quercus variabilis except for one (HKAS 38963)

under Pinus yunnanensis. June to August in Yunnan, China.

ADDITIONAL SPECIMENS EXAMINED: Inocybe caroticolor — CHINA, YUNNAN PROv.,

KUNMING: Kunming Botanical Garden, alt. 1900 m, under Quercus variabilis, 24 Jul

2011, Y.G. Fan 2011123 (HMJAU 24614; GenBank JX025773); 27 Aug 2010, Y.G. Fan

2010126c (HMJAU 24621); 27 Aug 2010, Y.G. Fan 2010126d (HMJAU 24622); 28 Aug

2010, Y.G. Fan 2010126e (HMJAU 24623); 26 Jun 2000, X.H. Wang 977 (HKAS 36740);

Qiongzhu temple, alt. 2100m, under Quercus variabilis, 26 Jul 2011 Y.G. Fan 2011139

(HMJAU 24615; GenBank JX025772); Wupine: Shizishan, under Pinus yunnanensis

Franch., 18 Aug 2000, EQ. Yu 168 (HKAS 38963).

Inocybe lutea — JAPAN, SHIGA PREF. Otsu city, Ishiyama-dera, on the ground in

forest, 29 Sep 1955, T. Hongo 1285 (TNS-F-237714); ISHIKAWA PREF., Tatsunokuchi

Hill Park, under Fagacea forest, 22 Jul 1995, G. Kazuo (TNS-F-32353).

“Astrosporina lutea” — PAPUA NEW GUINEA, Morose District, Bulolo,

Manki, alt. 1400 m, under Castanopsis and Lithocarpus, 28 Mar 1972, E. Horak (ZT

Myc11121 [= ZT 72/335]).

ComMENtTs: Inocybe caroticolor can be easily recognized in the field by its carrot

color, distinct aromatic odor, and entirely pruinose stipe. Microscopically,

the basidiospores are nodulose, hymenial cystidia are thick-walled, and

caulocystidia descend to the base of stipe. The basidiospore outline is variable.

Certain collections or individuals predominately possess weakly nodulose

basidiospores (HMJAU 24615, and one basidioma from HMJAU 24614,

as in Fic. 1c). However, some collections or individual basidiomata possess

predominately strongly nodulose basidiospores with 7-9 nodules (as in the

remaining basidiomata of HMJAU 24614, HKAS 38963, and HKAS 36740; see

Fic. le). Anintermediate phenotype also exists (HMJAU 23271, HMJAU 24621,

HMJAU 24622 and HMJAU 24623; see Fic. 1d). However, the ITS sequences

from HMJAU 24615: GenBank JX025772 and the strongly nodulose-spored

material of HMJAU 24614: JX025773 are identical with that from the holotype

(HMJAU 23271: JX025774) except for a single base difference in HMJAU

24614. Furthermore, no distinct morphological difference was found among

these collections. We consequently interpret the basidiospore shape divergence

as intraspecific variation.

Few nodulose-spored Inocybe species resemble I. caroticolor. Inocybe

bresadolae Massee has a similar basidiospore shape and sweet smell, but its

pileus and stipe have only an indistinct orange tint that is found only in older

specimens (Hobart & Tortelli 2009). Furthermore, I. bresadolae has a more

robust habit and a context that reddens after cutting (Massee 1904, Horak

Inocybe caroticolor sp. nov. (China) ... 173

Fic. 2. Inocybe caroticolor (HMJAU 23271, holotype [except for c & e]). a. Basidiomata. b. Pileipellis.

c. Basidiospores (HMJAU 24615). d. Basidiospores. e. Basidiospores (HMJAU 24614). f. Pleuro-

cystidia. g. Basidia and sterile cells. h. Cheilocystidia, paracystidia, and basidium. i. Caulocystidia,

paracystidia, and hypha. Scale bars: a = 10 mm, b-e = 10 um, f-i = 20 um.

174 ... Fan & Bau

1979). An East Asian taxon, I. umbratica f. aurantiaca Takah. Kobay., which

resembles the new species to some extent, has a marginate stipe base, fungoid or

grassy to spermatic odor, and its type was collected from a Abies mariesii, Pinus

koraiensis, and Betula ermanii forest (Kobayashi 2002). Inocybe polycystidiata

Kobayasi, described from Japan, also resembles the new species but differs by

its subviscid pileus (Kobayasi 1952), recurved pileus scales (Kobayashi 2002),

shorter basidia, and smaller basidiospores (Kobayasi 1952 = 5-6.5 x 4.5-5 um;

Kobayashi 2002 = 5.4-7.3 x 4.2-5.9 um, Q = 1.1-1.5).

Another East Asian species, I. lutea, also resembles the new species. We have

examined the authentic materials of I. lutea identified by Hongo (TNS-F-32353,

TNS-F-237714), who helped describe the species. Our examinations reveal that

it can be distinguished by the following features: (1) the pileipellis hyphae are

hyaline with only a slight yellow tinge; (2) the hymenial cystidia are broadly

fusiform and slightly thick-walled (1-2 um thick); and (3) caulocystidia are

present only at the stipe apex. An examination of the syntype and additional

collections of I. lutea also reveal that caulocystidia are restricted to the stipe

apex (Kobayashi 2002). Furthermore, Kobayasi (1952) originally described

I. lutea as having a yellow fibrillose stipe that tapers upwards and an “iodine-

like” odor.

Lastly, I. lutea as described by Horak (1979, as “Astrosporina lutea” (Kobayasi

& Hongo) E. Horak) based on materials from Papua New Guinea is very close

to I. caroticolor in basidioma color and entirely pruinose stipe but differs by

smaller basidiospores (5.5-8 x 5-6 um), a burnt horn smell, a marginate stipe

base, and a fibrillose pileus with brown fibrils only. Our examinations of a

voucher specimen of the Papua New Guinea material (ZT Myc11121) reveal

pileipellis hyphae identical to those of I. caroticolor and hymenial cystidia that

are predominately utriform usually with a truncate and strikingly thickened

base. The Papua New Guinea “Astrosporina lutea” collections appear to share

more similarities with I. caroticolor than with East Asian materials of I. lutea.

However, “A. lutea” differs from both sets of materials at least by its unique

odor.

BLASTn results of our I. caroticolor ITS sequences show one match (95%

similarity) with unidentified environmental sequence (Wilson et al. 2008).

Also, one sequence by Ryberg et al. (2008) from Estonian material labeled I. aff.

grammata reaches 95% similarity.

Inocybe olivaceonigra (E. Horak) Garrido, Biblioth. Mycol. 120: 177, 1988. Fics 3-6

= Astrosporina olivaceonigra E. Horak, Persoonia 10(2): 194, 1979.

BASIDIOMATA small, slender. PiLEus 11-23 mm diam., conical at first,

then convex to applanate with a blunt umbo, usually rimose or occasionally

uplifted or recurved at margin in age, mostly silky-smooth, rarely scaly,

Inocybe caroticolor sp. nov. (China) ... 175

Vege

Fic. 3. Inocybe olivaceonigra basidiomata (HMJAU 24616).

entirely deep green (27A7-8) at first, then fading to olivaceous (28b7-8) with

fuliginous tinge towards center, ochraceous (1B6-8) outwards, the olivaceous

color stays at least around the center finally. LAMELLAE 2.5-4.5 mm broad in

mid-radius, adnexed, crowded, when young cream white (1A1-2) when young,

then grayish white (1B3) and discoloring to brown (4B1) with whitish edge in

age; edge not smooth. STIPE 28-50 x 1.5-3.0 mm, equal with a swollen base,

solid, longitudinally pruinose over the entire length except for the base, pale

cream color (2A2-3), whitish at the apex and base. CONTEXT fleshy, in pileus

2.5-3.5 mm thick under the umbo, white (1A1), greenish near the pileipellis,

odor spermatic; in stipe shiny, white (1A2), pinkish (5A2-3) near the surface,

striate.

BASIDIOSPORES = (8.7—)9.0-10.0(-10.6) x (5.8-)6.0-7.5(-7.8) um,

conspicuously nodulose, brown, Q = (1.20)1.24-1.55(1.58). Basip1a 22-31 x

7-10 um, clavate, with 4 sterigmata, clamp connections present at the base.

PLEUROCYSTIDIA abundant, 46-82 x 13-21 um, thick-walled (2-3.5 um thick),

yellow, crystalliferous at apex, mostly fusiform to narrowly fusiform, at times

with a short to long neck, tapered in lower part, with clamp connections at

base. CHEILOCYSTIDIA 41-70 x 10-18 um, abundant, similar to pleurocystidia;

PARACYSTIDIA abundant, often catenate, thin-walled, almost hyaline, terminal

cells clavate to obovoid, rarely subglobose, 9-23 x 5-12 um. CAULOCYSTIDIA

37-62 x 8-21 um, descending to the base of stipe, similar to pleurocystidia but

176 ... Fan & Bau

— PR os .

Fi ee Pes ae f y—

lel of) a ;,

Fic. 4. Inocybe olivaceonigra in situ in a Yunnan Castanopsis forest.

with thinner walls; CAULOPARACYSTIDIA 12-22 x 6-13 um, obovoid or ellipsoid.

HYMENIAL TRAMA composed of subregularly arranged, hyaline, inflated hyphae

12-18 um wide. PILEIPELLIs a cutis, duplex, the upper layer indistinct pinkish

in color, up to 130 um thick, composed of subregular, cylindric hyphae 3-5 um

in diam., almost hyaline; the subtending layer golden yellow, up to 68 um in

thickness, composed of smooth to incrusted, inflated hyphae 8-20 um in diam.

CLAMP CONNECTIONS present in all tissues, but not at every septum.

HABITAT AND ECOLOGY: scattered and common on the ground in forests

dominated by Castanopsis. July to September in Yunnan, China. Type recorded

from Castanopsis forests at high elevations in Papua New Guinea.

SPECIMENS EXAMINED: CHINA, YUNNAN PRov., KUNMING: Qiongzhu temple, alt.

2100 m, under Castanopsis, 26 Jul 2011, T. Bau & Y.G. Fan 2011131 (HMJAU 24616;

GenBank JX025775); 26 Jul 2011, X. Jin 2011131a (HMJAU 24617); 26 Jul 2011, Q.X.

Guo 2011131b (HMJAU 24618); 26 Jul 2011, S.S. Yang 2011131c (HMJAU 24619); 26

Jul 2011 T. Bau 2011131d (HMJAU 24620; GenBank JX025776); 21 Sep 2006, Y.C. Li

717 (HKAS 51154). PAPUA NEW GUINEA, MoroBE DIsTRICT, Bulolo, Heads Hump,

alt. 1200 m, under Castanopsis, 9 Mar 1972, E. Horak (ZT Myc11128 [= ZT 72/192],

holotype).

Comments: The authors have examined the holotype of Astrosporina

olivaceonigra (ZT Myc11128). The dried material is in good condition with two

and a half basidiomata. The pilei are olivaceous-green around the center, paler

outwards; the lamellae are apricot to yellowish brown, and the stipes are buff.

Inocybe caroticolor sp. nov. (China) ... 177

Fic. 5. Inocybe olivaceonigra (HMJAU 24620). a. Basidiomata; b. Pileipellis; c. Basidiospores;

d. Basidia and sterile cell; e. Pleurocystidia; f. Cheilocystidia and paracystidia; g. Caulocystidia and

cauloparacystidia. Scale bars: a = 10 mm; b-c = 10 um; d-g = 20 um.

178 ... Fan & Bau

O00)

BEGTEL

Fic. 6. Inocybe olivaceonigra (ZT Myc11128, holotype). a. Hymenial cystidia; b. Basidiospores.

Scale bars: a = 20 um; b = 10 um.

Microscopically, the basidiospores (Fic. 6a) are nodulose with 9-11 rounded

nodules ((7.7—)8.1-9.7(-10.7) x (5.3-)5.8-7.0(-7.8) um, Q = (1.21-)1.34-

1.55(-1.58)); hymenial cystidia (Fic. 6b) are fusoid (33-54 x 11-15 um) with

walls up to 4.5 um; caulocystidia are similar to those of hymenial cystidia, but

have thinner walls. Although Horak (1979) emphasized the conspicuous brown

intra-hyphal pigments (in KOH) in cuticle hyphae in his type description, we

observed that the hyphae in upper pileipellis layer in the holotype are identical

to those of Chinese materials, and the subtending layer is pale yellow and

composed of inflated and incrusted hyphae. The Chinese basidiospores exhibit

a slightly more prominent knob at the apex, and most Chinese basidiomata

possess hymenial cystidia with a thinner wall and a slender appearance (as

in Fic. 5), but HMJAU 24616 and two basidiomata of HMJAU 24617 possess

hymenial cystidia that agree well with the holotype (as in Fic. 6a). In view of

the highly similar gross morphology, habitat, and ecology, we prefer to ignore

any divergence at the species level. The two new ITS sequences (HMJAU 24616:

GenBank JX025775; HMJAU 24620: JX025776) differ by a single base.

Inocybe glaucodisca Buyck & Eyssart., originally described from Zambia and

which greatly resembles I. olivaceonigra in the glaucous grayish umbo, an entirely

pruinose stipe, nodulose basidiospores, and thick-walled cystidia, differs by a

finely squamulose pileus disc, a stipe base with a marginate bulb, and different

habitat and ecology (Buyck & Eyssartier 1999). A recently described species,

I. pyriformis Takah. Kobay. & S. Kurogi, which also has a smooth pileus, entirely

pruinose stipe with a non-marginate base, and nodulose basidiospores differs

in its dull red- to brown-colored stipe and pyriform to ventricose pleurocystidia

(Kobayashi 2009). Inocybe insignis A.H. Sm., originally described from North

Inocybe caroticolor sp. nov. (China) ... 179

America is similar in having an entirely pruinose stipe, stellate basidiospores,

and thick-walled cystidia; it shares the greenish color only after bruising or

cutting. However, I. insignis has a larger habit, an aromatic odor, and larger

stellate basidiospores (Smith 1941). Inocybe viridiumbonata Pegler differs by

its robust habit, thin-walled cheilocystidia, slightly thick-walled pleurocystidia,

and larger basidiospores. In addition, I. viridiumbonata occurs in a degraded

xerophytic forest in the Caribbean (Pegler 1983).

BLASTh results of our I. olivaceonigra ITS sequences show 10 matches (94%

similarity) labeled I. suaveolens D.E. Stuntz based on materials from North

America, among which the holotype sequence of I. suaveolens (HQ222010)

produced by Matheny is also included and three matches (93%-94% similarity)

labeled I. umbratica Quél. based on materials from both North America and

Europe. Morphologically, I. suaveolens (Smith & Stuntz 1950) and I. umbratica

(Kobayashi 2002) can be easily distinguished from I. olivaceonigra by their lack

of olivaceous colors on the pileus. The three species, however, do share the

silky-smooth pileus, the entirely pruinose stipe, nodulose basidiospores, and

thick-walled hymenial cystidia.

Acknowledgments

We are sincerely grateful to Dr. Takahito Kobayashi (Hokkaido University Museum)

for examining one specimen and providing valuable references and suggestions, to Jukka

Vauras (Biological Collections of Abo Akademi University, Finland) for reviewing the

manuscript, and to Dr. Brandon Matheny (University of Tennessee) for twice reviewing

the manuscript, and also to Dr. Egon Horak for giving valuable suggestions. Thanks are

also given to Dr. Reinhard Berndt (the Joint Herbarium of the University of Zurich and

the ETH Zurich), Dr. Kentaro Hosaka (National Museum of Nature and Science, Japan)

and Dr. H. Peng & Dr. L.S. Wang (the Herbarium of Cryptogams, Kunming Institute of

Botany, Chinese Academy of Sciences) for kind help in forwarding specimens. We also

thank Dr. Z.L. Yang and Dr. N.K. Zeng (Kunming Institute of Botany, Chinese Academy

of Sciences) for kindly helping in field work. This study was supported by the National

Natural Science Foundation of China (Nos. 31093440, 31270063) and Program for

Changjiang Scholars and Innovative Research Team in University (No. IRT1134).

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MY COTAXON

http://dx.doi.org/10.5248/123.183

Volume 123, pp. 183-191 January-March 2013

Stipitate hydnums of the southern United States 1:

Phellodon mississippiensis sp. nov.

RICHARD E. Barrp’, LisA E. WALLACE’, & GERALD BAKER?

' Dept. of BCH-EPP, Mississippi State University &

? Dept. of Biological Sciences, Mississippi State University

Mississippi State, MS 39762, USA

* CORRESPONDENCE TO: rbaird@plantpath.msstate.edu

ABSTRACT—During a general survey of fleshy fungi from Mississippi, an undescribed

Phellodon species was observed in the Tombigbee National Forest at the same location

over a two-year period. No other studies of the stipitate hydnums are available for the

midsouthern states that directly compare with the new species. Molecular sequence data of

P. mississippiensis were distinct from other Phellodon spp. of the southeastern United States.

The highly fused (concrescent) basidiomata of P. mississippiensis are found under mixed

hardwoods and associated pines. The light tan to light rust (orange) or salmon color on the

marginal spines and along the margin of the small (<5 cm broad) stout spongy tomentose

basidiomata makes this species distinct. The presence of rare clamp connections was observed

in the trama hyphae. However, identification of P. mississippiensis can be complicated by its

concrescent nature and the indeterminate growth forms caused by environmental conditions.

Morphological characters and results from the phylogenetic analysis are included in the

species description.

Key worps—white spored, mycorrhizal, early summer, fenugreek odor

Introduction

The stipitate hydnums in the southeastern United States have been studied

extensively either directly or in larger monographs (Banker 1906, 1913a,b,c;

Coker 1919, 1926, 1927, 1939, 1942; Jones 1935; Coker & Beers 1951; Baird

1986a,b, 1987; Baird & Kahn 1986, 1987). However, no specific studies of the

group have been conducted for the mid-south, including Mississippi. David

Lewis (Gulf States Mycological Society, pers. comm.) reports that many species

of fleshy fungi have been identified and new species recorded in numerous

published research papers from the state. Lewis stated that FS. Earle & L.M.

Underwood spent time along the Gulf Coast in the Ocean Springs, MS, area,

collecting primarily Amanita and Boletus spp. Many of these species were

184 ... Baird, Wallace & Baker

sent to W.A. Murrill, who published many as new. More recently W.G. Cibula

extensively documented fleshy fungi for Mississippi (Cibula 1979, Cibula &

Weber 1996), sending most collections sent to the Field Museum, Chicago,

IL. Finally, of the three stipitate hydnums cited in Dukes’ (2000) book on

Mississippi fleshy fungi, two were correctly identified to genus as Hydnellum

and another listed as Phellodon was incorrectly identified and was actually a

wood decay fungus.

A study of the stipitate hydnums from southern Appalachian Mountains

represents the most recent monograph on stipitate hydnums from the southern

region (Baird 1986a). During that research, a thorough type study was also

conducted with worldwide implications (Baird 1986b). More recently revision

and phylogenetic analyses (unpublished data) of stipitate hydnums have shown

that nine Phellodon spp. from Great Smoky Mountains and the surrounding

southern Appalachian Mountains are distinct.

During a general survey of macrofungi in northern Mississippi, an unknown

Phellodon sp. (Fic. 1) was collected from the same location during 2009 and

2010. The collections were found under mixed hardwood with conifers near

the site. Morphological comparisons with all other known Phellodon spp.

from southeastern United States indicated that none of the general herbarium

collections or from previous type studies were similar (Baird 1986b).

Furthermore, a BLAST search of molecular sequences from other Phellodon

spp. from the southeastern United States and GenBank revealed no matches,

supporting the species as undescribed. We present here a detailed description

of the new species and its phylogeny (Fie. 3).

Materials & methods

In 2009 and 2010, collections were obtained from the North Trail sect. D of Noxubee

Crest Mountain Bike Trail System of Tombigbee District of Tombigbee National

Forest. Survey of associated tree species was conducted within a 30 m radius of the

basidiomata.

All macro- and microscopic data were analyzed according to Baird & Kahn (1986).

Colors were determined with Kornerup & Wanscher (1978). Digital images were taken

of slide mounts of basidiospores and basidia using a Jenoptik ProgRes 3.3 megapixel

digital camera mounted on a Olympus BX50 compound microscope. Basidiospore,

basidium, and sterigma sizes were measured from these images using image-analysis

software (IMT i-Solution Lite), and 20-40 measurements were made for each of three

variables per collection. Scanning electron microscope (SEM) photographs were made

of dried specimen material mounted on aluminum stubs with double-sided sticky

carbon tape, coated with 15 nm platinum, and examined with a JEOL JSM 6500F SEM.

The accelerating voltage was 5 KV and the images were digitally recorded.

Molecular examination

The nrITS DNA region from the new species was sequenced for phylogenetic

comparisons with other Phellodon species. Genomic DNA was extracted from dried

Phellodon mississippiensis sp. nov. (United States) ... 185

basidiomata tissue using DNeasy, Plant Mini Kit by Qiagen (Valencia, CA) following

procedures discussed in the brochure (08/2000) instructions. The ITS primers ITS1F

(Gardes & Bruns 1993) and ITS 4 (White et al. 1990) were used to amplify the internal

transcribed spacer region ITS 1-5.8S-ITS 2. Amplification reaction for ITS used

procedures previously described (Lickey et al. 2007). The PCR products were purified

with Qiagen PCR columns (Qiagen, Chatsworth, CA) and sequences by Eurofins-

MWG/Operon (Huntsville, AL). Resulting electropherograms were viewed and edited

using DNAstar of Lasergene (Madison, WI), and multiple sequences per taxon were

collapsed due to lack of variation. When electropherograms were unreadable due to

multiple sequences or insertion-deletions that could not be resolved, PCR products

were cloned using pGEM-T vector system and SM 109 competent cells following

manufacturers protocol (Promega). Sequences generated include Phellodon sp. 1

(TENN65891 = JN135199), P alboniger (Peck) Banker (TENN65868 = JN135206),

P. brunneoolivaceus (Coker & Beers) R.E. Baird (TENN65882, JN135201), P. confluens

(Pers.) Pouzar (TENN65874= JN135198), P ellisianus Banker (TENN65884= JN135204),

P. fuligineoalbus (Fr.) Baird (in press) (TENN65893= JN135196), P. fibulatus K.A. Harrison

(TENN65865= JN135205), P. melaleucus (Sw. ex Fr.) P. Karst. (TENN65896= JN135197),

P. niger (Fr.) P. Karst. (TENN65876= JN135202), P putidus (G.F. Atk.) Banker (TENN65883=

JN135200), and P tomentosus (Fr.) Banker (TENN65889= JN135203). Sequences of

Sarcodon atroviridis (Morgan) Banker (TENN65871= JN135190), S. imbricatus (L.) P.

Karst. (TENN65890= JN135194), and S. scabripes (Peck) Banker (TENN65894= JN135191)

were designated as outgroup. All sequences were subjected to GenBank program BLAST

to detect similar sequences, resulting in the inclusion of six additional sequences in the

data set: Phellodon sp. 352-363 (4B509722), P. nothofagi McNabb ppp89898 (Gu222318),

P. nothofagi PDD89880 (GU222316), P. atratus K.A. Harrison (HQ650766, EN185793), and

P. sinclairii (Berk.) G. Cunn. (GU222291). Sequences of the new species were submitted

to GenBank and accession numbers JN247563 (MS-1) and JN247564 (MS-3) were

designated.

The sequences were aligned using the web version of MAFFT (Katoh et al. 2005) with

the G-INS-i option, which is provided by the CBRC (http://mafft.cbrc.jp/alignment/

server/). The resulting alignment of 1,105 characters was used directly in jModeltest

(Posada 2008) to determine TrN+G as the best fitting model of molecular evolution

under the AICc. A Bayesian phylogenetic analysis was performed on the data set using

MrBayes v. 3.1.2 (Ronquist & Huelsenbeck 2003). The three Sarcodon sequences were

designated as outgroup. Using a GITR+G model the MrBayes analysis was conducted for

1 million generations, after which the split standard deviation reached 0.006. A burn-

in of 2,500 trees was used prior to determining the posterior probability of the trees

with the highest likelihood. The consensus tree is reported with posterior probability

indicated as support for clades.

Taxonomy

Phellodon mississippiensis R.E. Baird, L-E. Wallace & G. Baker sp.nov. — Fras 1-2

INDEX FUNGORUM IF550077

Differs from Phellodon putidus and Phellodon tomentosus by its salmon colored spines,

very stout fused pilei or stipes, and presence of clamp connections.

186 ... Baird, Wallace & Baker

Type: United States of America. Mississippi, Noxubee County, Tombigbee National

Forest, North Trail sect. D of Noxubee Crest Mountain Bike Trail System, (35°44.350'N

83°25.415'E), June 15, 2010, P. Scott, MS-3 (holotype, TENN65879; GenBank,

JN247564).

Erymo.ocy: The epithet refers to the State in which the type was collected.

Basip1omarTa single, usually highly concrescent, <5.0 cm broad, stout, convex

to plane or subdepressed or rarely colliculose; margin blunt in immature

specimens to thin or irregular in age; spongy-tomentose becoming matted or

appressed at disc, smooth to pitted or irregular at disc in aged tomentum, rarely

radially rugulose, white (5A1) to orange white (5A2) or light orange (5A4)

nearest margin, light brown (6D5), cinnamon (6D6) to burnt umber (6F6) or

dark brown (6F7) at disc; underside of pileus margin and immature spines white

(5A1) to pale orange (5A3) or light orange (5A5); context <3.0-4.0 mm thick

nearest stipe, evidence of zones, concolorous with pileus. STIPES <1.5 x 2.0 cm

concrescent or <1.0 x 1.0 cm single, central to eccentric, flattened or irregular on

a side, tomentose becoming matted in age, bruised surface irregular, cylindrical

to subattenuate above, at lower third entire base often fused forming a mycelial

mass, lower portion of mycelial mass buried forming a thick network in the

duff layer, white (5A1) nearest base to pale orange (5A3) or dark brown (6F6);

context <1.5 cm thick, not obviously duplex, azonate, concolorous with pileus

context. SPINES up to 2.0 mm long, subcrowded, subdecurrent, white (5A1)

to light orange (5A5) nearest margin. Taste none; odor moderate to strong

fenugreek increasing upon drying. CHEMICAL REACTION in KOH light to dark

brown on pileus context tissues over short period of time.

PILEUS TRAMA HYPHAE $5.0 um diam, uninflated, generally interwoven

in subsurface layer, parallel below, unclamped; rare occurring gloeophorous-

like hyphae <7.0 um diam, cylindrical to highly irregular-shaped. STIPE

HYPHAE <4.5 um diam, uninflated, interwoven in subsurface layer to parallel

below, unclamped, rarely observed gloeophorous-like hyphae <9.0 um diam,

when present. SPINE TRAMA HYPHAE subparallel to parallel, <4.0 um diam,

rare clamp connections in subhymenial hyphae. BAsip1osporEs 16.0-22.0 x

5.0-6.0 um (mean = 17.9 + 1.7 x 5.8 + 0.5 um), subglobose to globose, hyaline;

ornamentation echinulate, spinules not prominent; hilar appendage oblique.

BASIDIA 16.0-22.0 x 5.0-6.0 um (mean = 17.9 + 1.7 x 5.8 + 0.5 um), clavate,

unclamped, 4-spored; sterigmata <4.0 um long (mean = 3.6 + 0.39 um). Clamp

connections absent in pileus and stipe contact tissues, rare in subhymenial

hyphae of the spines.

ADDITIONAL SPECIMEN EXAMINED: UNITED STATES of AMERICA. Mississippi,

Noxubee County., Tombigbee National Forest, North Trail sect. D of Noxubee Crest

Mountain Bike Trail System, (35°44.350'N 83°25.415’E), May 28, 2010, P. Scott, MS-1

(TENN65879; GenBank JN247563).

Phellodon mississippiensis sp. nov. (United States) ... 187

Mississippi State

Mmsion WA

Fic. 1. Phellodon mississippiensis. a: Basidiomata showing upper portion of basidiomata;

b: Basidiomata side view of fused pilei and stipes with thickened mycelial base. Scale bars = 5.0 cm.

188 ... Baird, Wallace & Baker

Fic. 2. Phellodon mississippiensis. a: Basidiospore with echinulate ornamentation;

b: Scanning electron microscope (SEM) micrograph of basidiospores; c: Gloeophorous-

like hyphae present in pileus and stipe; d: Clamp connection in the subhymenial hyphae;

e-f: Basidia and sterigmata. Scale bar = 10 um.

Phellodon mississippiensis sp. nov. (United States) ... 189

Hasirat: The type location contained the following tree species (percent

densities) within 17.4 m of the collections: Pinus taeda L. (20% = 5 trees),

Carya glabra (Mill.) Sweet (16%), Acer rubrum L. (16%), Fraxinus americana L.

(12%), Cornus florida L. (8%), Oxydendrum arboretum (L.) DC. (8%), Ostrya

virginiana (Mill.) K. Koch (1%), Quercus velutina Lam. (1%), Q. rubra L. (1%),

and Nyssa sylvatica Marshall (1%). The site (6.0% slope), which was 3.0 m from

a small intermittent stream, also contained 30 year-old pines planted >25.0 m

from the collections that were under the hardwoods.

Discussion

The phylogenetic analysis strongly supports P. mississippiensis as a member of

the Phellodon clade, which is distinct from the outgroup Sarcodon species (100%

PB; Fic. 3). The two P. mississippiensis samples are identical and unique within

Phellodon, further supporting its separation as a new species. Within Phellodon

however, the evolutionary relationship of P. mississippiensis with other species

is unclear. Current phylogenetic analyses clearly place P mississippiensis within

Phellodon but are unable to resolve most sister species relationships within the

genus (FIG. 3).

57 Pheliodon alboniger JN135206

Phellodon sp. 1 KC571747

65 400 Phellodon atratus HQ650766

Phellodon atratus FN185793

Phellodon niger JN135202

100 Phellodon putidus JN135200

75 Phellodon tomentosus JN135203

Phellodon fuligineoalbus JN135196

100 Phellodon confluens JN135198

* 53 Phellodon sp. AB509722

Phellodon melaleucus JN135197

400 Phellodon mississippiensis MS-3 JN247564

88 Phellodon mississippiensis MS-1 JN247563

100 Phellodon fibulatus JN135205

100 Phellodon brunneoolivaceus JN135201

Phellodon ellisianus JN135204

400 Phellodon sinclairii GU222291

99 Phellodon nothofagi GU222318

Phellodon nothofagi GU222316

100 Sarcodon imbricatus JN135194

Sarcodon scabripes JN135191

Sarcodon atroviridis JN135190

0.07

Fic. 3. Bayesian consensus phylogeny resulting from analysis of nrITS sequences. Values on

branches are posterior probabilities and indicate support for nodes.

190 ... Baird, Wallace & Baker

Molecular sequence data and morphological differences separate this species

from all other Phellodon spp. known to occur in the southeastern United States,

including Florida (Baird 1986a,b; Baird & Kahn 1986). Morphologically the

basidiomata may be confused with several species based on indeterminate

growth forms and concrescent nature, in particular, P fibulatus, P. confluens,

P. tomentosus, P. radicatus R.E. Baird and P. putidus. Phellodon putidus has

marginal colors overlapping those of P mississippiensis but is much larger

(<11 cm broad) and lacks the thickened fused stipe bases that can occur in

P. mississippiensis. Basidiomata of P. fibulatus (with abundant clamp connections)

and P. tomentosus (with clamp connections scattered randomly on stipe

hyphae) are much thinner than P mississippiensis. Unlike P. mississippiensis,

P. tomentosus occurs only under conifers. Phellodon confluens has very thin

basidiomata and lacks the light orange or salmon colors seen on the margin of

P. mississippiensis. Phellodon radicatus, reported from Florida, has a blackish

colored context and radicating stipe bases not present in P mississippiensis,

which is instead characterized by the thick mycelial mass formed from fused

stipes and buried in the humus layer. Also, basidiospores of P. radicatus are 1.0

um larger than in the new species.

Previous studies in Florida and surveys in the midsouth and along the Gulf

Coast lead us to expect that new stipitate hydnum species will continue to be

identified during surveys of this region.

Acknowledgements

Special thanks to the curators for supplying specimens from NYBG, MICH, NCSC,

and TENN, and to Mississippi State University (MAFES publication number 12185)

and Highlands Biological Station for use of their facilities and equipment during a long-

term study of the Phellodon spp. from the southern United States.

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genera: Bankera, Hydnellum, Phellodon, Sarcodon. Bibliotheca Mycological04. 156 p.

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MY COTAXON

http://dx.doi.org/10.5248/123.193

Volume 123, pp. 193-204 January-March 2013

Two new species of Lignosus (Polyporaceae) from Malaysia —

L. tigris and L. cameronensis

CHON-SENG TAN ’, Szu-TING NG? & Ji TAN 3

"Malaysian Agricultural Research and Development Institute (MARDI)

PO. Box 12301, 50774 Kuala Lumpur, Malaysia

°LiGNO Biotech Sdn Bhd, 43300 Balakong Jaya, Malaysia

*Institute of Biological Sciences, University of Malaya, 50603, Kuala Lumpur, Malaysia

* CORRESPONDENCE TO: tanchonseng@gmail.com

ABSTRACT —Lignosus cameronensis and L. tigris are described as new based on collections

from the tropical forests of Pahang, Malaysia. Phenotypic and genotypic data support both as

new species. Morphological features are illustrated, and the associated Internal Transcribed

Region (ITS1 + 5.8S + ITS2) rDNA sequences have been deposited in the GenBank. Pore and

basidiospore sizes are the main characters distinguishing these two Lignosus species from

other members of the genus, with L. tigris distinguished from L. sacer by its larger pore size

and smaller basidiospores and L. cameronensis separated from L. ekombitii by its smaller

basidiospores. A key to the species of Lignosus is provided.

KEY worpDs — medicinal mushroom, sclerotia, taxonomy, phylogeny

Introduction

Lignosus Lloyd ex Torrend, a genus including six poroid species—L. dimiticus

Ryvarden, L. ekombitii Douanla-Meli, L. goetzii (Henn.) Ryvarden, L. hainanensis

B.K. Cui, L. rhinocerotis (Cooke) Ryvarden, L. sacer (Afzel. ex Fr.) Ryvarden—is

known from the paleotropics (Cui et al. 2011, Dai 2012, Douanla-Meli & Langer

2003, Ryvarden & Johansen 1980). Members of this genus characteristically

exhibit central stipitate basidiocarps, which rise from subterranean sclerotia.

The genus is microscopically characterized by dimitic to trimitic hyphal

systems, with dimitism currently reported only in L. dimiticus, which lacks

binding hyphae in the context, sclerotium and stipe (Ryvarden & Johansen

1980). Apart from L. goetzii, basidiospores are known from all Lignosus species

and are mostly hyaline, globose, isodiametric to ellipsoid depending on the

species (Cui et al. 2010, Douanla-Meli & Langer 2003, Ryvarden & Johansen

1980).

194 ... Tan, Ng & Tan

Molecular analysis has supplemented conventional taxonomic classification

of these macrofungi. Molecular markers used for Lignosus are the nuclear

large subunit (nLSU), RNA polymerase II second largest subunit (RPB2),

mitochondrial ATPase subunit six (ATP6) (Sotome et al. 2008), and the Internal

Transcribed Region (ITS1 + 5.8S + ITS2) ribosomal DNA (Cui et al. 2010, Tan

et al. 2010, Zhao et al. 2013, Tian et al. 2013). Easily amplified and possessing

highly variable non-conserved ITS1 and ITS2 regions, the ITS region has

proved useful for phylogenetic reconstruction of Lignosus species (Martin &

Rygiewicz 2005, White et al. 1990).

Lignosus species have been widely studied due to their potential medicinal

value. In Malaysia, locals regard the basidiocarps as important ingredients in

traditional medicine, particularly for treating coughs and asthma, as well as

for improving general well being (Chang & Lee 2004). Despite the promising

benefits of these mushrooms, taxonomic research of Lignosus in Malaysia

has been gradual and mediocre, with no significant records published since

Ryvarden’s discovery of L. rhinocerotis in the Penang island of Peninsular

Malaysia (Ryvarden 1972).

Recent expeditions to the rainforests of Peninsular Malaysia have led to the

discovery of two Lignosus species from the state of Pahang that conform to the

characters of the genus but do not match any known species. We describe here

these two new Lignosus based on morphological observations and molecular

data and infer their phylogenetic relationships within Lignosus.

Materials & methods

Lignosus tigris specimens K and T were collected froma tropical forest in Lata Iskandar

(4°17.46'N 101°34.41’E), Pahang, Malaysia on 12 June 2010; L. cameronensis specimens

T1 and T8 were collected from the same forest on 17 July 2010. A small portion (50 mg)

of sclerotia tissue was excised for maintaining a live culture and for DNA extraction; the

remaining specimen was oven-dried for preservation as a herbarium specimen. Lignosus

tigris K and L. cameronensis T1 herbarium specimens have been deposited at K(M)

(Mycological Herbarium, Royal Botanic Gardens, Kew, UK); collections L. tigris T and

L. cameronensis T8 are maintained in the personal herbarium of C.S. Tan, MARDI.

Morphological studies

Macroscopic characters are based on both fresh and subsequently dried material.

Colors are based on the RGB color model, denoted by three numbers in brackets

representing Red, Green, and Blue, respectively. Microscopic descriptions were carried

out predominantly on dried specimens using a Nikon Z200 microscope with phase

contrast illumination. Freehand sections were rehydrated in 70-95% ETOH prior

to mounting in H,O, 3% KOH, or Melzer’s reagent. Basidiospores were observed

and measured directly from the sections. Spore dimensions exclude the 5% extreme

measurements from each end of the range given in parentheses. Q represents the mean

length/width of the specified number of spores studied (n) (Largent 1977, Largent et al.

1977, Dai 2010).

Lignosus spp. nov. (Malaysia) ... 195

Molecular taxonomy

DNA specimens were ground with fine sand powder, and DNA was extracted using

a modified CTAB procedure (Cota-Sanchez et al. 2006). Polymerase Chain Reaction

(PCR) amplification of the ITS was conducted in 25ul reaction volumes using the primers

ITS4 (5'-TCCTCCGCTTATTGATATGC-3’) and ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3’)

suggested by White et al. (1990). The PCR parameters were as follows: initial denaturing

at 94°C for 5 min, followed by 20 cycles of (94°C for 45 sec, 45°C for 45 sec, 72°C for 1

min), and final extension at 72°C for 5 min. Amplicons were analyzed through staining

with ethidium bromide and 1% agarose gel electrophoresis, and subsequently purified

using a glass-milk matrix (Fermentas, USA) according to the manufacturer's protocol.

Purified PCR products were ligated into pGEM-T Easy vector (Promega, USA) prior to

transformation of the Escherichia coli strain JM109. Four clones were sequenced in both

directions using forward and reversed M13 sequencing primers via 1* Base Laboratories

Malaysia (ABI system).

The resulting DNA electropherograms of the ITS region were truncated and

assembled into consensus sequences via ChromasPro V1.41 (Technelysium Pty

Ltd). Multiple sequence alignments were generated with the incorporation of several

sequences from the GenBank, and subsequently encoded in a NEXUS format using

ClustalX V2.0 (Larkin et al. 2007). Details of samples used in this study are summarized

in TABLE 1

TABLE 1. Lignosus and Daedaleopsis samples used in molecular analyses.

SAMPLE LOCALITY GENBANK No. DNA souRCE

L. cameronensis T1 (holotype) Malaysia JQ409483 Sclerotium

T8 (isotype) Malaysia JQ409484 Sclerotium

L. ekombitii Cameroon (ex Douanla-Meli) Sclerotium

L. hainanensis China GU580884 Sclerotium

L. rhinocerotis Ryvarden 11324 Malaysia (ex Ryvarden) Pileus

CH2 Malaysia FJ380871 Sclerotium

CH31 Malaysia FJ899143 Sclerotium

L. sacer Ryvarden 11329 Zimbabwe GU001674 Pileus

K(M) 1037 Kenya GU001675 Pileus

L. tigris K (holotype) Malaysia JQ409481 Sclerotium

T (isotype) Malaysia JQ409482 Sclerotium

D. confragosa (Bolton) J. Schrét Germany FR686551 —

D. sinensis (Lloyd) Y.C. Dai — FJ627256 —

Maximum parsimony (MP) and maximum likelihood (ML) analyses were used in

the phylogeny construction via PAUP 4.0b10 (Swofford 2003). The MP tree was inferred

through a heuristic search procedure using 1000 bootstrapping replications, with 100

random sequence addition and tree bisection reconnection (TBR) branch swapping.

All characters were unordered and weighted equally. The best-fit HKY + G model was

selected using hLRT (a = 0.01) through Modeltest v3.7 (Posada & Crandall 1998), and

implemented for the ML analysis. Parameters were similar to the MP analysis, but with

100 ML bootstrap replicates instead.

Bayesian inference (BI) was conducted using Mr. Bayes 3.2.1 (Huelsenback et al. 2001,

Ronquist & Huelsenbeck 2003). Best-fit models were generated by Kakusan v.3 using

the Bayesian Information Criterion (BIC) (Schwarz 1978) (Tanabe 2007). BI analysis

196 ... Tan, Ng & Tan

was performed using the Markov Chain Monte Carlo (MCMC) method over a run of

2,000,000 generations, with a tree sampled every 100 generations. Chain stationarity was

determined graphically using Tracer v1.5 (http://tree.bio.ed.ac.uk/software/tracer/), and

the first 20,000 trees were conservatively discarded as burn-ins prior to the construction

of the final 50% majority rule consensus tree. The resulting phylogenetic inferences were

analysed and processed via Figtree (Morariu et al. 2008).

Taxonomy

Lignosus tigris Chon S. Tan, sp. nov. PLATE 1

MycoBank MB 800843

Differs from Lignosus sacer by its larger pores and smaller basidiospores.

Type: Malaysia. Pahang, Lata Iskandar, 4°17.46'N 101°34.41’E, in tropical rain forests,

12 June 2010, Tan CS ‘K’ (Holotype, K(M) 177826; GenBank, JQ409481). Tan CS “T”

(isotype, MARDI; GenBank, JQ409482).

EryMo_oey: tigris, from the local folklore that the species arose from milk that dripped

onto the ground while a tigress fed her cubs.

BASIDIOCARP annual, single, terrestrial, centrally stipitate with an isodiametric

pileus; pilei up to 11.3 cm in diameter and 0.5 cm thick at the center; tough,

without odor and taste. Pireus obtusely umbonate, slightly depressed,

concentrically zonate and cinnamon brown (169, 79, 5) to ochre (223, 161,

49), margin wrinkled, thin, slightly folded and becoming crisp when dry. Pore

surface cream (249, 218, 161) to pale brown (234, 152, 4), pruinose when fresh.

Pores roughly pentagonal or hexagonal, coherent. more or less honeycomb-

like, 1-2 per mm, gradually smaller and denser towards the margin. Tubes

layered, somewhat polygonal, apparent and deep, up to 4 mm depth; tube

walls gradually thinning, becoming corky or crisp upon drying. Pileal context

white (255, 245, 231), tough, up to 5 mm thick, turning cream (249, 218, 161)

when dry. STIPE single, caramel brown (186, 81, 2), up to 14 cm long, 1-1.8 cm

thick, slightly contorted; enlarged, rough and wrinkled at the base, gradually

thinning and smoother towards the pileus. Context from stipe white (255, 249,

171), turning cream (249, 218, 161) when dry. SCLEROTIUM irregular, light-

weight, wrinkled and soiled, up to 7 cm long and 6.5 cm broad. Context pure

white (236, 236, 236) to cream (249, 218, 161), wrinkled, floccose-cottony to

tomentulose around the core.

HyYPHAL SYSTEM trimitic; all hyphae negative in Melzer’s Reagent. Generative

hyphae with abundant clamp, diameter less than 1.4 um, hyaline and thin-

walled, interwoven with flexuous, aseptate skeletal hyphae, 2.6-3.1 um in

diameter, within the hymenium. Binding hyphae common, branched, ranging

from almost straight to tortuous, up to 2.3 um in diameter. Trama composed

predominantly of thick-walled skeletal hyphae, up to 3.4 um diameter, with

narrow lumen; and occasional binding hyphae, hyaline, branched, up to 1.9 um.

Lignosus spp. nov. (Malaysia) ... 197

O9000

AES do

10 um 100 um

PLaTE 1. Microscopic structures of Lignosus tigris (holotype). a External morphology. b Basidia

and basidioles. c Basidiospores and spore attachment. d Hyphae from trama. e Hyphae from stipe.

f Sclerids.

198 ... Tan, Ng & Tan

Stipe context dominantly occupied by binding hyphae, hyaline, infrequently

branched, diameter up to 1.9 um; overlapped or slightly intertwined with

sparse, slightly flexuous skeletal hyphae, hyaline, unbranched, diameter up

to 4.7 um, thick-walled with a narrow lumen. ScCLERIDs dense and abundant

within the sclerotium, subglobose, becoming somewhat oblong to irregular

upon germination or when collectively clumped, 23.4-42.7 x 12.2-29.9 um,

margins wrinkled, appearing layered or stratified. Skeletal hyphae dominant,

slightly sinuous, aseptate, hyaline, up to 3.5 um in diameter, unbranched and

thick-walled. Binding hyphae common, slightly flexuous, branched, interwoven,

diameter up to 2.2 um. Generative hyphae present, thin-walled, unbranched,

hyaline, up to 1.5 um in diameter.

Cystip1A and cystidioles absent. Basip1a clavate, subclavate, hyaline to

granular, 4.9-9.1 x 1.75-3.3 um; four sterigmata slender, generally 1.7-4.3 um

long, with some up to 4.3 um long; basidioles mostly subclavate. BASIDIOSPORES

mainly broadly ellipsoid to subglobose, infrequently globose and amygdaliform,

(2.3-)2.5-5.5(-5.6) x (1.7-)1.8-3.6(-3.7) um; Q = 1.34, (n = 44), thin-walled,

hyaline, smooth.

Lignosus cameronensis Chon S. Tan, sp. nov. PLATE 2

MycoBank MB 800844

Differs from Lignosus ekombitii by its smaller basidiospores.

Type: Malaysia. Pahang, Lata Iskandar, 4°17.46’N 101°34.41’E, in tropical rain forests,

17 July 2010, Tan CS “T1’ (Holotype, K(M) 177843, GenBank, JQ409483). Tan CS “T8’

(isotype, MARDI; GenBank, JQ409484).

ETYMOLOGY: cameronensis, referring to Cameron Highlands, Malaysia; the locality of

the type collection.

BASIDIOCARP annual, single, terrestrial, centrally stipitate with a more or less

circular pileus; pilei up to 13.5 cm in diameter and 0.4 cm thick at the center;

stout, without odor and taste. PrLEus orbicular, slightly convex, brown (83, 32,

1) to chestnut brown (140, 55, 2), concentrically zonate, sulcate, rugose, margin

deflexed, occasionally layered and overlapping; inrolled or incurved when dry.

Pore surface cream brown (206, 157, 70), pruinose when fresh. Pores hardly

visible, tiny and packed, isodiametric, 2-4 per mm. Tubes minute, dense,

angular and layered; shallow, up to 2 mm deep; tube walls gradually thinning

and becoming corky or crisp when dried. Tube layer up 4 mm thick. Context

white (255, 245, 231), tough, up to 3 mm thick. Stripe muddy brown (175,

149, 97), single, central, equal, ellipsoidal, up to 8.5 cm long and 1.5 cm thick;

woody hard and smooth when dry. Context from stipe cream (249, 218, 161).

SCLEROTIUM irregular, light, rugose and soiled, up to 6.5 cm long and wide.

Context pure white (236, 236, 236), wrinkled and floccose-cottony around the

center.

Lignosus spp. nov. (Malaysia) ... 199

PRS

a WN

ercnaicoononces: id fs 0 Sia AAS

2cm ey x :

ees

we, 2

ee,

- 9Q900 *

aver onaize!

P voare

10 um 100 um

PLATE 2. Microscopic structures of Lignosus cameronensis (holotype). a External morphology.

b Basidia and basidioles. c Basidiospores and spore attachment. d Hyphae from trama. e Hyphae

from stipe. f Sclerids.

200 ... Tan, Ng & Tan

HYPHAL SYSTEM trimitic; all hyphae negative in Melzers Reagent.

Generative hyphae with clamped connections, very irregular and contorted in

the hymenium, thin-walled and hyaline, 1.2-1.5 um in diameter; interwoven

with occasional skeletal hyphae. Skeletal hyphae dominant within the trama,

up to 3.6 um in diameter, aseptate, thick-walled (up to 1 um) with a narrow

lumen, slightly flexuous, with scarce branching; mixed with thin-walled, almost

straight to tortuous, aseptate binding hyphae, up to 2 um in diameter; generative

hyphae infrequent. Context of stipe, mainly of binding hyphae, integrating with

relatively sparse, slightly flexuous skeletal hyphae; branching scarce. Sclerotium

composed mainly of sclerids, originally subglobose or somewhat pear-shaped,

becoming cordate to irregular upon germination or when clumped with other

sclerids, 24.9-49.2 x 21.3-42.1 um. Sclerid margins uneven, and seemingly

stratified or striate. Skeletal hyphae dominant, thick-walled, hyaline, 1.3-4.1

um in diameter, often branching. Generative hyphae less common, thin-walled,

and unbranched, 1-1.6 um in diameter. Binding hyphae present, interwoven,

hyaline, often branching, 1.2-2.3 um in diameter.

Cystip1A and cystidioles absent. Basip1a clavate, hyaline to granular, 3.6-8.9

x 1.4-4.5 um; four sterigmata slender, generally 1.7-4.3 um long, with some up

to 5.7 um long; basidioles mostly subclavate. BAstDIosPoREs mainly ellipsoid

to subglobose, rarely globose, (2.2—)2.4-4.8(-4.9) x (1.8-)1.9-3.2(-3.4) um;

Q = 1.47, (n = 37), hyaline, smooth, thin-walled.

Molecular phylogeny

The ITS sequences were truncated at terminals to avoid ambiguous

nucleotides, resulting in a length of 646 bp. Within the dataset, a total of 144

characters were considered parsimony-informative, whereas 466 characters

remained constant. Parsimonious analyses of the ITS region resulted in two

most parsimonious trees, with a consistency index (CI) of 0.902 and a retention

index (RI) of 0.925. The nucleotide region encoding the 5.88 rRNA showed no

variation among the Lignosus congeners. Conversely, large variations, including

long indels were observed within the ITS1 and ITS2 regions. A genetic distance

of up to 7.1% was observed between L. tigris and L. cameronensis. Lignosus

rhinocerotis and L. ekombitii showed the highest genetic distance of up to 11.2%

within the genus. An intraspecific genetic difference of 1.5% was observed

between L. cameronensis T1 and T8. On the other hand, no intraspecific genetic

variance was observed within the ITS region for the two L. tigris specimens.

The resulting phylogenetic tree (PLATE 3) based on the ITS region indicates a

clear distinction between the Lignosus ingroups and the Daedaleopsis outgroup.

The suitability of this outgroup has been suggested and used in several studies

(Cui et al. 2010; David & Michael 1995; Sotome et al. 2008). The monophyly

(Clades A and B) of the Lignosus samples was inferred with high support

Lignosus spp. nov. (Malaysia) ... 201

FJ380871 L. rhinocerotis CH2 Malaysia

100/100/1.00 | — )g99143 L. rhinocerotis CH31 Malaysia A

L. rhinocerotis (Cooke) Ryvarden Malaysia

GU001674 L. sacer (Afzel. ex Fr.) Ryvarden

84/94/1.00

GU001675 L. sacer (Afzel. ex Fr.) Ryvarden

100/100/1.00

saa ea L. ekombitii Douanla-Meli Cameroon Bt

JQ409481 L. tigris K Malaysia B

100/99/1-00} 409482 L. tigris T Malaysia

100/100/1.00

JQ409483 L. cameronensis T1 Malaysia

92/59/1.00 B2

JQ409484 L. cameronensis T8 Malaysia

GU580883 L. hainanensis B.K. Cui China

400/100/4.00 FJ627256 Daedaleopsis sinensis

FR686551 Daedaleopsis confragosa

0.03

PLaTE 3. Maximum likelihood 50% majority-rule consensus tree based on the ITS region. Numeric

values at nodes are arranged in the order of ML bootstrap support/MP bootstrap support/Bayesian

posterior probabilities.

(ML = 100%; MP = 100%; BI = 1.00). The L. rhinocerotis sequences appeared

as a sister group (Clade A) to the remaining Lignosus specimens. Clade B

(ML = 92%; MP = 59%; BI = 1.00) is composed of L. cameronensis (Subclade

B1); L. tigris, L. sacer, and L. ekombitii (Subclade B2); and L. hainanensis. The

association between subclades B1 and B2 is currently unresolved. Lignosus

tigris specimens (K and T) and L. cameronensis specimens (T1 and T8) each

grouped with high support.

Discussion

Species currently placed in Lignosus generally have similar gross

morphologies. Variations in size and dimensions of either sclerotia or

basidiocarps are not characteristically unique to the species. Similarly, there

is little variation between the hyphal systems, sclerids, etc. on a microscopic

level. Currently, the sizes of the pores and basidiospores are the two characters

sufficiently reliable to be used for species identification (TABLE 2). Lignosus

tigris has strong similarities with L. sacer, and L. cameronensis with L. ekombitii.

202 ... Tan, Ng & Tan

TABLE 2. Pore and basidiospore sizes in Lignosus.

ceuitns PORES BASIDIOSPORES

(per mm) (um)

L. goetzii 0.5-2 6-9 x 5-8

L. tigris 1-2 2.5-5.5 x 1.8-3.6

L. sacer 1-3 5-7 x 3-4.5

L. cameronensis 2-4 2.4-4.8 x 1.9-3.2

L. ekombitii 2-4 8.1-9.3 x 2.5-3.8

L. hainanensis 3-4 4,9-6 x 2.2-2.9

L. dimiticus 6-8 3-4.5 x 2.5-3

L. rhinocerotis 7-8 3-3.5 x 2.5-3

Lignosus tigris is differentiated from L. sacer by its larger pores (1-2 pores per

mm) and smaller basidiospores (2.5-5.5 x 1.8-3.6 um), and L. cameronensis

differs from L. ekombitii by its smaller basidiospores (2.4-4.8 x 1.9-3.2 um).

The phylogeny clearly separates Lignosus rhinocerotis from the other

Lignosus species. All sampled L. rhinocerotis specimens clustered together with

minor dissimilarities (<0.2% genetic distance), possibly resulting from spatial

and time differences. The phylogenetic association between L. cameronensis,

L. sacer, L. ekombitii, and L. hainanensis within Clade B remains unresolved

with the ITS region molecular marker. This can eventually be addressed with

longer gene regions, better molecular markers, and more specimens. The two

L. sacer sequences (GenBank GU001674 and GU001675) differ slightly, with

only two nucleotide substitutions at sites 69bp and 603 bp of the ITS region.

The phylogenetic tree indicates that L. ekombitii is quite closely related, if not

conspecific, with L. sacer; more specimens are needed to verify the identity

of L. ekombitii phenotypically and genotypically. The high support for the

monophylies of L. tigris and L. cameronensis confirms the morphological data

that separate these from the other Lignosus species.

The molecular approach has thus far proved capable of inferring reliable

phylogenetic relationships within Lignosus. More efforts in sequencing

representatives of the remaining species and a broader sampling would help

further elucidate this interesting genus.

Key to Lignosus species

LS Pikens wariie Or OCHTACEOUS Tink, se. ten k seablin tematic thea tye ecat tas telah an Reena ne beeen 2

IPreusiioht brown to dabkebrown rtm bi Wmatle md Arde Ames oe eee ee 3

REY scr CEN 48 PLO pea er 13 01 ee eo L. goetzii

2-Rores sinall6=8 per wiariin i.°, gdh gti adinact zdinecr FAthace Binet Binet A L. dimiticus

PF POLES 27 PEL IUIM) 2g hea dhe ia fet tase ge ei seed bide dade: L. rhinocerotis

SEP OLES OsPCE MII ore, Fatale: Ba peleess Pa ptees Papi eae oa Dae oa Pugs te cae teat coke fel coke food tote. 4

4. Basidiospores broadly ellipsoid to subglobose .......... 0.0... cee eee eee eee 5

4. Basidiospores oblong ellipsoid to cylindrical ......... 0... eee eee 7

Lignosus spp. nov. (Malaysia) ... 203

5 Basidiospores 5= 7p IONS 8 e5 aN ee eG ae pels eee wwe le SES L. sacer

Di DASIMIOSPOLES, 20 St: LUN) LON Ge. seh eri tee aie Rue uM dey Ped ios Mein edn scandy sable taeiate 6

Ot POresaeS PERI se At BE Rite BIEN BIEL WE Nt ee Oe eT L. tigris

GRP ORES 2 SA POTTING, 5s erly co veneebeh ce nenee beck costo 8 pcenlenyt ¥ pcenlenyt poenlrt Seetleseegt L. cameronensis

7. DastdiOspOres 6 pn JONG. uk ae x wt iae x whee Behe Fels Hehe Be ea L. ekombitii

7.-BASTGIOS POLES: AO [LIONS 2 pines 2h psoas te, BAG BRAN e Mputde HMAC wRteade ok L. hainanensis

Acknowledgments

The authors would like to express gratitude to Dr. Leif Ryvarden for providing the

specimens of Lignosus rhinocerotis and L. sacer and Dr. Clovis Douanla-Meli for the ITS

DNA sequence of L. ekombitii. The authors thank Dr. Leif Ryvarden and Dr. Yu-Cheng

Dai for the critical reviewing of the manuscript and constructive comments. Thanks are

also due to Dr. Tan Swee Lian for the grammar checking and correction.

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MY COTAXON

http://dx.doi.org/10.5248/123.205

Volume 123, pp. 205-211 January-March 2013

Pionnotes, a synonym of Dacrymyces rather than Fusarium

KEITH A. SEIFERT

Biodiversity (Mycology & Botany), Agriculture & Agri-Food Canada,

Ottawa, ON, K1A 0C6 Canada

CORRESPONDENCE TO: keith. seifert@agr.gc.ca

AxstRaActT — The holotype of Fusarium capitatum, the type of the genus Pionnotes, was

re-examined. Although Pionnotes was historically considered a synonym of Fusarium, it

should henceforth be designated a synonym of Dacrymyces, with F. capitatum a synonym of

D. chrysospermus. The generic name Pionnotes and species epithet capitatum should be

evaluated further in future phylogenetic revisions of the Dacrymycetales, where most of

the genera as currently understood are polyphyletic, and many common species such as

D. chrysospermus may represent complexes of phylogenetic species.

Key worps — Hypocreales, Dacrymyces palmatus, Guepiniopsis, taxonomy

Introduction

‘Pionnotes’ is an obscure term in the mycological lexicon. As a noun, it is

mostly used in the taxonomy of Fusarium Link for colonies that have lost the

discrete sporodochia that characterize the wild type. Its use as a descriptive term

was probably proposed first in German by Appel & Wollenweber (1910: 28) as,

“lagerartige Konidienansammlungen, die keine bestimmte Form haben und als

Pionnotes mehr oder weniger ausgebreitete Schleime darstellen.” By the time of

the first international meeting of Fusarium taxonomists held at the University

of Wisconsin in Madison in August 1924, pionnote was firmly established in

the English terminology of Fusarium taxonomy (Wollenweber et al. 1925). In

the modern literature, the term is still often used although, surprisingly, rarely

explicitly defined (Booth 1971, Nelson et al. 1983, Gerlach & Nirenberg 1982,

Burgess et al. 1994, Leslie & Summerell 2006). A pionnote is a continuous layer

of slimy conidia, usually produced in culture but also sometimes in nature,

essentially a broadly spreading sporodochium that lacks a fixed margin. On

agar, such colonies usually lack aerial mycelium and are often considered

mutants, or symptoms of cultural degeneration.

206 ... Seifert

TABLE 1. Current status of other species attributed to Pionnotes, based on a review of the

literature. Fa = Fusicolla. Fm = Fusarium.

Pionnotes sp.

P. betae (Desm.) Sacc.

P. biasolettiana

(Corda) Sacc.

P. capillacea Sacc.

P cesatii Sacc.

P. ebulliens (Fr.) Sacc.

P. flava (Fr.) Sacc.

P. flavicans

Sacc. & D. Sacc.

P. navarrae Ay.-Sacca

P. pinastri P. Karst.

P. polysciadis Henn.

P. pseudonectria Speg.

P. rhizophila (Corda) Sacc.

P. sanguinea (Fr.) Sacc.

P. solani-tuberosi

(Desm.) Sacc.

P. uda (Berk.) Sacc.

P. vagans Speg.

P. violacea

Lambotte & Fautrey

P. viridis Lechmere

CURRENT CLASSIFICATION

= Fa betae (Desm.) Bonord.

?= Fa merismoides (Corda)

Grafenhan et al.

¢= Dialonectria ullevolea Seifert

& Grafenhan

?= Elsinoe ampelina Shear

?= Fm avenaceum (Fr.) Sacc.

type lost, identity unknown

¢= Fm graminearum

Schwabe sensu lato

Unknown

?= Linodochium hyalinum

(Lib.) Hohn.

= Cercosporella polysciadis

(Henn.) Hansf.

?= Microcera coccophila Desm.

?= Fa merismoides

¢= Fm avenaceum

?= Fm solani (Mart.) Sacc.

?= Fa merismoides

¢= Fm sambucinum Fuckel

?= Fm lateritium Nees

= Fm solani sensu lato

REFERENCE

Grafenhan et al. 2011

[lectotype K(M) 167520!]

Wollenweber & Reinking 1935

(as Fusarium)!

Wollenweber & Reinking 1935

[as Gloeosporium ampelophagum

(Pass.) Sacc.]

Wollenweber & Reinking 1935

Schroers et al. 2008

Wollenweber & Reinking 1935

no subsequent treatment

Wollenweber & Reinking 1935

Braun 1995 (lectotype, B!)

Wollenweber & Reinking 1935

(as Fusarium)

Wollenweber & Reinking 1935

Wollenweber 1916

(type illus., no. 418)

"The synonymies of Wollenweber & Reinking (1935) are considered tentative here, unless there is an

indication in the series Fusaria autographice delineata (Wollenweber et al. 1916, etc.) that the type

was examined.

The association of this term with Fusarium is not a coincidence; it is derived

from the generic name Pionnotes Fr. (Fries 1849), generally listed as a synonym

of Fusarium (e.g., Wollenweber & Reinking 1935). The generic name itself

was derived from the Greek word pidn, meaning ‘fat, referring to the texture

of such colonies in nature. The “Wollenweber system’ of Fusarium taxonomy,

while not accepting the genus, incorporated the name into section Eupionnotes

Pionnotes, a synonym of Dacrymyces ... 207

for several slow-growing species that typically produce such colonies. ‘This

section continued to be used in most taxonomic treatments of Fusarium until

its species were transferred to Fusicolla Bonord. and other genera by Grafenhan

et al. (2011).

The putative but unconfirmed synonymy of Pionnotes with Fusarium was

based partly on the origin of the basionym of the type species as F capitatum

Schwein. In his first treatment of hyphomycetes, Saccardo (1886) transferred

nine additional species to Pionnotes. Although he did not alter the generic

concept, he noted that, “Verisimiliter huc ducende sunt et alize Fusarii species.”

An additional eight species and one variety were added to Pionnotes by various

authors after this influential compilation. Taxa attributed to Pionnotes are

briefly reviewed in TABLE 1.

This paper presents observations on the holotype of F capitatum, leading

to the reconsideration of the generic name Pionnotes presented here. The

surprising result shifts this name to the Basidiomycota, where it will need to be

considered in phylogenetic reassessments of the generic concept of Dacrymyces

Nees. This synonymy was reported cursorily by Grafenhan et al. (2011) and

Seifert et al. (2011); the details are published and discussed here.

This is the second of a planned series of papers on the typification and

nomenclature of taxa that have been attributed to or confused with Fusarium

(Seifert & Grafenhan 2012).

Taxonomy

Pionnotes Fr., Summa Veg. Scand. 2: 481. 1849.

TYPE SPECIES: Pionnotes capitata (Schwein.) Fr., Summa Veg. Scand. 2: 481. 1849.

= Fusarium capitatum Schwein., Trans. Amer. Philos. Soc. 4(2): 302. 1832.

TYPE: ©3032 12 Syn. Fung., Fusarium capitatum Schwein. Penns.’ (Schweinitz Herbarium,

PH 01089966, holotype).

The protologue of E capitatum reads:

“3032. 12. F CAPITATUM, L. v. S., in ligno putrido Pini canadensis prope Factory,

Pennsyly.

E sporodochiis induratis, capitato-gyroso-expansis, (ut fere Tremella) ex

aurantio rubris, majusculis, subcompresso-stipitatis. Aquae immersa, omnino

solvuntur in sporidia majuscula, diaphana, teretia et vermiformia aut flexuosa,

apicibus obtusatis.”

In proposing Pionnotes, Fries (1849) paraphrased Schweinitz’s diagnosis, but

added no details, suggesting that he probably did not see a specimen himself:

“{LVI. Pionnotes. Fr.

Sporae majusculae, cylindricae, flexuosae, pellucidae in massam gelatinosam

rigescentem (aurantio-rubram) effiguratam junctae. Typus Fusar. capitatum

Schw. et pl. sp. exot.]”

208 ... Seifert

The type specimen of E capitatum is a small piece of wood with several dry,

hard, dark orange fruiting bodies of variable size (Fic. 1), the largest about 9

mm long and 4 mm wide. The smallest are more or less convex, but the larger

are lobed and were probably originally cerebriform. When rewetted, the fruiting

bodies are paler orange or yellow. Their surface is covered by what appears to

be a dense hymenium with clavate probasidia that sometimes start to become

bifurcate (Fics 2, 3). No mature basidia were seen. Abundant 7-septate spores

(Fic. 4) cover the surfaces of the fruiting body. They are 20-25.5 x 6-7.5 um,

cylindrical to oblong and usually curved, with the most acute curvature near

the base above a conspicuous papillate hilum.

The observations suggest that this is the common _basidiomycete

Dacrymyces chrysospermus Berk. & M.A. Curtis, as described by Kennedy

(1956) and McNabb (1973), among others. This is a well-known species in

eastern North America where the type of F. capitatum was collected, and it

commonly grows on recently dead, fallen or cut Tsuga canadensis, the substrate

reported by Schweinitz (as Pinus canadensis). When dry, the fruiting bodies of

D. chrysospermus collapse from their hydrated, gelatinous state to hard, orange

structures identical to those on the holotype of FE. capitatum.

Discussion

Observations of the type specimen of Fusarium capitatum confirm that it is

not a hyphomycete, but a basidiomycete identical to Dacrymyces chrysospermus.

The taxonomic status of this epithet, and the genus it typifies, Pionnotes, must

now be reevaluated.

In common with many classical fungal taxa, the pre-molecular generic

concepts in the Dacrymycetales are challenged by new information. Shirouzu

et al. (2007, 2009) used nuclear ribosomal large subunit (LSU) sequences to

show that Dacrymyces as monographed by McNabb (1973) is polyphyletic.

In their analyses, members of other morphologically defined genera, such

as Guepiniopsis Pat., Calocera (Fr.) Fr., and Dacryopinax G.W. Martin, are

intercalated in the Dacrymyces phylogeny. The clade including the type species of

Dacrymyces, D. stillatus Nees, is separated from that including D. chrysospermus

by Guepiniopsis buccina (Pers.) L.L. Kenn. The type species of Guepiniopsis,

G. tortus Pat., is as yet unsequenced and the taxonomic consequences of this

paraphyly are presently uncertain. Phylogenetic reevaluation of the generic

concepts in the Dacrymycetales is forthcoming, and Pionnotes must now be

considered as a potential name for the clade including D. chrysospermus, if it is

to be recognized as distinct at the generic rank. Additionally, these published

LSU phylogenies provide preliminary evidence for phylogenetic speciation

within many morphologically defined species in this order, including

D. chrysospermus.

Pionnotes, a synonym of Dacrymyces ... 209

Fics 1-4. Fusarium capitatum, holotype. 1. Basidiomata. 2, 3. Two probasidia becoming bifurcate.

4, Basidiospores (composite photograph). Scale bars: Fig. 1 = 1 mm. Fig. 4 (also 2, 3) = 10 um.

Dacrymyces chrysospermus was known for many years, and is still often

referred to in field guides, as D. palmatus Bres. The confusion over these two

names requires some explanation. Dacrymyces palmatus was based on Tremella

palmata Schwein. 1832, an illegitimate name (McNeill et al. 2012, ICN Art. 53.1)

because of the earlier Tremella palmata Schumach. 1803. Bresadola (Hoéhnel

1904) transferred the epithet palmata to Dacrymyces, a nomenclatural act that

legitimized it as a new name in Dacrymyces dating from 1904 rather than 1832

(ICN Art. 58.1). McNabb (1973) referred to both “T. palmata Schwein’? and

“D. palmatus (Schwein.) Bres.” [sic] as “nom. nud’, his designation for what

we now call illegitimate names. The next available name known to McNabb

(1973) was D. chrysospermus Berk. & M.A. Curtis, the binomial adopted in

210 ... Seifert

most subsequent academic literature. Considering the synonyms of Dacrymyces

chrysospermus presented by McNabb (1973), Fusarium capitatum would be

the earliest known legitimate name for this species. However, the epithet is

already occupied in Dacrymyces, ironically by another name proposed in the

same publication of Schweinitz (1832), D. capitatus Schwein. Thus, F. capitatum

must be considered as a potential species epithet for phylogenetically delimited

segregate species of the D. chrysospermus complex only if they are classified in

a genus other than Dacrymyces.

In the absence of broader species sampling and multigene phylogenies for

the Dacrymycetales, name changes would be premature and none are proposed

here. Indeed, the long association of Pionnotes with the Fusarium literature may

make it more prudent to reject this genus name as a potential source of error

in interpreting historical literature, in the spirit of ICN Art. 57.1. This decision

should be left to the taxonomic practitioners working with the Dacrymycetales

or the mycologists now considering the creation of lists of protected or rejected

names for inclusion in future Codes. This paper will serve its purpose by

bringing these possibilities to their attention.

Acknowledgments

Iam grateful to the curator of the Academy of Natural Sciences at Philadelphia (PH)

for the loan of the type specimen of Fusarium capitatum. This paper benefitted from

discussions with my colleagues Scott Redhead, Walter Gams, and Tom Grafenhan,

who also reviewed the manuscript. This paper is dedicated to the memory of my MSc

supervisor, R.J. Bandoni (1926-2009), who introduced me to the Dacrymycetales but

failed to deflect me away from the hyphomycetes.

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Arbeiten aus der Kaiserlichen anstalt fir Land- und Forstwirtschaft 8: 1-207.

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Braun U. 1995. A monograph of Cercosporella, Ramularia and allied genera (phytopathogenic

hyphomycetes). Vol. 1. IHW-Verlag, Eching bei Miinchen. 333 p.

Burgess LW, Summerell BA, Bullock S$, Gott KP, Backhouse D. 1994. Laboratory manual for

Fusarium research. 3™ ed. University of Sydney/Royal Botanic Gardens, Sydney, Australia.

134 p.

Fries EM. 1849. Summa vegetabilium Scandinaviae. Sectio posterior. pp. 259-572. Typographia

Academica, Uppsala.

Gerlach W, Nirenberg HI. 1982. The genus Fusarium - a pictorial atlas. Mitteilungen aus der

biologischen Bundesanstalt fiir Land- und Forstwirtschaft Berlin-Dahlem, 209: 1-406.

Grafenhan T, Schroers H-J, Nirenberg HI, Seifert KA. 2011. An overview of the taxonomy, phylogeny,

and typification of nectriaceous fungi in Cosmospora, Acremonium, Fusarium, Stilbella, and

Volutella. Studies in Mycology 68: 79-113. http://dx.doi.org/10.3114/sim.2011.68.04

Héhnel, F von. 1904. Mykologisches. Osterreichische botanische Zeitschrift 54: 425-439.

http://dx.doi.org/10.1007/BF01794052

Pionnotes, a synonym of Dacrymyces ... 211

Kennedy LL. 1956. Dacrymyces palmatus. Mycologia 48: 311-319.

http://dx.doi.org/10.2307/3755479

Leslie JE, Summerell BA. 2006. The Fusarium laboratory manual. Blackwell Publishing, Ames,

Iowa. 388 p.

McNabb RER. 1973. Taxonomic studies in the Dacrymycetaceae VIII. Dacrymyces Nees ex Fries. New

Zealand Journal of Botany 11: 461-524. http://dx.doi.org/10.1080/0028825X.1973.10430296

McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W, Hawksworth DL, Herendeen PS, Knapp

S, Marhold K, Prado J, Prud’ homme van Reine WF, Smith GF, Wiersema JH, Turland NJ. 2012.

International code of nomenclature for algae, fungi, and plants (Melbourne Code). Regnum

Vegetabile 154: 1-240. A.R.G. Gantner Verlag KG.

http://www.iapt-taxon.org/nomen/main.php

Nelson PE, Toussoun TA, Marasas WFO. 1983. Fusarium species, an illustrated manual for

identification. Pennsylvania State Univ. Press, University Park, London. 193 p.

Saccardo PA. 1886. Sylloge fungorum omnium hucusque cognitorum, vol. 4. Published by the

author, Padova. 807 p.

Schroers H-J, O'Donnell K, Lamprecht SC, Kammeye PL, Johnson S, Sutton DA, Rinaldi MG,

Geiser DM, Summerbell RC. 2008. Taxonomy and phylogeny of the Fusarium dimerum species

group. Mycologia 101: 44-70. http://dx.doi.org/10.3852/08-002

Schweinitz LD von. 1832. Synopsis fungorum in America boreali media degentium. Transactions

of the American Philosophical Society 4: 141-316. http://dx.doi.org/10.2307/ 1004834

Seifert KA, Grafenhan T. 2012. Lectotypification and characterization of the natural phenotype of

Fusarium bactridioides. Mycotaxon 120: 415-422. http://dx.doi.org/10.5248/120.415

Seifert KA, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series no. 9: 1-997.

Shirouzu T, Hirose D, Tokumasu S. 2007. Sequence analysis of the 28S gene D1/D2 region suggests

Dacrymyces (Heterobasidiomycetes, Dacrymycetales) is polyphyletic. Mycoscience 48: 388-394.

http://dx.doi.org/10.1007/s10267-007-0378-0

Shirouzu T, Hirose D, Tokumasu S. 2009. Taxonomic studies of Japanese Dacrymycetes. Persoonia

23: 16-34. http://dx.doi.org/10.3767/003158509X468443

Wollenweber HW. 1916. Fusaria autographice delineata [1]: nos 1-509. Self published, Berlin.

Wollenweber HW, Reinking OA. 1935. Die Fusarien, ihre Beschreibung, Schadwirkung und

Bekampfung. Paul Parey, Berlin. 355 p.

Wollenweber HW, Sherbakoff CD, Reinking OA, Johann H, Bailey AA. 1925. Fundamentals for

taxonomic studies of Fusarium. Journal of Agricultural Research 30: 833-843.

MY COTAXON

http://dx.doi.org/10.5248/123.213

Volume 123, pp. 213-220 January-March 2013

A new species of Stropharia from Western Ghats, India

GUNASEKARAN SENTHILARASU*? & SANJAY K SINGH?

National Facility for Culture Collection of Fungi, MACS’ Agharkar Research Institute,

G. G. Agarkar Road, Pune-411 004, India

CORRESPONDENCE TO”: 'senthilarasug@rediffmail.com & *singhsksingh@gmail.com

ABSTRACT—Stropharia rubrobrunnea, characterized by its caespitose habit, smooth

grayish red or reddish to violet brown hygrophanous pileus, annular stipe, and dimorphic

cheilocystidia, is described and illustrated as a new species. It is distinguished from

S. rugosoannulata by its stipe color and basidiospore size.

Key worps—Agaricales, Basidiomycota, dark-spored agarics, diversity, taxonomy

Introduction

The genus Stropharia (Fr.) Quél. (Strophariaceae Singer & A.H. Sm.) is

poorly known from India, where only ten species have been recorded (Manjula

1983; Natarajan et al. 2005; Farook et al. 2013; Kumaresan & Senthilarasu,

unpublished check list). Of these, S. bicolor Pegler, S. rugosoannulata Murrill

(Manimohan et al. 2007), and S. aurantiaca (Cooke) M. Imai (Natarajan

& Raman 1983) were recently recorded from southern India. Two excluded

species are S. pokhraensis Dhanch. & Bakhukh. (Dhancholia & Bakhukhandi

1992), poorly known, and S. semiglobata (Batsch) Quél. (Natarajan & Raman

1983; Vrinda & Pradeep 2011; Mohanan 2011), which lacks true acanthocytes

(Moncalvo et al. 2002) and has been transferred to Protostropharia semiglobata

(Batsch) Redhead et al. The other poorly known species earlier recorded

from India are Stropharia aureofulva (Berk.) Sacc. (as Agaricus aureofulvus),

S. gollanii Henn., S. mephistopheles (Cooke) Sacc. (no material or illustration at

Kew), S. psathyroides Henn., and S. pygmaea Henn. (Berkeley 1850, Hennings

1900). In the present paper, a new species collected from Western Ghats,

Stropharia rubrobrunnea, is described, illustrated, and compared with closely

related species.

Materials & methods

Thin handmade sections were made from dried specimens rehydrated with

alcohol were mounted in 10% KOH and stained in 3% phloxine or cotton blue prior

214 ... Senthilarasu & Singh

to microscopical examination. Approximately 50 basidiospores were measured; the

average spore range includes extreme values in parentheses. Colour terminologies follow

Kornerup & Wanscher (1978). The type specimens are deposited at Ajrekar Mycological

Herbarium (AMH), MACS’ Agharkar Research Institute, Pune, India.

Taxonomy

Stropharia rubrobrunnea Senthil. & S.K. Singh, sp. nov. PLATE 1-3

MycoBank MB 564359

Differs from S. rugosoannulata by its white, smooth stipe and smaller basidiospores.

Type: India, Maharashtra state, Sinhgad, 18°21'56.39"N 73°45'18.97"E, 04.09.2011, coll.

G. Senthilarasu (holotype, AMH 9447).

Erymo toy: ‘rubrobrunnea’ refers to the colour of the pileus.

PILEus 6-18 mm (closed pileus) to 30-100 mm (matured pileus), hemispheric to

broadly parabolic, not umbonate, slightly areolate, exposing white background

when young; becoming convex to plane, slightly depressed, finally uplifted,

broadly umbonate in the shallow depression; surface grayish red (8C4-9C4),

dull red (8D4), reddish brown (9E7), changing to violet brown (10E7-10E8) to

grayish red (11D5) when young, strongly hygrophanous, pinkish white (7A2-

11A2-11A3) on orange white (5A2) ground, becoming yellowish white (4A2),

moist, smooth; margin incurved, becoming decurved to plane, finally uplifted,

irregularly appendiculate, fugacious, not striate, entire, becoming eroded.

LAMELLAE adnexed to adnate, <5 mm wide, grayish red (11D4) to violet brown

(11F4-11F5), becoming blackish brown, crowded with numerous lamellulae,

eroded. Stipe central, 30-120 x 2-13 mm, terete, equal, slightly tapering

towards base; surface white, shiny, smooth, longitudinally striate, cartilaginous

to fibrous, solid, arising from white to yellowish white (4A2) rhizomorphs.

ANNULUS membranous, white, becoming vinaceous, rugulose, attached at the

one third of the stipe. PILEUS CONTEXT white, <3 mm thick.

BASIDIOSPORES (7-)7.5-8.5(-10) x (4.5-)5-5.5(-6.5) um, (8.01 + 0.36 x

5.47 + 0.28) um, Q = 1.46, ellipsoid, slightly rhomboid in face-view, brown with

thickened dark wall, apically truncated by a broad germ-pore, smooth. BASIDIA

17-21 x 7-8 um, cylindric clavate, tetrasporic; sterigmata <4 x 1.5 um. LAMELLA

EDGE Sterile with crowded dimorphic cheilocystidia: CHEILOCHRYSOCYSTIDIA

19-38 x 6.5-11 um, sublageniform with a short mucronate apex, with subhyaline

to yellowish contents, thin-walled. CHEILOLEPTOCYSTIDIA 20-52 x 6.5-10

um, clavate to cylindric with mucronate to rostrate apex; rostrum <30 x 2.5

um, sometimes sphaeropedunculate, branched, thin-walled. CarysoOcysTIDIA

abundant on the sides of the gills, 20-51 x 7-14 um, lageniform with short

mucronate apex, containing subhyaline to yellowish contents, thick-walled.

HYMENOPHORAL TRAMA regular, with thin-walled, hyaline hyphae, 3-20 um

diam. SUBHYMENIUM well developed, <20 um wide, pseudoparenchymatous.

Stropharia rubrobrunnea sp. nov. (India) ... 215

PLATE 1. Stropharia rubrobrunnea basidiomata under natural conditions: a. Young basidiomata.

b. Mature basidiome showing rugulose annulus. c. Gill view of mature basidiome. (Photo

Senthilarasu.)

PILEAL SURFACE a regular cutis of radially repent hyphae, 2-30 um diam.

PILEAL CONTEXT with hyaline hyphae 3-23 um diam., thin-walled with clamp

connections. STIPITIPELLIS with hyaline hyphae 2-17 um diam., thin-walled,

clamped. RH1IzZOMORPH with sphaerocytes and hyaline hyphae, <7 um diam,

clamped; sphaerocytes <96 x 82 um, globose to subglobose or ovate, hyaline,

thin-walled; acanthocytes with several outgrowths, <83 x 3.7 um, thick-walled,

<1.7 um thick, arising from a solid base, hyaline.

216... Senthilarasu & Singh

1 5 2 Oa

PLATE 2. Stropharia rubrobrunnea: a. Basidiospores. b. Basidia. c. Cheilochrysocystidia. d. Cheilo-

leptocystidia. e. Pleurochrysocystidia.

ECOLOGY & DISTRIBUTION: Solitary to caespitose to connate, on decayed

gunny bag, in mixed forest vegetation. Known only from type locality.

The diagnostic features of Stropharia rubrobrunnea are the grayish red, reddish

brown to violet brown pileus, white stipe with a membranous grooved annulus,

and spores <10 um long.

Recently, several Stropharia species have been described from the

neotropical region (Cortez & Coelho 2004, 2008, Bandala et al. 2005, Silva et

Stropharia rubrobrunnea sp. nov. (India) ... 217

PLATE 3. Stropharia rubrobrunnea rhizomorph: a. Sphaerocytes along with

acanthocytes. b. Hyphae with clamp connections. c. Acanthocyte.

218 ... Senthilarasu & Singh

al. 2006, 2009, Cortez & Silveira 2007, 2008, Cortez 2008a,b) and China (Bau

& Meng 2008). Stropharia rubrobrunnea resembles the most common species

S. rugosoannulata (Cortez & Silveira 2008) in similar sized basidiomes.

Although the pileus colour in S. rugosoannulata is highly variable, it has a

yellowish brown stipe covered with brownish fibrils and larger basidiospores

(9.5-14.5 x 6.5-8 um) and basidia (21-38 x 8-12 um).

Stropharia coronilla (DC.) Quél. and S. araucariae Cortez & R.M. Silveira

(Cortez & Silveira 2008) also have a fleshy grooved annulus. Although

S. araucariae also has leptocystidia and chrysocystidia at the gill edge, it differs

from S. rubrobrunnea in its umbonate dark grayish brown pileus and larger

basidiospores (10.5-13 um long). The basidiospore size (6.5-10.5 x 4.5-6.5

uum) in S. coronilla is similar to that in S. rubrobrunnea, but it is a grassland

inhabiting mushroom and differs morphologically in smaller basidiome size

(pileus = 16-55 mm diam.; stipe = 18-51 mm long), yellowish colored pileus,

and absence of cheilochrysocystidia.

The neotropical species S. venusta PS. Silva et al., known only from Araucaria

angustifolia forests of southern Brazil (Silva et al. 2009), also somewhat

resembles S. rubrobrunnea in the larger basidiome size, grayish red to reddish

brown pileus, and rugulose annulus. However, S. venusta clearly differs in its

greenish grey colored stipe covered with scattered white squamules, larger

basidiospores (11-15 x 6-10 um) and chrysocystidia (35-70 x 7-16 um), and

absence of cheilochrysocystidia.

Stropharia variicolor Desjardin & Hemmes (Desjardin & Hemmes 2001)

and S. cifuentesii Bandala et al. (Bandala et al. 2005) with their reddish to

reddish brown colored pilei somewhat superficially resemble S. rubrobrunnea.

Stropharia variicolor also has similar sized basidiospores (6.5-9 x 4.5-5.7 um)

but differs in smooth annulus, smaller cheiloleptocystidia (17.5-26 um) that

lack a rostrum, and an absence of cheilochrysocystidia. Stropharia cifuentesii

clearly differs in its smaller basidiome size (pileus = 8-30 mm diam.; stipe =

9-37 mm long), fibrillose to squamulose pileus, whitish to yellowish brown

lamellae, more slender stipe, absence of a fleshy annulus, smaller spores (5.5-7

um), and absence of cheilocystidia.

Acknowledgments

We thank Prof. Vagner G. Cortez and Dr. Gast6n Guzman for critically reviewing

the manuscript. GS personally thanks Prof. V. Cortez, Dr. V. Kumaresan and Dr. CK

Pradeep for providing literature on Stropharia. We greatly acknowledge Dr. Shaun

Pennycook and Dr. Lorelei Norvell for presubmission reviews. Sincere thanks to the

Director, Agharkar Research Institute, for providing all laboratory facilities. We thank

the Department of Science and Technology (DST), Government of India, New Delhi,

for providing financial support for the project National Facility for Culture Collection

of Fungi, Agharkar Research Institute, Pune, India.

Stropharia rubrobrunnea sp. nov. (India) ... 219

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new records. Fungal Diversity 20: 17-29.

Bau T, Meng TX. 2008. Strophariaceae of China (II) Stropharia. Journal of Fungal Research 6(1):

7-34.

Berkeley MJ. 1850. Decades of fungi. Decades XXV to XXX. Sikkim Himalayan fungi, collected

by Dr. J.D. Hooker. Hooker's Journal of Botany and Kew Gardens Miscellany 2: 42-51, 76-88,

106-112.

Cortez VG. 2008a. Type studies on South American Strophariaceae: 1. Pholiota varzeae from the

Brazilian Amazon. Mycotaxon 103: 137-140.

Cortez VG. 2008b. Type studies on South American Strophariaceae: 2. Pholiota trinitensis is

transferred to Stropharia. Mycotaxon 105: 7-10.

Cortez VG, Coelho G. 2004. The Stropharioideae (Strophariaceae, Agaricales) from Santa Maria,

Rio Grande do Sul, Brazil. Mycotaxon 89(2): 355-378.

Cortez VG, Coelho G. 2008. Occurrence of the rare agaric Stropharia melanosperma (Strophariaceae)

in Brazil. Bol. Soc. Micol. Madrid 32: 49-55.

Cortez VG, Silveira RMB. 2007. A new species of Stropharia with hymenial acanthocytes. Mycologia

99(1): 135-138. http://dx.doi.org/10.3852/mycologia.99.1.135

Cortez VG, Silveira RMB. 2008. The agaric genus Stropharia (Strophariaceae, Agaricales) in Rio

Grande do Sul State, Brazil. Fungal Diversity 32: 31-57.

Dhancholia S, Bakhukhandi D. 1992 [“1991”]. New records of agarics from India. Indian Journal of

Mycology and Plant Pathology 21(3): 248-250.

Desjardin DE, Hemmes DE. 2001. Agaricales of the Hawaiian Islands-7. Notes on Volvariella,

Mycena sect. Radiatae, Physalacria, Porpoloma and Stropharia. Harvard Papers in Botany 6(1):

85-103.

Farook AV, Khan SS, Manimohan P. 2013. A checklist of agarics (gilled mushrooms) of Kerala State,

India. Mycosphere 4(1): 97 - 131. http://dx.doi.org/10.5943/mycosphere/4/1/6

Hennings P. 1900. Fungi Indiae Orientalis. Hedwigia Beiblatter 39: 150-153.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3 ed. Eyre Methuen, London

243 p.

Manjula B. 1983. A revised list of the agaricoid and boletoid basidiomycetes from India and Nepal.

Proc. Indian Academy of Sci. (Pl. Sci.) 92: 81-213.

Manimohan P., Thomas AK, Nisha VS. 2007. Agarics on elephant dung in Kerala. Mycotaxon 99:

147-157.

Mohanan C. 2011. Macrofungi of Kerala. KFRI Handbook No. 27, Kerala Forest Research Institute,

Peechi, Kerala, India 597p.

Moncalvo JM, Vilgalys R, Redhead SA, Johnson JE, James TY, Aime MC, Hofstetter V, Verduin

SJW, Larsson E, Baroni TJ, Thorn RG, Jacobsson $, Clémengon H, Miller Jr. OK. 2002. One

hundred seventeen clades of euagarics. Molecular Phylogenetics and Evolution 23: 357-400.

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Natarajan K, Kumaresan V, Narayanan K. 2005. A checklist of Indian agarics and boletes

(1984-2002). Kavaka 33: 61-128.

Natarajan K, Raman N. 1983. South Indian Agaricales. A preliminary study on some dark spored

species. Bibliotheca Mycologica 89: 1-203.

Silva PS, Cortez VG, Silveira RMB. 2006. The mycobiota of Itapua Park, Rio Grande do Sul, Brazil.

I. Species of Strophariaceae (Agaricales). Mycotaxon 97: 219-229.

220 ... Senthilarasu & Singh

Silva PS, Cortez VG, Silveira RMB. 2009. New species of Stropharia from Araucaria angustifolia

forests of southern Brazil. Mycologia 101(4): 539-544. http://dx.doi.org/10.3852/08-097

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Mycopathological Research 49(2): 231-246.

MY COTAXON

http://dx.doi.org/10.5248/123.221

Volume 123, pp. 221-228 January-March 2013

Two new species of Endophragmiella from Spain

MARGARITA HERNANDEZ-RESTREPO’, JULIO MENA-PORTALES’,

JOSEP GUARRO’ & JOSEPA GENE**

'Unitat de Micologia, Facultat de Medicina i Ciéncies de la Salut, Universitat Rovira i Virgili,

43201 Reus, Tarragona, Spain

? Instituto de Ecologia y Sistematica, Ministerio de Ciencia, Tecnologia y Medio Ambiente de Cuba

(CITMA), A. P. 8029, Ciudad de la Habana 10800, Cuba

*CORRESPONDENCE TO: josepa.gene@urv.cat

ABSTRACT — Two new species of Endophragmiella are described and _ illustrated.

Endophragmiella bicolorata is characterized by (1-)2(-3)-septate conidia with a dome-

shaped hyaline to subhyaline apical cell and brown basal and middle cells, a pigmentation

pattern not previously described for species of this genus. Endophragmiella cantabrica is

distinguished from species with 1-septate conidia by oblong, ellipsoidal, or sometimes ovoid

conidia comprising two equal and uniformly pale brown cells.

KEY worpDs — anamorphic fungi, plant debris, taxonomy

Introduction

Sutton (1973) established Endophragmiella for two species: E. pallescens

B. Sutton, selected as type, and E. canadensis (Ellis & Everh.) B. Sutton, currently

considered a synonym of E. subolivacea (Ellis & Everh.) S. Hughes. Hughes

(1979) clarified the conidiogenesis in Endophragmiella, and amended the genus

definition. Subsequently, Kirk (1985), Holubova-Jechova (1986), and Wu &

Zhuang (2005) contributed to the knowledge of the genus with keys based on

morphological revisions of the species.

Endophragmiella is characterized by simple or branched conidiophores,

monoblastic percurrently proliferating conidiogenous cells, and conidia with

very variable morphology and septation, seceding rhexolytically. The species

are mostly saprobes occurring on rotten wood, dead branches, and decaying

leaves of different plants. Currently, more than 80 species are accepted in the

genus (Rifai 2008, Ma et al. 2011, Ren et al. 2011, Seifert et al. 2011).

During an extensive survey of anamorphic fungi on plant debris in the

Iberian Peninsula, two Endophragmiella species were found. We propose these

as new species based on conidial morphology, size, and pigmentation.

222 ... Hernandez-Restrepo & al.

Materials & methods

Samples of plant material were processed using the methodology described by

Castafieda-Ruiz (2005). Fungal specimens were studied and illustrated following the

procedure described by Hernandez-Restrepo et al. (2012). All attempts to isolate the

specimens in pure culture failed, despite trying different media and culture conditions;

therefore, only dried material is preserved. Specimens are deposited in the herbariums

of the CBS-KNAW Fungal Biodiversity Centre, Utrecht, the Netherlands (CBS) and

Faculty of Medicine, Reus, Spain (FMR).

Taxonomy

Endophragmiella bicolorata Hern.-Rest., J. Mena, Guarro & Gené, sp. nov.

MycoBank. MB 800594 FIGS 1, 2

Differs from Endophragmiella oblonga by the conidial cell pigmentation pattern and

dome-shaped apical cell.

TYPE: Spain, Valencia, Km 21 Road CV 425, Los Pedrones direction to Bufiol, 39°18'28"N

0°52'38"W, on dead wood, 15/3/2010, M. Hernandez-Restrepo & K. Rodriguez

(Holotype, CBS H-21042; Isotype, FMR 10965).

Erymo.oey: Latin bicolorata, referring to the pigmentation of conidia.

Co.tonies on the natural substratum effuse, hairy, black. Mycelium mostly

immersed, composed of septate, smooth, subhyaline to pale brown hyphae, 2-3

um wide. CONIDIOPHORES macronematous, simple, erect, straight or slightly

flexuous, smooth, septate, brown, paler towards the apex, up to 125 um long, 2-

5 um wide at the base, with up to 9 percurrent proliferations. CONIDIOGENOUS

CELLS monoblastic, integrated, terminal, percurrent, cylindrical, tapered to

a truncate apex. CONIDIA acrogenous, solitary, simple, ellipsoidal to oblong

ellipsoidal, sometimes obclavate or obovoid, (1-)2(-3)-septate, often constricted

at the septa, basal and middle cells brown, apical cell hyaline to subhyaline and

dome-shaped, smooth all over, 19-26.5 x 7.5-13 um, with a distinct subhyaline

basal frill, 1.5-4 x 2-3 um. TELEOMORPH unknown.

Note: This species resembles Endophragmiella oblonga (Matsush.) S. Hughes

in morphology and conidial size, but the E. oblonga conidia are less variable

in shape (Matsushima 1975, Hughes 1979). Although both species have a

dome-shaped apical cell, their pigmentation pattern distinguishes the two; in

E. oblonga the middle cell is brown and the end cells are pale brown (Hughes

1979, Melnik 2000).

Other Endophragmiella species with 2-septate conidia somewhat similar

to E. bicolorata are E. collapsa (B. Sutton) S. Hughes, E. ontariensis S. Hughes,

E. suboblonga W.P. Wu, and E. tripartita S. Hughes. However, the conidia of

E. collapsa are smaller (14.4-17(-20) x 7.2-8.3(-9) um) and have pale brown

to dark brown distal cells and a basal cell that is paler and sometimes collapsed

(Sutton 1973, Hughes 1979, Wu & Zhuang 2005). In E. ontariensis, the conidia

Endophragmiella spp. nov. (Spain) ... 223

Fic. 1. Endophragmiella bicolorata (CBS H-21042). a. Habit. b. Conidiophores and conidiogenous

cells producing conidia. c. Conidia. Scale bars: a = 100 um; b,c = 10 um.

are also smaller (18-22 x 8.5-11 um) and have pale brown to dark brown

distal cells and pale brown basal cells (Hughes 1978a, Wu & Zhuang 2005).

The conidia of E. suboblonga measure 18-20 x 9-13 um and have the middle

224 ... Hernandez-Restrepo & al.

Fic. 2. Endophragmiella bicolorata (CBS H-21042). a. Conidia. b. Conidiophores, conidiogenous

cells with conidia. Scale bars: 10 um.

Endophragmiella spp. nov. (Spain) ... 225

cell brown and larger and both apical and basal cells paler and smaller (Wu

& Zhuang 2005). The conidia of E. tripartita measure (16-)18-21.5(-23.5) x

8.1-9.5 um and have all cells brown to dark brown (Hughes 1979).

Endophragmiella cantabrica J. Mena, Hern.-Rest., Guarro & Gené, sp. nov.

MycoBank. MB 800595 FIGS 3, 4

Differs from all other bicellular Endophragmiella species by its conidial size.

Type: Spain, Cantabria, Los Tojos, Barcena Mayor, Saja-Besaya Natural Park.

43°06'42.81"N 4°12'13.48"W, 675 m.a.s.l., on dead wood, 12/7/2010, M. Hernandez-

Restrepo, J. Mena & J. Guarro (Holotype, CBS H-21043; Isotype, FMR 11857).

7 |

Fic. 3. Endophragmiella cantabrica (CBS H-21043). a. Habit. b. Conidiophores and conidiogenous

cells producing conidia. c. Conidia. Scale bars: a = 200 um; b,c =10 um.

226 ... Hernandez-Restrepo & al.

{ 4 _ : i

Fic. 4. Endophragmiella cantabrica (CBS H-21043). a. Conidiophore. b. Detail of conidiogenous

cells. c. Conidia. Scale bars: 10 um.

ErymMo.oey. Latin cantabrica, referring to the Spanish region where the fungus was

collected.

Cotontgs on the natural substratum effuse, hairy, brown. Mycelium mostly

immersed, composed of septate, smooth or verruculose, subhyaline to pale

brown hyphae, 1.5-3 um wide. CONIDIOPHORES macronematous, simple,

erect, straight or slightly flexuous, smooth, septate, brown, paler towards the

apex, up to 195 um length, 5-6 um wide at the base, with up to 20 percurrent

proliferations. CONIDIOGENOUS CELLS monoblastic, integrated, terminal,

percurrent, cylindrical, tapered to a truncate apex. CONIDIA acrogenous,

solitary, simple, oblong, ellipsoidal, sometimes ovoid, 1-septate, pale brown,

with both cells equal and uniformly pigmented, smooth, 10-14 x 5-6 um, with

a distinct subhyaline basal frill, 0.5-1.5 x 0.5-1.5 um. TELEOMORPH unknown.

Norte: Among the taxa of Endophragmiella with 1-septate conidia, E. cantabrica

has some similarities with E. arranensis P.M. Kirk, E. bogoriensis Rifai, E. ovoidea

P.M. Kirk and E. uniseptata var. pusilla Hol.-Jech. Endophragmiella arranensis

has very short conidiophores (<45 um long after successive percurrent

proliferations) and smaller conidia (6.5-9.5 x 4-5 um) with the basal cell paler

Endophragmiella spp. nov. (Spain) ... 227

than the apical one (Kirk & Spooner 1984). The other three species have more

pigmented and larger conidia: 12-18 x 7-9 um in E. bogoriensis, (10—)14-16 x

(5-)5.5-6.5 um in E. ovoidea, and (9-)15(-17) x (7-)9(-10) um in E. uniseptata

var. pusilla (Kirk 1981, Holubova-Jechova 1986, Mel'nik 2000, Rifai 2008).

Other Endophragmiella taxa with 1-septate conidia slightly similar to

E. cantabrica in conidial morphology are: E. angustispora S. Hughes, E. resinae

P.M. Kirk, and E. uniseptata (M.B. Ellis) S. Hughes var. uniseptata. Conidia

of E. angustispora, however, are navicular to ellipsoidal or narrowly ovoid,

14.4-20.5 x 4.5-5.4 um (Hughes 1978b); those of E. resinae are ovoid to

pyriform, 17-22 x 9-10.5 um (Kirk 1981); and those of E. uniseptata var.

uniseptata are ellipsoidal, obovoid to pyriform, broadly obtuse at the distal end,

13-27 x 9-12 um (Ellis 1959, Hughes 1979, Holubova-Jechova 1986).

Acknowledgments

The authors would like to thank Dr. Rafael F. Castafieda Ruiz, Dr. Cony Decock, and

Dr. David Minter for their critical review of the manuscript. This study was supported

by the Spanish Ministerio de Economia y Competitividad, grant CGL 2011-27185.

Literature cited

Castafieda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Ellis MB. 1959. Clasterosporium and some allied Dematiaceae. Phragmosporae I. Mycological

Papers 72: 1-75.

Hernandez-Restrepo M, Silvera-Simon C, Mena-Portales J, Mercado-Sierra A, Guarro J. & Gené

J. 2012. Three new species and a new record of Diplococcium from plant debris in Spain.

Mycological Progress 11: 191-199. http://dx.doi.org/10.1007/s11557-011-0741-6.

Holubova-Jechova V. 1986. Lignicolous hyphomycetes from Czechoslovakia 8. Endophragmiella

and Phragmocephala. Folia Geobotanica et Phyto-taxonomica 21: 173-198.

http://dx.doi.org/10.1007/BF02854666.

Hughes SJ. 1978a. Endophragmiella ontariensis. Fungi Canadenses 128: 1-2.

Hughes SJ. 1978b. Endophragmiella angustispora. Fungi Canadenses 123: 1-2.

Hughes SJ. 1979. Relocation of species of Endophragmia auct. with notes on relevant generic names.

New Zealand Journal of Botany 17: 139-188.

Kirk PM. 1981. New or interesting Microfungi I. Dematiaceous hyphomycetes from Devon.

Transaction of the British Mycological Society 76: 71-87.

http://dx.doi.org/10.1016/S0007-1536(81)80010-1.

Kirk PM. 1985. New or interesting Microfungi XIV. Dematiaceous hyphomycetes from Mt Kenya.

Mycotaxon 23: 305-352.

Kirk PM, Spooner BM. 1984. An account of the fungi of Arran, Gigha and Kintyre. Kew Bulletin

38: 503-597.

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.

Matsushima T. 1975. Icones Microfungorum a Matsushima lectorum. Published by the author.

Kobe, Japan.

Mel'nik VA. 2000. Classis Hyphomycetes Fasc 1 Fam. Dematiaceae. Nauka. Petropoli.

228 ... Hernandez-Restrepo & al.

Ren LG, Ma JJ, Zhang XG. 2011. A new species and new records of Endophragmiella from China.

Mycotaxon 117: 123-130. http://dx.doi.org/10.5248/117.123.

Rifai MA. 2008. Endophragmiella bogoriensis Rifai, spec. nov. Reindwardtia 12: 275-276.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. CBS-KNAW Fungal Biodiversity Centre. Utrecht.

Sutton BC. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycological Papers

132: 1-143.

Wu WP, Zhuang WY. 2005. Sporidesmium, Endophragmiella and related genera from China. Fungal

Diversity Research Series 15. Fungal Diversity Press. Hong Kong.

MY COTAXON

http://dx.doi.org/10.5248/123.229

Volume 123, pp. 229-231 January-March 2013

Sporisorium linderi, a new record for Asia

MUHAMMAD F1Az',, ABDUL NASIR KHALID? & HABIB AHMAD?

"Department of Botany, Hazara University Mansehra, Pakistan

*Department of Botany, University of the Punjab, Quaid-e-Azam Campus,

Lahore, 54590, Pakistan

*Department of Genetics, Hazara University Mansehra, Pakistan

* CORRESPONDENCE TO: muhammadfiazhu@gmail.com

ABSTRACT — Sporisorium linderi (Ustilaginomycetes), reported here from Pakistan, is a new

record for Asia. Digitaria ciliaris is a new host record for smut fungi in Pakistan.

KEY worps — KPK, Poaceae, taxonomy

Introduction

During exploration of smut fungi of Mansehra District, Khyber-

Pakhtunkhwa (KPK) Province, Pakistan, Digitaria ciliaris was found infected

with Sporisorium linderi.

The sampling site, located at 34°14’-35°11'N 72°49'-74°08’E, is bounded in

the north by Batagram and Kohistan, in the east by the Muzafarabad district

of Azad Jammu and Kashmir, in the south by Abbottabad and Haripur, and in

the west by Swat and Kala Dhaka. The mountain ranges that enter Mansehra

district from Kashmir are offshoots of the great Himalayan range. ‘The district

has a seasonal temperature range of 2-36 °C (Mustafa 2003).

Sporisorium Ehrenb. ex Link is the largest genus of smut fungi and is

represented by 327 species (Vanky 2011, Denchev et al. 2013), of which 18

species have been recorded previously from Pakistan (Ahmad et al. 1997,

Vanky 2007, Denchev et al. 2013).

Materials & methods

Freshly collected infected flowers/spikelets of Digitaria ciliaris were examined

under a stereomicroscope. For light microscopic observation spores were mounted

in lactophenol on glass slides and gently heated. Preparations were observed under a

NIKON YS 100 microscope and photographed with the help of Digipro-Labomed. Spore

230 ... Fiaz, Khalid & Ahmad

Fic. 1: Sporisorium linderi on Digitaria ciliaris. A. Infected inflorescence of the host plant. B. Spores

in SEM. Scale bars: A = 1.2 cm; B= 5 um.

measurements were taken by using an ocular Zeiss Eye Piece Screw Micrometer. For

SEM, the spores were critically point dried, attached to specimen holders by double-

sided adhesive tape, and coated with a 50 nm film of gold in a Polaron E5300 freeze

drier. The gold-coated stubs were observed and photographed in a Camscan 3-30BM

Scanning Electron Microscope.

Sporisorium linderi on Digitaria ciliaris (Pakistan) ...231

Taxonomy

Sporisorium linderi (Zundel) Vanky, Mycotaxon 73: 142, 1999. Fic. 1

Sori in most of the spikelets of the inflorescence, mostly the basal spikelets

are infected, sub-globoid to cylindrical, 1.7-3 mm long, initially covered by

a thick peridium of the host origin which flakes away irregularly, exposing

dark brown powdery spore mass and a well developed central columella,

branched at tip, spore balls composed of loosely arranged cells. Spores globose

to subglobose or ovoid to irregular, light olivaceous brown to somewhat darker,

7-9.2 x 8.2-10.4 um, wall up to 0.7 um thick, moderately to densely echinulate

in LM, sparsely echinulate in SEM. Sterile cells either occur singly among the

spores or in groups or chains, usually larger than the spores, hyaline, walls up

to 0.4 um thick, smooth.

SPECIMEN EXAMINED — On leaves of Digitaria ciliaris (Retz.) Koeler: PAKISTAN,

KHYBER-PAKHTUNKHWA, Mansehra, Durbund, ca. 600 m, 3 Sep. 2010, M. Fiaz FS-24

(HUP 304).

CoMMENTs: Sporisorium linderi is known from central and southern Africa on

several Digitaria spp., including D. ciliaris (Vanky et al. 2011). This is the first

record of its occurrence in Asia. Previously, ~50 % of the known Sporisorium

species have been reported from Asia. This is the first report of a smut fungus

on Digitaria ciliaris in Pakistan (Ahmad et al. 1997, Vanky 2011).

Acknowledgements

We sincerely thank Dr. Kalman Vanky for his help in identification of the fungi

and Dr. C.M. Denchev (Institute of Biodiversity and Ecosystem Research, Bulgarian

Academy of Sciences) and Dr. Najam-ul-Sahar Afshan (Centre for Undergraduate

Studies, University of the Punjab, Lahore) for their valuable suggestions to improve the

manuscript and acting as presubmission reviewers.

Literature cited

Ahmad §S, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Sultan Ahmad Mycological Society of

Pakistan, Lahore.

Denchev CM, Fiaz M, Denchev TT, Ahmad H, Khalid AN. 2013 [“2012”]. Additions to the smut

fungi of Pakistan. 1. Mycotaxon 121: 165-170. http://dx.doi.org/10.5248/121.165.

Mustafa G. 2003. Mansehra, an introduction. Gazetteer of the Hazara District.

Shah GM, Khan MA. 2006. Check list of medicinal plants of Siran Valley Mansehra - Pakistan.

Ethnobotanical Leaflets 10: 63-71.

Vanky K. 2007. Smut fungi of the Indian Subcontinent. Polish Botanical Studies 26: 1-265.

Vanky K, Vanky C, Denchev CM. 2011. Smut fungi in Africa - a checklist. Mycologia Balcanica

$: 1277.

Vanky K. 2011 [“2012”]. Smut fungi of the world. APS Press, St. Paul, Minnesota, USA, xvii +

1458 p.

MY COTAXON

http://dx.doi.org/10.5248/123.233

Volume 123, pp. 233-234 January-March 2013

New combinations, Scheffersomyces amazonensis and S. ergatensis

HECTOR URBINA & MEREDITH BLACKWELL*

Department of Biological Sciences, Louisiana State University,

Baton Rouge, Louisiana 70803

*CORRESPONDENCE TO: mblackwell@lsu.edu

ABSTRACT — On the basis of gene sequence analyses, two new combinations are proposed for

the genus Scheffersomyces (Saccharomycotina, Debaryomycetaceae).

KEY worps — Ascomycota, xylose-fermenting yeasts.

Introduction

Based on recent radical changes to the rules governing the nomenclature of

pleomorphic fungi (McNeill et al. 2012: Article 59), seven new combinations

were made in the Scheffersomyces clade (Saccharomycotina, Debaryomycetaceae)

supported by a multilocus phylogenetic analysis, and three new species were

described in Scheffersomyces (Urbina & Blackwell 2012). Here we propose two

additional new combinations in Scheffersomyces for species included in the

same phylogenetic analysis, Candida amazonensis and C. ergatensis.

Taxonomy

Scheffersomyces amazonensis (Cadete, M.A. Melo, M.R. Lopes, Zilli, M.J.S. Vital,

E.C.O. Gomes, Lachance & C.A. Rosa) Urbina & M. Blackw., comb. nov.

MycoBank MB 563718

= Candida amazonensis Cadete, M.A. Melo, M.R. Lopes, Zilli, M.J.S.

Vital, EC.O. Gomes, Lachance & C.A. Rosa, International Journal

of Systematic and Evolutionary Microbiology 62: 1439, 2012.

Scheffersomyces ergatensis (Santa Maria) Urbina & M. Blackw., comb. nov.

MycoBank MB 801388

= Candida ergatensis Santa Maria, Anales del Instituto Nacional de

Investigaciones Agrarias, Agricola, Madrid 1: 85, 1971.

234 ... Urbina & Blackwell

Discussion

An ex-type culture of Candida amazonensis UFMG-HMD-26.3' (= CBS

12363 = NRRL Y-48762) was included in the multilocus phylogenetic analysis

(Urbina & Blackwell 2012), but a new combination for this taxon was not

proposed because the species name had not yet been effectively published

at the time of our revision. A second taxon included in our analysis and in

the Scheffersomyces clade, Candida ergatensis NRRL Y-17652' (= CBS 6248)

was inadvertently omitted from our taxonomic revision (Urbina & Blackwell

2012).

Both S. amazonensis and S. ergatensis were isolated from rotted wood,

and they share the ability to ferment glucose, galactose, and the disaccharide

cellobiose, and assimilate other wood components such as D-xylose and D-

arabinose (Santa Maria 1971, Cadete et al. 2012).

Acknowledgments

We thank Heide-Marie Daniel for calling our attention to the omission of C. ergatensis

from the previous revision, as well as Scott Redhead and Cletus Kurtzman for

nomenclatorial advice and manuscript review.

Literature cited

Cadete RM, Melo MA, Lopes MR, Pereira GM, Zilli JE, Vital MJ, Gomes FC, Lachance MA, Rosa

CA. 2012. Candida amazonensis sp. nov., an ascomycetous yeast isolated from rotting wood in

the Amazonian forest. Int J Syst Evol Microbiol. 62: 1438-1440.

http://dx.doi.org/10.1099/ijs.0.036715-0

McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W, Hawksworth DL, Herendeen PS, Knapp

S, Marhold K, Prado J, Prud’ homme van Reine WF, Smith GF, Wiersema JH, Turland NJ. 2012.

International Code of Nomenclature for algae, fungi, and plants (Melbourne Code), adopted

by the Eighteenth International Botanical Congress Melbourne, Australia, July 2011. Regnum

Vegetabile 154. 240 p. http://www.iapt-taxon.org/nomen/main.php

Santa Maria J. 1971. Candida ergatensis nov. spec. Anales del Instituto Nacional de Investigaciones

Agrarias, Agricola, Madrid 1: 85-88.

Urbina H, Blackwell M. 2012. Multilocus phylogenetic study of the Scheffersomyces yeast clade and

characterization of the N-terminal region of xylose reductase gene. PLoS ONE 7(6): €39128.

http://dx.doi.org/10.1371/journal.pone.0039128

MYCOTAXON

http://dx.doi.org/10.5248/123.235

Volume 123, pp. 235-239 January-March 2013

Additional information on Lecanora loekoesii

Hat-YING WANG"™*, AN-NA GE!*, HONG-MEI LI?’ & ZUN-TIAN ZHAO?

'Key Laboratory of Plant Stress Research, College of Life Science, Shandong Normal University,

Jinan, 250014, P. R. China

College of Life Sciences, HeBei University, Baoding, 071002, P. R. China

*CORRESPONDENCE TO: “endolichen@gmail.com oR *geanna521@yahoo.com.cn

ABSTRACT — Sixty-eight specimens of Lecanora loekoesii from China were collected for

further study to supplement data derived from the South Korean type (previously the only

known specimen). The ascospores of L. loekoesii often appear to be 1-septate because the

protoplasm is divided. Four chemical types of L. loekoesii were discovered.

Key worps — Asia, taxonomy, lichen, fungi

Introduction

The multispored species of Lecanora include eleven members worldwide:

L. bruneri Imshaug & Brodo, L. cateilea (Ach.) A. Massal., L. japonica Mull. Arg.,

L. loekoesii, L. pleospora Mill. Arg., L. polysphaeridia Alstrup, L. praesistens Nyl.,

L. sambuci (Pers.) Nyl., L. strobilinoides Giralt & Gomez-Bolea, L. subjaponica

L. Li & H.Y. Wang, and L. weii L.F. Han & S.Y. Guo (Alstrup 1993, Guderley &

Lumbsch 1999, Han et al. 2009, Lt et al. 2011, 2012 ). Lecanora strobilinoides is

the only species where the ascospores are frequently 1-septate.

As a new species to science, L. loekoesii was described by only one specimen

from South Korea (Li et al. 2011). In the present paper, a more detailed and

complete description of this species is given based on 68 specimens from

China.

Materials & methods

The specimens studied are preserved in SDNU (Lichen Section of Botanical

Herbarium, Shandong Normal University). The morphologicaland anatomical characters

of the specimen were examined using a stereomicroscope (COIC XTL7045B2) and

polarizing microscope (OLyMpus CX41-32). Lichen substances were identified using

standardized thin layer chromatography techniques (TLC) with C system (Orange et al.

2010). Photos of the lichen were taken under OLympus SZX16 and BX61 with DP72.

236 ... Wang & al.

Results & discussion

Lecanora loekoesii Y. Joshi, L. Li, & J.S. Hur, Lichenologist 43(4): 321-329 (2011)

Fics 1-2

DESCRIPTION — The L. loekoesii specimens from China are characterized

by the following characters: corticolous habit; thin continuous crustose thallus

that is whitish gray to grayish green; soredia, prothallus, and pycnidium all

lacking; indistinct cortex with crystals; lecanorine apothecium; yellowish brown

apothecial discs with/without white pruina; amphithecium with indistinct

cortex and small K-soluble crystals; epihymenium with coarse K-soluble

granules; hyaline hymenium without oil droplets; hyaline hypothecium; simple

non-pigmented paraphyses; 12—16-spored asci; simple or 1-septate-like (due to

divided protoplasm) ellipsoid hyaline ascospores; and thallus spot tests of K+

yellow, KC+ yellow, C-, and P+ yellow.

Both the Chinese specimens and Korean holotype of L. loekoesii have the same

epihymenium crystals. Lui et al. (2011) described the crystals in epihymenium

of holotype as fine granules. However, we think these are better interpreted as

coarse granules. Half of the L. loekoesii specimens from China have 1-septate-

like ascospores because the protoplasm is divided, while the ascospores of the

holotype are simple. We observed the 1-septate-like ascospores often in the same

apothecium or even in the same ascus together with the simple ascospores.

We retested the lichen substances of the holotype by TLC; in addition to

atranorin, usnic acid, zeorin and norstictic acid, we found three unknown

lichen substances B, D, and E were found (Fic. 2). Of the 68 Chinese specimens

tested, 51 have the same chemicals as the holotype, 14 specimens lack norstictic

acid, two specimens have placodiolic acid, while one specimen has stictic acid

plus an additional unknown substance J.

DISTRIBUTION IN CHINA — Heilongjiang, Jilin, Liaoning, Hubei, Shaanxi,

Tibet.

SPECIMENS EXAMINED (except for the holotype, all conserved in SDNU): Chemical

type 1: SOUTH KOREA. GaNnGwon-bo, Taebaek-si, Mungoksodo-dong, Mt. Taebaek,

37°06'48-8"N 128°56'51-0"E, alt. 910 m, on Quercus bark, 13 Oct 2005, Lé6kés 050717

(holotype, KoLRI). CHINA. HEILONGJIANG, Wuchang, Mt. Datudingzi, alt. 1800 m,

21 Aug 2011, Y.L. Cheng 201222748; alt. 1300 m, 21 Aug 2011, Y.L. Cheng 20122316B;

alt. 1200 m, 21 Aug 2011, Y.L. Cheng 20122060A 1, 20122233B, 20122249, 20122249D1,

20122273B, 20122288C; alt. 1100 m, 21 Aug 2011, Y.L. Cheng 20122088B, 20121912,

20122047, 20122149B, 20122220D, 201223944; alt. 900 m, 21 Aug 2011, Y.L. Cheng

20127190; alt. 800 m, 21 Aug 2011, YL. Cheng 20122097, 20122155B, 20122155B1,

20122212B, 20122311A; Daxinganling, Mt. Daqing, alt. 800 m, 26 Aug 2011, Y.L. Cheng

20125519A; Heihe, 727 linchang, alt. 488 m, 16 Aug 2009, Q. Ren 20100966; Tahe, Mt.

Baikalu, alt. 700 m, 24 Aug 2011, Y.L. Cheng 20125087; Tieli, Taoshan national forest

park, alt. 900 m, 23 Aug 2011, Y.L. Cheng 20125116B, 20125738A, 20125154; alt. 800 m,

23 Aug 2011, Y.L. Cheng 20125355A1, 20125607B, 20125687E, 20125694B, 20125759B;

alt. 700 m, 23 Aug 2011, Y.L. Cheng 20125310, 20125348A, 20125628A, 20125648,

Lecanora loekoesii ... 237

and apothecia; D, F: Crystals of thallus and apothecia; G-H, J-L: Asci and ascospores; I. Iodine

reaction of ascus.

20125751, 20125775. JILIN, Helong, Mt. Zengfeng, alt. 1600 m, 19 Aug 2011, Y.L. Cheng

20119440C, 20119440D, 20119643A; alt. 1500 m, 19 Aug 2011, Y.L. Cheng 201202614,

20119686C, 20119841; alt. 1000 m, 19 Aug 2011, Y.L. Cheng 20119329A; Wangqing,

Dadoudigou, alt. 900 m, 23 Jul 2012, H.Y. Wang 20128203, 20128211. LIAONING,

Kuandian, Mt. Huabo, alt. 1000 m, 16 Aug 2011, Y.D. Wei 20120438C; alt. 900 m, 14

238 ... Wang & al.

After acid and heating

Sunlight 365 nm

, RE. tec -

Fic. 2. Lecanora loekoesii: TLC results and crystals in holotype apothecium. A: Atranorin; C: Usnic

acid; F: Zeorin; G: Norstictic acid; H: Placodiolic acid; I: Stictic acid; B, D, E, J: Unknown substances;

K, L: Section and crystals of apothecium of holotype. Chemical type 1: 52 specimens detected,

including holotype; Chemical type 2: 14 specimens detected; Chemical type 3: Two specimens

detected; Chemical type 4: one specimen detected.

Aug 2011, Y.D. Wei 20120449A, 20120450A: 16 Aug 2011, Y.D. Wei 20120547; alt. 800

m, 16 Aug 2011, Y.L. Cheng 20120514A.

Chemical type 2: CHINA. HEILONGJIANG, Wuchang, Mt. Datudingzi, alt. 1200

m, 21 Aug 2011, Y.L. Cheng 20122139, 20122261A7, 20122288D; alt. 1100 m, 21 Aug

2011, Y.L. Cheng 20122126B; alt. 900 m, 21 Aug 2011, Y.L. Cheng 20122243B; alt. 800

m, 21 Aug 2011, Y.L. Cheng 20122068A, 20122212A, 20122231B. Jrtin, Helong, Mt.

Zengfeng, alt. 1600 m, 19 Aug 2011, Y.L. Cheng 201194404; alt. 1500 m, 19 Aug 2011,

Y.L. Cheng 20119686B. Huse, Shennongjia, Mt. Jinhou, alt. 2190 m, 9 Nov 2008,

Z.T. Zhao 20101591. SHAANXI, Meixian, Mt. Taibai, alt. 2900 m, 17 Jun 2011, Y.L.

Cheng 20114386D. TrBeT, Nyingchi, Lulangzhen, alt. 2500 m, 17 Jul 2011, Y.L. Cheng

20119194, 20119213A.

Chemical type 3: CHINA. HEILONGJIANG, Tieli, Taoshan national forest park,

alt. 800 m, 23 Aug 2011, Y. L. Cheng 20125175; alt. 700 m, 23 Aug 2011, Y. L. Cheng

20125762.

Lecanora loekoesii ... 239

Chemical type 4: CHINA. HEILONGJIANG, Wuchang, Mt. Datudingzi, alt. 1100 m,

21 Aug 2011, Y.L. Cheng 20122106B.

ComMENTs — Lecanora loekoesii is similar to L. strobilinoides in morphology

and anatomy. The latter is the only multispored Lecanora species that frequently

has 1-septate ascospores. However, L. strobilinoides can be readily distinguished

by having only usnic acid and zeorin. In addition, L. loekoesii ascospores are not

truly 1-septate.

Acknowledgements

The project was financially supported by Program for Scientific Research Innovation

Team in Colleges and Universities of Shandong Province, the National Natural

Science Foundation of China (31070010, 31170187), and Science Foundation of Jinan

(201202024). We thank Prof. A. Aptroot (ABL Herbarium, Soest, The Netherlands) and

Dr. Shou- Yu Guo (State Key Laboratory of Mycology, Institute of Microbiology, Chinese

Academy of Sciences, Beijing, China) for presubmission reviews. We also thank Dr.

Alstrup (Institut for Okologisk Botanik, Oster Farimagsgade 2D, DK-1353 Kobenhavn

K) for providing the information on Lecanora polysphaeridia to us.

Literature cited

Alstrup V. 1993. News on lichens and lichenicolous fungi from the Nordic countries. Graphis

Scripta 5: 96-104.

Guderley R, Lumbsch HT. 1999. Notes on multispored species of Lecanora sensu stricto.

Lichenologist 31: 197-210.

Han LE Zhao JC. Guo SY. 2009. Lecanora weii, a new multispored species of Lecanora s. str. from

northeastern China. Mycotaxon 107: 157-161.

Lit L, Joshi Y, Elix JA, Lumbsch HT, Wang HY, Koh YJ, Hur JS. 2011. New and noteworthy species

of lichen genus Lecanora (Ascomycota; Lecanoraceae) from South Korea. Lichenologist 43:

321-329. http://dx.doi.org/10.1017/S0024282911000144

LiL, Zhang LL, Liu XL, Zhao ZT, Wang HY. 2012. Lecanora subjaponica, a new lichen from China.

Lichenologist 44: 465-468. http://dx.doi.org/10.1017/S002428291200014X

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

MY COTAXON

http://dx.doi.org/10.5248/123.241

Volume 123, pp. 241-249 January-March 2013

Three new species and one new record of Lobothallia from China

XING-RAN Kou, SHU-XIA LI, QIANG REN’

College of Life Science, Shandong Normal University Jinan, 250014, P. R. China

* CORRESPONDENCE TO: rengiang@sdnu.edu.cn*

ABSTRACT — A comprehensive analysis of morphological, anatomical, chemical and

molecular data indicates the presence of three additional species in the genus Lobothallia

(Megasporaceae, lichenized ascomycetes), which are described here as new to science under

the names L. crassimarginata, L. helanensis, and L. pruinosa. Lobothallia praeradiosa is newly

reported from China.

Key worps — lichen, taxonomy

Introduction

The lichen genus Lobothallia (Clauzade & Cl. Roux) Hafellner was originally

established for four species, L. alphoplaca, L. melanaspis, L. praeradiosa, and

L. radiosa (Hafellner 1991), distinguished from Aspicilia by their lobate thallus,

small spores, short conidia and chemistry. Recently, five additional species

L. farinosa, L. recedens (Nordin et al. 2010), L. cernohorskyana, L. chadefaudiana,

and L. cheresina (Roux 2012) have been included. As a consequence of this

extension, Lobothallia is characterized by immersed to appressed or constricted-

sessile apothecia, asci with an inamyloid tholus (Aspicilia-type), unbranched

paraphyses, simple, hyaline spores and mainly bacilliform conidia. Lobes are

distinct in some species, such as L. alphoplaca, L. melanaspis, L. praeradiosa,

and L. radiosa, while other species have indistinct lobes, such as L. farinosa and

L. recedens. In phylogenetic studies, Lobothallia is the sister group of the other

genera within Megasporaceae (Nordin et al. 2010).

During our study on Megasporaceae of China, we discovered three new

species of Lobothallia, described here as L. crassimarginata, L. helanensis, and

L. pruinosa. Lobothallia praeradiosa is reported for the first time from China.

Materials & methods

The specimens studied were collected in Northern and Western China, and are

preserved in SDNU (Lichen Section of the Botanical Herbarium, Shandong Normal

242 ... Kou, Li & Ren

TABLE 1. Specimens used in the phylogenetic analyses. New sequences are in bold.

SPECIES ORIGIN VOUCHER GENBANK No.

Aspicilia cinerea Sweden UPS: Nordin 6213 JF703115

A. epiglypta Sweden UPS: Nordin 6105 HQ259262

Lobothallia alphoplaca China, IM SDNU: 20117646 JX476025

China, IM SDNU: 20117616 JX499233

L. crassimarginata China, IM SDNU: 20122565 (holotype) JX476026

China, IM SDNU: 20122583 KC007439

L. helanensis China, IM SDNU: 20122791 JX476031

China, IM SDNU: 20122517 (holotype) JX476030

L. melanaspis Sweden UPS: Nordin 6622 HQ259272

Norway UPS: Owe-Larsson 8943a JF825524

L. praeradiosa China, XJ SDNU: 20126683 JX499229

China, XJ SDNU: 20126355 JX499230

China, XJ SDNU: 20126314 JX499232

China, XJ SDNU: 20126613 JX499234

L. pruinosa China, IM SDNU: 20123630 JX476027

China, IM SDNU: 20123278 (holotype) JX476028

China, IM SDNU: 20123909 JX499231

L. radiosa Sweden UPS: Nordin 5889 JF703124

L. recedens Sweden UPS: Nordin 6035 HQ406807

*Province abbreviations: IM = Inner Mongolia; XJ = Xinjiang.

University). Specimen morphology and anatomy were examined using a dissecting

microscope (Olympus SZ51) and a compact light microscope (Olympus CX41). Lichen

substances were identified using standardized thin layer chromatography techniques

(TLC) with solvent C (Orange et al. 2010). Photos of the thalli were taken using an

Olympus SZX16 with DP72.

SAMPLING: Nuclear ITS1-5.8S-ITS2 rDNA sequences of 18 specimens representing

9 species were used in the molecular study. New sequences were produced from 13

specimens and six sequences were downloaded from GenBank (Tas. 1). Aspicilia cinerea

and A. epiglypta were used as outgroup.

EXTRACTIONS, PCR AMPLIFICATIONS, AND SEQUENCING: Total DNA was extracted

from the samples using the SanPrep Colum DNA Gel Extraction Kit. Lichen sample

vouchers of the new sequences are listed in TaBLE 1. To amplify the ITS1-5.8S-ITS2

regions, the primers ITS1-F (Gardes & Bruns 1993) combined with LR7 (Vilgalys &

Hester 1990) or ITS4 combined with ITS5 (White et al. 1990) were used. The PCR

ran for 37 cycles (1 min. at 95°C, 30 sec. at 53°C, 30 sec. at 72°C) using Tiangen Taq.

Sequencing reactions were carried out by BGI (www.genomics.cn) with an ABI 3730 XL

DNA Analyzer.

PHYLOGENETIC ANALYSES: The data set was processed with the Minimum Evolution

method (Rzhetsky & Nei 1992). The percentages of replicate trees (1000 replicates) in

which the associated taxa clustered together in the bootstrap test (Felsenstein 1985) are

shown next to the branches (Fic. 1). The evolutionary distances were computed using

Lobothallia spp. nov. (China) ... 243

97 Lobothallia praeradiosa JX499229

90 jt Lobothallia praeradiosa JX499234

100 | Lobothallia praeradiosa JX499232

65 Lobothallia praeradiosa JX499230

100 ¢ Lobothallia alphoplaca JX476025

Lobothallia alphoplaca JX499233

94 100 Lobothallia melanaspis HQ259272

Lobothallia melanaspis JF825524

100 Lobothallia pruinosa JX499231

97 Lobothallia pruinosa JX476027

109 |! Lobothallia pruinosa JX476028

100 Lobothallia crassimarginata JX476026

100 Lobothallia crassimarginata KC007439 a

74 100 Lobothallia helanensis JX476030

Lobothallia helanensis JX476031 |

Lobothallia radiosa JF703124

99 Lobothallia recedens HQ406807

Aspicilia cinerea JF703115

Aspicilia epiglypta HQ259262

———

0.02

FiGureE 1. The ME tree inferred from ITS data. Bootstrap values greater than 60% (1000 replicates)

are shown next to the branches.

the Maximum Composite Likelihood method (Tamura et al. 2004). There were a total of

571 positions in the final dataset. All positions containing alignment gaps and missing

data were eliminated only in pairwise sequence comparisons (Pairwise deletion option).

Phylogenetic analyses were conducted in MEGA4 (Tamura et al. 2007).

Results & discussion

New Species

Lobothallia crassimarginata X.R. Kou & Q. Ren, sp. nov. Fic. 2

MycoBank MB 801753

Differs from all other Lobothallia spp. by its persistently prominent apothecial margin

and thick thallus.

Type: China. Inner Mongolia, Mt. Helan, on rock, alt. 1500 m, 19 Aug 2011, H.Y. Wang

20122565 (Holotype, SDNU; Genbank, JX476026).

ErymMo.Loey: Latin crassus (= thick) + marginatus (= margined), referring to the

apothecial margins.

THALLUS placodioid, up to 2-5 mm thick centrally, tightly adnate, areolate;

AREOLES 1-1.5 mm in diam., discrete, rounded, plane to somewhat convex;

LOBES 1-2 mm long, 0.5-0.8 mm across, 0.5-0.7 mm or more thick, usually

distinctly elongated, radiating and separate, plane to more often strongly

convex; UPPER SURFACE ashy gray, sometimes tinted ochraceous or rosy,

244 ... Kou, Li & Ren

Ficure 2. Lobothallia crassimarginata (Holotype). A. Thallus; B. Lobes; C. Apothecia; D. Apothecial

anatomy; E. Ascus and spores; F. Conidia. Scale bars: A = 2 mm; B = 1 mm; C = 250 um; D = 100 um;

E, F = 10 pm.

usually epruinose, smooth to somewhat wrinkled. APoTHECIA lecanorine,

usually solitary, 0.6-1.5 mm in diam., appressed-sessile to distinctly sessile;

pisc black, plane or convex, epruinose; MARGIN remaining strongly raised

and inflexed, 0.2-0.5 mm wide, concolorous with the thallus; EptIHyYMENIUM

brown, the pigment fading in K, N+ lightly green; HyMENIUM hyaline, 70-90

um tall; PARAPHYSES separating in KOH, submoniliform; suBHYMENIUM and

HYPOTHECIUM hyaline, 50-70 um tall together, with algae layer below; Asc1

Aspicilia-type, 8-spored; AscosporEs hyaline, simple, subglobose to globose,

9-10 x 8-9 um; conip1a bacilliform, 5.4-7.6 x 1-1.2 um.

SPOT TESTS — medulla K+ yellow then red, C-, I-, P+ orange.

SECONDARY METABOLITES — norstictic, stictic and constictic acids.

SUBSTRATE — Calciferous rock.

ADDITIONAL SPECIMENS EXAMINED (all conserved in SDNU) — CHINA. INNER

MoneotiiA, Mt. Helan, on rock, alt. 1500 m, 19 Aug 2011, H.Y. Wang 20122418B,

20123084B, 20123089C; D.B. Tong 20122513, 20122583 (GenBank, KC007439),

20122518.

Comments — Lobothallia crassimarginata differs from the other members of

this genus by its persistently prominent apothecial margin and thick thallus.

Both Lobothallia alphoplaca and L. praeradiosa have raised apothecial margins

Lobothallia spp. nov. (China) ... 245

only when young. Three other Lobothallia species with a thick thallus are

L. alphoplaca, L. helanensis, and L. recedens. Lobothallia alphoplaca differs from

L. crassimarginata by its intact removable thallus with longer lobes, L. helanensis

by having a rough thallus, a thin apothecial margin, and lacking secondary

metabolites, while L. recedens has a pruinose thallus and shorter conidia (3-5

x 1 um; Smith et al. 2009). In the phylogenetic tree, these four species lie in

different clades. Evolutionary distances between L. crassimarginata and other

three species range from 0.074 to 0.111.

Lobothallia helanensis X.R. Kou & Q. Ren, sp. nov. FIG. 3

MycoBank MB 801755

Differs from Lobothallia recedens by its thallus having a gray to white-gray upper surface

with a brown tinge, and by its larger conidia.

Type: China. Inner Mongolia, Mt. Helan, on rock, alt. 1500 m, 19 Aug 2011, D.B. Tong

20122517 (Holotype, SDNU; Genbank, JX476030).

ErymMo_oey: Referring to the type locality.

THALLUS crustose, rimose to areolate, gradually thicker towards the central part,

sometimes up to 7 mm thick; AREOLES 0.7-1.4(-2) mm in diam., contiguous,

Figure 3. Lobothallia helanensis (Holotype). A. Thallus; B. Lobes; C. Apothecia; D. Apothecial

anatomy; E. Ascus and spores; F. Conidia. Scale bars: A = 2 mm; B = 1 mm; C = 500 um; D = 50 um;

E, F= 10 um.

246 ... Kou, Li & Ren

angular to irregular, +convex; UPPER SURFACE gray to white-gray, with a brown

tinge, distinctly rough. APoTHECIA 0.5-1.3(-2) mm in diam., aspicilioid,

rounded or angular; thalline margin thin, slightly elevated, sometimes with

slightly dentate incision, inner part brown, outer part concolorous with the

thallus; pisc black, without pruina; EPIHYMENIUM brown, K+ brown, N+ weakly

green; HYMENIUM hyaline, I+ blue, 70-80 um tall; PARAPHYSES separating

in KOH, submoniliform to moniliform; SsUBHYMENIUM colourless to weakly

brown, I+ blue, +50 um tall; ayPOTHECIUM colourless, I+ blue, 30 um tall, algae

continuous or grouped below the hypothecium. Ascr clavate, Aspicilia-type,

8-spored; AscosporEs hyaline, simple, ellipsoid, 10-12.5 x 5.5-6.5 um; CONIDIA

bacilliform, 5.5-6.4(-8) x 1.2-1.4 um.

SPOT TESTS — medulla K-, C-, I-, P-.

SECONDARY METABOLITES — none detected by TLC.

SUBSTRATE — Calciferous rock.

ADDITIONAL SPECIMENS EXAMINED (all conserved in SDNU) — CHINA. INNER

MoneoiiA, Mt. Helan, on rock, alt. 1500 m, 17 Aug 2011, D.B. Tong 20122791

(GenBank, JX476031); 19 Aug 2011, H.Y. Wang 20122986, 20123043, 20122780A; P.M.

Wang 20123301, 20123308A, 20123709.

Comments — Lobothallia helanensis resembles most closely L. recedens, which

shares a thick thallus and the absence of secondary metabolites but has a white

sheen resembling pruina and smaller conidia. Lobothallia alphoplaca, another

species with a thick thallus, differs by its loosely adnate thallus containing

norstictic, constictic, or salazinic acids. In the phylogenetic tree, Lobothallia

helanensis JX476030 and Lobothallia helanensis JX476031 form a clade

supported by 99% bootstrap value. These three species lie in different clades.

The evolutionary distances between L. helanensis and the other two species

range from 0.091 to 0.132.

Lobothallia pruinosa X.R. Kou & Q. Ren, sp. nov. Fic. 4

MycoBank MB 801756

Differs from Lobothallia recedens by its thin and pruinose thallus, its more sparse

apothecia, and its smaller ascospores.

Type: China. Inner Mongolia, Mt. Helan, on rock, alt. 1500 m, 19 Aug 2011, H.Y. Wang

20123278 (Holotype, SDNU; Genbank, JX476028).

ErymMo_oey: Referring to the pruinose thallus and apothecial discs.

THALLUS placodioid to squamulate, up to 0.5-1.0 mm thick, tightly attached,

areolate, at the margin radiate; AREOLES 0.5-1 mm wide, contiguous, plane;

LOBES plane, contiguous, confluent, tightly attached, 1-2 mm long, 0.7-1 mm

wide, 0.15-0.2 mm thick; UPPER SURFACE usually whitish gray to brownish

gray, often pruinose especially at the margins of the areoles. APOTHECIA

solitary, round, 0.7-1.2 mm in diam., somewhat higher than the areoles; Disc

Lobothallia spp. nov. (China) ... 247

Figure 4. Lobothallia pruinosa (Holotype). A. Thallus with white pruina; B. Lobes and apothecia;

C. Apothecial anatomy; D. Asci and spores; E. Conidia. Scale bars: A = 2 mm; B = 500 um; C = 50 um;

D, E= 10 um.

dark brown, usually pruinose; asci Aspicilia-type, 8-spored; EPIHYMENIUM

brown, the pigment fading in K, N+ weakly green; HyMENIvUM hyaline, I+ blue,

80-120 um tall, PARAPHYSES separating in KOH, submoniliform to moniliform;

SUBHYMENIUM and HYPOTHECIUM colourless, I+ blue, 50-70 um tall together,

algae continuous or grouped below the hypothecium; ascosporgs broadly

ellipsoid, 12.5-15 x 8.5-10 um; conip1a bacilliform, 5-7 x 1-1.3 um.

SPOT TESTS — medulla K+ yellow, C-, I-, P+ orange.

SECONDARY METABOLITES — norstictic and constictic acids.

SUBSTRATE — siliceous rock intermingled with calciferous granules.

ADDITIONAL SPECIMENS EXAMINED (all conserved in SDNU) — CHINA. INNER

Moneco tia, Mt. Helan, on rock, alt. 1500 m, 19 Aug 2011, H.Y. Wang 20123091,

20123630 (GenBank, JX476027); 1600 m, 19 Aug 2011, H.Y. Wang 20123575. Mt.

Huhebashige, on rock, alt. 1600 m, 16 Aug 2011, H.Y. Wang 20123447B, 20123408; 17

Aug 2011, H.Y. Wang 20123909 (GenBank, JX499231); X.R. Kou 20123915, 20123950;

P.M. Wang 20123678.

ComMMENTs — Another species with distinct pruina is L. recedens, which

differs from L. pruinosa by its thick thallus, numerous apothecia and somewhat

larger spores. The two species lie in different clades. The evolutionary distances

between them range from 0.130 to 0.139.

248 ... Kou, Li & Ren

: » ‘ a ‘i : er.

a he ' to. - “te ‘aa

: >

. “ * #

_ ™/ . Fe

t. om” - ~

Figure 5. Lobothallia praeradiosa, 20126613 (SDNU). Scale = 1 mm.

New Record

Lobothallia praeradiosa (Nyl.) Hafellner, Acta Bot. Malac. 16: 138 (1991). Fic. 5

THALLUuS placodioid, up to 0.5-1.0 mm thick, tightly attached, areolate;

AREOLES 0.5-1 mm wide, contiguous, elongate to irregular, plane to convex;

LOBES contiguous, confluent, plane, broad, sometimes imbricate, loosely

attached, 3-6 mm long, 0.5-1.5 mm wide, 0.2-0.5 mm thick; UPPER SURFACE

usually green gray to orange brown, often radiate. APOTHECIA solitary to

numerous, round, 0.5-1.5 mm in diam., adnate, somewhat higher than other

areoles; pisc: dark brown, usually epruinose, Asci Aspicilia-type, 8-spored;

EPIHYMENIUM brown, K+ brown, N-; HYMENIUM hyaline, I+ blue, 75-100

um tall, PARAPHYSES separating in KOH, submoniliform; suBHYMENIUM and

HYPOTHECIUM colourless, I+ blue, 40-50 um tall together; algae continuous or

grouped below the hypothecium; ascosporss ellipsoid, 12.5-15 x 5-7.5 um.

SPOT TESTS — medulla K+ yellow, C-, I-, P+ orange or yellow.

SECONDARY METABOLITES — norstictic acid.

SUBSTRATE — Siliceous rock.

ADDITIONAL SPECIMENS EXAMINED (all conserved in SDNU) — CHINA. XINJIANG, Mt.

Tian, on rock, alt. 1910 m, 23 Aug 2011, Z.L. Huang 20126613 (GenBank, JX499234),

Z.L. Huang 20126355 (GenBank, JX499230); L. Li 20126314 (GenBank, JX499232); Mt.

Nan, on rock, alt. 1900 m, 28 Aug 2011, Z.L. Huang 20126683 (GenBank, JX499229).

Lobothallia spp. nov. (China) ... 249

Acknowledgements

The project was financially supported by the National Natural Science Foundation

of China (31100011). We are grateful to Dr. A. Nordin (Museum of Evolution, Botany,

Uppsala University, Uppsala, Sweden), Dr. H.Y. Wang (College of Life Sciences,

Shandong Normal University, Jinan, China) and Dr. Mohammad Sohrabi (Iranian

Research Organization for Science and Technology, Tehran, Iran) for the professional

advice and unselfish help during this study, to D.F. Jiang and L. Hu (College of Life

Sciences, Shandong Normal University, Jinan, China) for the help with DNA extraction.

The authors thank Dr. H.J.M. Sipman (Botanischer Garten und Botanisches Museum,

Freie Universitat, Berlin, Germany) and Dr. Shou-Yu Guo (Institute of Microbiology,

Chinese Academy of Sciences, Beijing, China) for presubmission reviews.

Literature cited

Felsenstein J. 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution

39: 783-791. http://dx.doi.org/10.2307/2408678

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes - application

to identification of mycorrhizae and rusts. Mol. Ecol. 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Hafellner J. 1991. Die Gattung Aspicilia, ihre Ableitungen nebst Bemerkungen tiber cryptolecanorine

Ascocarporganisation bei anderen Genera der Lecanorales (Ascomycetes lichenisati). Acta

Botanica Malacitana 16: 133-140.

Nordin A, Savi¢ S, Tibell L. 2010. Phylogeny and taxonomy of Aspicilia and Megasporaceae.

Mycologia 102: 1339-1349. http://dx.doi.org/10.3852/09-266

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society, London. 101 p.

Roux C. 2012. Liste des lichens et champignons lichénicoles de France. Bulletin de la Société

linnéenne de Provence, Numéro spécial 16: 1-220.

Rzhetsky A, Nei M. 1992. A simple method for estimating and testing minimum evolution trees.

Molecular Biology and Evolution 9: 945-967.

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA (eds). 2009. The

lichens of Great Britain and Ireland. Natural History Museum Publications, in association with

The British Lichen Society.

Tamura K, Nei M, Kumar S. 2004. Prospects for inferring very large phylogenies by using the

neighbor-joining method. Proceedings of the National Academy of Sciences (USA) 101:

11030-11035. http://dx.doi.org/10.1073/pnas.0404206101

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics

Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24: 1596-1599.

http://dx.doi.org/10.1093/molbev/msm092

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.

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 Applications. Academic Press, Inc., New York.

MY COTAXON

http://dx.doi.org/10.5248/123.251

Volume 123, pp. 251-253 January-March 2013

A new halotolerant species of A/ternaria

from Qinghai-Tibet Plateau, China

FANG WANG, AN-NA L1, DONG-MEI DAI, XIAO-XUE XU & DUO-CHUAN LI*

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

Key Laboratory of Agricultural Microbiology, Shandong Province, Taian, 271018, China

* CORRESPONDENCE TO: lidc20@sdau.edu.cn

AsBsTRACT — A new halotolerant species of Alternaria, A. xiaochaidanensis, is described and

illustrated. The specimen was collected from a salt lake on Qinghai-Tibet Plateau. The type

specimen is deposited in the Herbarium of the Department of Plant Pathology, Shandong

Agricultural University (HSAUP).

KEY WORDS — taxonomy, saline, extremophiles

Introduction

Salt tolerant fungi have evolved special physiological mechanisms and

metabolic products to survive in the extreme environment of salt-water lakes.

In recent decades, more attention has been paid to salt tolerant fungi - both

halotolerant and halophilic fungi. Cronin & Post (1977) described the first

halophilic filamentous fungus, Cladosporium sp., from a hypersaline lake in the

Great Salt Lake, USA. To date, more than 80 halotolerant fungal species and six

halophilic fungi have been reported (Gunde-Cimerman et al. 2005, 2009).

Qinghai-Tibet Plateau region has the world’s highest altitude salt lake. It

is a potentially huge treasure trove of genetic resources with its diversity of

water chemistry characteristics and different biotic and abiotic anti-positive

gene. Thus, it is important to study microorganisms of Qinghai-Tibet Plateau,

especially halophilic and halotolerant microorganisms. Here, we describe a new

species of Alternaria that was isolated from Xiaochaidan, a salt lake on Qinghai-

Tibet Plateau. This is the first report of a halotolerant Alternaria species.

Alternaria xiaochaidanensis F. Wang, A.N. Li, D.M. Dai, X.X. Xu & D.C. Li _ sp. nov.

MycoBAnk MB 800882 FIG.1

Differs from Alternaria chlamydospora and A. mouchaccae by its larger usually beaked

conidia and its different conidial shape.

252 ... Wang & al.

Fic. 1. Alternaria xiaochaidanensis (ex holotype).

Conidia, conidiophores and conidiogenous cells. Left: drawings. Right: photomicrographs.

(Bar = 50 um).

Type: China, Qinghai, Xiaochaidan, from a lake soil, altitude 2800-2840 m, July 2011,

E Wang (Holotype, HSAUP II 011501; GenBank, JX235958).

EryMoOLocy: in reference to the type locality.

Alternaria xiaochaidanensis grows rapidly, colonies on potato carrot agar at

28°C reach 6.5-7.5 cm diam. in 7 days, effuse, circular, pale brown; reverse

fuscous-black. Mycelium superficial or immersed; hyphae hyaline, grey—white.

Conidiophores either solitary, not branched, 28-54 (-68 ) x 3-5 um, or arising

as lateral branches from superficial hyphae and up to 170 um long. Conidia

produced from conidiophores or directly from hyphae, solitary or in short

chains of 2-5, rostrate or non-rostrate, 13-74 x 6-6lum. Juvenile conidia

ovoid or obclavate, pale brown; mature conidia symmetrical or asymmetrical,

irregular in shape and size, brown, smooth, walls thickened, with transverse ,

longitudinal and oblique septa, often constricted at septa.

ComMENnTs -This species may be confused with A. mouchaccae (Simmons

1981) and A. chlamydospora (Mouchacca 1973). However, conidia of those

species are smaller: A. mouchaccae (25-40(-50) x 12-20(-22) um) and

A. chlamydospora (26-70 x 8-48 um). Although sometimes found in chains

of 2-5, conidia in A. xiaochaidanensis are usually single; in A. chlamydospora

conidia are often in chains of 5-8 while conidia of A. mouchaccae are rarely

in chains. In contrast to A. xiaochaidanensis, conidia of A. mouchaccae and

A. chlamydospora have no beak. A conspicuous characteristic of the new species

is that mature conidia are symmetrical or asymmetrical and have specific

shapes that obviously differ from those of A. chlamydospora. Conidiophores of

Alternaria xiaochaidanensis sp. nov. (China) ... 253

A. xiaochaidanensis are longer than those of A. chlamydospora (15-18 x 3 um,

up to 150 um) and shorter than those of A. mouchaccae (40-50 x 3-4 um).

A BLAST search comparison of its sequence in GenBank places

A. xiaochaidanensis into Alternaria but with no previously described species

close to it.

Gunde-Cimerman et al. (2005, 2009) characterized fungi that can grow

in vitro at 3 M salt concentration and are regularly isolated from global

environments at salinities above 1.7 M as halophilic, whereas isolates that

can grow in vitro sporadically at 3 M NaCl are considered halotolerant.

Species isolated above 1.7 M NaCl grow regularly on PDA but slowly at 3 M.

According to this definition, the authors consider Alternaria xiaochaidanensis

a halotolerant fungus.

Acknowledgments

The authors are grateful to Dr. Eric McKenzie, Prof. Y.L. Guo, and Dr. Shaun

Pennycook 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 (31000007), China Postdoctoral Science Foundation (20100471565) and the

Chinese National Special Fund of Sea Renewable Energy Sources (SDME2011SW01).

Literature cited

Cronin AE, Post FJ. 1977. Report of a dematiaceous hyphomycetes from the Great Salt Lake, Utah.

Mycologia 69: 846-847. http://dx.doi.org/10.2307/3758878

Gunde-Cimerman N, Frisvad JC, Zalar P, Plemenitas A. 2005. Halotolerant and halophilic fungi.

68-128, in: SK Deshmukh, MK Rai (eds). Biodiversity of Fungi - Their Role in Human Life.

Oxford & IBH Publishing, New Delhi.

Gunde-Cimerman N, Ramos J, Plemenitas A. 2009. Halotolerant and halophilic fungi. Mycological

Research 113: 1231-1241. http://dx.doi.org/10.1016/j.mycres.2009.09.002

Mouchacca J. 1973. Deux Alternaria des sols arides d Egypte: A. chlamydosporum sp. nov. et

A. phragmospora van Emden. Mycopathologiae et Mycologia Applicata 50(3): 217-225.

http://dx.doi.org/10.1007/BF02053370

Simmons EG. 1981. Alternaria themes and variations. Mycotaxon 13: 18-21.

MY COTAXON

http://dx.doi.org/10.5248/123.255

Volume 123, pp. 255-260 January-March 2013

Additions to the smut fungi of Pakistan. 2

CVvETOMIR M. DENCHEV’, MUHAMMAD FIAZ?, TEODOR T. DENCHEV,'

ABDUL NASIR KHALID? & HABIB AHMAD*

‘Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences,

2 Gagarin St., 1113 Sofia, Bulgaria

?Department of Botany, Hazara University Mansehra, Pakistan

*Department of Botany, University of the Punjab, Lahore, Pakistan

‘Department of Genetics, Hazara University Mansehra, Pakistan

* CORRESPONDENCE TO: cmdenchev@yahoo.co.uk

ABSTRACT — Two species of smut fungi, Sporisorium lingii on Themeda anathera and Urocystis

oryzopsidis on Piptatherum microcarpum, are reported for the first time from Pakistan. Both

smut fungi were found on new host plants. Chrysopogon serrulatus is recorded from Pakistan

as a new host of Anthracocystis chrysopogonis.

Key worps — Poaceae, taxonomy, Ustilaginomycetes

Introduction

Three smut fungi of poaceous hosts are described in this third contribution

to the smut fungi of Pakistan based on material collected by M. Fiaz in 2009-10,

in Mansehra District (Khyber Pakhtunkhwa Province) (Denchev et al. 2012,

Fiaz et al. 2013). Two species, both on new hosts, Sporisorium lingii on Themeda

anathera and Urocystis oryzopsidis on Piptatherum microcarpum, are reported

for the first time from Pakistan. Anthracocystis chrysopogonis is recorded on a

new host, Chrysopogon serrulatus.

Material & methods

Dried specimens from the Hazara University Herbarium (HUP), Pakistan were

examined under light (LM) and scanning electron (SEM) microscopes. For LM

observations, spores were mounted in lactophenol solution on glass slides, gently heated

to boiling point to rehydrate the spores, and then cooled. Spore measurements are given

in the form: min-max (mean + 1 standard deviation). For SEM, spores were attached

to specimen holders by double-sided adhesive tape and coated with gold with an ion

sputter. The surface structure of spores was observed at 10 kV and photographed with a

JEOL SM-6390 scanning electron microscope.

256 ... Denchev & al.

. is a na

Fics 1-4. Anthracocystis chrysopogonis on Chrysopogon serrulatus (HUP 307).

Spores in LM and SEM. Scale bars: 1, 2 = 10 um; 3, 4 =5 um.

Taxonomy

Anthracocystis chrysopogonis (Vanky) McTaggart & R.G. Shivas,

Persoonia 29: 120, 2012. FIGs 1-4

= Sporisorium chrysopogonis Vanky, Mycotaxon 18: 327, 1983.

Sort destroying the whole inflorescence, partly hidden by the leaf sheath, up

to 22 mm in length and 2 mm in width; initially covered by an yellowish brown

Sporisorium & Urocystis spp. new for Pakistan ... 257

peridium which later ruptures from its apex exposing semi-agglutinated, dark

reddish brown mass of spore balls and numerous filiform columellae. SPORE

BALLS subglobose, broadly ellipsoidal, ovoid or slightly irregular, 23-66 x

20-45 um, dark to middle reddish brown, composed of 8 to tens of spores that

separate by pressure. Single STERILE CELLS very rarely present; subglobose,

broadly ellipsoidal, slightly irregular, sometimes collapsed, 8.5-13 x 8-12 um,

hyaline; wall 1.2-2 um thick, smooth. Spores dimorphic, globose, subglobose,

broadly ellipsoidal, ovoid or slightly irregular. Outer spores 8-12(-13) x

7-10 (10.0 + 0.9 x 8.4 + 0.6) um (n = 50), middle reddish brown; wall uneven,

0.7-1.3 um thick, smooth on the contact surface, verruculose-echinulate on

the free surface, spore profile slightly affected. In SEM verruculose-echinulate,

finely punctate between the main ornaments. Inner spores (7—)7.5-11 x 6.5-9

(8.6 + 0.8 x 7.6 + 0.5) um (n = 50), light yellowish brown; wall evenly thickened,

0.4-0.7 um thick, smooth to very finely punctate.

SPECIMEN EXAMINED — On Chrysopogon serrulatus Trin.: PAKISTAN, KHYBER

PAKHTUNKHWA PROVINCE, Mansehra District, Chulandrian (Oghi), August 2010, leg.

M. Fiaz, no. FS-23 (HUP 307).

DISTRIBUTION — ‘This species has been previously known on Chrysopogon

fulvus (Spreng.) Chiov. (= C. montanus Trin.) and Chrysopogon sp. from Sri

Lanka and Pakistan (Vanky 2005, 2011). Chrysopogon serrulatus is a new host

record for this smut fungus.

CoMMENTS — The spore balls of this specimen are a little more loose compared

with those of the isotype (on Chrysopogon fulvus, Sri Lanka, North Central

Province, Polonnaruwa Distr., Habarane, alt. 250 m, 18 March 1974, K. Vanky,

in Vanky, Ustilag. exs., no. 407, SOMEF), studied by us.

Sporisorium lingii Vanky, Mycotaxon 51: 160, 1994. Fics 5-8

Sor! in all of the spikelets of the raceme, all racemes of an inflorescence

affected, 5-12 x 1-2 mm, cylindrical, initially covered by an yellowish brown

peridium that soon flakes away exposing a single, branched columella,

surrounded by a dark brown mass of readily disintegrating spore balls, spores,

and sterile cells. SPORE BALLS loose, middle to dark reddish brown, composed

of tens to hundreds of spores. STERILE CELLS in irregular groups, subglobose to

irregular, often collapsed, 9-19 x 8.5-18 um, hyaline to subhyaline; wall 0.9-1.9

um thick, smooth. Sporss globose, subglobose, broadly ellipsoidal or slightly

irregular, 8.5-12.5 x 7.5-11 (10.3 + 0.7 x 9.3 + 0.6) um (n= 100), middle reddish

brown; wall evenly thickened, 0.6-0.9 um thick, smooth to very finely punctate.

In SEM very finely echinulate, densely punctate between the spinules.

SPECIMEN EXAMINED — On Themeda anathera (Nees ex Steud.) Hack.: PAKISTAN,

KHYBER PAKHTUNKHWA PROVINCE, Mansehra District, Oghi, Khabbal Paien, August

2010, leg. M. Fiaz, no. FS-21 (HUP 309).

258 ... Denchev & al.

Fics 5-8. Sporisorium lingii on Themeda anathera (HUP 309).

Spores and sterile cells in LM and SEM. Scale bars: 5, 6 = 10 um; 7, 8 = 5 um.

DISTRIBUTION — This species has been previously known on Themeda triandra

Forssk. and Themeda sp. from S. Asia and Australia (Vanky 2011). Themeda

anathera is a new host record for this species.

Sporisorium & Urocystis spp. new for Pakistan ... 259

Urocystis oryzopsidis Padwick & Azmatullah, Mycological Papers 10: 1, 1944.

FIGS 9-12

Sor! in leaves, as long striae between the veins, at first lead-coloured and

covered by the epidermis which later ruptures and disclosing the spore balls.

SPORE BALLS MASS powdery, dark reddish brown. SpoRE BALLS subglobose,

Fics 9-12. Urocystis oryzopsidis on Piptatherum microcarpum (HUP 315).

Spore balls in LM and SEM. Scale bars: 9, 10 = 10 um; 11, 12 = 5 um.

260 ... Denchev & al.

broadly ellipsoidal, ellipsoidal or irregular, composed of 1-5(-6) central spores

[Do 6:19650.25 = 279905 342.3%, A, = 16:99 55:89, 6 = Os = 409),

usually completely invested by a layer of sterile cells; 15-25.5 x 15-20.5 um

[with 1 spore], 17.5-31 x 15.5-20.5 um [with 2 spores], 21-34.5 x 17-27.5

um [with 3 spores], 26-39.5 x 18.5-32.5 um [with 4 spores]. STERILE CELLS

variable in shape, often collapsed, 6-16 um long, light yellowish brown; wall

0.8-1.5 um thick, smooth. Spores subglobose, broadly ellipsoidal, ovoid,

slightly irregular, sometimes flattened on the contact sides, 10.5-17(-18) x

8-13 (13.7+1.6 x 10.8+1.0) um (n = 50), medium reddish brown; wall 0.8-1.3

uum thick, smooth.

SPECIMEN EXAMINED — On Piptatherum microcarpum (Pilg.) Tzvelev (= P. vicarium

(Grigorj.) Roshev.): PAKISTAN, KHYBER PAKHTUNKHWA PROVINCE, Mansehra

District, Jaba, 3 September 2009, leg. M. Fiaz, no. FS-12 (HUP 315).

DISTRIBUTION — This smut fungus has been previously known only from the

type locality (India, Kashmir) on Piptatherum munroi (Stapf ex Hook. f.) Mez

(as Oryzopsis munroi).

Acknowledgements

We gratefully acknowledge Prof. M. Kakishima (University of Tsukuba, Japan) and

Prof. Lin Guo (Key Laboratory of Systematic Mycology and Lichenology, Institute of

Microbiology, Beijing, China) for critically reading the manuscript and serving as pre-

submission reviewers.

Literature cited

Denchev CM, Fiaz M, Denchev TT, Ahmad H, Khalid AN. 2012. Additions to the smut fungi of

Pakistan. 1. Mycotaxon 121: 165-170. http://dx.doi.org/10.5248/121.165

Fiaz M, Khalid AN, Ahmad H. 2013. Sporisorium linderi (Ustilaginomycetes) on Digitaria ciliaris,

a new record for Asia. Mycotaxon 123: 229-231. http://dx.doi.org/10.5248/123.229

Vanky K. 2005. The smut fungi (Ustilaginomycetes) of Chrysopogon (Poaceae). Fungal Diversity 18:

177-187.

Vanky K. 2011[“2012”]. Smut fungi of the world. APS Press, St. Paul, Minnesota, USA. xvii + 1458

MY COTAXON

http://dx.doi.org/10.5248/123.261

Volume 123, pp. 261-264 January-March 2013

Yunchangia, a new genus of smut fungi (Ustilaginaceae)

from China

LIn Guo’* & BIAO XU?

'State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

?Tarium University, Xinjiang, Aral 843300, China

* CORRESPONDENCE TO: guol@im.ac.cn

ABSTRACT — Yunchangia puccinelliae gen. et sp. nov. on Puccinellia glauca, is described. The

fungus was collected from Xinjiang Uygur Autonomous Region, China.

Key worps —Ustilaginomycetidae, Ustilaginales, taxonomy

Introduction

The Taklimakan desert located in the southern Xinjiang Uygur Autonomous

Region is the second largest shifting sand desert in the world, covering 337,600

km’. During a mycological expedition to the margin of the western desert

in 2010, a smut species on Puccinellia glauca was collected. The sorus of the

smut causes a short, longitudinal leaf streak. It is covered by the epidermis and

simultaneously contains fungal hyphae and single ustilospores. The fungal

hyphae lie longitudinally along the sides of the sorus. Attempts to germinate

ustilospores from Puccinellia glauca were carried out several times at 24°C on

PDA (potato dextrose agar), but failed. To date 269 species of smut fungi have

been reported in China (Guo 2000, 2011, He et al. 2011).

Material & methods

Ustilospores and hyphae were mounted in lactophenol and examined by light

microscopy (LM), after heating to boiling point. For scanning electron microscopy

(SEM), dried ustilospores and hyphae were dusted onto double-sided adhesive tape,

fixed on specimen stubs, sputter coated with gold, ca. 20 nm thick, and studied with a

FEI Quanta 200 electron microscope.

262 ... Guo & Xu

Taxonomy

Yunchangia L. Guo & B. Xu, gen. nov.

FUNGAL NAME FN570049

Differs from Ustilago by its sorus simultaneously containing fungal hyphae and

ustilospores.

TYPE SPECIES: Yunchangia puccinelliae L. Guo & B. Xu

Erymo.Loecy: The genus is dedicated to Professor Wang Yunchang, an outstanding

Chinese mycologist on taxonomy of smut and rust fungi.

Sori on host plant in the Poaceae forming short streaks mainly on leaves,

covered by the epidermis. Ustilospores single, pigmented. The fungal hyphae lie

longitudinally along the sides of the sorus. Hyphae are composed of pigmented

cells.

ComMENTs: Yunchangia is similar to Ustilago (Pers.) Roussel, which differs by

absence of fungal hyphae in the sorus (Vanky 2002). Its characters demonstrate

that the genus Yunchangia is a member of the family Ustilaginaceae.

Yunchangia puccinelliae L. Guo & B. Xu, sp. nov. Fics 1-6

FUNGAL NAME EN 570050

Differs from Ustilago striiformis by its slightly larger ustilospores and with fungal hyphae

in sorus.

Type: China, Xinjiang, Wuqia, Baykurt, alt. 2600 m, on Puccinellia glauca (Regel)

V.I.Krecz. ex Drobow (Poaceae), 30.VIII.2010, YE Zhu & L. Guo 231 (HMAS 243467,

holotype; HMUT 1228, isotype).

Erymo.ocy: The epithet refers to the host plant genus.

Sori in leaves as short streaks, sparsely in pedicels and inflorescences, 0.5-3.5

mm long, covered by the epidermis which later ruptures exposing powdery,

dark brown spore mass. Ustilospores subglobose, ellipsoidal, ovoid or irregular,

9-19 x 7-12.5 um, reddish brown; wall 0.5-0.8 um thick, echinulate as seen by

LM, finely verruculose between the spines as seen by SEM. Fungal hyphae are

composed of cells; cells cylindrical, ovoid or irregular, 5-10.5 x 4-8 um, reddish

brown; wall 0.5-0.8 um thick, smooth as seen by LM, rough or verruculose as

seen by SEM.

ComMENTs: Yunchangia puccinelliae is similar to Ustilago striiformis (Westend.)

Niessl, which differs by its slightly smaller ustilospores [9-15(-16) x 8-12(-13)

um] and the absence of fungal hyphae in the sorus (Vanky 1994).

Fics. 1-6. Yunchangia puccinelliae (HMAS 243467, holotype). 1. Sori in leaves. 2-4. Ustilospores

and fungal hyphae (LM). 5-6. Ustilospores and fungal hyphae (SEM).

Yunchangia puccinelliae gen. & sp. nov. (China) ... 263

iad ig X j a? _:* - ao Te ts? > re

WD Mag — ‘Oum— : WD Mag HV Det Spot- 20.0um

9 ann 11.8 mm 3000x 10.0 kVETD 3.0

264 ... Guo & Xu

Acknowledgements

The authors would like to express their deep thanks to Dr. K. Vanky (Tubingen,

Germany) for suggestion to describe the new genus, to Drs. Eric H.C. McKenzie

(Auckland, New Zealand) and Cvetomir M. Denchev (Sofia, Bulgaria) for serving as

pre-submission reviewers, to Dr. Shaun Pennycook (Auckland, New Zealand) for

nomenclatural review, and to Dr. Wenli Chen (Institute of Botany, Chinese Academy

of Sciences) for identifying the host plant. This study was supported by the National

Natural Science Foundation of China (No. 30960019).

Literature cited

Guo L. 2000. Flora fungorum sinicorum vol. 12 Ustilaginaceae. Science Press, Beijing. 124p.

Guo L. 2011. Flora fungorum sinicorum vol. 39 Tilletiales Urocystales etc. Science Press, Beijing.

124p.

He SQ, Li HX, Chang XY, Wang SR. 2011. Anthracoidea maquensis, a new species of smut fungi

from China. Mycol Progr. 10:53-55. http://dx.doi.org/10.1007/s11557-010-0672-7

Vanky K. 1994. European smut fungi. Gustav Fischer Verlag Stuttgart, Jena, New York. 570p.

Vanky K. 2002. Illustrated genera of smut fungi. APS Press, St. Paul, Minnesota. 238 p.

MY COTAXON

http://dx.doi.org/10.5248/123.265

Volume 123, pp. 265-270 January-March 2013

Peltaster fructicola, a newly recorded species from China

associated with sooty blotch and flyspeck

CHEN CHEN’, Liu GAo’', MEIYAN QU’, XIAOYONG WEI’,

WENHUAN LI’, RONG ZHANG', GUANGYU SUN’ & MARK L. GLEASON?

'State Key Laboratory of Crop Stress Biology in Arid Areas and College of Plant Protection,

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 — Peltaster fructicola is first reported and described from China. This fungus was

isolated from peels of crabapple and hawthorn collected from Liaoning and Shaanxi Provinces

and exhibited the punctate mycelial type of sooty blotch and flyspeck on both hosts. Our

isolates were identified based on morphological characteristics and DNA sequence analysis.

Key worps — Rosaceae, taxonomy, morphology, phylogeny

Introduction

Sooty blotch and flyspeck (SBFS) is a commonly occurring disease complex

in humid and temperate regions worldwide, causing blemishes on apple, pear,

banana, and other cultivated crops that result in economic losses (Williamson

& Sutton 2000, Batzer et al. 2005, Gleason et al. 2011). The SBFS fungi colonize

the waxy cuticle of plants and display various mycelial types. The term “sooty

blotch” refers to colonies on hosts that consist of a dark mycelial mat with or

without sclerotium-like bodies, whereas “flyspeck” denotes a cluster of black

dots without a visible mycelial mat. “Sooty blotch” was attributed to a single

fungal species for more than 75 years (Colby 1920, Williamson & Sutton

2000) until the late 1990s, when Johnson et al. (1996, 1997) found three “sooty

blotch” species based on morphological examination of sooty blotch isolates

from apples in USA. One newly described species from that work was Peltaster

fructicola (Johnson et al. 1996).

During a recent survey of SBFS complex on fruit of rosaceous hosts in China,

crabapples and hawthorns were found to display colonies of SBFS. Based on

analysis of morphology and DNA sequences of nuclear ribosomal ITS region,

our isolates were identified as P fructicola.

266 ... Chen & al.

Materials & methods

Isolates and morphology

Crabapple (Malus xmicromalus) and hawthorn (Crataegus pinnatifida) fruit

with SBFS signs were collected from Liaoning and Shaanxi Provinces. Two isolates

were obtained from hawthorns in Liaoning and Shaanxi Provinces, and three from

crabapples in Liaoning Province. Peels displaying SBFS were examined, photographed

and preserved. Thalli were transferred directly from peels to potato-dextrose agar

(PDA) and cultured at 25°C in the dark (Sun et al. 2003). After one month, pure isolates

were transferred to PDA and water agar (WA, 2% agar) for morphological examination.

Colony descriptions were based on cultures on PDA after 3 weeks in the dark at 25°C.

Morphological descriptions were made on WA and synthetic nutrient-poor agar (SNA)

after 3 weeks using the method of Zhang et al. (2009). Specimens and representative

dried culture were deposited in the Fungal Herbarium of Northwest A&F University

(HMUABO), Yangling, Shaanxi Province, China.

DNA extraction, PCR and sequencing

Genomic DNA was extracted from fungal mycelium according to the protocol of

Barnes et al. (2001). The primers ITS1-F (Gardes & Bruns 1993) and ITS4 (White et

al. 1990) were used to amplify the internal transcribed spacer (ITS) region of nuclear

ribosomal DNA. The PCR conditions followed the methods of Zhuang et al. (2010).

PCR products were sequenced by Sangon Biotech (Shanghai), China.

Sequence alignment and phylogenetic analysis

Sequences generated in this study were added to other sequences of Peltaster

spp., as well as the outgroup species Mycosphaerella lateralis and Schizothyrium pomi

obtained from GenBank. Preliminary alignments were performed using CLUSTAL

X (Thompson et al. 1997), and then manually adjusted using BioEdit v. 5.0.9.1 (Hall

1999). Phylogenetic analysis of aligned sequences was performed using PAUP v. 4.0b

10 (Swofford 2003). The heuristic search option was 1000 random taxa addition and

tree bisection-reconnection (TBR) as the branch-swapping algorithm. The robustness of

clades and internal branches was evaluated by 1000 bootstrap replications. Tree length

(TL), consistency index (CI), retention index (RI) and rescaled consistency index (RC)

were also calculated. Sequences generated in this study were deposited in GenBank.

Alignment and the representative tree were deposited in TreeBase (http://purl.org/

phylo/treebase/phylows/study/TB2:S13540).

Results

Phylogenetic analysis

The manually adjusted ITS alignment contained 27 sequences (including

the outgroup sequences). Of the 544 characters used for the phylogenetic

analysis, 153 were parsimony-informative, 95 were variable and parsimony-

uninformative, and 296 were constant. The first 100 equally most parsimonious

trees were obtained from the parsimonious analysis, and the first of which is

shown in Fic. 1 (TL = 403 steps, CI = 0.8734, RI = 0.9373, RC = 0.8187). The

Peltaster species formed a strongly supported clade with a bootstrap value of

Peltaster fructicola, new to China ... 267

Peltaster fructicola FJ808757

Peltaster fructicola JN573671

Peltaster fructicola JN573670

Peltaster fructicola JN573669

Peltaster fructicola AY598887

Peltaster fructicola AY598886

Peltaster fructicola JQ358792

LNHT1405a

Peltaster fructicola FJ438382

LNHT1802

SNSZ17

LNSZ1301a

Peltaster fructicola FJ808756

LNHT1506

Peltaster sp. 65rap JN573668

Peltaster sp. 1 HJS-2011 JN573682

Peltaster sp. ZalMj1-4 HQ386245

1001 peltaster sp. 1 HJS-2011 JN573672

Peltaster sp. 1 HJS-2011 JN573677

Peltaster sp. P2.2 AY598890

Peltaster sp. WLE15 HQ386246

Peltaster sp. P2.2 AY598889

91) Peltaster sp. P2.1 AY598888

Peltaster sp. P2.3 FJ438383

Peltaster sp. P2.5 JQ347532

Schizothyrium pomi GU570543

Mycosphaerella lateralis AY725551

Fic. 1. One of 100 equally most parsimonious trees obtained from a heuristic search with 1000

random taxon additions of the ITS sequence alignment. The bootstrap support values (50%)

based on 1000 replicates are shown at the nodes. ‘The tree was rooted to Mycosphaerella lateralis

and Schizothyrium pomi. The scale bar shows 10 changes. Strains investigated in this paper are

presented in bold.

100%, which further divided into two subclades. Our five strains clustered

together with Peltaster fructicola within one of the subclades with a high

bootstrap value of 100%, indicating that they might represent the same species

(Fic. 1).

Taxonomy

Peltaster fructicola Eric M. Johnson, T.B. Sutton & Hodges,

Mycologia 88(1): 120, 1996 Fic. 2

HypHAE smooth, septate, branched, hyaline and turning to pale brown

as culture aged, 1.5-2.0 um wide. CONIDIOPHORES absent. CONIDIOGENOUS

CELLS hyaline, monoblastic, intercalary, formed directly on undifferentiated

hyphae. Conidiogenous loci inconspicuous, stub-shaped, concolorous with

conidiogenous cells. Conip1A solitary, unicellular, hyaline, elliptic to ovoidal,

produced in basipetal order from the conidiogenous loci and aggregated in

268 ... Chen & al.

masses near the conidiogenous cells, 4-6.5(-7) x 1.5-2.5 um. Microcyclic

conidiation observed in culture, with expansion and germination of conidia.

Conidia on WA slightly smaller in size than those on SNA, 4-6(-7) x 1.5-2.0 um.

CULTURAL CHARACTERISTICS — On PDA erumpent, compact, with smooth,

crenate margin and sparse to no aerial mycelium, surface strongly folded, outer

zone light gray, center greenish gray to olivaceous-gray, reverse black with

yellow masses of conidia at the bottom of the colony; reaching (13-)14(-16)

mm diam after 21 d at 25°C in the dark.

On peels of crabapple fruit, showing punctate mycelial type, visible mycelial

mat with dull brown, flattened, circular or irregular sclerotium-like bodies,

68-145 um diam. On peels of hawthorn fruit similar, sclerotium-like bodies

62-223 um diam.

SPECIMENS EXAMINED (all conserved in HMUABO): CHINA, LIAONING PROVINCE:

Huludao City, Suizhong County, 40°19'32"N 120°17'28"E, on fruit surface of crabapple

(Malus xmicromalus Makino), Sept. 2010, Zhuang JL LNHT1506 (GenBank JX961608),

LNHT1802 (GenBank JX961609), LNHT1405a (GenBank JX961610); on fruit surface of

hawthorn (Crataegus pinnatifida Bunge), Sept. 2010, Zhuang JL LNSZ1301a (GenBank

JX961607). SHAANXI PROVINCE: Shangluo City, Shangnan County, on fruit surface of

hawthorn (Crataegus pinnatifida), 21 Oct. 2011, Chen C & Dang JL SNSZ17 (GenBank

JX961611).

Discussion

The genus Peltaster was established by Sydow & Sydow in 1917; the type

species is Peltaster hedyotidis Syd. & P. Syd. Sutton and his colleagues described

P. fructicola (Johnson et al. 1996) as one of the pathogens of sooty blotch and

later associated it with punctate mycelial type on apple (Williamson et al. 2004).

Since then, this fungus has been found widely in the eastern and midwestern

United States (Johnson et al. 1997, Batzer et al. 2005, Diaz Arias et al. 2010).

As the etiology of SBFS is studied worldwide, P fructicola has been reported

to cause sooty blotch in many other countries and regions, including Serbia,

Montenegro, and Poland (Ivanovic et al. 2010, Mirzwa-Mroz & Winska-Krysiak

2011), indicating that P fructicola is among the most prevalent SBFS species.

Based on phylogenetic analysis of the ITS region and morphological

comparison, we identified our isolates as Peltaster fructicola, a newly recorded

species for China. To our knowledge, this is also the first report of SBFS on

crabapple and hawthorn caused by P. fructicola.

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, Private Bag 92170, Auckland,

Peltaster fructicola, new to China ... 269

Fic. 2. Peltaster fructicola (LNHT 1506). A. Signs on crabapple. B. Colony on PDA. C, D. Conidia and

conidiogenous cells. E-G. Conidiogenous cells giving rise to conidia, with visible conidiogenous

loci. H-J. Conidia expanding and germinating and microcyclic conidiation. Scale bars: A = 0.2 mm;

C-E, H-J = 10 um; KF G=5 um.

New Zealand) and Professor Zhongyi Zhang (College of Plant Protection, Yunnan

Agricultural University, Kunming, Yunnan, China) for reviewing the manuscript.

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midwestern United States. Phytopathology 100: 345-355. http://dx.doi.org/10.1094/PHYTO-

100-4-0345

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes - application

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Gleason ML, Batzer JC, Sun GY, Zhang R, Diaz Arias MM, Sutton TB, Crous PW, Ivanovic

M, McManus PS, Cooley DR, Mayr U, Weber RWS, Yoder KS, Del Ponte EM, Biggs AR,

Oertel B. 2011. A new view of sooty blotch and flyspeck. Plant Disease 95: 368-383.

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Mirzwa-Mroéz E, Winska-Krysiak M. 2011. Diversity of sooty blotch fungi in Poland. Acta

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Zhuang JL, Zhu MQ, Zhang R, Yin H, Lei YP, Sun GY, Gleason ML. 2010. Phialophora

sessilis, a species causing flyspeck signs on bamboo in China. Mycotaxon 113: 405-413.

http://dx.doi.org/10.5248/113.405

MY COTAXON

http://dx.doi.org/10.5248/123.217

Volume 123, pp. 271-276 January-March 2013

A new greenish gilled species of Marasmius (Agaricales)

from Hainan Island, China

Y1-Hua YANG'”, CHUN- YING DENG? & TAI-HUI LI”

'Chuxiong Institute of Forestry Sciences & Chuxiong Extending Station of Forestry Technology,

Chuxiong 675005, China

? Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application,

Guangdong Open Laboratory of Applied Microbiology, State Key Laboratory of Applied

Microbiology (Ministry-Guangdong Province Jointly Breeding Base), South China, Guangdong

Institute of Microbiology, Guangzhou 510070, China

° Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource, Hainan University,

Haikou, Hainan 570228, China

“ CORRESPONDENCE TO: mycolab@263.net

AsBstTRACT —Marasmius subviridiphyllus is described as a new species from China supported

by macro- and microscopic characters. It is classified in sect. Marasmius and characterized

by its collariate greenish lamellae, developed rhizomorphs, slender fruiting bodies, dull red

sulcate pileus with a small greenish white disc, middle sized basidiospores (8.8-10.6 x 3.9-5

um), Siccus-type cheilocystidia, and pileipellis broom cells. The holotype (GDGM 26402) is

deposited in the Fungal Herbarium of Guangdong Institute of Microbiology (GDGM).

Key worps — Basidiomycetes, Marasmiaceae, morphology, taxonomy

Introduction

Marasmius (Basidiomycota, Marasmiaceae) is a large genus with complex and

diverse morphological characters, for which six sections are accepted (Wilson &

Desjardin 2005). Representatives of sect. Marasmius are characterized by their

collariate lamellae, insititious stipe, and hymeniform pileipellis with Rotalis- or

Siccus-type broom cells. More than 200 species belong to this section (Singer

1964, 1976; Desjardin & Horak 1997, Desjardin & Ovrebo 2006, Antonin 2003,

2007; Antonin & Buyck 2006, Antonin & Noordeloos 2010).

Twenty-two species of Marasmius sect. Marasmius have been recorded in

China (Bi et al. 1985, 1993, 1997; Chang & Mao 1995, Keissler & Lohwag 1937,

Li & Bau 2003, Li et al. 1994, Mao 1998, Shao & Xiang 1997, Tai 1979, Teng

1936, Zhuang 2005). During a survey in Hainan Province, we discovered a new

species belonging to sect. Marasmius, which we describe and illustrate here.

272 ... Yang, Deng & Li

Materials & methods

Specimens were annotated and photographed in the field, dried in an electric drier

at a temperature of 70°C, and then preserved in the herbarium. Spacing of lamellae

spacing is accompanied by the number of lamellae reaching the pileus margin from

the stipe, and the series number indicates the number of lamellulae between lamellae.

Color terms and notations follow those of Kornerup & Wanscher (1978). Fungal tissues

were mounted in 5% KOH and Melzer’s reagent for microscopic examination. Spore

statistics include: x, , the arithmetic mean of the spore length by spore width (+ standard

deviation) for n spores measured in a single specimen; Q, the quotient of spore length

by spore width in any one spore, indicated as a range of variation in n spores measured;

Q_. the mean of Q-values in a single specimen; n, the number of spores measured per

specimen; s, the number of specimens involved. Specimens are deposited in Fungal

Herbarium of Guangdong Institute of Microbiology (GDGM). Authors of fungal names

are cited according to the International Plant Names Index Authors website (http://www.

ipni.org/ipni/authorsearchpage.do).

Taxonomy

Marasmius subviridiphyllus Chun Y. Deng, Yi H. Yang & T.H. Li, sp. nov. Fics 1, 2

MycoBank MB800428

Differs from Marasmius nigrobrunneus by its dull red pileus, pale green lamellae, and

narrower basidiospores.

Type: China, Hainan Province, Ledong County, Jiaxi Provincial Nature Reserve, 22 July

2009, Chun Y. Deng (Holotype, GDGM 26402).

ErymMo oey: sub- = almost, viridis = green, phyllum = lamella; referring to the greenish

lamellae.

PILEus 3-7 mm, hemispherical to convex when young, umbilicate to depressed

at the center, deeply sulcate, glabrous, mostly reddish gray to dull red (8B2-3;

9B3), with a minute papilla at a small greenish white to greenish (29A2-3) disc.

LAMELLAE collariate near to the stipe, or subfree with the broken collar when

mature, distant (13-14), with 0-1 lamellulae, greenish white (29A2, 30A2) to

pale green (29A3). STIPE 30-60 mm long, 0.2-0.5 mm thick, deep chestnut,

brownish gray (8F2-3) to fuliginous or black, shiny, glabrous, insititious, equal,

arising from black rhizomorphs.

BASIDIOSPORES 8.8-10.6 x 3.9-5 um, [x = 9.3 x 4.8 um, Q = 1.68-2.4, Q

= 1.9, n = 20 spores, s = 1 specimen], ellipsoid, smooth, thin-walled, hyaline,

inamyloid. Basrp1a 26-32 x 6-8 um, clavate, 2- or 4- spored, thin-walled,

hyaline, inamyloid. BAstpIOLEs 15-23 x 6-8 um, fusoid, clavate, thin-walled,

hyaline, inamyloid. LAMELLAE EDGE sterile. CHEILOCysTIDIA 10-30 x 6-11

um, clavate, subcylindrical or irregular in outline, broom cells of the Siccus-

type, with several apical cylindrical to conical appendages or branched setulae

1-7 x 1-1.5 um, thin- or thick-walled. PLEUROcysTIDIA absent. HYPHAE of

hymenophoral trama and the pileus trama thin-walled, hyaline, dextrinoid,

Marasmius subviridiphyllus sp. nov. (China) ... 273

Scale bar = 1 cm. (Photo: Chun Y. Deng).

non-gelatinized. STIPITIPELLIS consisting of parallel, thin-walled, brown,

dextrinoid hyphae. PILEIPELLIs a hymeniform layer of Siccus-type broom cells:

main body 23-32 x 6-10 um, cylindrical, subglobose to clavate, thin- to thick-

walled, hyaline, inamyloid; apical setulae 2-7 x 1-2 um, cylindrical to conical,

rarely forked or warted, thin- to thick-walled, light brown in KOH, inamyloid.

CAULOCYSTIDIA absent. CLAMP CONNECTIONS present in all tissues.

ECOLOGY & DISTRIBUTION—Saprotrophic, gregarious on dead branches,

sticks, etc. of dicotyledonous plants. In mixed forest, China (Hainan). July.

CoMMENTS— ‘The main characters of Marasmius subviridiphyllus are the

reddish gray to dull red, glabrous pileus, slender and shiny stipe, collariate pale

green lamellae, black rhizomorphs, and Siccus-type pileipellis broom cells, and

cheilocystidia. It belongs in sect. Marasmius subsect. Sicciformes according to

Antonin (1991) and Wilson & Desjardin (2005). The pale green lamellae are

distinctive within subsect. Sicciformes, because almost all known species in the

subsection have white, yellowish white, or cream lamellae.

Among the species with tiny dark basidiomata, the new species resembles

M. nigrobrunneus (Pat.) Sacc. and its provisional form M. nigrobrunneus

“f. cinnamomeus,” M. ruforotula Singer, and M. subruforotula Singer. Marasmius

274 ... Yang, Deng & Li

() om WH

Ke; 7

Fic. 2: Marasmius subviridiphyllus (Holotype, GDGM 26402).

A. Cheilocystidia; B. Basidiospores; C. Pileipellis. Scale bars = 10 um.

nigrobrunneus is separated by a grayish brown pileus and broader basidiospores

(5-6 um) (Patouillard 1893, Singer 1958, 1976; Pegler 1983, 1986; Antonin

2007), and M. nigrobrunneus “f. cinnamomeus” differs in its brown to orangish

brown pileus and the presence of smooth cells in the pileipellis and fewer

broom cells (Wannathes et al. 2009); M. subruforotula differs in having smaller

Marasmius subviridiphyllus sp. nov. (China) ... 275

broom cells in the pileipellis and cheilocystidia and coralloid cells among apical

setulae (Singer 1964, Antonin & Buyck 2006). Marasmius ruforotula differs in

closer (<17) and marginate lamellae, longer stipe, absence of rhizomorphs, and

inamyloid pileus and lamellar trama (Singer 1948, Wannathes et al. 2007)

Marasmius bekolacongoli Beeli, M. grandiviridis Wannathes et al., and

M. galbinus T.H. Li & Chun Y. Deng resemble the new species in having green

lamellae, but all three have a hymeniform pileipellis composed of smooth

cells and belong to Marasmius sect. Globulares (Beeli 1928, Antonin 2007,

Wannathes et al. 2009, Deng & Li 2011).

Acknowledgments

The authors unaffectedly thank Dr. Vladimir Antonin (Moravian Museum, Brno,

Czech Republic), Dr. Machiel E. Noordeloos (University of Leiden, The Netherlands)

and Jadson J.S. Oliveira (Biodiversidade Vegetal e Meio Ambiente, Instituto de Botanica,

Sao Paulo, Brazil) for reviewing the manuscript. We also thank Dr. Genevieve M. Gates

(University of Tasmania) for reviewing the English grammar. The study was supported

by the National Natural Science Foundation of China (No.30770004, 31260011), the

Natural Science Foundation of Guangdong (No. A06020222, E05202480), The Central

Financial Forestry Science and Technology to Promote the Demonstration Fund Projects

of China (No. [2009] TQYN004), and High-level Personnel Training Project of Yunnan

Province (No. 2010CI104).

Literature cited

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section Marasmius and a key to European species of Marasmius section Marasmius. Acta Mus.

Moraviae, Sci. Nat. 76: 145-147.

Antonin V. 2003. New species of marasmioid genera (Basidiomycetes, Tricholomataceae) from

tropical Africa I. Sect. Epiphylli, Fusicystides, Globulares, Hygrometrici and Neosessiles.

Mycotaxon 85: 109-130.

Antonin V. 2007. Monograph of Marasmius, Gloiocephala, Palaeocephala and Setulipes in tropical

Africa. Fungus flora of tropical Africa I. Meise, National Botanic Garden (Belgium).

Antonin V, Buyck B. 2006. Marasmius (Basidiomycota, Marasmiaceae) in Madagascar and the

Mascarenes. Fungal Diversity 23: 17-50.

Antonin V, Noordeloos M. 2010. A monograph of marasmioid and collybioid fungi in Europe.

Eching, IHW Verlag.

Beeli M. 1928. Contribution a létude de la flore mycologique du Congo. Fungi Goossensiani V.

Bull. Soc. Roy. Bot. Belg. 60: 153-174.

Bi ZS, Zheng GY, Liang JQ, Li C, Li TH. 1985. Taxonomic studies on Marasmius from Dinghu

Mountain of China. Acta Mycol. Sinica 4(1): 41-50. (in Chinese)

Bi ZS, Zheng GY, Li TH. 1993. The macrofungus flora of China's Guangdong province. Hong Kong

(China), The Chinese University Press.

Bi ZS, LiTH, Zhang WM, Song B. 1997. A preliminary agaric flora of Hainan Province. Guangzhou,

Guangdong Higher Education Press. (in Chinese)

Chang ST, Mao XL. 1995. Hong Kong mushrooms. Hong Kong, The Chinese University press Hong

Kong. (in Chinese)

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Deng CY, Li TH. 2011. Marasmius galbinus, a new species from China. Mycotaxon 115: 495-500.

http://dx.doi.org/10.5248/115.495

Desjardin DE, Horak E. 1997. Marasmius and Gloiocephala in the South Pacific Region: Papua New

Guinea, New Caledonia and New Zealand taxa. Part 1: Papua New Guinea and New Caledonia

taxa. Part 2: New Zealand taxa. Biblioth. Mycol. 168: 1-152.

Desjardin D, Ovrebo CL. 2006. New species and new records of Marasmius from Panama. Fung.

Divers. 21: 19-39.

Keissler K, Lohwag H. 1937. Fungi. In: H Handel-Mazzetti (ed.). Symbolae Sinicae 2. 83 p.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3 ed. London. Methuen.

Li TH, Bi ZS, Zheng GY, Zhang WM. 1994. Species of Marasmius from Guangdong and Hainan

provinces. Acta Mycol. Sinica 13(4): 249-254. (in Chinese)

Li Y, Bau T. 2003. Mushrooms of Changbai Mountains. Beijing, Science Press. (in Chinese)

Mao XL. 1998. Economic fungi of China. Beijing, Science Press. (in Chinese)

Patouillard N. 1893. Quelques champignons du Thibet. J. Bot. Morot. 7: 343-344.

Pegler DN. 1983. Agaric flora of the Lesser Antilles. Kew Bull. Addit. Ser. 9: 668 p.

Pegler DN. 1986. Agaric flora of Sri Lanka. Kew Bull. Addit. Ser. 12: 519 p.

Shao LP, Xiang CT. 1997. Forest mushrooms of China. Harbin, Northeast Forestry University Press.

(in Chinese)

Singer R. 1948. Diagnoses fungorum novorum agaricalium. Sydowia 2: 26-42.

Singer R. 1958. Studies toward a monograph of the South American species of Marasmius. Sydowia

12: 54-148.

Singer R. 1964. Marasmius. Flore Iconographique Champignons Congo, Fasc. 14: 253-278,

planches XLIV-XLVI. Jardin Botanique de l Etat, Ministére de [Agriculture Bruxelles.

Singer R. 1976. Marasmieae (Basidiomycetes—Tricholomataceae). Flora Neotropica 17: 1-347.

Tai FL. 1979. Sylloge fungorum Sinicorum. Beijing, Science Press. (in Chinese)

Teng SC. 1936. Additional fungi from China II. Sinensia 7: 490-527.

Wannathes N, Desjardin DE, Lumyong S. 2007. Mating studies, new species, and new reports of

Marasmius from Northern Thailand. Mycol. Res. 111: 985-996.

http://dx.doi.org/10.1016/j.mycres.2007.06.013

Wannathes N, Desjardin DE, Hyde KD, Perry BA, Lumyong S. 2009. A monograph of Marasmius

(Basidiomycota) from Northern Thailand based on morphological and molecular (ITS

sequences) data. Fung. Diver. 37: 209-306.

Wilson AW, Desjardin DE. 2005. Phylogenetic relationships in the gymnopoid and marasmioid

fungi (Basidiomycetes, euagaric clade). Mycologia 97(3): 667-679.

http://dx.doi.org/10.3852/mycologia.97.3.667

Zhuang WY. 2005. Fungi of Northwestern China. Mycotaxon Ltd., Ithaca. 430 p.

MYCOTAXON

http://dx.doi.org/10.5248/123.277

Volume 123, pp. 277-280 January-March 2013

A new species and new record of Chloridium

from the Qinghai-Tibet Plateau Area, China

YUE-MING Wu & TIAN-YU ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

Key Laboratory of Agricultural Microbiology, Shandong Province, Taian, 271018, China

*CORRESPONDENCE TO: tyzhang1937@yahoo.com.cn

ABSTRACT — Anew species, Chloridium xigazense, and anew record for China, C. phaeosporum,

are described and illustrated. Specimens (dried cultures) and living cultures are deposited in

the Herbarium of Shandong Agricultural University, Plant Pathology (HSAUP). Duplicates

are kept in the Herbarium of Institute of Microbiology, Academia Sinica (HMAS).

Key worps — dematiaceous hyphomycetes, soil fungi, taxonomy

Introduction

During the course of a survey of soil dematiaceous hyphomycetes in China,

two Chloridium species were isolated. One represents a new species and the

other is a new record for China. They are both described and illustrated from

cultures grown on corn meal agar (CMA; Matsushima 1975).

The genus Chloridium was established by Link (1809) and is characterized

by proliferating macronematous conidiophores. The conidiogenous cells are

monophialidic, integrated, terminal, usually percurrent, and more or less

cylindrical. Conidia, which are formed in slimy masses, are simple, ellipsoidal

or subspherical, 0-septate, hyaline, and smooth. Index Fungorum (2012) lists 69

taxa; however, many of these are infraspecific, and Seifert et al. (2011) estimate

that the genus may contain only 20 valid species.

Chloridium xigazense Y.M. Wu & T.Y. Zhang, sp. nov. FIG. 1

MycoBank MB 801786

Differs from Chloridium smithiae by its larger conidia and from C. phaeosporum by its

frequently proliferating conidiogenous cells and hyaline conidia.

TyPE: China, Tibet, Xigaze, from a grassland soil, altitude 3600 m, 7 Sept. 2007, Y.M. Wu

(Holotype HSAUP II ,.0874; isotype HMAS 196268).

ETyMOLoGey: in reference to the type locality.

278 ... Wu & Zhang

Fic. 1. Chloridium xigazense (ex holotype).

Conidia, conidiophores and conidiogenous cells. Scale bar = 25 um.

CoLonigs on CMA broadly effuse, brown to dark brown. Mycelium mostly

immersed, composed of branched, septate, smooth, subhyaline to light

brown, 2-3 um wide hyphae. ConrpropHoREs brown, paler towards the apex,

macronematous, mononematous, solitary or in groups, erect, septate, smooth,

80-140 um long, 2-3 um wide. CONIDIOGENOUS CELLS monophialidic,

terminal, constricting abruptly and expanding in a flaring collarette, thereafter

frequently proliferating 1-4 times. Conip1 produced singly but often adhering

Chloridium xigazense sp. nov. (China) ... 279

in chains, 0-septate, smooth, hyaline, ellipsoidal to oblong, obtuse at the apex,

with a dark hilum at the subtruncate base, 3-5 x 2-2.5 um.

Comments - Morphologically, C. xigazense resembles C. smithiae R.C. Sinclair

& Eicker (Sinclair & Eicker 1985) and C. phaeosporum (Gams & Holubova-

Jechova 1976), but C. smithiae has smaller conidia (2-3 x 1-1.7 um) while

C. phaeosporum shows infrequent proliferation of the conidiogenous cells and

its conidia are pigmented.

Chloridium phaeosporum W. Gams & Hol.-Jech., Stud. Mycol. 13: 27,1976 Fic. 2

CoLonigs on CMA effuse, olivaceous brown, velvety. Mycelium superficial

or immersed; hyphae branched, septate, smooth, pale to mid brown, 1.5-2.5 um

wide. CONIDIOPHORES mid brown, paler towards the apex, macronematous,

Fic. 2. Chloridium phaeosporum (HSAUP II 1037).

Conidia, conidiophores and conidiogenous cells. Scale bar = 25 um.

280 ... Wu & Zhang

mononematous, solitary or in groups, erect, septate, smooth, 70-120 um long,

2.5-3.5 um wide. CONIDIOGENOUS CELLS monophialidic, terminal, constricting

abruptly and expanding in a flaring collarette, rarely proliferating. CoNnrp1A

aggregated in heads, 0-septate, smooth, pale brown, ellipsoidal to oblong,

obtuse at the apex, with a dark hilum at the subtruncate base, 3-6 x 1.5-2.5

um.

SPECIMEN EXAMINED: CHINA, TIBET, Xigaze, altitude 3000 m, from a mountain soil, 10

Sept. 2007, Y.M. Wu (HSAUP II 1037, HMAS 196269).

Chloridium phaeosporum is reported for the first time from China. Compared

with the morphological characters of the species as described by Gams &

Holubova-Jechova (1976), both collections have pale brown conidia that are

ellipsoidal to oblong, 0-septate, obtuse at the apex, with a dark hilum at the

subtruncate base, and measure 3-6 x 1.5-2.5 um. We believe they are the same

species. Chloridium phaeosporum is most similar to C. virescens (Pers.) W. Gams

& Hol.-Jech. var. virescens (Gams & Holubova-Jechova 1976) in conidial shape,

but the latter species has hyaline conidia without a dark basal hilum.

Acknowledgments

The authors are grateful for pre-submission comments and suggestions provided by

Dr. Eric McKenzie, Prof. Y.L. Guo, and Dr. Shaun Pennycook. This project was supported

by the National Science Foundation of China (no. 30970011 & 30499340).

Literature cited

Gams, W, Holubova-Jechova V. 1976. Chloridium and some other dematiaceous hyphomycetes

growing on decaying wood. Stud. Mycol. 13: 1-99.

Index Fungorum. 2012. http://www.indexfungorum.org/Names/Names.asp - accessed 16 Oct

2012.

Link HF. 1809. Observationes in ordines plantarum naturales. Mag. Ges. Natur. Freunde, Berlin

3: 3-42.

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum, Kobe. 209 p. + plates.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series, vol. 9. 997 p.

Sinclair RC, Eicker A. 1985. A new species of Chloridium from South Africa. Trans. Br. Mycol. Soc.

84: 566-568. http://dx.doi.org/10.1016/S0007-1536(85)80029-2

MY COTAXON

http://dx.doi.org/10.5248/123.281

Volume 123, pp. 281-284 January-March 2013

A new species of Heliocephala from Vietnam

VADIM A. MEL’NIK', RAFAEL F. CASTANEDA-RUIZ?

& MILAGRO GRANADOS?*

'V. L. Komarov Botanical Institute of the Russian Academy of Sciences,

Prof. Popov Street 2, 197376, St. Petersburg, Russia

?Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

°Centro de Investigaciones en Proteccién de Cultivos (CIPROC), Universidad de Costa Rica

*CORRESPONDENCE TO: milagranados@gmail.com

ABSTRACT —Heliocephala vietnamensis sp. nov., a hyphomycetous fungus collected on

decaying leaves of an unidentified plant, is described and illustrated. It is distinguished by

determinate apical cluster of conidiogenous cells and obclavate to sub-navicular, 3-septate,

pale brown conidia.

KEY worpDs — anamorphic fungi, systematics, leaf litter

Several samples of dead plant material colonized by anamorphic fungi were

collected in a dense monsoon tropical forest in Vietnam. Among these

samples was a conspicuous Heliocephala species, which is herein described and

illustrated.

Taxonomy

Heliocephala vietnamensis Melnik & R.F. Castafieda, sp. nov. Figs 1,2

MycoBaNnkKMB801539

Differs from Heliocephala elegans and H. triseptata by conidial shape, number of septa

and size.

TyPeE: Vietnam, Dong Nai, Cat Tien National Park, right bank of the Dong Nai River, near

to Ficus sp. plot, a dense monsoon tropical forest, 11°26'N 107°25’E, on decaying leaves

of unidentified plant, 2 January 2011, coll. Yu. Novozhilov, (Holotype: LE 261853).

ErymMo_oey: Latin, vietnamensis, refers to the country where the fungus was found.

Co.LoniEs on the natural substrate effuse, hairy, amphigenous, brown. Mycelium

mostly superficial composed of septate, branched, dark brown, smooth-walled

282 ... Melnik, Castafteda-Ruiz & Granados

Fic 1. Heliocephala vietnamensis (ex holotype LE 261853). A. Conidia. B. Conidiophore,

conidiogenous cell and attached conidia. Scale bars = 10 um.

hyphae, 1.8-2.8 um diam. Conip1oPHoREs distinct, single, cylindrical, erect,

straight, 210-340 x 6-8 um, 14-16 um wide at the base, 7-12-septate, bearing

a conidiogenous apparatus at the apex consisting of 3-6 irregular-campanulate

Heliocephala vietnamensis sp. nov. (Vietnam) ... 283

Fic 2. Heliocephala vietnamensis (ex holotype LE 261853).

Conidiogenous cells and conidia. Scale bar = 10 um.

to irregular-doliiform or cuneiform primary and secondary branches (metula-

like), pale brown, 3-4 x 3.5-4.0 um. CONIDIOGENOUS CELLS monoblastic,

discrete, determinate, delicate, lageniform, 1.7-3.0 x 1.5-2.5 um, pale brown,

smooth, forming a compact cluster at the ends of the branches. CONIDIAL

SECESSION schizolytic. Conrp1A solitary, acrogenous, obclavate to navicular,

3-septate, pale brown, 14-17 x 2.8-3.8 um, smooth-walled, dry. Teleomorph

unknown.

Note: The genus Heliocephala V. Rao et al. (Rao et al. 1984) was based on

H. proliferans, which was distinguished by the production of compact clusters of

discrete, lageniform, monoblastic conidiogenous cells arising more less radially

or closely fasciculate from short secondary or tertiary branches (metula-like) at

the apex of the conidiophores. The conidia are obclavate, long or short rostrate,

septate, pale brown, with an unusual germination of the apical cell to form a

secondary cluster of conidiogenous cells.

284 ... Melnik, Castafteda-Ruiz & Granados

Heredia et al. (2011), who emended Heliocephala after molecular and

morphological studies of some Heliocephala species and the closely similar

Holubovaniella R.F. Castafieda (Castafieda 1985), considered Holubovaniella a

synonym of Heliocephala. They provided a key to the species and expanded

the generic concept of Heliocephala to accommodate taxa with indeterminate

conidiophores with several extensions of the main axis and solitary conidia

produced by the same kind of conidiogenesis. Following Heredia et al. (2011),

Heliocephala comprised six species: H. elegans (R.F. Castafieda) R.F. Castafieda

& Unter., H. gracilis (R.F. Castafieda) R.E Castafieda & Unter., H. natarajanii

Kumaresan & M. Sriniv., H. proliferans V. Rao et al., H. triseptata Heredia et al.,

and H. zimbabweensis Decock et al.

Among the previously described Heliocephala species, only H. elegans and

H. triseptata are similar to H. vietnamensis, but H. elegans (Castafieda-Ruiz

1985) has indeterminate conidiophores with several clusters of short branches

and intercalary conidiogenous cells on the axis before each extension and

olivaceous conidia that are obclavate to lecythiform with a shortly rostrate

apical cell, mostly 2-septate, 8-25 x 3-4 um. H. triseptata (Heredia et al. 2011)

has conidia that are obclavate or navicular to broadly fusiform, rostrate, 15-27

x 3.5-4.5 um with a 10-14 um long rostrum. Neither species can be confused

with H. vietnamensis.

Acknowledgments

The authors express their sincere gratitude to Dr. Bryce Kendrick and Dr. De-Wei Li

for their critical review of the manuscript. We are deeply indebted to Dr. Yuri Novozhilov

for providing the specimen collected. In part this study has been done in frame of

Research Program of the Russian-Vietnamese Tropical Research and Technological

Center is also appreciated. We also thank Dr. Gabriela Heredia for facilities during

microscopic observations and photographs. The authors are deeply indebted to the

Russian Academy of Sciences and the Cuban Ministry of Agriculture for facilities. We

also thank Mirtha Caraballo and Beatriz Ramos for technical assistance. We acknowledge

the facilities provided by Dr. P.M. Kirk and Drs. V. Robert, G. Stegehuis and A. Decock

through the maintenance of the Index Fungorum and Mycobank websites. Dr. Lorelei L.

Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclatural review are greatly

appreciated.

Literature cited

Castafieda-Ruiz RF. 1985. Deuteromycotina de Cuba. Hyphomycetes. 3. Instituto de Investigaciones

Fundamentales en Agricultura Tropical “Alejandro de Humboldt”. La Habana, Cuba. 72 p.

Heredia Abarca G, Castafieda-Ruiz RF, Arias Mota RM, Becerra Hernandez CI, Gémez S, Bogale

M. Untereiner WA. 2011. A new species of Heliocephala from Mexico with an assessment of the

systematic positions of the anamorph genera Heliocephala and Holubovaniella. Mycologia 103:

631-640. http://dx.doi.org/10.3852/10-230

Rao V, Reddy KA, de Hoog GS. 1984. Heliocephala, a new genus of dematiaceous hyphomycetes.

Persoonia 12: 239-242.

MY COTAXON

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Volume 123, pp. 285-287 January-March 2013

Lecanora gansuensis sp. nov. (subfusca group)

from China

LEI LU', QIANG REN’, DAI-FENG JIANG’,

Hat-YING WANG” ’& ZUN-TIAN ZHAO?"

'Shandong Provincial Key Laboratory of Microbial Engineering, School of Food and Bioengineering,

Shandong Polytechnic University, Jinan, 250353, China

College of Life Sciences, Shandong Normal University, Jinan, 250014, China

* CORRESPONDENCE TO: “endolichen@gmail.com & * ztzhao@sohu.com

ABSTRACT -- Lecanora gansuensis is described as new to science from China and South

Korea. It is characterized by flexuous apothecial margins and the presence of stictic acid,

norstictic acid, and connorstictic acid in addition to atranorin.

Key worps -- East Asia, fungi, lichen, stictic acid complex, taxonomy

Introduction

The large genus Lecanora Ach. has been divided into several groups. The

subfusca group is characterized by having atranorin, calcium oxalate crystals in

the apothecial margin, and (usually) reddish brown apothecia (Brodo 1984). The

subfusca group has received considerable interest during recent decades (e.g.,

Brodo 1984, Miyawaki 1988, Brodo et al. 1994, Upreti 1998, Guderley 1999,

Lumbsch et al. 2004, Ryan et al. 2004). A further study of Lecanora specimens

collected from China revealed a new species belonging to the subfusca group,

which is described here.

Materials & methods

The specimens studied are housed in SDNU (Lichen Section of Botanical Herbarium,

Shandong Normal University) or KoLRI (Korean Lichen Research Institute). Thalli were

examined and measured under a stereo—microscope (COIC XTL7045B2) and apothecial

anatomy was observed under a polarizing microscope (OLYMPUS CX41-32). Photos of

the thalli were taken with an OLympus SZX12 with DP70. Chemical analysis using spot

tests and thin layer chromatography followed standard methods (Orange et al. 2010),

mainly using solvent system C.

286 ... Li & al.

C. Granules in amphithecium and epihymenium; D. Ascospores.

Taxonomy

Lecanora gansuensis L. Li & H.Y. Wang, sp. nov. PLATE 1

MycoBANnk 563751

Differs from Lecanora kansriae by its crowded flexuous apothecia and smaller spores.

Type: China, Gansu, Wen County, Qiujiaba, 2300 m, on twigs, 3 Aug 2006, Yang et al.

061482-2 (spNnu holotype).

Erymo_oey: the epithet refers to the holotype locality, Gansu.

Thallus dark brown, well-developed, continuous or rimose-areolate, margin

definite, prothallus absent. Apothecia sessile, crowded, 0.5-1.0 mm in diam.,

discs red brown, epruinose, margins gray-white, prominent, entire, thick,

strongly flexuous. Cortex is distinct, thin, gelatinous, uniform below (15-23

um). Amphithecium containing small crystals (campestris—type) insoluble

in KOH. Epihymenium yellowish brown, with fine granules (pulicaris-type),

12.5-15 um tall. Hymenium hyaline, 50-60 um tall, paraphyses simple, to

be slightly branched at the tip, neither thickened apically nor pigmented.

Hypothecium hyaline, 45-55 um thick. Asci clavate, 8-spored. Ascospores

simple, hyaline, ellipsoid, 8.5-10 x 5-5.5 um.

CHEMISTRY — atranorin (major), stictic acid (major), norstictic acid

(minor), and connorstictic acid (minor).

Lecanora gansuensis sp. nov. (China) ... 287

ADDITIONAL SPECIMENS EXAMINED: CHINA. Gansu: Wen County, Qiujiaba, 2350 m,

on bark, 3 Aug. 2006, Yang et al., 061643; 2600 m, Yang et al., 062075-2. GuIzHOU:

Kaili City, Mt. Leigong, 1700m, on bark, 23 Aug. 2010, Wang 20103351; Libo County,

Maolan Nature Reserve, 350m, on bark, 2 Nov. 2009, Wang 20102329 (SDNU). SOUTH

KOREA. JEOLLANAM-DO: Gurye-gun, Sandong-myeon, Mt. Jiri, 1085 m, on bark, 16

Jun. 2006, Hur 060165-3 (KoLRI).

DISTRIBUTION AND SUBSTRATE — At present, L. gansuensis is known from

China (Gansu and Guizhou) and South Korea on bark or wood.

COMMENTS — Few species of Lecanora have a pulicaris-type epihymenium

with a campestris-type amphithecium, and even fewer contain stictic acid as

a main lichen product. Lecanora kansriae described from Thailand agrees in

all these respects but has much larger spores (17-20 x 10-12 um) and does

not have crowded apothecia with flexuous margins (Papong & Lumbsch 2011).

Lecanora gansuensis morphologically resembles L. allophana (Ach.) Ach. and

L. imshaugii Brodo. However, L. allophana has a glabrata-type epihymenium and

different substances (atranorin alone), while L. imshaugii differs in chemistry,

producing atranorin and zeorin as major substances (Brodo 1984).

Acknowledgements

The project was financially supported by the National Natural Science Foundation

of China (31070010) and Shandong Provincial Natural Science Foundation

(ZR2010CQ038). The authors are grateful to Professor Hur (KoLRI) for providing access

to specimens deposited in KoLRI and Associate Professor Zhang Xuejie (Shandong

Normal University) for providing access to facilities.

Literature cited

Brodo IM. 1984. The North American species of the Lecanora subfusca group. Beihefte zur Nova

Hedwigia 79: 63-185.

Brodo IM, Owe-Larsson B, Lumbsch HT. 1994. The sorediate saxicolous species of the Lecanora

subfusca group in Europe. Nordic Journal of Botany 14: 451-461.

http://dx.doi.org/10.1111/j.1756-1051.1994.tb0063 1.x

Guderley R. 1999. Die Lecanora subfusca-Gruppe in Siid- und Mittelamerika. Journal of the

Hattori Botanical Laboratory 87: 131-257.

Lumbsch HT, Elix JA. 2004. Lecanora. 11-62, in: PM McCarthy, K Mallett (eds). Flora of Australia,

Volume 56 A, Lichens 4. ABRS, CSIRO Australia, Melbourne.

Miyawaki H. 1988. Studies on the Lecanora subfusca group in Japan. Journal of the Hattori Botanical

Laboratory 64: 271-326.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

Papong, K, Lumbsch HT. 2011. A taxonomic survey of Lecanora sensu stricto in Thailand

(Lecanoraceae; Ascomycota). The Lichenologist 43(4): 299-320.

Ryan BD, Lumbsch HT, Messuti MI, Printzen C, Sliwa L, Nash TH III. 2004. Lecanora. 176-286,

in: TH Nash II et al. (eds). Lichen Flora of the Greater Sonoran Desert Region, Vol. 2. Lichens

Unlimited, Arizona State University, Tempe, Arizona.

Upreti DK. 1998. Notes on saxicolous species of the Lecanora subfusca group in India. The

Bryologist 101: 256-262. http://dx.doi.org/10.2307/32442.

MY COTAXON

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Volume 123, pp. 289-292 January-March 2013

A new species of Gautieria from China

TOLGOR Bau?” & Yu Liv?

"Institute of Mycology/Engineering Research Center of Chinese Ministry of Education for Edible

and Medicinal Fungi, Jilin Agricultural University, Changchun 130118, China

?Institute of Mycological Science and Technology, Ludong University/Shandong Key Laboratory

of Edible Mushroom Technology, Yantai 264025, China

* CORRESPONDENCE TO: junwusuo@126.com

ABSTRACT —Gautieria xinjiangensis, collected from Xinjiang, China, is described and

illustrated as new to science.

Key worps —Gomphaceae, new taxon, morphology, taxonomy

Introduction

The diagnostic characteristics of Gautieria Vittad. (Gomphaceae,

Basidiomycota) are a subglobose to reniform fruit body with a thin and easily

evanescent peridium, a cartilaginous gleba with simple or branched columella

and a coralloid type of development, often elongated, labyrinthiform cavities, a

gelatinous-cartilaginous tramal plate and basidiospores with longitudinal ribs

(Cunningham 1944, Zeller 1948, Rauschert 1975, Beaton et al. 1985). About 36

species and varieties have been described, with nine species known from China

(Liu 1998, Ying 1984, 1995).

Recently, we have encountered an additional species, which is proposed here

based on morphological characters.

Materials & methods

Materials were collected at Tianshan Mountains of Xinjiang, northwestern China

and were deposited in the Herbarium of Mycology of Jilin Agricultural University

(HMJAU). Morphological characters of the fungus were described and illustrated with

traditional taxonomic methods. 5% KOH solution and Melzer’s reagent were used when

examining the microscopic characters.

Taxonomy

Gautieria xinjiangensis T. Bau, sp. nov. PLATES 1-2

MycoBank MB 801448

290 ... Bau & Liu

PLaTE 1. Basidiomata of Gautieria xinjiangensis (holotype).

Differs from Gautieria morchelliformis by its larger basidiomata and its broadly ellipsoid

to orbicular ovate basidiospores.

Type: China, Xinjiang Autonomous Prefecture, Urumchi City, Tianshan Mountains, on

ground in forest dominated by Picea schrenkiana Fisch. & C.A. Mey., 5 September 2005,

Tolgor Bau (Holotype, HMJAU6009; GenBank, JX860192).

ETYMOLOGY: xinjiangensis refers to the type locality.

MACROCHARACTERS — Basidioma emergent, 3-5.5 x 3-4 cm, subglobose to

ovoid in outline, surface brain-like, yellow-brown to light rust; basal rhizomorph

whitish, distinct. Peridium practically absent, or barely visible. Gleba yellow-

brown to yellow-tan; columella absent; locules irregularly shaped, labyrinthine;

inner surface covered with yellow-brown or yellow-tan hymenium.

MICROCHARACTERS — Basidiospores (13—)15-19(-21) x (8—)8.5-10(-11)

um, Q = 1.8-2, broadly ellipsoid to orbicular-ovate, yellow-brown to pale

brown in KOH, with 9-11 longitudinal ribs, sometimes with branches or cross

veins between ribs; apex broadly round; apiculus long, 2-3 x 1.5-2 um. Basidia

20-30 x 7-10 um, clavate, with granular or oil drops, 4- or 2-spored; sterigmata

2-3 um long. Cystidia 30-40 x 8-12 um, clavate, thin-walled to slightly thick-

walled, hyaline, colorless. Peridium traces can be seen under a microscope,

and composed of pseudo-parenchyma of large, subglobose to polygonal cells,

usually 30-40 um in diam, thin-walled, smooth, hyaline, frequently septate

(cells 12-25 um long). Tramal hyphae gelatinized, interwoven, frequently

septate and branching, 3-7 um diam., smooth, hyaline. Clamps absent in all

tissues.

Gautieria xinjiangensis sp. nov. (China) ... 291

PLATE 2. Microscopical characters of Gautieria xinjiangensis (holotype). a: young basidiospores;

b: mature basidiospore in polar view; c: mature basidiospores; d: hymenium; e: pseudo-parenchyma

cells of peridium traces; f: cystidia; g: basidium with young basidiospores; h: tramal hyphae.

Scale bar = 10 um.

Discussion

The main characteristics of G. xinjiangensis are the relatively large basidioma

with a white basal rhizomorph, the absence of a columella, and the broadly

ellipsoid basidiospores with 9-11 longitudinal ribs.

The nine Gautieria species reported from China (Liu 1998, Ying 1984, 1995)

are all very different from the new taxon according to the authors’ review of

292 ... Bau & Liu

the literature and examination of voucher specimens. Gautieria hubeiensis

K. Tao et al. and G. morchelliformis Vittad. are distinguished by their lemon-

shaped to long-ellipsoid basidiospores, and G. globispora K. Tao et al. by

its globose basidiospores (Liu et al. 1996, Pegler et al. 1993, Tao et al. 1996,

Vittadini 1831, Zeller & Dodge 1918). The other six species (G. pallida Harkn.,

G. shennongjiaensis K. Tao et al., G. macrospora G. Cunn., G. sinensis J.Z. Ying,

G. chengdensis J.Z. Ying, G. gautierioides (Lloyd) Zeller & C.W. Dodge) differ

from G. xinjiangensis by their smaller basidiomata (Liu 1998). In addition,

G. xinjiangensis is associated with Picea schrenkiana, a plant endemic to

Tianshan Mountains in China.

The ITS sequence of the holotype of G. xinjiangensis (Genbank JX860192)

is highly (<99%) similar to sequences of USA material of G. morchelliformis

(GenBank AF377066, AF377067), but the two species clearly differ in their

basidiospore shape and basidioma size.

Acknowledgments

We thank Dr. Z.L. Yang (Key Laboratory of Biodiversity and Biogeography, Kunming

Institute of Botany, Chinese Academy of Sciences, China) and Dr. R.H. Petersen

(Department of Biology, University of Tennessee, USA) for their presubmission reviews

of the manuscript. Financial support for this work was provided by Changjiang Scholars

and Innovative Research Team in University (No. IRT1134).

Literature cited

Beaton G, Pegler DN, Young TWK. 1985. Gasteroid Basidiomycota of Victoria State, Australia. 3.

Cortinariales. Kew Bulletin 40: 167-204. http://dx.doi.org/10.2307/4108269

Cunningham GH. 1944. The gasteromycetes of Australia and New Zealand. John MclIndoe,

Dunedin. 236 p. [1979 facsimile edition: Bibliotheca Mycologica 67.]

Giachini AJ, Hosaka K, Nouhra E, Spatafora J, Trappe JM. 2010. Phylogenetic relationships of the

Gomphales based on nuc-25S-rDNA, mit-12S-rDNA, and mit-atp6-DNA combined sequences.

Fungal Biology 114: 224-234. http://dx.doi.org/10.1016/j.funbio.2010.01.002

Liu B. 1998. Flora fungorum sinicorum, vol. 7 Hymenogastrales, Melanogastrales et Gautieriales.

Science Press, Beijing (in Chinese).

Liu B, Tao K, Chang MC, Xu AS. 1996. New species and new records of hypogeous fungi from

China VII. Journal of Shanxi University (Natural Science Edition) 19(3): 319-323.

Pegler D N, Spooner B M, Young T W K. 1993. British truffles. A revision of British hypogeous

fungi. Royal Botanic Gardens, Kew. 216 p.

Rauschert R. 1975. Die Gattung Gautieria (Gasteromycetes) in der DDR. Hercynia 12: 217-227.

Tao K, Chang MC, Liu B. 1996. New species and new records of hypogeous fungi from China V.

Journal of Shanxi University (Natural Science Edition) 19(1): 82-87.

Vittadini C. 1831. Monographia Tuberacearum. 88 p.

Ying JZ. 1984. A new species of genus Gautieria Vitt. from China. Acta Mycologica Sinica 3: 89-91.

(in Chinese).

Ying JZ. 1995. Gautieria sinensis sp. nov. (gasteromycetes) from China. Mycotaxon 54: 299-302.

Zeller SM. 1948. Notes on certain gasteromycetes, including two new orders. Mycologia 40:

639-668. http://dx.doi.org/10.2307/3755316

Zeller SM, CW Dodge. 1918. Gautieria in North America. Annals of the Missouri Botanical Garden

5: 133-142. http://dx.doi.org/10.2307/2990107

MY COTAXON

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Volume 123, pp. 293-299 January-March 2013

Typification of Tuber formosanum

(Tuberaceae, Pezizales, Ascomycota) from Taiwan, China

PENG Qiao *’, PeI-Gui Liu ‘+, HUNG Tao Hu? & YUN WANG *3""

" Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany,

Chinese Academy of Science , Lanhei Road 132, Kunming 650201, Yunnan, China

’ School of Forestry and Resource Conservation, National Taiwan University,

Taipei, Taiwan, China

> The New Zealand Institute for Plant & Food Research Ltd.,

Private Bag 4704 Christchurch 8140, New Zealand

* CORRESPONDENCE TO: “giaopeng@mail.kib.ac.cn * Yun. Wang@plantandfood.co.nz

ABSTRACT—When Hu described Tuber formosanum in 1992 as a new species based on

collections from Dung-Pu, Nantou County, Taiwan, he neglected to designate a type, so that

the name was not validly published. The current authors re-describe, illustrate, and validate

T. formosanum based on a type selected from recent collections from the same region.

Tuber formosanum is closely related to the indicum complex but differs in morphological,

phylogenetic, and other respects and should be treated as a separate taxon.

Key Worps—truffle, taxonomy, ITS

Introduction

A few ascocarps of Tuber species were collected from the natural forest of

Cyclobalanopsis glauca (Thunb.) Oerst. (Fagaceae) in Dung-Pu, Nantou County,

Taiwan, in late October 1987, mid-December 1988, early February 1990, and

early January 1991. They grew in calcareous soils (pH 7.0) under Cyclobalanopsis

glauca trees at 1300 m elevation. Hu (1992) described Tuber formosanum as a

new species based on these collections. Because Hu did not designate a type in

his description, the new species was not validly published (McNeill et al. 2006:

Art. 37.1). Recent phylogenetic and morphological analyses of T. formosanum

collections confirm that T: formosanum is an independent species, although

closely related to the T. indicum complex. In addition, it has different host

plants and is restricted to Taiwan. Consequently, T: formosanum should be

treated as a separate taxon (Huang et al. 2009). In this paper, we re-describe,

294 ... Qiao & al.

Taiwan Strait

pu ee mee) He Ho She | |

sap u )

ay [Duna Pu

~ Ohsinrig

43 Q

PLATE 1. Map of Taiwan showing where the type of Tuber formosanum was collected

[adapted from Huang et al. 2009].

illustrate, and validate T: formosanum based on a type newly selected from

specimens collected from the Ho-She district of National Taiwan University

Experimental C. glauca Plantation, Nantou County, Taiwan. The plantation was

established with trees inoculated by T: formosanum from the natural C. glauca

forests in Dung-Pu, Nantou County. The distance between Ho-She plantation

(23°35'01"N 120°51'36"E) and the original natural C. glauca forest in Dung-Pu

(23°30'27"N 120°53'45"E) is 9.21 km (PL.1).

Tuber formosanum, validated (Taiwan) ... 295

TABLE 1: The ITS sequences of Tuber species used in this study.

TAXON VOUCHER OR CODE GEOGRAPHICAL ORIGIN GENBANK

T. melanosporum A71 Spain AF106877

Tm13 Vaucluse, France AF132501

MelW Unknown AF300825

T. sinense Tsin-hd01 Huidong, Sichuan, China DQ375526

Tsin-hd02 Huidong, Sichuan, China DQ375527

KUN-HKAS44319 Huidong, Sichuan, China GU979061

IFS89925 Huidong, Sichuan, China GU979062

T. indicum KUN-HKAS44999 Kunming, Yunnan, China GU979066

KUN-HKAS49746 Yimen, Yunnan, China GU979067

KUN-HKAS41312 Haikou, Yunnan, China GU979069

KUN-HKAS30261 Panzhihua, Sichuan, China GU979050

KUN-HKAS30262 Panzhihua, Sichuan, China GU979051

T. formosanum KUN-HKAS48268 Ho-She, Nantou, Taiwan GU979048

KUN-HKAS49707 Ho-She, Nantou, Taiwan GU979049

KUN-HKAS62628 Ho-She, Nantou, Taiwan JN655530

(holotype)

T. aestivum E5 Molise, Italy AF516791

Materials & methods

The macroscopic and microscopic characters were described based on the type

selected from new specimens collected from Nantou County, Taiwan in 2005. The

methods followed those of Yang & Zhang (2003). Sections were made with a razor blade,

mounted in a 5% KOH solution and examined under a Nikon E400 microscope. For

scanning electron microscopy (SEM), spores were scraped from the dried gleba onto

doubled-sided tape, mounted directly on an SEM stub, coated with gold-palladium,

and examined and photographed with a JEOL, JMS-5600LV SEM. The new specimens

were deposited at the Herbarium of Cryptogams, Kunming Institute of Botany, Chinese

Academy of Sciences (KUN-HKAS).

DNA was extracted from ascomata using a modified CTAB protocol (Doyle 1987). The

primers ITS1F (Gardes & Bruns 1993) and ITS4 (White et al. 1990) were used to amplify

the rDNA ITS region. PCR reaction solution and cycling parameters used by Chen & Liu

(2007) were adopted and modified when necessary. The PCR reaction was performed

on a Takara TP100 thermal cycler. Amplification products were electrophoresed on a

1% agarose gel, and purified with Sangon’s purification kit. Sequencing was performed

with a BigDye® Terminator v3.1 Cycle Sequencing Kit on an ABI 3730XL automatic

sequencer. Sixteen partial internal transcribed spacer ribosomal DNA (ITS-rDNA)

sequences of Tuber species, T: formosanum, T. melanosporum, T. sinense, T. indicum and

T. aestivum were used for analysis. Tuber aestivum was selected as outgroup (TABLE 1).

Software and methods used in sequence alignment and phylogenetic analysis followed

Chen & Liu (2007).

296 ... Qiao & al.

Taxonomy

Tuber formosanum H.T. Hu & Y. Wang, sp. nov. PLATE 2

MycoBank MB 563693

“Tuber formosanum” H.T. Hu, Quart. J. Exp. Forest Nat. Taiwan Univ.

6: 79-86 (1992), nom. inval. [No holotype designated].

Differs from T: indicum by its asci with short stipitate and spiny-reticulate ascospores,

and its association with Cyclobalanopsis glauca.

Type: Taiwan, China: Nantou County, Ho-She, (E120°51', N23°35’), in a Cyclobalanopsis

glauca plantation, alt. 1200 m, 2005, H.D. Hu (Holotype, KUN-HKAS 62628; GenBank

, JN655530).

EryMo_oey: from the Latin formosanum referring to Taiwan, the location of the type

collection.

AscoMarta globose to subglobose and slightly lobed, firm, dark reddish brown

to dark grayish brown, up to 8 cm in diameter, with low pyramidal warts,

pentagonal, having 4-5 ridges, up to 1 mm high. Opor slightly aromatic

when mature. PERIDIUM 215-815 um thick, variable, pseudoparenchymatous,

composed of two layers: outer layer (50-—)135-300(-465) um thick, composed

of angular or irregular cells (4—)8-39(-50) x (4—)5-15(-20) um in diam., with

thick dark brown walls of 0.5-2 um; inner layer (50-)100-135(-560) um thick,

composed of hyaline to yellowish, narrow polygonal cells, merging with glebal

tissue of interwoven hyphae. GLEBA solid, whitish, light yellowish when young,

becoming brown to dark purplish brown at maturity, marbled with distinct,

whitish, meandering veins merging at many points and sometimes penetrating

through the peridium. Asc1 globose or broadly elliptic, (37—)45-65(-85) x

(26-)30-55(-69) um (n = 160), with short stipitate, 1-4(-5) spored, randomly

dispersed in glebal tissue. Ascospores ellipsoid, whitish to yellowish when

young, becoming dark brown at maturity, (27-)29-45(-48) x 20-32(-35)

um (1-spored), (26—)27-36(-39) x (18-)19-24(-28) um (2-spored), 24-34 x

(16—)18-23(-25) um (3-spored), (25-)26-32(-33) x (17-)18-22 um (4-spored)

(excluding ornamentation, n = 40), Q = (1.17-)1.27-1.62(-1.70) (n = 170),

somewhat spiny-reticulate, spines up to 2-5(-6) um (n = 170) tall, wall (2-)3-5

(-6) um (n = 160) thick.

EcoLoGy & DISTRIBUTION: Hypogeous in calcareous soils with pH 7.0

under C. glauca trees at 1200-1300 m a.s.L., fruiting from late October to early

February. Known only from Nantou County, Taiwan, China.

ADDITIONAL SPECIMENS EXAMINED: TAIWAN, CHINA. NantTou County, Ho-She,

23°35'N 120°51’E, in Cyclobalanopsis glauca plantation, alt. 1200 m, 2005, HD. Hu

(KUN-HKAS48268, KUN-HKAS49707).

PLATE 2. Tuber formosanum (holotype, KUN-HKAS62628). A. Ascoma; B. Warts on peridium

surface; C. Part of a section of peridium; D. Sectioned ascoma showing the gleba; E. Ascus with two

ascospores inside; F. Ascospore showing the ornamentation; G, H. Scanning microscopic photos

of ascospores (bars = 10um).

Tuber formosanum, validated (Taiwan) ... 297

298 ... Qiao & al.

T.melanosporum AF 106877

Ue T.melanosporum AF132501 | Subclade I

T.melanosporum AF300825

T.sinense DO375526

T.sinense DO375527

90 CLADE I

T.sinense GU979061

98 T.sinense GU979062 Subclade IT

T. indicum GU979066

is 32 T. indicum GU979067

T. indicum GU979069

T.formosanum GU979048

28 T.formosanum GU979049__| SubcladelII

100 T.formosanum JN655530 CLADE II

T.indicum GU979050

95 Subcladel

T.indicum GU979051

T.aestivum AF516791

PLATE 3: Strict consensus of equally parsimonious trees derived from the analysis of ITS1/5.8S/

ITS2 sequence data (Length = 259steps; CI = 0.942; RI = 0.964). Numbers above branches indicate

bootstrap support above 50%.

Phylogenetic analysis

The ITS phylogenetic tree revealed two major clades. Clade I divided into

two subclades: subclade I corresponded to T. melanosporum and subclade II to

T. indicum from Huidong, Sichuan and Yunnan prov. with high bootstrap

support (298%). Clade II also divided into two subclades; subclade III

corresponded to T: formosanum and subclade IV to two T: indicum isolates from

Panzhihua, Sichuan prov. The three sequenced specimens of T: formosanum

formed a subclade with bootstrap support of 76% (PL.3).

Discussion

The Taiwanese truffle Tuber formosanum is closely related to other Asian

black truffle species, such as T: indicum Cooke & Massee and T: sinense K. Tao

& B. Liu (Chen 2007; Huang et al. 2009; Chen et al. 2011). However, recent

phylogenetic and morphological analyses of these taxa show that although

T. formosanum is closely related to T: indicum and T. sinense, it should still be

Tuber formosanum, validated (Taiwan) ... 299

treated as a distinct species indigenous to Taiwan (Huang et al. 2009; Chen et al.

2011; Kinoshita et al. 2011). Tuber formosanum differs from T. indicum by its asci

with short-stipitate spiny-reticulate ascospores. Most T. indicum asci lack stipes

and its ascospore surfaces are more spike-like. Tuber formosanum associates

naturally only with Cyclobalanopsis glauca, while T. indicum associates with

both coniferous and broad-leaved trees, such as pine and chestnut. Because the

original type specimen on which T! formosanum was based does not exist, we

designate as type KUN-HKAS62628 from among specimens newly collected

at the Ho-She district of National Taiwan University Experimental C. glauca

Plantation, Nantou County, Taiwan. The plantation was established with the

trees inoculated by T: formosanum gathered from the natural forests in Dung-

Pu, Nantou County, Taiwan. The results of morphological and molecular

analyses of samples from KUN-HKAS62628, KUN-HKAS48268 and KUN-

HKAS49707 (PLATE 3) were identical to those of Huang et al. (2009).

Acknowledgements

The authors are grateful for the help of Dr. Shannon Birch and Dr. Ian R. Hall, who

critically reviewed the manuscript and provided invaluable suggestions. This study

was financed by the Yunnan Program of Innovation to strong provinces by Science &

Technology (No. 2009AC013), the National Science Foundation of China (No.30470011

and 31270075), the Joint Funds of the National Science Foundation of China and

Yunnan Province Government (No.U0836604 and U1202262), and Key Laboratory

of Biodiversity and Biogeography, Kunming Institute of Botany, Chinese Academy

of Sciences (No.0806361121 and Y0205111L1), as well as the Knowledge Innovation

Program of the Chinese Academy of Sciences (No.KSCX2-YW-G-025), and The New

Zealand Institute for Plant & Food Research Ltd.

Literature Cited

Chen J. 2007. Taxonomy and phylogeny of the genus Tuber in China (Pezizales, Ascomycetes,

Ascomycotina). PhD Thesis of Graduate School of Chinese Academy of Sciences, China (in

Chinese).

Chen J, Guo SX, Liu PG. 2011. Species recognition and cryptic species in the Tuber indicum

complex. PLoS ONE 6(1): e14625. http://dx.doi.org/10.1371/journal.pone.0014625

Hu HT. 1992. Tuber formosanum sp. nov. and its mycorrhizal associations. Journal of the

Experimental Forest of National Taiwan University 6: 79-86.

Huang JY, Hu HT, Shen WC. 2009. Phylogenetic study of two truffles, Tuber formosanum and Tuber

furfuraceum, identified from Taiwan. FEMS Microbiol. Lett. 294: 157-171.

Kinoshita A, Sasaki H, Nara K. 2011. Phylogeny and diversity of Japanese truffles (Tuber spp.)

inferred from sequences of four nuclear loci. Mycologia 103(4): 779-794.

http://dx.doi.org/10.3852/10-138

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.

Yang ZL, Zhang LE. 2003. Type studies on Clitocybe macrospora and Xerula furfuracea var. bispora.

Mycotaxon 88: 447-454.

MY COTAXON

http://dx.doi.org/10.5248/123.301

Volume 123, pp. 301-319 January-March 2013

Three new /nocephalus species with cuboid basidiospores

from New South Wales and Queensland, Australia

Davip L. LARGENT?, SARAH E. BERGEMANN?, SANDRA E. ABELL-DAVIS?,

KERRI L. KLUTING’, & GRIFFIN A. CUMMINGS?

‘Biological Sciences, Humboldt State University 1 Harpst St, Arcata CA 95521 USA

"Biology Department, Middle Tennessee State University

PO Box 60, Murfreesboro TN 37132 USA

*School of Marine and Tropical Biology, Australian Tropical Herbarium and Centre for Tropical

Environmental & Sustainability Science, James Cook University,

PO Box 6811, Cairns QLD 4870 AU

* CORRESPONDENCE TO: mrp@humboldtl.com

ABSTRACT — Three new Inocephalus species with cuboid spores are described from central

New South Wales and northern Queensland, Australia. Inocephalus plicatus is diagnosed by its

yellow pileus becoming plicate-striate and yellowish orange when mature, pungent odor, bitter

taste, and pseudocystidia on all lamellar surfaces. Inocephalus hypipamee is differentiated by

its brown umbonate pileus, white lamellae and stipe, obclavate cheilocystidia, subisodiametric

basidiospores and clampless hyphae. Inocephalus parvisporus possesses small basidiospores

and a Leptonia-like stature.

Key worps — Basidiomycota, Entolomataceae, LSU, mtSSU, RPB2

Introduction

The Entolomataceae Kotl. & Pouzar (Agaricomycetes, Basidiomycota)

is a highly diverse euagaric family that includes more than 1500 species

(Noordeloos & Gates 2012). Three genera are typically recognized: Clitopilus

(Fr. ex Rabenh.) P. Kumm., Rhodocybe Maire, and Entoloma (Fr.) P. Kumm.

sensu lato. Species of Entolomataceae are difficult to diagnose to genus and

the proposed concepts of classification typically recognize either 13 subgenera

within a single genus Entoloma sensu lato (Noordeloos 1992, 2004) or 13

genera including Entoloma sensu stricto (Largent 1994). Several phylogenetic

studies were recently undertaken to produce a framework for placing species

into genera or subgenera (Co-David et al. 2009, Baroni et al. 2011, Baroni &

Matheny 2011). Although these studies have provided an important context for

302 ... Largent & al.

framing ongoing debates of evolutionary relationships among species, most of

the monophyletic subgroupings lack statistical support.

Without a stable system of classification, newly described species of

Entolomataceae are classified into genera or subgenera using a combination

of morphological characters. Here, we follow the concept of Largent (1994)

and Baroni & Halling (2000) for Inocephalus (Noordel.) P.D. Orton based on

these morphological features: a mycenoid or collybioid habit, a non-glabrous

innately fibrillose to squamulose pileus, an entangled pileipellis often nearly a

trichoderm on the disc, a pileipellis that intergrades imperceptively with the

pileal trama, abundant oleiferous hyphae and lipoid bodies, cheilocystidia and/

or pseudocystidia, and basidiospores with 4-7 distinct angles, some of which

are cuboid.

Cuboid basidiospores, unique to the Entolomataceae, have interested

investigators ever since Entoloma virescens (Sacc.) E. Horak ex Courtec.

was described from the Bonin Islands in 1857 (Horak 1976). Their history,

distribution, systematics, taxonomic significance, and development within

the Entolomataceae have been thoroughly reviewed (Baroni & Halling 2000;

Eyssartier et al. 2001; Horak 1976, 1977; Pegler & Young 1978; Romagnesi

1941). In Australia, two species with a cuboid spore-type have been reported:

Inocephalus virescens (Sacc.) Largent & Abell-Davis (= E. virescens) (Young

2005, Largent & Abell-Davis 2011) and E. procerum G. Stev. (May & Wood

1997, Ratkowsky & Gates 2002, Noordeloos & Gates 2012). Herein, we describe

three new species with cuboid basidiospores: Inocephalus plicatus, I. hypipamee,

and I. parvisporus.

Materials & methods

Macromorphological and micromorphological features

Specimens were collected during February—April 2009-11 from various localities

within the Wet Tropics Bioregion throughout northeastern Queensland and during

April 2010-11 from temperate rainforests in central New South Wales. General and

technical names indicated by quotations marks in the descriptions are from the Colour

Diagrams Section of Kornerup & Wanscher (1978: 196-225).

The factors determined from mathematical analyses in the descriptions include: the

arithmetic means (x) of spore lengths and spore widths + standard deviation in n objects

measured; the quotient of spore length by spore width (E) indicated as a range variation

in n objects measured; the mean of E-values (Q) + standard deviations. The sample size

(n) is equal to the total number of microscopic structures measured (x) and the number

of basidiomata studied (y) and is given in the format n = x/y. The Q values are used

to define general basidiospore shape with the following types recognized: isodiametric

with Q = 1.0-1.15; subisodiametric with Q = 1.16-1.27; heterodiametric Q >1.27

(Largent 1994). Details of macroscopic and microscopic features and measurements

were described in a previous paper (Largent & Abell-Davis 2011).

Inocephalus spp. nov. (Australia) ... 303

All collections for New South Wales cited in the ‘Additional collections examined’

were deposited in The Plant Pathology Herbarium, Orange Agricultural Institute

(DAR), while collections made in Queensland were split with duplicates deposited

in the Australian Tropical Herbarium (CNS) and The Queensland Herbarium (BRI).

All holotype and isotype collections are deposited in the herbaria designated using

acronyms from Thiers (2012).

DNA sequences

The details of protocols for DNA extractions, PCR amplification of partial sequences

of the mitochondrial small subunit of the ribosomal DNA (mtSSU), variable domains

(D1, D2) of the nuclear large subunit (LSU) and sequencing protocols are described

in previous publications (Largent et al. 2011a, b). PCR primers used for amplification

of the second largest subunit of the RNA polymerase gene (RPB2) from Inocephalus

were performed with either of one of two forward primers, rpb2-EntF1 (5' to 3’- Gaa

GGT CAA GCT TGY GGT C) or rpb2-EntF2 (5’ to 3’- GAA GGY CAR GCY TGY GGT C) or

with either one of two reverse primers, rpb2-EntR3 (TGG ATY TCR CAR TGC GTC CA) or

rpb2-EntR5 (TGR ATY TCR CAR TGG GTC CA). PCR amplifications of the RPB2 gene were

performed in 25 uL reactions containing 1x GoTaq Flexi Buffer (Promega, Madison WI

USA), 2mM MgCl, 0.2 mM dNTPs, 500 nM forward and reverse primers, 0.2 mg/mL

bovine serum albumin, 1 M betaine, 0.025 U Taq polymerase, and 1-3 uL of undiluted

template DNA. PCR cycling conditions were as follows: 95°C for 2 min, followed by 40

cycles of 95°C for 30 sec, 52°C or 54°C or 56°C for 1 min, and 72°C for 1 min, followed

by a final extension of 72°C for 10 min. Forward and reverse sequences for each locus

were generated on an Applied Biosystems 3130xl Genetic Analyzer at Middle Tennessee

State University following the protocols outlined in Largent et al. (201 1a, b).

Results

We obtained partial sequences for the mitochondrial small subunit rRNA

gene (mtSSU), nuclear large subunit (LSU), and the second largest RPB2 subunit

for a subset of collections of I. plicatus and I. hypipamee (TABLE 1). Notes on

the morphology of the basidiomata exist for seven I. plicatus collections and

TABLE 1. Inocephalus specimens included in this study.

SPECIES COLLECTION IDENTIFIER GENBANK ACCESSION NUMBERS

mtSSU LSU RPB2

I. hypipamee DL Largent 9649 JQ624602 JQ624607

DL Largent 9981 JQ624603 JQ624608 -

DL Largent 10071 JQ624604 JQ624609 JQ624616

I. plicatus DL Largent 9691 JQ624605 JQ624610 JQ624617

DL Largent 9885 - JQ624611 JQ624618

DL Largent 10083 - JQ624612 JQ624619

DL Largent 10091 - JQ624613 JQ624620

DL Largent 10096 - - JQ624621

DL Largent 10124 - JQ624614 JQ624622

DL Largent 10216 JQ624606 JQ624615 JQ624623

304 ... Largent & al.

the three I. hypipamee collections with sequences. These sequences were used

to formulate the descriptions and develop the diagnostic characters for both

species.

Sequences of I. parvisporus were omitted after several attempts to sequence

PCR products from the three loci failed.

Taxonomy

Inocephalus plicatus Largent, sp. nov. PLATES 1-2

MycoBank MB 564924

Differs from Inocephalus pseudomurrayi by its larger basidiomata, yellow pileus maturing

to plicate-striate and yellowish orange, pungent odor, latently unpleasant taste, and

pseudocystidia on lamellar edge and face.

Type — Australia, New South Wales, central Hunter District, Strickland State Forest,

Lower Parking Lot Track, within 20m of 33°22'47"S 151°19'32"E, 16 April 2010, DL

Largent 9885 (holotype DAR).

EryMoLocy — from the Latin plicatus referring to the pleated (plicate) pileal surface.

Piteus 17-40 mm broad, 5-8 mm high, conic to convex at first, becoming

broadly convex to broadly campanulate with maturity, with a rounded to

mammillate umbo, glabrous or atomate to the eye, or under 4x or 20x

magnification, minutely appressed-fibrillose except for the minutely roughened

center, dull, opaque, not hygrophanous, at first even but with maturity quickly

becoming striate and then plicate-striate from the margin to near the center,

at first pale yellow to light yellow; (2-3A5, 3-4A4), often with a slightly darker

center (3A6, 4A5-4B6, 445-6) becoming greyish yellow to greyish orange when

picked, drying or fading (4-5A-B3-4, 5B4-5, 7E-F8); margin decurved at all

times, entire and with age becoming broadly lobed and crenulate and finally

eroded. Taste mild at first and then latently bitter to unpleasant. Opor mild

and then pungent. LAMELLAE 6-20 mm long, adnexed, subdistant, moderately

broad (3-8 mm), pale yellow at first (3A3) becoming orange white to near

pale orange with spore maturity (4-5A2-3, 4A3-4, 5A3-4, 5A5-6); margin

smooth and concolorous to the eye, cystidiate under 4x or 20x magnification;

3 lamellulae (2 medium short; 1 medium long) in two layers between the

lamellae. StrpE 35-85 mm long, 3-5 mm broad at apex, 3.5-8 mm broad at

base, at times flattened and longitudinally grooved and then 3 x 4.5 mm broad

at apex, 4 x 5 mm broad at base, typically clavate, rarely twisted, glabrous except

minutely roughened to pruinose at the apex, pale yellow to light yellow (2A4

to 3-4A2-4) darkening to pale orange (5A3-5, 5B4) and/or discoloring to light

brown (4-5D7, 5D7) stuffed at first, becoming hollow, sturdy; basal tomentum

scarce to absent. BRUISING REACTIONS pileus frequently staining brownish

orange (6C8) to dark brown with a reddish orange tint.

Inocephalus spp. nov. (Australia) ... 305

—_—— y ’

PiaTE 1. Inocephalus plicatus (DLL 10091). A: Basidiomata (2.5x); B: Pileus surface (2x); C: Stipe

apex (8x).

306 ... Largent & al.

BASIDIOSPORES in profile view versiform, cuboid, rhomboid, and at times

rectangular, often with one angle enlarged, rarely 3-angled and then trapezoid-

like, sometimes 5-angled, isodiametric to heterodiametric in dorsal or ventral

view, 4-5-angled and isodiametric in polar view, 7.7-11.9 x 6.6-10.4 um

(x = 9.6 + 0.9 x 8.5 + 0.8 um; E = 1.0-1.4; Q = 1.2 + 0.1 (subisodiametric);

n = 169/7). Basrp1a typically long and clavate, tapered to the base, the base

typically enlarged, 40.5-73.2 x 9.5-18.1 um (x = 55.5 + 6.9 x 12.4 + 1.5 um;

E = 3.2-7.4;Q =4.5 + 0.8; n = 53/6); 4-sterigmate, sterigma 3.7-6.2 um; base

of basidium 1.9-5.2 um. ABORTED BASIDIA absent. CHEILOCYSTIDIA typically

forming a sterile layer, at times scattered, colorless to opaque, clavate to cylindro-

clavate eventually becoming strangulated to somewhat contorted, tapering to

the apex, 66.5-144.4 x 4.8-12.2 um (n = 14/2). PseUDOCysTIDIA scattered to

abundant on the lamellar edge and face, filled with granular refractive material

yellowish in 3% KOH, cylindro-clavate, typically contorted, 21.8-87.3 x 3.2-9.1

um (n = 29/4). HYPHAE OF THE LAMELLAR TRAMA Subparallel and quite long,

136-1119 x 3.7-22.7 um (n = 37/5). PILEIPELLIS an entangled, semi-erect layer

of hyphae, entirely erect at first, with maturity remaining so in the center but

a cutis elsewhere. PILEOCysTIDIA cylindro-clavate, 66.5-144.4 x 4.9-21.5 um

(n = 22/3). HYPHAE OF THE PILEAL TRAMA quite long, 43.6-507.0 x 6.1-26.9

um (n = 11/3). STIPITIPELLIS at the apex an entangled layer of semi-erect

caulocystidia. CAULOCYSsTIDIA cylindro-clavate, similar to the pileocystidia,

37.6-167.6 x 4.6-11.2 um (n = 30/5). HYPHAE OF THE STIPE TRAMA not studied.

LIPOID GLOBULES abundant in all tissues and obscuring all microscopic features

when viewed in 3% KOH. OLEIFEROUS HYPHAE abundant in the lamellar

trama, in the subhymenium, and the pileal trama. BRILLIANT GRANULES absent.

PIGMENTATION cytoplasmic and yellowish in the hyphae of the pileipellis and

stipitipellis in 3% KOH. CLAMP CONNECTIONS present in all tissues.

ECOLOGY AND DISTRIBUTION — Solitary, scattered, or gregarious in sandy

organic rich soil, in deep leaf humus or protected by logs and woody debris,

warm temperate gallery rainforest near Ceratopetalum apetalum D. Don,

Backhousia myrtifolia Hook., and Syncarpia glomulifera (Sm.) Nied. (Strickland

State Forest, New South Wales). In other localities, found in warm, temperate,

PLATE 2. Inocephalus plicatus. A: 4-angled basidiospores (2500x), rectangular in profile view (left),

nearly square in dorsi-ventral view (center), and becoming rhomboid in profile view (right) (DLL

10091); B: Pseudocystidium to left of basidium (2000x) (SSF 12); C: Stipitipellis with cylindro-

clavate caulocystidia (50x) (SSF 12); D: Pileipellis with clavate to cylindro-clavate pileocystidia

(125x) (SSF 12); E: Pseudocystidium from lamellar face (450x) (DLL 10091); F: Strangulated to

somewhat contorted cheilocystidia, 2-sterigmate basidium (lower left corner), and oleiferous

hyphae (darker) (350x) (DLL 9885, holotype); G: Section of lamellae with abundant oleiferous

hyphae, lipoid globules, and showing origin of pseudocystidium (100x) (DLL 9885, holotype).

307

Inocephalus spp. nov. (Australia) ...

308 ... Largent & al.

subtropical gallery rainforests (New South Wales) and wet tropical rainforests

(northeastern Queensland).

ADDITIONAL COLLECTIONS EXAMINED — AUSTRALIA. NEw SouTH WALES, central

Hunter District, Barrington Tops National Park, Jerusalem Creek Track, 25 April 2010,

S Moore 59; Strickland State Forest, Lower Parking Lot, 33°22'47"S 151°19'27"E, 23

March 2009, SSFO7, SSF12; 19 April 2010, DL Largent 9903; 33°22'47"S 151°19'31”E,

14 April 2011, DL Largent 10091, 10096; 33°22'48”S 151°19'28"E, 19 April 2011, DL

Largent 10124; 33°22'47"S 151°19'33"E, 28 April 2011, DL Largent 10165; 33°22'47"S

151°19'29"E, 7 May 2011, DL Largent 10216; Watagans National Park, Boardinghouse

Dam Area, 32°59'56"S 151°24'19"E, 12 April 2011, DL Largent 10083. QUEENSLAND,

Cook Region, Herberton National Park, 17°27'07.0"S 145°28'32.8"E, 24 March 2009, DL

Largent 9691; 17°27'14.8"S 146°28'34.2"E, 16 March 2010, DL Largent 9786; Mt. Lewis

National Park, 16°16'10.1"S 147°16'11.0"E, 20 March 2010, DL Largent 9803; Mossman

Gorge National Park, 16°28'16.9"S 145°19'51.6"E, 25 February 2010, DL Largent 9741,

9742.

ComMENtTs — Inocephalus plicatus is distinguished from other Inocephalus

taxa by the following set of diagnostic features: yellow basidiomata when

young with a pileus that is greyish yellow to greyish orange, plicate-striate and

mammillate umbonate at maturity and that stains brownish orange or darker

with handling; pileus tissue producing a latently bitter to unpleasant taste and

a latently pungent odor; mostly cuboid or rhomboid basidiospores averaging

<10 x 9.0 um; contorted and refractive pseudocystidia on lamellar face and

edge as well as abundant, contorted or strangulated and opaque cheilocystidia;

a pruinose stipe apex composed of cylindro-clavate caulocystidia; abundant

oleiferous hyphae in the lamellar and pileal trama; and abundant clamps.

Morphologically, I. plicatus resembles Entoloma pseudomurrayi Eyssatt. et

al., E. cuspidatum Sacc., and I. murrayi (Berk. & M.A. Curtis) Rutter & Watling

(= E. murrayi (Berk. & M.A. Curtis) Sacc.).

Entoloma pseudomurrayi from New Caledonia shares many similar

morphological features with I. plicatus but is differentiated by the smaller

basidiomata, the citrine yellow, non-plicate, non-staining pileus that does

not darken to greyish yellow or greyish orange, the mild odor and taste, the

lanceolate to fusiform and moniliform cheilocystidia, and pseudocystidia

found only on the lamellar edge (Eyssartier et al. 2010).

Entoloma cuspidatum (= Agaricus cuspidatus Peck, nom. illegit., non A.

cuspidatus Bolton) is morphologically differentiated from I. plicatus by its conic

to campanulate, cuspidate, pale yellow, non-striate pileus and basidiospores

measuring 9-13 x 8-12 (-12.5) um. The basionym of Inocephalus murrayi (=

Agaricus murrayi Berk. & M.A. Curtis) is morphologically differentiated from

I. plicatus by its non-plicate pileus that turns reddish brown on drying and

basidiospores measuring 9-11 x 9.5-11 um.

Inocephalus spp. nov. (Australia) ... 309

The synonomy of Entoloma cuspidatum and Inocephalus murrayi

Entoloma cuspidatum is considered a synonym of I. murrayi by Baroni &

Halling (2000) and of E. murrayi by Hesler (1963, 1967), Horak (1976, 1980),

and Singer (1942). This synonymy should be re-evaluated because of the

following discrepancies.

Peck’s (1872: 64-65) protologue description of A. cuspidatus differs from that

of A. murrayi (Berkeley & Curtis 1859: 289). Agaricus cuspidatus has a conic to

campanulate, non-striate pileus pale yellow throughout, an elongated papilla or

cusp, and basidiospores 1/2000 inch (= 12.7 um) in diameter. Agaricus murrayi

has a convex regularly striate pileus that is straw yellow turning rich red brown

when dry and basidiospores 1/2500 inch (= 10.16 tm) in diameter. Using these

features, Peck considered the two species related but different (Noordeloos

2008: 96).

Studies of the holotype collections indicate the basidiospores of A. murrayi

measure 9-11 x 9.5-11 um (Singer 1942) and are indeed a little smaller than

those measured for A. cuspidatus Peck (9-13 x 8-12 um, Hesler 1963; 11-13.5

(-14) x (9.5-) 10-11.5 (-12.5) um, Baroni pers. comm.; 9-11.7 x 8.0-11.7 um,

Noordeloos 2008).

Australasian species morphologically similar to Inocephalus plicatus

Several other species from Australasia morphologically resemble I. plicatus

in yellow coloration and cuboid basidiospores. Entoloma pallidoflavum (Henn.

& E. Nyman) E. Horak from Java, Borneo, and New Guinea is differentiated

by the larger pileus (30-80 mm) and stipe (<130 mm long), an indistinctive

or radish-like odor, and coarsely dentate-serrate lamellae. Entoloma albogracile

E. Horak from Papua New Guinea and E. avilanum (Dennis) E. Horak from

Venezuela have tetrahedral-cuboid basidiospores. Entoloma gracilius E. Horak

from Papua New Guinea has smaller basidiospores (5.5-7.0 um; Horak 1976).

Inocephalus luteus (Peck) TJ. Baroni (= Entoloma luteum Peck) from eastern

North America is morphologically differentiated by the smoky yellow, at

times greenish yellow, distinctly fibrillose pileus as typified by the protologue

description and illustration (Baroni, pers. comm.).

Inocephalus hypipamee Largent, sp. nov. PLATES 3-4

MycoBank MB 564925

Differs from Entoloma flavotinctum by its larger basidiomata, non-hygrophanous pileus,

white then yellowish white stipe, white context, and larger basidiospores.

Type — Australia. Queensland, Cook Region, Mt. Hypipamee National Park, within

20 m of 17°25'35.2"S 145°29'14.6"E, 25 March 2011, DL Largent 10071 (holotype BRI,

isotype CNS).

EryMoLocy — from the Australian Aborigine hypipamee, referring to the frequent

occurrence of this species in Mt. Hypipamee National Park.

310 ... Largent & al.

PiLEus 9-70 mm broad, 4-23 mm high, when young narrowly parabolic, conic,

or convex, entirely and densely matted-tomentose to tomentulose, quickly

developing an acute to broad umbo; upon expansion and maturity becoming

broadly conic, broadly convex, campanulate-convex, or campanulate, always

remaining densely matted-fibrillose to tomentulose on the apex and umbo

but becoming densely appressed-squamulose elsewhere, eventually becoming

matted-fibrillose-squamulose and somewhat rimulose at or near the margin,

overly mature specimens becoming fibrillose and rimose everywhere except

on or near the umbo; when very young or in basidiomata protected by

overhanging logs, the fibrils, hairs, and tomentum greyish orange to brownish

orange (6B-C2-3) on and around the umbo and white to orange white (6A-B1-

2) elsewhere, upon exposure and in young exposed basidiomata brown to dark

brown (6F4-5) everywhere, in maturing specimens remaining so on or near

the umbo, elsewhere fading to a lighter brown (between 5E3-4 and 6E3-4) then

becoming brownish orange (between 5C-D3-4 and 6C-D3-4) from the margin

to the umbo, in very old specimens the fibrils become rimose and thus the

orange white to greyish orange (6A-B2-3) background color becomes visible;

surface opaque (not hygrophanous, not translucent-striate); margin decurved

and entire, eventually becoming plane or uplifted, fibrillose, crenulate, and

eroded; context up to 1.25 mm above the stipe, <1 mm midway to margin, and

nearly non-existent at the margin, white to off-white. Taste mild becoming,

at most, latently slightly unpleasant. Opor mild or indistinct. LAMELLAE 6-32

mm long, 1-9 mm deep, when young yellowish white (3A2) then very pale

yellow (4A3) and with spore maturity between orange white and pale orange

(5A2 or 5A3), narrowly adnate or broadly adnexed at all times but often with

a decurrent tooth, at first close to crowded and narrow then subdistant and

moderately broad to sigmoid; margin smooth and concolorous to the eye but

under the dissecting microscope abundantly cystidiate; 3 lamellulae/2 lamellae.

STIPE 18-68 mm long, 1-7 mm broad at apex, 2-8 mm broad at base, equal

in young specimens, but with maturity typically enlarged at the base, white or

off-white (3A2) becoming yellowish white (4A2) with age, to the eye glabrous

except pruinose at the apex, under the dissecting microscope suggestively

appressed-fibrillose, hollow but solid or sturdy; basal tomentum scarce to

absent; context unchanging when cut. BRUISING REACTIONS slightly brownish

on the stipe.

BASIDIOSPORES in profile view typically cuboid or rectangular, sometimes

5-angled, isodiametric to heterodiametric in profile and dorsiventral views,

isodiametric in polar view, 7.1-10.4 x 6.2-8.6 um (x = 8.8 + 0.8 x 7.4 + 0.6 um;

E = 0.9-1.4; Q= 1.2 + 0.1 (subisodiametric); n = 58/2). BAsip1a mostly clavate,

typically long and tapered to the base, 37.9-49.3 x 9.0-12.0 um (x = 43.2 + 3.24 x

10.1 + 0.79; E = 3.4-5.3; Q = 4.1 + 0.5; n = 20/2); 4-sterigmate, sterigma 2.5-4.1

Inocephalus spp. nov. (Australia) ... 311

PLATE 3. Inocephalus hypipamee. A: Basidiomata stature (2x) (DLL 10035); B: Pileus surface,

young (left), and mature (right) (2x) (DLL 10035); C: Lamellae and lamellulae (2x) (DLL 9999).

312 ... Largent & al.

um long; base of basidia 3.5-5.4 um wide. ABORTED BASIDIA rare to scattered,

embedded in the hymenium. HYMENIAL cysTip1A abundant and obvious,

present as PsEUDOCYSTIDIA, typically filled with a yellowish oily material,

becoming reddish brown in 3% KOH, those on the lamellar edge clavate to

obclavate or fusoid, 38.3-46.2 x 4.4-12.8 um (n = 7/1); those on the lamellar

face similar in shape but longer than those on the lamellar edge, originating

from the outer layers of the lamellar trama, 47.6-106.5 x 5.0-19.5 um (x =

76.6 + 18.4 x 12.7 + 3.7 um; E = 4.3-9.6; Q = 6.3 + 1.6; n = 14/2). HYPHAE

OF THE LAMELLAR TRAMA Subparallel, moderately long to long, 107.7-582.3 x

4.2-14.7 um (n = 8/2). PILEIPELLIs in fresh specimens composed of semi-erect

and laterally agglutinated hyphae with cylindro-clavate to clavate terminal

cells, in dried specimens also composed of entangled and laterally agglutinated

hyphae but with cylindro-clavate terminal cells often collapsed on top of clavate

ones. PILEOCYSTIDIA cylindro-clavate or clavate 30.5-51.6 x 5.9-14.6 um

(n = 6/1). HYPHAE OF THE PILEAL TRAMA above the lamellar trama 55.0-173.8

x 9.8-15.0 um (n = 9/1). STIPITIPELLIS a cutis except at the apex with clusters of

basidioles or small clavate caulocystidia. CAULOCYsTIDIA 31.0-41.2 x 7.1-10.7

um (n = 3/1). HYPHAE OF THE STIPE TRAMA parallel, 55.0-188.0 x 9.8-26.3

um (n = 9/1). LrpoID GLOBULES abundant in all tissues, often obscuring other

microscopic features. OLEIFEROUS HYPHAE abundant throughout the lamellar

and pileal trama. BRILLIANT GRANULES absent. PIGMENTATION composed

of abundant dark brown, cytoplasmic plaque-like areas in the hyphae of the

pileipellis, absent elsewhere. CLAMP CONNECTIONS absent in all tissues.

ECOLOGY AND DISTRIBUTION — Typically solitary but at times gregarious

with clusters of 2-4 basidiomata, in granitic soil and leaves beneath various

species of Lauraceae and Myrtaceae in a complex notophyll vine forest of cloudy

and moist to wet highlands. In Mt. Hypipamee National Park, I. hypipamee

produced 50-100 basidiomata throughout the same forested slope above

Dinner Falls from 16 February to 5 April 2011.

ADDITIONAL COLLECTIONS EXAMINED: AUSTRALIA. QUEENSLAND, Cook Region,

Mt. Hypipamee National Park, 17°25'35.4S”" 145°29'12.4"E, 14 March 2009, DL Largent

9649; 17°25'34.5"S 145°29'16.6"E, 16 February 2011, DL Largent 9981, 9981A, 9982;

17°25'35.4"S 145°29'10.9"E, DL Largent 9984; 17°25'58.6"S 145°29'27.4"E, 21 February

2011, DL Largent 9988; 17°25'34.8"S 145°29'17.0"E, 28 February 2011, DL Largent

9992; 17°25'37.1"S 145°29'15.2"E, 5 March 2011, DL Largent 9998; 17°25'35.2"S

145°29'14.6"E, DL Largent 9999; 17°25'36.7"S 145°29'15.7"E, 17 March 2011, DL Largent

10035; 17°25'35.2"S 145°29'14.6"E, 29 March 2011, DL Largent 10074; 17°25'36.7"S

145°29'15.7"E, 5 April 2011, DL Largent 10081; Mt. Lewis National Park, 16°34'15.3"S

145°15'54.9"E, 20 March 2010, DL Largent 9804.

ComMENtTs — Inocephalus hypipamee is diagnosed by the mammillate, broadly

conic to broadly convex to convex-campanulate pileus with dark brown

tomentum in the center and dark brown fibrils and squamules elsewhere, the

Inocephalus spp. nov. (Australia) ... 313

PLATE 4. Inocephalus hypipamee. A: Basidiospores in profile view (1700x) (DLL 9649); B: Pileipellis

composed of entangled hyphae with cylindro-clavate pileocystidia (1000x) (DLL 9988); C: Caulo-

cystidia at apex of stipe (1000x) (DLL 9988); D: narrowly obclavate pseudocystidium originating

from outer portion of lamellar trama, nearby basidioles (in focus), and mature basidium (out of

focus) (150x) (DLL 9649); E: pseudocystidia on face of lamella staining reddish-brown in 3% KOH

(1000x) (DLL 9988); F: Obclavate pseudocystidium, and basidia (1000x) (DLL 9649).

314... Largent & al.

white to off-white lamellae and stipe, the cuboid to rectangular, subisodiametric

basidiospores measuring 7.1-10.4 x 6.2-8.6 um, the clavate to obclavate or

fusoid pseudocystidia found on the lamellar edge and face, and the absence of

clamp connections.

Because of its brown pileus, white lamellae and stipe, cuboid basidiospores,

pseudocystidia and absence of clamp connections, E. flavotinctum E. Horak

& Corner from Brazil is the only species remotely similar to I. hypipamee.

Entoloma flavotinctum is morphologically differentiated by its smaller

basidiomata, hygrophanous pileus, a stipe that becomes yellowish or greenish

ochraceous, a white context that turns yellowish on cutting and bruising, and

smaller quadrate basidiospores (6-8(-9) um; Horak 1982).

Inocephalus parvisporus Largent, sp. nov. PLATES 5-6

MycoBank MB 564926

Differs from Entoloma neocaledonicum by the Leptonia-like pileus with a small broad

umbo in a depression, white lamellae, longer cheilocystidia, and smaller basidiospores.

Type — Australia, Queensland, Cook Region, Danbulla National Park, Kauri Creek

Track, within 20m of 17°07'42.3"S 145°25'55.0"E, 25 March 2009, DL Largent 9829

(holotype BRI, isotype CNS).

EryMoLocy —from the Latin words parvi + sporus, referring to the small basidiospore

size.

PitEus 10-32 mm broad, 5-15 mm high, when young dark brown with a

reddish tinge (near 8F4) on the disc and ornamentations with a slightly lighter

dark brown (8E4) background, parabolic or campanulate-convex, or convex,

entirely minutely tomentulose, and typically with a small, broad umbo and an

even, incurved margin, when maturing and expanding remaining dark brown

(8F4) on the disc becoming reddish white to pale red (7A-B2-3) elsewhere,

becoming broadly convex to broadly campanulate-truncate with a depression

developing, the umbo often disappearing, remaining tomentulose in the center

becoming minutely squamulose to appressed-fibrillose towards the margin,

and appressed-fibrillose at the margin, with the margin becoming decurved,

crenulate-lobed and then eroded, typically opaque, at times becoming

hygrophanous in the disc, not striate nor translucent-striate, dull when dry,

shiny when moist. TasTE indistinct or mild. ODor indistinct or mild. LAMELLAE

4-19 mm long, 1.75-5 mm deep, narrow then quickly moderately broad,

adnate, close to subdistant, white when young with spore maturity remaining

light on the edge but becoming flesh-colored elsewhere; margin smooth and

concolorous, entire at all times; up to 5 lamellulae between lamellae, in 3 tiers

(2 short, 2 medium, 1 medium long). Stipe 27-45 mm long, 1.25-4 mm broad,

equal, pruinose at the apex, glabrous with hygrophanous streaks elsewhere,

white (6A1) but bruising reddish grey (8B-C2), hollow with some solidity

Inocephalus spp. nov. (Australia) ... 315

. 4 er : seh

Pate 5. Inocephalus parvisporus (DLL 9829, holotype). A: Basidiomata stature (2x); B: Lamellae

and pruinose stipe apex (2x); C: Pileus surface (2x).

316 ... Largent & al.

A : as

PiatE 6. Inocephalus parvisporus (DLL 9829, holotype). A: Basidiospores (2700x); B: Basidium

(1000x); C: Cylindro-clavate to clavate cheilocystidia (300x); D: Pileipellis just off disc with

cylindro-clavate pileocystidia (200x); E: Cylindro-clavate caulocystidia at stipe apex (900x).

but ultimately becoming fragile; basal tomentum moderate becoming scarce.

BRUISING REACTIONS absent.

BASIDIOSPORES cuboid, square to rectangular, rarely rhomboidal in profile

view, 5.3-8.1 x 4.5-7.1 um (x = 6.5 + 0.6 x 5.6 + 0.7 um; E = 1.0-1.5; Q = 1.2

+ 0.1 (subisodiametric); n = 30/1). BAsip1A clavate and tapered, 33.5-41.4 x

10.4-13.1 um (x = 38.9 + 2.3 x 11.2 + 0.9 um; E = 2.9-3.9; Q = 3.5 + 0.3; n = 9/1),

4-sterigmate. ABORTED BASIDIA absent. CHEILOCYSTIDIA cylindro-clavate to

clavate, abundant, portions of some lamellar edge completely sterile, similar

in size to the caulocystidia. PLEUROCysTIDIA and PsEUDOCYSTIDIA absent.

HYPHAE OF THE LAMELLAR TRAMA subparallel 92.2-398.5 x 4.4-10.5 um (n =

6/1). PILEIPELLIS a trichodermal palisade, with the hyphae erect but entangled

on the disc, semi-erect and entangled towards the margin, and repent near and

at the margin; PILEOCySTIDIA 48.3-148.6 x 4.9-9.3 um (n = 6/1), long and

cylindro-clavate, resembling the cheilocystidia. HYPHAE OF THE PILEAL TRAMA

357.8-491.3 x 13.4-16.6 um (n = 3/1). STIPITIPELLIs with abundant cylindro-

Inocephalus spp. nov. (Australia) ... 317

clavate cystidioid caulocystidia at the apex, a cutis elsewhere. CAULOCYSTIDIA

clavate, similar to the cheilocystidia, 52.4-66.5 x 4.6-7.2 um (n = 4/1). HYPHAE

OF THE STIPE TRAMA not studied. L1poIp GLOBULEs abundant, particularly in

the lamellar and pileal trama. OLEIFEROUS HYPHAE abundant, particularly in

the lamellar and pileal trama. BRILLIANT GRANULES absent. PIGMENTATION

with dark brown cytoplasmic particles in the hyphae of the pileipellis, absent

elsewhere. CLAMP CONNECTIONS absent in all tissues.

ECOLOGY AND DISTRIBUTION — Scattered to gregarious on leaves in humus

along side of Kauri Creek Track, near the end of the track as it drops down to

Kauri Creek, Danbulla National Park; late March; known only from the type

locality.

ComMENTS — Unique features of Inocephalus parvisporus include its Leptonia-

like habit and basidiomata with dark brown, umbonate, opaque pilei with

slight reddish tones, the small basidiospores, the presence of cheilocystidia,

small cuboid basidiospores, and no clamps. In the field, it resembles a species

of Leptonia (Fr.) P. Kumm., although the abundant oleiferous hyphae, abundant

lipoid bodies, and cuboid basidiospores suggest Inocephalus.

Because of their brown basidiomata and cuboid basidiospores, Entoloma

griseoalbum E. Horak from Papua New Guinea and E. neocaledonicum E. Horak

from New Caledonia share similar morphological features with I. parvisporus.

However, both E. neocaledonicum and E. griseoalbum are morphologically

differentiated by their mycenoid basidiomata with a conical pileus. In addition,

E. griseoalbum has a grey brown striate pileus and clavate cheilocystidia often

with finger-like protrusions, and E. neocaledonicum has pale wax yellow

lamellae, broadly clavate cheilocystidia, larger basidiospores, and clamp

connections (Horak 1976, 1977).

Acknowledgments

Fieldwork in Australia was supported by the Largent family trust and we are

particularly grateful for the support of Pamela Largent. Fieldwork and logistical support

were provided by the Australian Tropical Herbarium and the School of Marine and

Tropical Biology, James Cook University. The DNA sequences generated in this study are

based upon work supported by the National Science Foundation under Grant No. DRI

0922922 awarded to SE Bergemann. Comments by the two reviewers, Dr. Timothy J.

Baroni and Dr. Genevieve Gates, and by the nomenclature editor Dr. Shaun Pennycook,

were also helpful. We wish to thank Peter Newling for sharing his collections from

northern Queensland and for providing the identification of vascular plants and habitat

descriptions for taxa collected in Mt. Hypipamee National Park, and Pam O’Sullivan

as well as Skye Moore for sharing their collections and descriptions from New South

Wales. We also wish to thank Dr. Baroni for information provided of his type study of

Agaricus cuspidatus and the identification of Inocephalus luteus.

318 ... Largent & al.

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MYCOTAXON

http://dx.doi.org/10.5248/123.321

Volume 123, pp. 321-326 January-March 2013

New Lecanora, Lecidea, Melaspilea, Placynthium, and Verrucaria

records for Turkey and Asia

ALI ASLAN’ & KENAN YAZICI *?

"Biology Department, Kazim Karabekir Education Faculty, Atatiirk University,

Erzurum, Turkey

Biology Department, Faculty of Science, Karadeniz Technical University,

61080, Trabzon, Turkey

* CORRESPONDENCE TO: kcagri_1997@yahoo.com

ABSTRACT — Five species of lichenized fungi - (Lecanora invadens, Lecidea promiscua,

Melaspilea interjecta, Placynthium garovaglioi, and Verrucaria bryoctona) —- are reported as

new to Turkey. Three of them, M. interjecta, P. garovaglioi, and V. bryoctona, are also new to

Asia. Descriptions are presented, including geographic distribution, substrate, chemistry, and

comparisons with morphologically similar taxa.

Key worps — Ascomycota, biodiversity, Igdir, Van, Burdur

Introduction

The lichen flora of Turkey is still incompletely known, as it is in many parts

of the world. Although many recent studies contributed to the lichen flora of

Turkey (Halici & Cansaran-Duman 2008, Karagéz et al. 2011, Kinalioglu 2010,

Halici et al. 2010a,b, Osyczka et al. 2011, Candan & Halici 2011, Oran & Oztiirk

2012, Senkardesler 2010, Yazic1 & Aslan 2009, Yazici et al. 2008, 2010a,b,

2011a,b), there are more unexplored regions than explored ones in the country.

Therefore, more studies are needed to complete the lichen flora of Turkey.

Here we report five new records for Turkey, of which three are new for

Asia, based on our fieldwork in the regions of Van, Igdir and Burdur in eastern

Turkey.

The Van region has a climate characterized by cold and snowy winters and

hot dry summers, with a temperature range of -26.9-36.0 °C, a mean annual

rainfall of around 370-570 mm, and mean annual humidity of 51-66% (Akman

1999). The examined area is dominated by steppe vegetation and lacks trees

completely. Three localities were surveyed: Kap1 village in Saray district near

the Iranian border is an open windswept area with gently sloping rocky terrain;

322 ... Aslan & Yazici

the Catak district is a partly well lit high elevation sloped terrain with a waterfall

and humid rocks; and Bahcesaray, at the border of the Van district, follows

the road through a well-lit windswept weakly sloping land with a stream and

humid rocks (Baytop & Denizci 1963).

The climate of the Igdir region is similar to Van, with a mean annual

temperature of 11.6 °C, mean annual rainfall of 257.6 mm, and average

humidity of 63%, Precipitation is generally moderate in the summers and

heavy during winter (Akman 1999). The Igdir region is dominated by steppe

with few forested areas, although Elaeagnus, Populus, Prunus, Pyrus, and Salix

trees are seen occasionally in the visited locality, Kula village (Tuzluca) (Baytop

& Denizci 1963). Kula is a well-lit, high elevation, windswept, treeless area

with gently sloping terrain containing streams, grass, and many calcareous and

siliceous rocks.

Burdur has a continental Mediterranean climate with cold, snowy winters

and very hot, long, dry summers with a mean annual temperature of 15°C and

the temperature range of -16-39 °C, mean annual rainfall of about 468 mm,

and average humidity of 51.2% (Akman 1999). The visited area, Bucak district,

is mountainous with much forest dominated by Abies, Cedrus, Ficus, Fraxinus,

Juniperus, Olea, Pinus, Prunus, Quercus, Rhus, and Pistacia alternating with

streams and lakes (Baytop & Denizci 1963).

Materials & methods

Lichens were collected from five localities in the districts Igdir, Burdur and Van

in Turkey in July 2009. Air-dried samples were examined with a Nikon SMZ1500

stereomicroscope and a Nikon Eclipse 80i compound light microscope. For the

identifications relevant keys consulted (Czeika & Czeika 2007, Hertel 2006, Magnusson

1952, Orange 1991, Sliwa 2007, Smith et al. 2009). Vouchers are stored in the Herbarium

of the Biology Department, Karadeniz Technical University, Trabzon, Turkey (KTUB)

and in the Herbarium of the Biology Department, Kazim Karabekir Education Faculty,

Atatiirk University, Erzurum, Turkey (ATAKKEF). The descriptions are based on

Turkish specimens and supplemented by data from the indicated literature.

Species recorded

Lecanora invadens H. Magn., Lichens Central Asia: 87. 1940.

Thallus crustose, + superficial or evanescent, indistinct, to 0.6 cm in diam.,

more evident around apothecia, + grey to greenish grey, often with distinct

bluish pigment. Apothecia mostly clustered in groups, sessile, or constricted

at the base and raised, + flat when mature, to 0.6 mm in diam.; disc plane to

slightly convex, + smooth, dark brown to slightly black and slightly pruinose,

rarely epruinose; margin level + with the disc + rough, entire, pruinose, paler

than the thallus, white. Amphithecium 155-175 um thick, corticate, with algae

mostly dense below the cortex; cortex mostly delimited, slightly thicker at the

Lichens newly recorded for Turkey ... 323

base, 16-55 um thick, composed of adglutinated hyphae, thin and bluish-

green at the very top of the margin; epithecium yellowish-brown, granular;

granules usually sparse, between the paraphysis tips; hymenium hyaline, 50-65

um high; hypothecium hyaline or yellow, 60-100 um high. Paraphyses simple

or branched at the tips, slender or + thickened, + pigmented at the tips. Asci

clavate, 8-spored; ascospores hyaline, simple, + broadly ellipsoid, 9-12 x 6-8

um. Pycnidia absent. Apothecial margin K+ yellow, C-, KC+ yellow, P-; disc

K+ yellow, C-, KC+ yellow, P-.

Lecanora invadens grows on non-calcareous or weakly calcareous rock

and is mostly ‘parasitic’ on other lichens (e.g., Aspicilia, Collema, Endocarpon,

Lecanora, Verrucaria). It is rarely found on man-made substrata. New to Turkey

and the Middle East, where it was accompanied by Aspicilia calcarea, Collema

tenax, Endocarpon sp., and Fulgensia fulgens. Also known from Europe, Asia,

and North America (Sliwa 2007).

SPECIMEN EXAMINED: TURKEY. Burpur: Bucak, surroundings of Karaca6ren Dam,

37°21'35.14"N 30°50'05.80"E, 300 m, on Endocarpon sp., Collema tenax, and soil,

25.06.2012, leg. K. Yazici. (KTUB-2336).

REMARKS— Lecanora invadens resembles L. semipallida but has dark brown to

slightly black and slightly pruinose apothecia with a yellow-brown epithecium,

and a more distinct thallus.

Lecidea promiscua Ny]. Flora 57: 357. 1874.

Thallus thin, indistinctly areolate-rimose and sordid-white; prothallus not

developed or indistinct; areoles flat, irregular and indistinct; medulla white, I+

intensely violet. Apothecia sessile, with markedly or strongly constricted base,

to 2.5(-4) mm in diam.; disc black, flat to + convex, emarginate, epruinose or

slightly pruinose; margin black, persistent, dull or + shiny; exciple black, blue-

green to brown-black in a thin, peripheral rim, 60-150 um wide; epithecium

dark blue-green to green-brown-black, 12-18 um thick; hymenium hyaline

or faintly green, 46-58 um tall; paraphyses simple, occasionally branched, lax

or contiguous, especially towards the apical ends; hypothecium dark-brown

below, lens-shaped, hyaline above; asci clavate 8-spored, 40-50 x 13-17um;

ascospores hyaline, simple, oblong to oblong-ellipsoid, (7-)9-12.5 x (2.5-)

3-4.5 um. Pycnidia semi-immersed, 90-150 um in diam.; conidia bacilliform

to filiform, straight, (9-)9.5-14 x 1-1.3 um. C-, K-, KC-, Pd-, UV-.

Lecidea promiscua is a circumpolar, arctic-alpine to boreal-montane lichen,

growing on granite boulders close to the ground and on siliceous pebbles in

wind-exposed and sunny situations widespread in the Alps. New to Turkey

and the Middle East, where it accompanied Aspicilia desertorum, A. cinerea,

Acarospora fuscata, Caloplaca saxicola, Immersaria athroocarpa, and Lecanora

muralis. Also known from Europe and Central Asia (Hertel 2006).

324 ... Aslan & Yazici

SPECIMEN EXAMINED: TURKEY. VAN: SARAyY-KaPIKOy, vicinity of Iran border,

38°29'57"N 44°17'33"E, 2276 m, on siliceous rock, 28.09.2009, leg. A. Aslan.

(ATAKKEF-1450).

REMARKS— Lecidea promiscua resembles L. promiscens, which is distinguished

by a thick white thallus.

Melaspilea interjecta (Leight.) A.L.Sm., Monogr. Brit. Lich. 2: 228. 1911.

Thallus thin, pale white-green-grey to pale fawn or brown; photobiont cells

5-16 um in diam., trebouxioid. Apothecia 0.4-1.2 x 0.2-0.3 mm, lirellate, simple

or forked, at with a black, slit-like, later widening disc and a tumid margin,

later developing additional slits or becoming subgyrose-contorted; true exciple

well developed, reddish brown-black, K+ olivaceous; epithecium pale to dark

brown, K+ olivaceous; hymenium 70-85 um tall, colourless, I+ yellowish, K/I+

blue; hypothecium concolorous with exciple; paraphyses 1.5-2 um wide, richly

branched and anastomosing. Asci 47-53 x 21-26 um, 8-spored; apical dome

with narrow ocular chamber. Ascospores 17-23 x 7-9 um, 1-septate, sickle-

shaped, constricted and easily broken at the septum, colourless but old spores

brown and finely warted.

Melaspilea interjecta grows on vertical, smooth surfaces of slate rocks and

mica-schist rocks. New to Turkey and Asia. Also known from Europe (Smith

et al. 2009).

SPECIMEN EXAMINED: TURKEY. VAN: CaTAKk, vicinity of waterfall, 38°03'10"N

43°02'59"E, 1689 m, on Aspicilia calcarea on calcareous rock, 16.09.2009, leg. A. Aslan.

(ATAKKEF-1451).

REMARKS— Melaspilea interjecta has usually the appearance of an Opegrapha,

while aged specimens with subgyrose apothecia are likely to be confused with

Polysporina simplex. The bilocular ascospores are distinctive.

Placynthium garovaglioi (A. Massal.) Malme, Lichenes Suecici Exs.: 734. 1918.

Thallus thick, brown in the centre, with green-brown lobe tips, small, 0.7

— 1.5 cm wide, with at least part of the margin obscurely lobed; thallus lobes

very slender; lobes 0.1-0.12 mm wide, 0.4-1 mm long; marginal lobes, when

present, are very slender; thallus centre is uneven; thallus with dense, flat to

convex, nodular-scurfy squamules 0.08-0.25 mm in diam.; the entire thallus

powdery with dense, bluish-white pruina, + dull grey-brown or + green-brown

when wet; blue-black prothallus occasionally visible at tips and margins of the

lobes. Apothecia not seen.

Placynthium garovaglioi grows in patches along fissures on sheltered,

vertical to overhanging, dry, and on steeply inclined, sunny calcareous rocks

with some water seepage. New to Turkey and Asia, where it accompanied

Aspicilia, Calcarea, Caloplaca lactea, Caloplaca variabilis, Candelariella aurella,

Lichens newly recorded for Turkey ... 325

Dermatocarpon miniatum, and Verrucaria calciseda. Also known from Europe

(Czeika & Czeika 2007).

SPECIMENS EXAMINED: TURKEY. VAN: BAHGESARAY, exit of district, 38°06'08"N

42°48'41"E, 1587 m, on calcareous rock, 14.09.2009, leg. A. Aslan, (ATAKKEF-1452).

IGpir: Tuzituca, Kula village, roadside, 40°51'4.25"N 43°25'53.48"E, 1175 m, on

calcareous rock, 12.07.2010, leg. K. Yazici. (KTUB-2337).

REMARKS— Placynthium garovaglioi is similar to P hungaricum, but has

appressed marginal lobes and central parts with tightly packed squamules

instead of appressed, and central parts have branched isidioid lobules as in

P. hungaricum. In addition, the ascospores in P hungaricum are 1-septate.

Verrucaria bryoctona (Th. Fr.) Orange, Lichenologist 23: 3. 1991.

Thallus superficial, 0.9-1.4 cm in diam, yellow-green, granular-verrucose,

composed of goniocyst-like units 15-40 um in diam., without pigment in cell

walls. Photobiont green, cells isodiametric to broadly rectangular, 5.5-10 x

4-7.5 um. Perithecia spherical to broadly ovoid, 0.12-0.28 mm in diam., black,

immersed in the substratum, ostiole pale grey, exciple pigmented throughout;

involucrellum absent. Excipulum 20-40 um thick, of textura angularis, darkly

pigmented or slightly paler at base, + reddish-brown, K+ dark grey-brown.

Periphyses up to 25-35 x 2-2.5 um, branched, sparingly anastomosing at base.

Asci clavate, 60-75 x 12-17 um, fissitunicate. Ascospores narrowly ellipsoid,

1(-3)-septate when mature, 19.5-26.5 x 5-7 um, apices rounded or + truncate

with a small gelatinous appendage 2-3.5 um wide and 1 um long. Pycnidia

absent.

Verrucaria bryoctona grows on slightly to strongly basic soil, usually with

acrocarpous mosses, sometimes, overgrowing dead moss, in dry grassland.

It occurs occasionally in semi-natural and man-made habitats such as dunes,

waste ground, spoil heaps, and wall tops. New to Turkey and Asia, where it was

accompanied by Bilimbia sabuletorum, Cladonia pocillum, and Collema tenax.

Also known from Europe and North America (Orange 1991).

SPECIMEN EXAMINED: TURKEY. Burpur: Bucak, surroundings of Karacaéren dam,

37°21'35.14"N 30°50'05.80"E, 300 m, on soil and mosses, 25.06.2012, leg. K. Yazici.

(KTUB- 2335).

REMARKS— Verrucaria bryoctona is similar to V. xyloxena, whichis distinguished

by less elongated ascospores, the absence of a small gelatinous appendage, and a

brown pigment in the thallus.

Acknowledgements

We are grateful to Dr. Harrie Sipman and Dr. Michele D. Piercey-Normore for

revisions and helpful comments on an earlier draft of this manuscript, and also to Dr

André Aptroot for identifications of the lichen species. This study was supported by

TUBITAK (Projects 107T035, 108T566 and 111T857).

326 ... Aslan & Yazici

Literature cited

Akman Y.1999. Climate and bioclimate (The methods of bioclimate and climate types of Turkey).

1“ Edn., Kariyer Matbaacilik Ltd., Sti, Ankara. 350 p.

Baytop A, Denizci R. 1963. Tirkiyenin Flora ve Vejetasyonuna Genel Bir Bakis. Ege Univ. Fen Fak.

Monografiler Ser. 1, Ege Univ. Mat., Izmir. 43 p.

Candan M, Halici MG. 2011. New Cercidospora records for Turkey. Turkish Journal of Botany 35:

625-629.

Czeika H, Czeika G. 2007. Placynthium in den Alpen und Karpaten sowie in benachbarten

Gebieten. Herzogia 20: 29-51.

Halici MG, Cansaran-Duman D. 2008. A New Record for Asia: Abrothallus tulasnei M. Cole & D.

Hawksw. (Dothideomycetes, Ascomycota) from Turkey. Turkish Journal of Botany 32: 325-328.

Halici1 MG, Akata I, Kocakaya M. 2010a. New records of lichenicolous and lichenized fungi from

Turkey. Mycotaxon 114: 311-314. http://dx.doi.org/10.5248/114.311

Halici MG, Hawksworth DL, Candan M, Ozdemir -Tiirk A. 2010b. A new lichenicolous species of

Capronia (Ascomycota, Herpotrichiellaceae), with a key to known lichenicolous species of the

genus. Fungal Diversity 40: 37-40.

Hertel H. 2006. World distribution of species of Lecidea (Lecanorales) occurring in Central

Europe. 19-74, in: A. Lackovicova et al. (eds). Central European Lichens — Diversity and Threat.

Mycotaxon Ltd., Ithaca, and Institute of Botany, Slovak Academy of Sciences, Bratislava.

Karago6z Y, Aslan A, Yazici K, Aptroot A. 2011. Diplotomma, Lecanora, and Xanthoria lichen species

new to Turkey. Mycotaxon 115: 115-119. http://dx.doi.org/10.5248/115.115

Kinalhioglu K. 2010. Five new records for the lichen biota of Turkey. Mycotaxon 112: 371-375.

http://dx.doi.org./10.5248/112.371

Magnusson AH. 1952. Key to the species of Lecidea in Scandinavia and Finland. I. Saxicolous

Species. Svensk Botanisk Tidskrift 46 (2): 313-323.

Oran S, Oztiirk S. 2012. Epiphytic lichen diversity on Quercus cerris and Q. frainetto in the Marmara

region (Turkey). Turkish Journal of Botany 36: 175-190.

Orange A. 1991. Notes on some Terricolous Species of Verrucaria. Lichenologist 23(1): 3-10.

http://dx.doi.org/10.1017/S002428299100004X

Osyczka P, Yazici A, Aslan A. 2011. Note on Cladonia species (lichenized Ascomycota) from

Ardahan province (Turkey). Acta Societatis Botanicorum Poloniae 80: 59-62.

Senkardesler A. 2010. New lichen records from Turkey. Mycotaxon 111: 379-386.

http://dx.doi.org/10.5248/111.379

Sliwa L. 2007. A revision of the Lecanora dispersa complex in North America. Polish Botanical

Journal 52(1): 1-70.

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA, Orange A.

2009. The lichens of Great Britain and Ireland. The British Lichen Society, London. 1046 p.

Yazic1 K, Aslan A. 2009. Lichen species new to Turkey and Asia. Mycotaxon 108: 463-466.

http://dx.doi.org/10.5248/108.463

Yazic1 K, Aptroot A, Etayo J, Aslan A, Guttova A. 2008. Lichens from the Batman, Mardin,

Osmaniye, and Sivas regions of Turkey. Mycotaxon 103: 141-144.

Yazici K, Aptroot A, Aslan A, Etayo J, Spier L, Karagoz Y. 2010a. Lichenized and lichenicolous fungi

from nine different areas in Turkey. Mycotaxon 111: 113-116.

http://dx.doi.org/10.5248/111.113

Yazici K, Elix JA, Aslan A. 2010b. Some parmelioid lichens new to Turkey and Asia. Mycotaxon

111: 489-494. http://dx.doi.org/10.5248/111.489

Yazic1 K, Etayo J, Aslan A. 201la. A note about lichenicolous fungi from Ardahan (Turkey).

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http://dx.doi.org/10.5248/117.145.

MY COTAXON

http://dx.doi.org/10.5248/123.327

Volume 123, pp. 327-334 January-March 2013

Ophiodothella angustissima comb. nov.,

anew name for Acerviclypeatus poriformans and O. vaccinii

RICHARD T. HANLIN * & MARIA C. GONZALEZ ”

"Mycology Herbarium, Museum of Natural History Annex, University of Georgia,

4435 Atlanta Highway, Bogart, GA 30622 USA

*Departamento de Botanica, Instituto de Biologia, Universidad Nacional Autonoma de México,

Ciudad de México, DF 04510 México

* CORRESPONDENCE TO: rhanlin@uga.edu

ABSTRACT —A common leaf spot disease on Vaccinium arboreum in the southeastern United

States has been attributed to Ophiodothella vaccinii. The leaves are infected in early spring

and during the summer abundant acervular conidia are produced. In addition to O. vaccinii,

two names have been applied to this fungus in the literature, Septoria angustissima and

Acerviclypeatus poriformans. A study was undertaken to determine which name should be

applied to this fungus, resulting in our proposing a new combination. On the basis of priority,

O. angustissima comb. nov. is the correct name for the taxon.

KEY worpDs — anamorph, Ascomycota, Phyllachoraceae, sparkleberry, taxonomy

Introduction

Vaccinium arboreum Marshall (sparkleberry) is a common understory shrub

in the southeastern United States. The leaves of this plant are frequently infected

by Ophiodothella vaccinii, which causes a leafspot commonly known as “fly

speck” disease. In her original description of the species, Boyd (1934) described

the anamorph as an acervulus, but without assigning it to a particular genus.

Hanlin (1990a) published illustrations of the species and based on the unique

morphology of the anamorph, he concluded that it could not be accommodated

in any current anamorph. He subsequently assigned it to the new genus

Acerviclypeatus as A. poriformans (Hanlin 1990b). Barr (1990), however, in her

review of pyrenomycetous genera, indicated that Septoria angustissima (Peck

1911) is an earlier name for O. vaccinii. Consequently, a study was undertaken

to determine the correct name for this fungus.

Materials & methods

Type specimens of A. poriformans, O. vaccinii and S. angustissima were examined and

compared visually. Because of the sparseness of material in the type of S. angustissima,

328 ... Hanlin & Gonzalez

no mounts were made, especially since the original description matches the later

descriptions in Boyd (1934) and Hanlin (1990a), which were confirmed by Barr

(1990).

Results

After close examination, it is clear that the three examined type specimens

Acerviclypeatus poriformans, Ophiodothella vaccinii, and Septoria angustissima

represent the same fungus.

The label on the package containing the holotype specimen of Septoria

angustissima is shown in PLATE 1A. Of significance here is the comment, “On

Maclura pomifera (Osage orange), leaves.” ‘The label on the type packet (PLATE

1B) contains the following information, “= Ophiodothella angustissima (Pk.)

n. comb. [in pencil], Herbarium N. Y. State Museum [printed], TYPE Septoria

angustissima Peck [in ink] Aberdeen, Miss. on lvs. of Osage orange [in ink]

(host is Vaccinium arboreum) [in pencil), T.C. Frye, 28 Aug. 1906 [in ink].”

Inside the type packet of S. angustissima are three dried leaves of the

specimen, two intact and the other with a portion missing (PLATE 2A). Also in

the packet are two additional leaves and several notes:

(1) A handwritten note in ink (PLATE 2B) bears the notation, “on leaves of

Osage orange growing wild in river bottoms, Aberdeen, Miss. Coll. T.C. Frye.

No. 26. Aug. 28, 1906.” Above this is a penciled note, “I’m at a loss to understand

why Peck called this a Septoria. (HDH)-

(2) A typewritten note (PLATE 3A) states, “The host is not Toxylon (a loose

leaf of Toxylon, with its acuminate apex enclosed), but seems to match the

smaller leaf enclosed which is Vaccinium arboreum March. [sic] (Batodendron

arboreum Nutt.). Do you agree?” Two leaves are included with this note. Toxylon

is a synonym of Maclura, the genus of Osage orange.

(3) A handwritten 8.5 x 8.5 cm note (PLATE 3B) says, “I compared these

with mulberry leaves. The upper surfaces are similar but the cuticles below

are different. The edges are similar. The fungus is immature. Can you collect

again about the 1* of May?” On the reverse side of this note (PLATE 4A) are the

comments:

“T share your wonder why this was described as a Septoria. The housing is not a

typical pycnidium. The raphe-like sheaves beside the globose cells remind one

of the conidia of some Entylomata. They may not be reproductive organs.

“The units seem to consist of a carbonous clypeus over an acervulus.

“T have retained the leaf I was working on to seek it among the [unclear] of

the heaths” over initials [unclear].

4) A handwritten note (PLATE 4B) is: “Conidial stage of Ophiodothella

vaccinii Boyd M. E. Barr Bigelow May 1963”.

Ophiodothella angustissima comb. nov. ... 329

NEW YORK STATE MUSEUM TYPE COLLECTION

Septoria angustissima Peck

Holotype

Leg. T.C.Frye (comm. F.D. Kern)

Aberdeen MS USA

On Maclura pomifera (Osage orange), leaves

Type database entry#: 0204

r¥?

0‘ —

eos. ; ane Bava a |

y 4 4 pd LS. ii fe

z “— oa i? -_ bil ak

PLATE 1. Septoria angustissima (holotype collection). A: Label. B: Front of packet.

Discussion

Although the leaves of Maclura and Vaccinium resemble one another

superficially in being broadly elliptical and dark green in color, the leaves of

Maclura are larger (7.5-13 cm long x 5-7.5 cm wide) and end in an acuminate

tip, whereas the leaves of Vaccinium are smaller (3-7 cm long x 2-4 cm wide),

with an obtuse apex. The plants also differ greatly in size; Maclura is a large tree,

330 ... Hanlin & Gonzalez

Vin ofa brea Th vnPucTind wh ah called

this @ Safin , HAD

PLATE 2. Septoria angustissima.

A: Type specimen. B: Note with collection information for the holotype.

whereas Vaccinium is a shrub or occasionally a small tree. The leaves of the type

specimen are clearly those of V. arboreum.

The structure of the conidioma, which Peck termed a perithecium, is an

acervulus, whereas in Septoria the conidia are formed in a well-defined

Ophiodothella angustissima comb. nov. ... 331

EO —— EEE eee SEE

The host is not Toxylon (a loose leaf

of Toxylon, with its acuminate atex enclosed),

but seems to match the smller leaf enclosed

which is Vaccinium arboreum Merch (Batodendron

arboreum Nutt.) Do you agree ?

PLATE 3. A: Note comparing a leaf of Maclura (=Toxylon) pomifera (upper) with one of Vaccinium

arboreum (lower). B: Note comparing mulberry leaves (not shown) with the leaves in 3A.

pycnidium (Sutton 1980). Although the conidia in both O. vaccinii and Septoria

are hyaline and filiform, Septoria conidia are usually septate, whereas the conidia

of O. vaccinii are aseptate. Also, conidia in Septoria tend to be larger then those

in Ophiodothella; Sprague (1944) showed that conidia of Septoria from grasses

332 ... Hanlin & Gonzalez

he tincks ttm bo Sonacat of &..

ft, icheumores ebesjrasea CREA. mar, 2 LL Bt wk sea.

WE. Sarr T3x9.cbow) Vag , 176 >

PLATE 4. Septoria angustissima (holotype collection). A: Note with comments regarding

identification of host leaves. B: Annotation by M. E. Barr Bigelow.

Ophiodothella angustissima comb. nov. ... 333

varied in size from 22-98 um long x 1.4-—2.6 um wide, compared with 18-24 x

1 um in Ophiodothella. This is implied by Peck (1911) in his description of the

species as, “Remarkable for its very narrow spores.’ In addition, the Vaccinium

anamorph has an unusual method of pore formation, as shown in a study of

ontogeny of the conidioma (Hanlin 2003). This served as the basis for assigning

it to the genus Acerviclypeatus (Hanlin 1990b).

The penciled annotation at the top of the type packet, “ = Ophiodothella

angustissima (Pk.) n. comb.,” is interesting, as it indicates that the connection

of the specimen with Ophiodothella was recognized early. When and by whom

this annotation was made is unknown, but the name appears to have not

been published. In Index Fungorum (CABI 2012), S. angustissima is listed as

“anamorphic Mycosphaerella” but this refers only to the generic anamorph/

teleomorph connection between Septoria and Mycosphaerella. Ophiodothella

vaccinii also has been mistakenly identified as Rhytisma vaccinii (Schwein) Fr.

due to the similarity in appearance of the acervuli with the apothecia of certain

species of Rhytisma, such as R. punctatum (Pers.) Fr. Rhytisma vaccinii was

described on V. frondosum from North Carolina.

Prior to the description of the genus Acerviclypeatus a search of the literature

was made to look for similar fungi reported on V. arboreum. Alfieri et al. (1984)

reported Septoria albopunctata Cooke on V. arboreum from Florida, and Farr et

al. (1989) list the same species from South Carolina and Texas. The report of S.

angustissima was overlooked because of the misidentification of the host.

Based on the above results, a new combination is proposed for the fungus

causing the leaf spot disease on Vaccinium arboreum.

Ophiodothella angustissima (Peck) Hanlin & M.C. Gonzalez, comb. nov. PL 1-4

MycoBank MB 803907

= Septoria angustissima Peck, Bull. N. Y. St. Mus. 150: 62. 1911.

= Ophiodothella vaccinii Boyd, Mycologia 26: 465. 1934.

= Acerviclypeatus poriformans Hanlin, Mycotaxon 37: 381. 1990.

SPECIMENS EXAMINED, UNITED STATES. Georata: Clarke Co., Athens, on leaves

of Vaccinium arboreum, April 1933, Sophie Boyd (GAM 7026 [=M 7091], holotype of

Ophiodothella vaccinii). Madison Co., on leaves of Vaccinium arboreum, 10 September

1989, RT Hanlin (903) (GAM 12745, holotype of Acerviclypeatus poriformans);

Mississippi: Monroe Co., Aberdeen, on leaves of “Osage orange” [actually V. arboreum],

28 August 1906, TC Frye (NYS 0204, holotype of Septoria angustissima).

On the basis of the priority of its epithet, O. angustissima is the correct name

for the taxon.

Acknowledgments

We thank the curators of NYS for loan of the type of S. angustissima. The senior

author is grateful to the late Margaret Barr for calling his attention to her remarks

regarding this fungus. We thank Sabine Huhndorf and Amy Rossman for their helpful

reviews of the manuscript.

334 ... Hanlin & Gonzalez

Literature cited

Alfieri SA Jr., Langdon KR, Wehlberg C, Kimbrough JW. 1984. Index of plant diseases in Florida.

Florida Dept. Agric. & Consum. Serv. Bull. No. 11 (revised). 389 p.

Barr ME. 1990. Prodromus to nonlichenized, pyrenomycetous members of class Hymenoascomycetes.

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MY COTAXON

http://dx.doi.org/10.5248/123.335

Volume 123, pp. 335-341 January-March 2013

The genus Wrightoporia in Korea

YEONGSEON JANG’, SUNG WOOK LEE’, YOUNG WOON LIM’,

Jin SuNG LEE’, TsuTOMU HATTORI‘ & JAE-JIN Kim’*

‘Division of Environmental Science & Ecological Engineering,

College of Life Sciences & Biotechnology, Korea University,

5-1 Anam-dong, Seongbuk-gu, Seoul, 136-701, Korea

School of Biological Sciences, Seoul National University, Seoul, 151-747, Korea

°National Institute of Biological Resources, Environmental Research Complex,

Incheon, 404-708, Korea

‘Kansai Research Center, Forestry and Forest Products Research Institute,

Biodiversity Research Group, Nagai-Kyutaro 68, Momoyama-cho, Fushimi-ku,

Kyoto, 612-0855 Japan

*CORRESPONDENCE TO: jae-jinkim@korea.ac.kr

ABSTRACT —The genus Wrightoporia (Hericiales, Basidiomycota) and two species, W. japonica

and an undetermined Wrightoporia sp. are reported as new to Korea. Wrightoporia luteola is

treated as a synonym of W. japonica. Illustrated descriptions and a key to Korean Wrightoporia

species are provided.

Key worps — ITS, nuc-LSU, phylogeny, polypore, taxonomy

Introduction

Wrightoporia Pouzar is a genus of wood-rotting fungi. The current generic

concept is broad and the genus includes species with: (1) resupinate to pileate

basidiocarps; (2) poroid hymenophore; (3) smooth to ornamented and amyloid

basidiospores; and (4) diverse hyphal characters such as monomitic or di-

trimitic hyphal system, generative hyphae with or without clamp connections,

and skeletal hyphae with or without a dextrinoid reaction. Some species

also have gloeoplerous hyphae (Hattori 2008). A total of 38 Wrightoporia

species have been described so far according to Index Fungorum databases

(http://www.indexfungorum.org/Names/Names.asp), and they have been

found in both tropical to temperate areas. Of the 18 species reported from

Asia, some have been described from Asia (Hattori 2008), but none have been

recorded in Korea.

336 ... Jang & al.

During studies of indigenous fungi, especially polypores, we found three

Wrightoporia specimens. Our examinations and sequence analysis established

that they represented W. japonica and an undetermined Wrightoporia sp. We

also determined that W. luteola is a taxonomic synonym of W. japonica.

Materials & methods

Collection and morphological examination

Basidiocarps were collected from Mt. Guryong and Mt. Dobong in Seoul, Korea,

and Mt. Songni in Chungcheongbuk-do, Korea, in 2011. They were dried completely

overnight using an air drier at 40°C. Slide preparations were made from dried specimens

mounted in 5% (w/v) KOH, 1% (w/v) phloxine and Melzer’s reagent (Largent et al. 1977)

using an Olympus BX51 light microscope (Olympus, Tokyo, Japan). The abbreviations

used in the text are as follows: IKI- = inamyloid, L = mean spore length, W = mean

spore width. Munsell soil color charts (Munsell Color 2000) was used as the color

standard. The voucher specimens were deposited at the National Institute of Biological

Resources, Incheon, South Korea (KB) with the acronym KUC (Korea University

Culture Collections, Korea University, Seoul, South Korea).

Phylogenetic analysis

The genomic DNAs were extracted according to Jang et al. (2012a). PCR amplification

reactions were performed with the primers LROR/LR3 or LROR/LR5 for nuclear large

subunit ribosomal DNA (nuc-LSU) region (Vilgalys & Hester 1990) and ITS1F/ITS4

for internal transcribed spacer (ITS) region (Gardes & Bruns 1993, White et al. 1990)

according to Jang et al. (2012a). The DNA sequencing was performed at the DNA

sequencing center (Macrogen, Korea). The obtained sequences were deposited in

GenBank. They were aligned with the reference sequences selected by BLASTn search

of GenBank using MAFFT 6.885, L-INS-i alignment method (Katoh & Toh 2008). Each

dataset was manually edited using MacClade 4.08 (Maddison & Maddison 2005). The

aligned nuc-LSU dataset of 20 taxa comprised 911 characters, and the aligned ITS dataset

of 19 taxa comprised 656 characters. Gaps were treated as missing. GTR + I + G model

was applied for both of the datasets via MrModeltest 2.3 under the AIC (Nylander 2004).

Bayesian analysis of each dataset was performed with MrBayes 3.2.1 (Ronquist et al.

2012). A 50% majority-rule consensus tree for each dataset was constructed according

to Jang et al. (2012b). Two runs with 1,000,000 generations were performed and every

100th generation was sampled. Among them, the last 75% sampled trees were used. The

tree was viewed using FigTree 1.3.1 (http://tree.bio.ed.ac.uk/software/figtree/).

Taxonomy

Wrightoporia japonica Nufez & Ryvarden, Fungal Diversity 3: 119. 1999. Fic. 1

= Wrightoporia luteola B.K. Cui & Y.C. Dai, Nova Hedwigia 83: 160. 2006.

BASIDIOCARPS annual, resupinate to effused-reflexed, adnate, ca. 13.5 cm or

more in longest dimension, ca. 5.5 cm in widest dimension in Korean specimens.

Sterile margin narrow, obtuse, felty-corky, very pale brown (10YR8/4), less than

1 mm wide. Pileus elongated, up to 6 mm wide in Korean specimens; pileus

Wrightoporia in Korea ... 337

B OOO000, (\

majeral 10 um

Fic. 1. Wrightoporia japonica. A. Basidiocarp (KUC20111019A-29). Scale bar = 1 cm. B. Microscopic

features (a—e, KUC20110908-13; f, KUC20111019A-29). a, basidiospores; b, basidia; c, cystidioles;

d, generative hyphae from trama; e, skeletal hyphae from trama; f, generative hyphae from context.

surface glabrous, dull, brownish yellow (10YR6/6), often darker near the base;

pileus margin thin, entire, somewhat darker than the sterile margin, less than 1

mm wide. Pores round to angular, 7-8/mm in Korean specimens; dissepiments

thin and entire; pore surface very pale brown (10YR8/4) to brownish yellow

(10YR6/8) in dried condition, slightly shiny when turned in incident light.

Subiculum felty-corky, concolorous to the pore surface, up to 1 mm thick.

Tubes felty-corky, pale yellow, up to 1 mm deep.

HYPHAL SYSTEM dimitic; generative hyphae with clamp connections and

skeletal hyphae.

SUBICULUM generative hyphae with clamp connections, hyaline, thin to

slightly thick-walled, 1.5-4.5 um in diameter; skeletal hyphae hyaline to

yellowish, thick-walled, occasionally branched, straight to sinuous, 3-6 um

in diameter, most with a narrow lumen, some hyphae covered with crystals,

interwoven, IKI-.

TUBES generative hyphae with clamp connections, hyaline, thin to slightly

thick-walled, 1.5-3 um in diameter; skeletal hyphae abundant, hyaline to

yellowish, thick-walled, unbranched, straight to sinuous, 2.5-5 um in diameter,

mostly with a narrow lumen, some hyphae covered with crystals, interwoven,

partly dextrinoid especially in the dissepiments; cystidia none, fusoid cystidioles

present in one specimen (KUC20111019A-29); basidia oblong ellipsoid to

clavate, 4 sterigmate, 9-12.5 x 4—-5.5 um.

BasipiosPorss broadly ellipsoid to subglobose, hyaline, thin-walled, with

finely asperulate ornamentations, amyloid, 2.6-3.5 x 1.6-2.6 um, L = 3.25 um,

W = 2.28 um.

338 ... Jang & al.

TYPE OF ROT— white rot.

SPECIMENS EXAMINED: KOREA, SEOUL, Mt. Guryong, 37°28'43"N 127°04'04"E,

on fallen wood branch, 08 September 2011, Yeongseon Jang (KB, KUC20110908-13;

GenBank KC166692). CHUNGCHEONGBUK-DO, Mt. Songni, 36°38'07"N 127°25'49"E,

on fallen wood branch, 19 October 2011, Yeongseon Jang, (KB, KUC20111019A-29;

GenBank KC166693). JAPAN, OKINAWA PREF., Iriomote Isl., riverside of Shiira River,

19 June 1994, M. Nujfiez (isotype of W. japonica, TFM F-20724). CHINA. ANHUI PRov.,

Huangshan County, Yellow Mts., 13 October 2004, B.K. Cui (isotype of W. luteola, TFM,

ex Dai 6199); Fuy1AN Prov., Wuyishan, the Virgin Forest Park, Longfenggu Scenicspot,

19 October 2005, Y.C. Dai (TFM, ex Dai 7221).

DISTRIBUTION — China, Japan and Korea.

REMARKS — Wrightoporia japonica was first described by Nuftez & Ryvarden

(1999), and Hattori (2008) gave a more detailed description based on ten

specimens including the isotype. An examination of the W. luteola type showed

it to be a resupinate form of W. japonica, which is morphologically similar to

W. gillesii A. David & Rajchenb. (Hattori 2008). Further study is desirable to

reveal whether they represent two distinct species.

Wrightoporia sp. FIG. 2

BASIDIOCARPS annual, resupinate, adnate, and confluent. Pores angular,

5-6/mm, dissepiments thin and entire, partly eroded; pore surface very pale

brown (10YR8/4). Marginal sterile zone indistinct to membranous, very pale

brown (10YR8/2), up to 2 mm wide. Subiculum very thin, less than 0.5 mm.

Tubes concolorous to the pore surface, up to 1 mm deep.

HYPHAL SYSTEM dimitic; generative hyphae with clamp connections and

skeletal hyphae.

SUBICULUM generative hyphae with clamp connections, hyaline, thin to

slightly thick-walled, 1.5-5 um in diameter; skeletal hyphae hyaline to yellowish,

thick-walled, occasionally branched, straight to sinuous, 2.5-6 um in diameter,

mostly with a narrow lumen, interwoven, IKI-.

TUBES generative hyphae with clamp connections, hyaline, thin to slightly

thick-walled, 1.5-3.5 um in diameter; skeletal hyphae abundant, hyaline to

yellowish, thick-walled, unbranched, straight to sinuous, 3-6 um in diameter,

mostly with a narrow lumen, interwoven, partly dextrinoid especially in the

dissepiments; cystidia none, fusoid cystidioles scattered; basidia clavate, 4

sterigmate, 10.5-12.5 x 3.5-5 um.

BASIDIOSPORES broadly ellipsoid to subglobose, hyaline, thick-walled, finely

asperulate, amyloid, 3.2-4 x 2.4-3.2 um, L = 3.62 um, W = 2.85 um.

TYPE OF ROT — white rot.

SPECIMEN EXAMINED: KOREA, SEOUL, Mt. Dobong, on fallen wood branch, 22

September 2011, Yeongseon Jang (KB, KUC20110922-37; GenBank KC166694).

Wrightoporia in Korea ... 339

B ©0009,

UL 00),

Fic. 2. Wrightoporia sp. (KUC20110922-37). A. Basidiocarp. Scale bar = 1 cm. B. Microscopic

features. a, basidiospores; b, basidia; c, cystidioles; d, generative hyphae from trama; e, skeletal

hyphae from trama; f, generative hyphae from context; g, skeletal hyphae from context.

REMARKS — For the time being, we refer this to Wrightoporia sp. because

the species outline is ambiguous with only a single specimen available. The

specimen is similar to W. japonica but is distinguished by the purely resupinate

basidiocarps with a very thin subiculum, partly eroded dissepiments, slightly

larger spores (3.2-4 um long), and parallel hyphae in the trama.

Phylogeny

Bayesian analyses of nuc-LSU and ITS regions were performed for the better

understanding of phylogenetic relationships with other Wrightoporia and

allied species available in GenBank. As shown in Larsson & Larsson (2003),

Wrightoporia species are polyphyletic in both trees (Fic. 3; LSU tree not shown).

Our specimens of W. japonica (KUC20010908-13 and KUC20111019A-29)

have the same nuc-LSU and ITS sequences and form monophyletic clades in

both trees. The ITS analysis (Fic. 3) clusters W. luteola (Dai 7221 from China)

with W. japonica with high posterior probability value (1.0 p.p.), separated in

the aligned ITS dataset by only two base-pair differences and one gap. This

result supports our morphological observation that W. luteola is identical

with W. japonica. Wrightoporia sp. KUC20110922-37 was monophyletic

with W. lenta (Overh. & J. Lowe) Pouzar KN150311 from Jamaica in both

trees, but this was only weakly supported (nuc-LSU, 0.54 p.p.; ITS, 0.65 p.p.).

Wrightoporia sp. KUC20110922-37 is morphologically similar to W. japonica,

and the Wrightoporia sp.- W. lenta clade clustered with W. japonica in the nuc-

LSU tree (not shown). The ITS tree does not resolve the relationships among

340 ... Jang & al.

Wrightoponia luteola Dai 7221 (FJ644289)

Wrightoporia sp. KUC20110922-37 (KC166694)

Wrightoporia lenta KN150311 (AF506489)

Wrghtoporna cylindrospora PRM915962 (GU594161)

Dentipellis leptodon GB011123 (EU118625)

,pLeucophleps spinispora F4674 (AY621 794)

Leucophleps spinispora F3527 (AY621754)

Albatrellus flettii DAVFP:27659 (JF 899544)

ogsflericium americanum CBS 493.63 (AY534581)

Hericium coralloides CBS 447.85 (AF173216)

an ane Creolophus cirrhatus RBG Kew K(M)125827 (EU784260)

Hericium abietis CBS 243.48 (AY534579)

Hericium erinaceum CBS 204.76 (DQ185924)

Creolophus cirrhatus RBG Kew K(M)135658 (EU784261)

Hericium alpestre CBS 539.90 (AY534580)

Laxitextum bicolor NO-7316-Sp (AM269787)

Neoaleurodiscus fujif Wu0807-40 (FJ799921)

0.75

0.1

Fic. 3. ITS region 50% majority-rule consensus tree of Wrightoporia spp. and allied species. For the

construction of consensus tree, 15,000 trees resulting from Bayesian analysis were used. Posterior

probabilities 20.50 are shown above branches. GenBank Accession numbers of the sequences are

shown in parentheses.

Wrightoporia sp.—W. lenta clade, W. japonica, W. cylindrospora Ryvarden, and

Dentipellis leptodon (Mont.) Maas Geest. (Fic. 3). Further study with more

Wrightoporia samples is required to determine the species relationships and

the identity of our undetermined Wrightoporia sp.

Acknowledgments

This research was supported by Basic Science Research Program through the National

Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-

2013R1A1A2A10011390) and was funded by the project on survey and excavation of

Korean indigenous species of NIBR under the Ministry of Environment, Republic of

Korea. TH wishes to express his thanks to Prof. Y.C. Dai (Beijing Forestry University)

for the deposit of W. luteola specimens in TFM. We are grateful to Dr. Leif Ryvarden and

Dr. Ota Yuko for their valuable suggestions on the manuscript.

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MY COTAXON

http://dx.doi.org/10.5248/123.343

Volume 123, pp. 343-352 January-March 2013

Three new cercosporoid fungi from the Brazilian Atlantic Forest

ANDRE L. FIRMINO, DANILO B. PINHO & OLINTO L. PEREIRA*

Departamento de Fitopatologia, Universidade Federal de Vicosa,

Vicosa, Minas Gerais, 36570-000, Brazil

* CORRESPONDENCE TO: oliparini@ufv.br

ABSTRACT — Three new cercosporoid hyphomycetes were found associated with leaf

spots on three host families in a stretch of Atlantic Forest in Vicosa, Minas Gerais, Brazil:

Pseudocercospora aurelianae sp. nov. on Aureliana velutina (Solanaceae), P. vicosae sp.

nov. on Eupatorium sp. (Asteraceae), and Passalora rolliniae sp. nov. on Rollinia sylvatica

(Annonaceae). These three species are fully described, illustrated, discussed, and compared

with allied species in the present work.

Key worps — Capnodiales, Dothideomycetes, Mycosphaerellaceae, taxonomy, tropical fungi.

Introduction

Brazil is considered one of twelve countries with so-called mega-diversity.

Covering the major biomes, the Atlantic Forest is regarded as one of the greatest

repositories of biodiversity on the planet with ca. 20,000 plant species, of

which 8000 are endemic (Myers et al. 2000). For example, in a single region in

southern Bahia state, 450 angiosperms were reported per hectare (Varjabedian

2010). Because the Atlantic Forest has been reduced to 7.5 % of its original

extent (Myers et al. 2000), the study of mycobiota in this endangered biome

has priority.

The lack of knowledge of the mycobiota associated with Atlantic Forest

plants can best be improved by comprehensive examination of certain fungal

groups (Pinho et al. 2012). We decided to study cercosporoid hyphomycetes,

as the knowledge of the diversity of this fungal group in tropical environments

is still sparse. In addition, cercosporoids are among the largest groups of

anamorphic fungi and are commonly associated with leaf spot diseases on

almost all families of flowering plants worldwide (Crous & Braun 2003). First

studies of cercosporoids in Brazil were made by Viégas and Batista (Viégas

1945; Silva & Minter 1995).

344 ... Firmino, Pinho & Pereira

During the last decades, several papers have dealt with cercosporoid fungi

from Brazil (Crous et al. 1997, 1999, 2011; Crous & Camara 1998; Barreto et

al. 1999). The majority of species have been reported in other biomes, mainly

cerrado (Dianese & Camara 1994; Medeiros & Dianese 1994; Inacio et al. 1996;

Dianese et al. 1998, 1999; Braun et al. 1999; Furlaneto & Dianese 1999; Inacio &

Dianese 1998, 1999; Braun & Freire 2002, 2004, 2006; Dornelo-Silva & Dianese

2003; Dornelo-Silva et al. 2007; Hernandez-Gutiérrez & Dianese 2008, 2009;

Freire & Braun 2009). Sporadic works on cercosporoid fungi in Atlantic Forest

fragments revealed some new, undescribed species, such as the first reports

of cercosporoid fungi on hosts of the family Siparunaceae and a revision of

cercosporoids on Piperaceae (Pereira et al. 2003, 2006, 2007; Vieira & Barreto

2004; Pereira & Barreto 2005, 2006; Soares & Barreto 2005; Rocha et al. 2007,

2008, 2010; Silva & Pereira 2007, 2008, 2010, 2011; Alves et al. 2008; Macedo

& Barreto 2008; Almeida et al. 2010; Alves & Barreto 2010; Machado et al.

2012; Silva et al. 2012). In this paper, we introduce three new species, one on

Solanaceae, one on Annonaceae, and one on Asteraceae.

Material & methods

Small samples of the fungi were removed from fresh leaf spots and mounted in

lactophenol. Observations, measurements and drawings were prepared using a Carl

Zeiss Standard W light microscope (Géttingen, Germany). Representative specimens

of the fungi were deposited in the herbarium of the Universidade Federal de Vi¢osa

(Herbarium VIC).

Taxonomy

Pseudocercospora aurelianae Firmino & O.L. Pereira, sp. nov. PLATE 1

MycoBank 800141

Differs from Pseudocercospora carolinensis and P. marcelliana by its light brown conidia,

and from P. modesta by its shorter conidiophores and narrower conidia.

Type: Brazil, Minas Gerais, Vicosa, Estagao Experimental Mata do Paraiso, on leaves

of Aureliana velutina Sendtn. (Solanaceae), 25 May. 2009, A. L. Firmino (VIC 31847,

holotype).

ETyMmo_ocy: referring to the host genus Aureliana.

Leaf spots epiphyllous, circular or irregular, 4-15 mm diam., necrotic, light

brown, with distinct dark brownish margin. Colonies epiphyllous, pale brown.

Stromata well developed, intraepidermal, 30-37.5 x 37.5-45 um, brown.

Conidiophores in dense fascicles, arising from stromata, straight, cylindrical

to slightly attenuated towards a truncate tip, aseptate, rarely 1-septate, not

geniculate, not branched, 8-20.5 x 2.5-4 um, pale brown, uniform in color,

thin-walled, smooth. Conidiophores reduced to conidiogenous cells (one-

celled) or conidiogenous cells integrated, terminal, straight to slightly curved,

Pseudocercospora and Passalora spp. nov. (Brazil) ... 345

PLATE 1. Pseudocercospora aurelianae.

A: Conidia. B: Stomata with conidiophores. Bar = 10 um.

346 ... Firmino, Pinho & Pereira

pale brown. Conidiogenous loci inconspicuous or only visible as truncate tip or

lateral shoulder in geniculate conidiogenous cells after sympodial proliferation,

but always unthickened and not darkened. Conidia solitary, straight to slightly

curved, obclavate-subcylindrical to fusiform, 27-100.5 x 2.5-4 um, 2-7-septate,

light brown, smooth, apex obtuse to subacute, obconically truncate at the base,

with hilum unthickened and not darkened.

COMMENTS — TWENTY-ONE Pseudocercospora species are known to associate

with the Solanaceae (Chupp 1954; Crous & Braun 2003). The only species

reported from Brazil are Pseudocercospora atromarginalis on Capsicum annuum,

P. daturina on Brugmansia suaveolens, P. solani-asperi on Solanum asperum,

and P. venezuelae on S. argenteum and S. gilo (Crous & Braun 2003). The new

species is the first cercosporoid fungus reported on Aureliana.

Pseudocercospora aurelianae is morphologically close to P. carolinensis

(Tharp) U. Braun & Crous and P. marcelliana (Chupp) U. Braun & Crous in the

lack of superficial mycelium and the presence of well-developed stromata with

short conidiophores forming sporodochial conidiomata, but P. carolinensis and

P. marcelliana differ in their hyaline to subhyaline conidia (Chupp 1954). Like

P. aurelianae, P. modesta (Syd.) Deighton has large stromata, but differs by its

longer conidiophores (15-75 um) and much broader conidia (4-6 um; Chupp

1954).

Pseudocercospora vicosae Firmino & O.L. Pereira, sp. nov. PLATE 2

MycoBank 800142

Differs from Pseudocercospora eupatorii by its shorter conidia.

Type: Brazil, Minas Gerais, Vicosa, Estagao Experimental Mata do Paraiso, on leaves of

Eupatorium sp. (Asteraceae), 4 May. 2009, A. L. Firmino (VIC 31848, holotype).

EryMo_oey: referring to the type locality.

Leaf spots amphigenous, circular or irregular, 2-11 mm diam., never

coalescent, dark brown. Colonies mainly epiphyllous, light brown. Stromata

well-developed, intraepidermal, 47.5-70 x 55-97.5 um diam., brown.

Conidiophores loosely fasciculate, arising from stromata, unbranched, flexuous

to curved, cylindrical, 18.5-56 x 3-4 um, 1-2-septate, light brown, paler at the

apex, thin-walled, smooth. Conidiogenous cells integrated, terminal, flexuous

to curved, pale. Conidiogenous loci inconspicuous or only visible as truncate

tip or subdenticulate lateral shoulder formed after sympodial proliferation,

but always unthickened and not darkened. Conidia solitary, straight to slightly

curved, obclavate to fusiforme, 20-91 x 2.5-4 um, 1-8-septate, light brown,

thin-walled, smooth, apex obtuse to subacute, obconically truncate at the base,

hilum unthickened, not darkened.

Pseudocercospora and Passalora spp. nov. (Brazil) ... 347

PLATE 2. Pseudocercospora vicosae.

A: Conidia. B: Stomata with conidiophores. Bar = 10 um.

348 ... Firmino, Pinho & Pereira

CoMMENTS — Forty-two Pseudocercospora species are known to associate with

the Asteraceae (Chupp 1954; Braun & Castafieda 1991; Crous & Braun 2003).

The only two other species reported from Brazil are P elephantopodicola on

Elephantopus sp. (Braun & Freire 2006) and P. plunkettii on Mikania micrantha

(Barreto & Evans 1995). Among the forty-two species of Pseudocercospora

described in association with plants of this family, only P. aciculina (Chupp)

U. Braun & Crous, P ageratoides (Ellis & Everh.) Y.L. Guo, P. eupatorii (Peck)

U. Braun & Castafieda and P eupatorii-formosani U. Braun & Bagyan. are

associated with Eupatorium spp. (Crous & Braun 2003).

Pseudocercospora vicosae is morphologically close to the North American

P. eupatorii in the lack of superficial mycelium and the presence of well-

developed stromata, but P. eupatorii differs in its shorter, rarely septate

conidiophores (10-35 um), and longer conidia (50-190 um; Chupp 1954).

Passalora rolliniae Firmino & O.L. Pereira, sp. nov. PLATE 3

MycoBank 800144

Differs from Passalora miliusae by its much longer and more septate conidiophores, its

solitary or catenate light brown conidia, and its annonaceous host plant.

Ho.orype: Brazil, Minas Gerais, Vicosa, Estagao Experimental Mata do Paraiso, on

leaves of Rollinia sylvatica (A. St.-Hil.) Mart. (Annonaceae), 21 January. 2009, A. L.

Firmino (VIC 31846, holotype).

EryMmo_oecy: referring to the host genus Rollinia.

Leaf spots hypophyllous, circular or irregular, 1-9 mm diam., brown. Colonies

mainly hypophyllous. Stroma absent, Conidiophores simple or mostly branched,

straight to somewhat curved, 107-361 x 2.5-4.5 um, 3-7-septate, solitary, erect

olivaceous, smooth, thin-walled,. Conidiogenous cells integrated, terminal or

intercalate, monoblastic or polyblastic (1-2 loci), olivaceous. Conidiogenous

loci conspicuous, somewhat thickened and slightly darkened. Conidia solitary

or catenate, straight to flexuous, cylindrical to clavate-obclavate, 6-37 x 2-4.5

um, 0-3-septate, without constrictions at the septa, obconically attenuated at

the base with a thickened and darkened hilum, apex rounded or attenuated,

without hilum in solitary conidia or with a single or occasionally two hila in

catenate conidia, light brown, thin-walled, smooth.

COMMENTS — Six Passalora species are known to associate with the Annonaceae

(Chupp 1954; Crous & Braun 2003). The only other species reported from

Brazil are Passalora annonigena on Annona spp. and P. xylopiae on A. dioica

and Xylopia aromatica (Braun & Freire 2002; Crous & Braun 2003). Passalora

rolliniae is the first cercosporoid hyphomycete on Rollinia.

Passalora rolliniae morphologically resembles Passalora miliusae U. Braun

& Crous (= Mycovellosiella indica P. Kumar & Kamal), which differs by its host

plant (Miliusa, family Rubiaceae), its growth habit on the leaf hairs, its much

Pseudocercospora and Passalora spp. nov. (Brazil) ... 349

PLATE 3. Passalora rolliniae.

A: Conidia. B: Conidiophores. Bar = 10 um.

350 ... Firmino, Pinho & Pereira

shorter and less septate conidiophores, and its larger, mostly solitary, subhyaline

conidia [14.5-50 x <5.5 um] (Kumar & Kamal 1982; Crous & Braun 2003).

Acknowledgments

The authors wish to thank Drs. Rafael Castafieda Ruiz (Instituto de Investigaciones

Fundamentales en Agricultura Tropical) and Uwe Braun (Martin-Luther- Universitat),

for reviewing the manuscript.

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MY COTAXON

http://dx.doi.org/10.5248/123.353

Volume 123, pp. 353-361 January-March 2013

A new species of Entocybe (Entolomataceae, Agaricomycetes)

from Québec, Canada

TIMOTHY J. BARONI” & YVES LAMOUREUX?

"Department of Biological Sciences, State University of New York - College at Cortland

PO Box 2000, Cortland, NY 13045 USA

Cercle des mycologues de Montréal, Jardin botanique de Montréal,

4101 rue Sherbrooke Est, Montréal, Québec, Canada H1X 2B2

* CORRESPONDENCE TO: tim.baroni@cortland.edu

AsBstTRAcT — Entocybe melleogrisea sp. nov. is described from a subboreal forest type in

Québec, Canada. ‘The stature of this species is robust and tricholomatoid and thus is very

different from the mostly collybioid species placed in the genus. The distinctive basidiospore

morphology, basidiospore histochemistry, and prolific, obvious clamp connections on all

hyphae of the basidiomata place this species in Entocybe. Preliminary data on ITS sequences

support this placement.

KEY worps — Agaricales, mushrooms, North America, key to species

Introduction

A fair number of publications are available on systematics of the

Entolomataceae Kotl. & Pouzar in North America (Atkinson 1902, 1909; Baroni

1981; Baroni & Petersen 1987; Hesler 1967; Largent 1977, 1994; numerous

publications by Peck from 1872 to 1908 [see Murrill 1917 and Noordeloos

2008]; Mazzer 1976; Murrill 1917), but as yet there is no comprehensive

monograph of this large and morphologically diverse group of agarics with

attached lamellae and flesh-pink angular spores. Although the genus Entoloma

(Fr.) P. Kumm. s.l. is much better known in Europe (Noordeloos 1992, 2004),

even there new species are still being discovered (Noordeloos & Hausknecht

2000; Noordeloos & Morozova 2010). The Entolomataceae in North America

is a seriously understudied mushroom family. With a minimum of at least 500

species already described from North America (mostly summarized in Hesler

1967 and Largent 1994), it is not surprising that newly discovered taxa are still

being found. This report documents one of these distinctive new species.

354 ... Baroni & Lamoureux

Baroni et al. (2011) erected the genus Entocybe T.J. Baroni et al. in the

Entolomataceae to recognize a group of species with several unique basidiospore

features and a distinctive DNA signature. The basidiospores of Entocybe taxa are

reminiscent of basidiospores in the genus Rhodocybe, i.e., they are subglobose

(isodiametric), 6-12 angled in polar view, often obscurely angled in profile view,

and the basidiospore surface is adorned with scattered pustules with broken or

partial ridging. In addition, all taxa produce obvious clamp connections on the

hyphae in all tissues. Taxa with this suite of characters are now confirmed to

belong to a basal clade in the Entolomataceae using molecular DNA evidence

(Baroni et al. 2011; Co-David et al. 2009). The morphological characters of

this new species from North America clearly indicate it belongs in the genus

Entocybe.

Materials & methods

Colors given are general color hues or matched to plates in Kornerup & Wanscher

(1978). All microscopic structures were measured in mounts using 10% aqueous

ammonia (household, non-sudsing, non-scented) or 3% KOH. Structures were also

examined in mounts of Congo Red/ammonia, Melzer’s Reagent and Cotton Blue/

lactic acid to determine histochemical reactions of basidiospore and hyphal walls. For

basidiospore measurements, the hilar appendix or apiculus was excluded. In those

measurements Q refers to the length divided by the width of an individual spore. The

notation n = 21 indicates that 21 individual spores were measured from a single sample.

Means for length and width are given with their standard deviations, as is the mean of

Q values, with the mean of the length divided by the width of all basidiospores measured

being designated as Q™.

Descriptive statistical analysis of basidiospores was determined using EXCEL 5.0.

All light microscopic images were made with an Olympus BX 50 transmitted light

microscope using DIC or bright field optics and captured with a Diagnostic Instruments,

Inc. Insight Spot 3-shot color digital camera.

Scanning electron micrographs were captured on an ISI DS 130c scanning electron

microscope run at 15 or 25 Kev. Tissue samples for SEM were prepared using a modified

procedure originally described in Baroni (1981). Dried lamella fragments were soaked

in 95% ETOH for 1 min, transferred to dH,O for 1 min, then 3% KOH for 1 min, and

finally washed three times in dH,O for 1 min each. These rehydrated lamella fragments

were then sandwiched between two filter papers in a modified Beem capsule and placed

in 10% ETOH for a minimum of 2 minutes. The samples were dehydrated successively

in 30, 50, 70, 80, 90, 95% ETOH solutions for 2 min each, and finally washed 2 more

times in ice cold 100% ETOH before critical point drying in liquid CO, in a Tousimis

Samdri PVT-3B critical point drier.

Results

The following new species is described from a single collection of

multiple fruiting bodies found in the province of Québec, Canada. The

large tricholomatoid stature and grayish yellow colors (Fic. 1) make the

Entocybe melleogrisea sp. nov. (Canada) ... 355

oy,

| q Wy,

Fic. 1. Entocybe melleogrisea (holotype). Basidiomata. Scale bar = 10 mm

taxon distinctive in the field and unlike any other known species in North

America. The subglobose or isodiametric basidiospores with obscure angles

in profile and face views, the minute angles in polar view (Fic. 2), and the

evenly cyanophilic walls of the basidiospores with the characteristic undulate-

pustulate ornamentation (Fic. 3) coupled with obvious clamp connections in

all tissues (Fic. 3) place this taxon in Entocybe (Baroni et al. 2011). The broken

reticulum-like ornamentation (Fic. 4) as viewed under the scanning electron

microscope is also a diagnostic feature for this group of temperate species of

Entolomataceae (Baroni et al. 2011).

ITS (KC701386) and nLSU (KC701387; KC701388) sequences for our

collection do not match any species in blast searches in GenBank. However

the taxa that come closest in blast searches at 81-82% similarity are: Entoloma

nitidum (AF335449.1 and AY228340.1), Rhodocybe trachyospora (JF899553.1),

and Entoloma turbidum (FJ824815.1). These taxa are all considered members

of Entocybe (Baroni et al. 2011), thus supporting our placement of this new

species in that genus.

Entocybe melleogrisea T.J. Baroni & Y. Lamoureux, sp. nov. Fics 1-5

MycoBank MB 800629

Differing from other Entocybe species by the tricholomatoid stature, the grayish cream,

opaque, non-hygrophanous, felted or fibrillose-squamulose pileus surface, the cream

then pale brown stipe with a white, subbulbous, radicating base.

356 ... Baroni & Lamoureux

Fics. 2-3. Entocybe melleogrisea (holotype). 2. Basidiospores. Scale bar = 10 um. 3. Basidiospores

and clamp connection. Scale bar = 5 um

TyPeE: Canada, Québec, City of Saint-Céme, scattered on soil and moss with dead leaves

of Betula papyrifera, Populus tremuloides, and Acer saccharum or some fruiting directly

from very rotten wood in a typical subboreal Abies balsamea-Betula papyrifera forest

type of approximately 75 year old class, and also present in the area were Picea spp.

and Pinus strobus, 18 August 2010, Yves Lamoureux 4150 (Holotype, CMMF; isotype,

CORT). GenBank Accessions: KC701386 (ITS), KC701387 (nLSU, haplotype 1),

KC701388 (nLSU haplotype 2).

EryMo_oey: from the distinctive grayish cream pileus and stipe colors dominating or in

later development as ground color.

BASIDIOMATA tricholomatoid on soil or humus, subboreal forest.

PitEus 40-50 mm, grayish yellow, grayish cream, blond or grayish beige

(4B-C3-6), or medium brown to dark blonde (5D4) over grayish cream ground

color, sometimes paler toward margin or other areas (irregularly pigmented),

convex to campanulate, surface not consistently smooth but irregular on some,

becoming plane with no or a very small rounded umbo, surface finely felted,

then finely fibrillose-squamulose toward margin, not viscid, not hygrophanous,

but a few watery spots sometimes present, not striate, but margin clearly with

fine raised lines on one basidioma (similar to species of Leucopaxillus Boursier

and some of the brown Tricholoma (Fr.) Staude species); CONTEXT white, very

thin (1 mm), fragile; LAMELLAE whitish then pinkish, adnexed then sinuate,

close, moderately thick, very broad (up to 5 mm), edge even; STIPE 35-60 x

4—6(-8 at base) mm, cream then pale brown but white over base, striate and with

appressed fibrils, equal but with base slightly bulbous and obliquely radicating,

solid becoming stuffed, then hollow; Opor and TasTE not distinctive or absent.

SPORE PRINT dull pink (pale brownish pink).

BASIDIOSPORES (6.4—)7.2-8(-8.8) x (5.6-)6.4-7.2(-8) um (x = 7.5 +

0.54 x 6.9 + 0.53 um, Q = 1-1.23, Q™ = 1.09 + 0.07, n = 21) isodiametric or

Entocybe melleogrisea sp. nov. (Canada) ... 357

Fics. 4-5. Entocybe melleogrisea (holotype). 4. SEM of basidiospores (note broken ridges and

isolated pustules). Scale bar = 5 um. 5. Pileipellis with pilocystidia and intracellular pigments.

Scale bar = 10 um

subisodiametric, obscurely angular in profile view with 6-9 facets, isodiametric

and obscurely angular in face view with 5-7 facets, minutely angled in polar

view with 6-9 facets, undulate-pustulate under the light microscope in all views,

walls evenly cyanophilic on smaller, younger spores and larger mature spores

358 ... Baroni & Lamoureux

with breached wall structure, otherwise wall of the larger spores not absorbing

the dye, ornamentation composed of broken ridges and isolated pustules under

an SEM; Basip1a (2-)4-sterigmate, long clavate, hyaline, filled with refractive oil

bodies, 39.7-44 x 10.5-12 um; HYMENIAL CysTIDIA absent; LAMELLA TRAMA

a hyaline layer of parallel, cylindrical or inflated hyphae, 3-13.8 um in diam;

PILEIPELLIS (Fic. 5) a brown layer of + repent, cylindrical hyphae producing

scattered mounds or clusters of + ascendant, entangled cylindrical, subclavate

or clavate pilocystidia, 6-16 um in diam, brown pigments intracellular and

often coagulated into discrete masses of various sizes; PILEUS CONTEXT hyaline,

composed of radially arranged cylindrical and inflated hyphae, 6-24 um in

diam; STIPITIPELLIS similar to pileipellis, a compact brown layer of repent,

cylindrical hyphae, 2-6 um in diam, producing scattered mounds or clusters

of entangled, ascendant cylindrical, subclavate or clavate end cells similar to

the pileipellis, brown pigments intracellular, frequently coagulated; CLamp

CONNECTIONS present in all tissues, large and obvious.

Discussion: The distinctive basidiospore morphology and presence of obvious

large clamp connections on hyphae of the basidiomata place this taxon in

Entocybe. From the senior author's experience, large obvious clamp connections

are not typical in the Entoloma s.. clade and thus seem to be another diagnostic

feature that can be used to recognize Entocybe species. The brownish pink

isodiametric basidiospores that are obscurely angled in profile view but possess

6-9 minute angular facets in polar view and pustule-like ornamentation are

diagnostic for Entocybe (Baroni et al. 2011). Also for Entocybe species the

ultrastructure of the basidiospore ornamentation is consistent and composed of

a bumpy pustulate ornamentation partially interconnected with an incomplete

network of ridges connecting at least some pustules. This incomplete reticulum

is best seen under a scanning electron microscope (Fia. 4). Finally, as noted in

the Results section, ITS sequences also appear to support the placement of E.

melleogrisea with other like taxa in Entocybe and confirm nothing like it occurs

in GenBank.

Molecular analyses (Baroni et al. 2011, Co-David et al. 2009) show

that Rhodocybe species in the Rhodocybe/Clitopilus clade that also have

clamp connections present on the hyphae, e.g., Rhodocybe nitellina and

R. melleopallens, produce very differently shaped and ornamented basidiospores.

The basidiospores are ellipsoid or amygdaliform and with scattered non-

connected pustules or rounded bumps. Entocybe (Baroni et al. 2011) belongs in

the Entoloma clade, not the Rhodocybe/Clitopilus clade of the Entolomataceae,

and these morphological features emphasized for the genus are consistent.

Entocybe melleogrisea is somewhat similar to E. turbida (Fr.) T.J. Baroni et al.

and E. pseudoturbida (Romagn.) T.J. Baroni et al. because of the tricholomatoid

habit. Both E. turbida and E. pseudoturbida differ from E. melleogrisea in

Entocybe melleogrisea sp. nov. (Canada) ... 359

having brown or grayish brown colors on the pileus and stipe, lacking yellow or

cream hues, and the pilei for both are hygrophanous and thus changing color

with loss of moisture. Entocybe turbida has a translucent-striate moist and

mostly glabrous pileus surface except for the disc, which can be subtomentose,

while E. pseudoturbida may be translucent-striate at the margin, but the pileus

is subviscid and opaque when moist. Both species also produce farinaceous

odors and taste (or a rancid taste in the case of E. turbida; Noordeloos 1992).

A few collections of E. turbida, or something very similar to that taxon,

have been found in southern Québec and in the southeastern US; however

E. pseudoturbida has not yet been reported for North America.

Due to the felted opaque pileus surface and tricholomatoid habit,

E. melleogrisea superficially reminds one of a member of the genus Trichopilus

(Romagn.) P.D. Orton. However, for Trichopilus species the pileus surface is

distinctly tomentose and not felted, the basidiospores are ornamented with

interconnected ridges, not broken ridges, and the pileipellis is a trichodermium,

not a cutis. Members of Trichopilus frequently have hymenial cystidia, and these

are often lecythiform on the lamella edge, whereas none of the taxa placed in

Entocybe have been found to produce hymenial cystidia.

Key to Entocybe species

Of the 12 Entocybe taxa documented from the northern hemisphere, most occur in

North America (Baroni et al. 2011). We are aware of at least two other undescribed

taxa in North America that appear to belong in this genus, but at this time we have

inadequate materials to produce scientific descriptions.

1a. Basdiomata with blue or bluish fuscous or violaceous or vinaceous colors on the

piletiswlaniellac-anid tor Stipes wplcawty hater tyleds aed ae cage aah AY aged Ms 2

1b. Basidiomata with brown, gray, yellowish brown or grayish yellow or grayish cream

colors on the pileus and often also the stipe, although the stipe may be white, but

constantly lacking any blue, violaceous or vinaceous hue..................06. 5

2a (1a). Pileus grayish blue, stipe blue or concolorous with pileus except for yellow

hues in the base, lamellae white at first (Noordeloos 1992) ............ E. nitida

2b. Pileus grayish brown or dark purplish brown or reddish brown or fuscous purplish,

lacking yellow hues in the base of the stipe, lamellae more highly colored ...... >

3a (2b). Pileus purplish brown to reddish brown at first, lamellae bluish gray at first,

stipe dark blue to bluish gray, pileus flesh deep blue or bluish gray or purplish

blue orviolaceous orayie 05% seo Wien ee hen E. trachyospora var. purpureoviolacea

3b. Pileus grayish brown, lamellae pallid or grayish brown not with blue colors, pileus

flesh not purplish or violaceous but may be concolorous with pileus surface... .4

4a (3b). Stipe surface, lamellae and pileus context with vinaceous

FTG erie 3 tea 8 Gch: 2G a aas 8 gatas ee LIE 9) rane dls E. trachyospora var. vinacea

4b. Vinaceous colors lacking in these tissues, pileus dark grayish brown but

hygrophanous and becoming paler, lamellae pallid or grayish brown at first,

CONICS RE DIEE so es ees eo teas es aw ote 5 Mee 8 E. trachyospora var. griseoviolacea

360 ... Baroni & Lamoureux

SA WED) DISMICOLOUG:...5 25 S805 Ap ey Ay cata a catacn poy Slee Solano len ey ney la Se OS 6

5b. On humus, mosses, leaf litter, but not lignicolous .................... 0c eee eee vs

6a (5a). Pileus tan or yellowish brown, stipe yellow or pale orange, end cells in

pileipellis variously shaped as pilocystidia, hyphae of lamella trama and pileus

COMLEXE CEMUMITOI. Ait cate he ta Nets Cuan testi N Skee nee eae ed E. speciosa

6b. Pileus “off white to pale buff” becoming a uniform tan, stipe off white, lacking

pilocystidia, hyphae of lamella trama and pileus context not dextrinoid (Largent

NC gal Gece ergs Ore Peet Rt OAL At RAM RARE CER odie Reo E. lignicola

7a (5b). Stature tricholomatoid, medium to large with pileus diameters reaching 50-70

mm, lamellae adnexed or deeply emarginated to almost free or sinuate ........ 8

7b. Stature collybioid, medium to small basidiomata, pileus diameters reaching 40

mm, lamellae attachment more variable, finely adnexed with decurrent tooth or

EMAroi Male GRIN UAL OT AAMAS besa 9 bd hts Fed ds Pek be feb bce povee a chy fe sd vin foto 10

8a (7a). Pileus grayish cream or grayish yellow or dark blond to medium brownish,

but with grayish cream ground colors, dry, not hygrophanous, surface

felted or minute fibrillose-squamulose, stipe concolor, odor and taste

AD SOMES suse the dete tn ota a Ac PR eee Se SMe Ee a aoa nna E. melleogrisea

8b. Pileus gray-brown or sepia or reddish brown or dark umber brown, hygrophanous

and becoming paler, stipe gray-brown or nearly concolorous with the pileus,

odor farinaceous or absent, taste farinaceous or rancid or mild ............... 9

9a (8b). Pileus moist, obscurely to distinctly translucent striate to mid pileus, grayish

brown or sepia or reddish brown, pallescent after losing moisture, smooth,

glabrous or subtomentose on disc when dry, stipe pale grayish brown and often

with yellow hues over base especially when bruised, odor absent or somewhat

farinaceous, taste mild or distinctly rancid (Noordeloos 1992)....... E. turbida

9b. Pileus subviscid when moist, not translucent striate or only at very margin, dark

umber brown or sepia but slightly pallescent after losing moisture, radially

fibrillose or subrugulose on drying, stipe gray or grayish brown, odor and taste

strongly farinaceous (Noordeloos, 1992) ................0008 E. pseudoturbida

10a (7b). Stipe grayish over apex, yellowish downwards, distinctly tapered, + rooting

ALC OTEN TEXUOLIS. 9S Feet Petes Fkars Nene tne kee trees mek E. vinacea

(if stipe entirely grayish E. vinacea var. fumosipes, if stipe violaceous then E. vinacea

var. violeipes - Noordeloos 1992)

10b. Stipe off white, orange gray or brownish orange or hazel or vinaceous buff,

enlarged at base or equal and tapering upward, not rooting, not flexuous ..... 11

11a (10b). Stipe off white, orange gray or brownish orange with age and

handling, lubricous, pileus grayish brown or dark yellowish brown or dark

brown, lubricous, caulocystidia often sphaeropedunculate or pedicellate-

COPY ALC Sra ete Hee ae Ie Pk tae ea ie aed E. trachyospora var. trachyospora

11b. Stipe hazel or vinaceous buff becoming buff, dry, pileus umber or brown

vinaceous on the disc, pale umber or hazel or grayish sepia elsewhere, dry,

caulocystidia mostly clavate or cylindrical ....................0000. E. priscua

Entocybe melleogrisea sp. nov. (Canada) ... 361

Acknowledgments

TJB would like to thank the Hesler Visiting Professor Fund at the University of

Tennessee for supporting work in North and South Carolina that allowed collecting

of Entolomataceae and especially the collection of Entocybe turbida. We are very

grateful to Dr. Karen Hughes, University of Tennessee, for producing the ITS and nLSU

sequences we used for comparison. We also thank Drs. D.L. Largent and S.A. Redhead

for providing valuable reviews and suggestions to improve the manuscript, especially

DLL. Dr. $.R. Pennycook kindly provided a further review and helpful suggestions of

the nomenclature, formatting, and presentation.

Literature cited

Atkinson GF. 1902. Preliminary notes on some new species of fungi. J. Mycol. 8: 110-119.

http://dx.doi.org/10.2307/3752544

Atkinson GF. 1909. Preliminary notes on some new species of Agaricaceae and Clavaria Annales

Mycologici 7(4): 365-376.

Baroni TJ. 1981. A revision of the genus Rhodocybe Maire (Agaricales). Beih. Nova Hedwigia 67:

1-194.

Baroni TJ, Petersen RH. 1987. A new genus in the Entolomataceae. Mycologia 79: 358-361.

http://dx.doi.org/10.2307/3807456

Baroni TJ, Hofstetter V, Largent DL, Vilgalys R. 2011. Entocybe is proposed as a new genus in

the Entolomataceae (Agaricomycetes, Basidiomycota) based on morphological and molecular

evidence. North American Fungi 6(12): 1-19. http://dx.doi:10.2509/naf2011.006.012

Co-David D, Langeveld D, Noordeloos ME. 2009. Molecular phylogeny and spore evolution of

Entolomataceae. Persoonia 23: 147-176. http://dx.doi.org/10.3767/003158509X480944

Hesler LR. 1967. Entoloma in southeastern North America. Beih. Nova Hedwigia 23: 1-196.

Kornerup A, Wanscher JH. 1978. Methuen handbook of color. Third edition. Methuen & Co., Ltd.,

London. 252 p., 30 pl.

Largent DL. 1977. The genus Leptonia on the Pacific Coast of the United States. Bibliotheca

Mycologica 55: 1-286.

Largent DL. 1989. A new lignicolous species of Entoloma (Entolomataceae, Agaricales) from

California. Mycotaxon 34: 129-131.

Largent DL. 1994. Entolomatoid fungi of the western United States and Alaska. Mad River Press.

Eureka, California, USA. 516 p.

Mazzer SJ. 1976. A monographic study of the genus Pouzarella. Bibliotheca Mycologica 46: 1-191.

Murrill WA. 1917. Agaricales, Agaricaceae, Agariceae, Subtribe Pluteanae. North American Flora

10(2): 77-144.

Noordeloos ME. 1992. Entoloma s. |. Fungi Europaei, vol. 5. Giovanna Biella, Italy. 760 p.

Noordeloos ME. 2004. Entoloma s. |. Fungi Europaei, vol. 5a. Edizione Candusso, Italy. pp.

761-1378.

Noordeloos ME. 2008. Entoloma in North America 2: the species described by C.H. Peck - type

studies and comments. Osterr. Z. Pilzk. 17: 87-151.

Noordeloos ME, Hausknecht A. 2000. Tre nuove Entolomataceae (Agaricales) dall’Italia. Il Bolettino

Gruppo Micologico G. Bresadola 43(3): 23-33.

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

MY COTAXON

http://dx.doi.org/10.5248/123.363

Volume 123, pp. 363-367 January-March 2013

A new species of Miriquidica from China

XIN ZHAO, Lu-Lu ZHANG & ZUN-TIAN ZHAO*

College of Life Sciences, Shandong Normal University, Jinan, 250014, P. R. China

* CORRESPONDENCE TO: ztzhao@sohu.com

ABSTRACT — A new lichen species, Miriquidica yunnanensis, is described from China.

Conspicuous features are its yellowish white to grayish white thallus, black-green

epihymenium, colorless to light brown hypothecium, and a unique chemistry. A key to the

seven similar Miriquidica species is also provided.

Key worps — Yunnan, Lecanorales, saxicolous, taxonomy

Introduction

The generic concept of Miriquidica (Lecanoraceae) was first formulated by

Hertel & Rambold (1987). It is characterized by lecideine apothecia, Lecanora-

type asci, simple ascospores without a perispore, filiform conidia, and the

occurrence of miriquidic acid in most species. This widely distributed genus

presently includes 26 species worldwide and usually occurs on acidic rocks

(Hertel & Rambold 1987; Rambold & Schwab 1990; Timdal 1993; Owe-Larsson

& Rambold 2001; Hertel & Andreev 2003; Andreev 2004; Nash et al. 2004;

Fryday 2008; Fryday & Coppins 2008; Coppins & Aptroot 2008; Lendemer &

Knudsen 2008; Ovstedal et al. 2009). In China, only one species, M. complanata,

has been reported (Wei 1991).

During our study of the lichen flora of Mt. Laojun in Yunnan province,

China, a species of Miriquidica new to science was found. Here we present a

brief diagnosis, an extended description, and a key to seven similar species in

the genus.

Materials & methods

The specimen was collected from Yunnan, China, and is preserved in the Lichen

Section of Botanical Herbarium, Shandong Normal University (SDNU). The

morphological and anatomical characters of the specimen were examined using a

stereo microscope (NIKON SMZ 745T) and a polarizing microscope (OLyMpus CX41).

Lichen substances in the specimen cited were identified using the standardized thin

364 ... Zhao, Zhang & Zhao

layer chromatography techniques (TLC) with C system (Orange et al. 2010). The

morphological and anatomical photos of the specie were taken under an OLYMPUS

SZX16 stereomicroscope and an OLymMpus BX61 compound microscope with DP72

respectively.

Taxonomic description

Miriquidica yunnanensis Lu L. Zhang & X. Zhao, sp. nov. Fics 1-2

MycoBank MB 802482

Differs from all other Miriquidica spp. by its yellowish white to grayish white thallus,

blackish-green epihymenium, colorless to light brown hypothecium, small ascospores,

and unique combination of chemical characters.

Type: China. Yunnan province, Mt. Laojun, alt. 4000 m, on rock, 7 Nov. 2009, H.Y. Wang

20121132 (Holotype, SDNU).

Erymo.oey: The specific epithet “yunnanensis” refers to Yunnan province, where this

species was found.

THALLUS crustose, areolate, dispersed on a conspicuous black prothallus;

AREOLES roundish, slightly convex to convex, small, 0.2-0.6(-0.8) mm;

UPPER SURFACE yellowish white to grayish white, due to pigmented hyphae

254nm 365nm = 365nm_~—s sunlight ~=mark

before acid after acid and heating

Figure 1. Miriquidica yunnanensis (Holotype: SDNU (Wang 20121132)). A, B. Thallus; C. TLC

of chemical components (At = atranorin; M = miriquidic acid; N = norstictic acid; U = unknown

substance).

Miriquidica yunnanensis sp. nov. (China) ... 365

FIGURE 2. Miriquidica yunnanensis (Holotype: SDNU (Wang 20121132)). A. Section of apothecium;

B. Paraphyses, after treatment with K; C. Ascus, after treatment with K/I.

of a lichenicolous fungus appearing dark gray, esorediate; MEDULLA I-, with

some granules; PHOTOBIONT chlorococcoid; algal cells of 7-12.5(-15) um in

diam. APOTHECIA: sessile, 0.4-0.9 mm in diam.; pisc black, flat to convex,

usually immarginate; EXCIPLE blackish green to black; EPIHYMENIUM blackish

green; HYMENIUM hyaline, 60-70 um; PARAPHYSES occasionally branched and

anastomosing, 1.5-3 um, apically 2-3.5 um thick; HyPOTHECIUM hyaline to

pale brown; asci of Lecanora-type s.., 8-spored; ascosporEs hyaline, simple,

ellipsoid to broadly ellipsoid, 8-11 x 5-6 um. Pycnip1a immersed, blackish;

conipiA filiform, 10-15 um long.

CHEMISTRY — Atranorin, miriquidic acid, norstictic acid, and unknown

substances present.

DISTRIBUTION AND SUBSTRATE — Known only from Laojun Mountain,

growing on siliceous rock at ca. 4000 m alt.

CoMMENTs — Atranorin is not common in Miriquidica; only M. atrofulva

(+) and M. pycnocarpa contain it in addition to M. yunnanensis. However,

M. atrofulva is distinguished by a rusty colored thallus with dark blue-gray to

blackish soralia and pale brown to olive-brown epihymenium and contains

366 ... Zhao, Zhang & Zhao

stictic acid as a major compound; M. pycnocarpa differs in its I+ violet medulla,

small apothecia (0.15-0.3 mm in diam.), and dark brown hypothecium and

contains only atranorin.

Because Miriquidica yunnanensis is similar in morphology and anatomy to

M. disjecta, M. griseoatra, M. leucophaeoides, M. limitata, and M. paanaensis,

and M. complanata has been reported in China, we provide a key to these seven

species.

Key to seven similar Miriquidica species

Ibelackine-timarprolocettarte acid ns ¢xtees Aah G a Stat ic iRG at leat Bes CROLL EY 2,

25 OFLA OPAULATIOLIN Aa 6 5 lyhey x wt & wpiage ¥ siege ¥ Mipdage ¥ iggy Feline e M. yunnanensis

2D AC ANS AL ATI OED So ae neti se nierlh spree b iecenont ¥ nnilye ¥ isenent ¥ wvereng? SgAeniayt Uedttenoy’ sacttenenst exalec 3

DAs ONEATMINS MORSTICIC ACE oa asco. teg dh vro tng dys pete eh pebon hrs bebeg duh debe grb fekg drh belw dye en ds 4

SDs PACKING MOUSLIGUC ACES are. Suck oes Tonk te tet artTane tice Tags thseatigee heat Ta hse Pa heseatu ceasing 5

4a. Thallus white, yellowish, or pale brown, glossy................. M. leucophaeoides

4b. Thallus pale gray to gray, tinged yellowish ...................00.. M. paanaensis

Savthallus white-to-purely-wiHite. a2. soe set tegs alesis ¥ siletage B aetage elute # e doge M. disjecta

5b. Thallus gray, gray-brown or brownish yellow to dark bluish gray, matt........... 6

6a. Apothecia 0.2-0.7 mm diam., immersed.................00eee eee M. complanata

6b. Apothecia:03=1:2 mimi-diam.,-Sessiles t.75 f.0e4eF fel we hed the! M. griseoatra

Acknowledgments

The project was financially supported by the National Natural Science Foundation of

China (31070010, 31170187) and the Program for Scientific Research Innovation Team

in Colleges and Universities of Shandong Province. The authors thank Dr. A. Aptroot

(ABL Herbarium, Soest, The Netherlands), Prof. Shou-yu Guo (Chinese Academy of

Sciences, Beijing, China) and Prof. G. Rambold (Dept. Of Mycology, University of

Bayreuth, Germany) for presubmission reviews.

Literature cited

Andreev MP. 2004. Notes on the lichen genus Miriquidica (Lecanorales, Lecanoraceae) in Russia.

Bibliotheca Lichenologica 88: 15-42.

Coppins BJ, Aptroot A. 2008. New species and combinations in the lichens of the British Isles.

Lichenologist 40(5): 363-374. http://dx.doi.org/10.1017/S0024282908008165

Fryday AM. 2006. New and interesting North American lichen records from the alpine and sub-

alpine zones of Mt. Katahdin, Maine. Bryologist 109(4): 570-578.

http://dx.doi.org/10.1639/0007-2745(2006)109[570:NAINAL]2.0.CO;2

Fryday AM. 2008. Three new species of lichenized fungi with cephalodia from the southern New

Zealand shelf islands (Campbell Plateau). Lichenologist 40(4): 283-294.

http://dx.doi.org/10.1017/S002428290800769X

Miriquidica yunnanensis sp. nov. (China) ... 367

Fryday AM, Coppins BJ. 2008. Ameliella, a new genus of lichen-forming fungi from north-west

Europe and western Canada. Lichenologist 40(5): 387-397.

http://dx.doi.org/10.1017/S0024282908007913

Hertel H, Andreev MP. 2003. On some saxicolous lecideoid lichens of the Beringian Region

and adjacent areas of Eastern Siberia and the Russian Far East. Bryologist 106(4): 539-551.

http://dx.doi.org/10.1639/0007-2745(2003)106[539:O0SSLLO]2.0.CO;2

Hertel H, Rambold G. 1987. Miriquidica genus novum Lecanoracearum (Ascomycetes lichenisati).

Mitt. Bot. Staatssamml. Miinchen 23: 377-392.

Lendemer JC, Knudsen K. 2008. Studies in lichens and lichenicolous fungi: further notes on North

American taxa. Mycotaxon 103: 75-86.

Nash TH, Ryan BD, Diederich P, Gries C, Bungartz F. 2004. Lichen flora of the Greater Sonoran

Desert Region, vol. 2. 742 p.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

@Ovstedal DO, Tonsberg T, Elvebakk A. 2009. The lichen flora of Svalbard. Sommerfeltia 33:

I—393;

Owe-Larsson B, Rambold G 2001. The sorediate species of the lichen genus Miriquidica (Lecanorales,

Lecanoraceae). Bibliotheca Lichenologica 78: 335-364.

Rambold G, Schwab AJ. 1990. Rusty coloured species of the lichen genus Miriquidica (Lecanoraceae).

Nord. J. Bot. 10: 117—121. http://dx.doi.org/10.1111/j.1756-1051.1990.tb01757.x

Timdal 1993. Miriquidica ventosa comb. nov., a rediscovered lichen. Bryologist 94(4): 616-618.

Wei JC. 1991. An enumeration of lichens in China. International Academic Publishers, Beijing.

313 p.

MY COTAXON

http://dx.doi.org/10.5248/123.369

Volume 123, pp. 369-373 January-March 2013

Lenzitopsis oxycedri (Thelephoraceae, Basidiomycota):

newly recorded for the Balkan Peninsula

MITKO KARADELEV' , KATERINA RUSEVSKA! & OLIVER AVRAMOVSKI”

'Ss Cyril and Methodius University, Faculty of Natural Science and Mathematics,

Institute of Biology, Arhimedova 5, 1000 Skopje, Macedonia

*Galicica National Park, Galicica bb, 4600 Ohrid, Macedonia

* CORRESPONDENCE To: mitkok@pmf.ukim.mk

ABSTRACT — Lenzitopsis oxycedri is reported from the Balkan Peninsula for the first time,

growing in a Mediterranean juniper forest in the Republic of Macedonia. This very rare

species was collected on living branches of Juniperus foetidissima from Galichica National

Park, in the southwestern part of the country.

KEY worpDs — wood-decay fungus, Thelephorales, stinking juniper

Introduction

Malencon & Bertault (1963) described Lenzitopsis as a monotypic genus

in the Thelephoraceae, where it has an isolated position. Lenzitopsis oxycedri

is characterized by perennial, resupinate to effuse reflexed basidiocarps, a

monomitic hyphal system, and globose to subglobose spores that are finely

warted and yellow to pale brown in KOH. Recently Zhou & Koljalg (2013)

described a second species in the genus, Lenzitopsis daii L.W. Zhou & Koljalg,

which differs by its annual basidiocarps, amyloid spores, and growth exclusively

on Juniperus chinensis L.

The original description of L. oxycedri is based on a specimen growing

on Juniperus oxycedrus L. in Morocco (Malencon & Bertault 1963). The first

European records are from Guadalajara, Spain, where the species was found

growing as a saprobe on J. oxycedrus and J. thurifera L. (Garcia-Manjon &

Moreno 1981). Ryvarden (1991: 174) proposed Lenzitella and L. malenconii as

replacement names for Lenzitopsis and L. oxycedri, which he mistakenly believed

to be invalidly published; but because he included the valid Malencon & Bertault

names in synonymy, both Lenzitella and L. malenconii are superfluous and

370 ... Karadelev, Rusevska & Avramovski

‘S

)

FIGURE 1. Geographical distribution of Lenzitopsis oxycedri.

illegitimate. Ryvarden’s (1991) designation of a Spanish collection as “holotype”

(to replace the original Moroccan material that is apparently lost) constitutes

a neotypification of L. oxycedri. Lenzitopsis oxycedri has been reported from

China (Dai et al. 2007), Italy (Bernicchia 2000), and Turkey, where it was

collected twice in Thermessos National Park, Antalya, on branches of Juniperus

foetidissima Willd. (Dogan et al. 2007). This paper extends the distribution to

Macedonia, where L. oxycedri occurs on J. foetidissima.

Materials & methods

Collections were made during springtime (May) 2010 from Juniperus—Quercus

forests in Galichica National Park, Republic of Macedonia. The forest primarily consists

of J. foetidissima mixed with Quercus spp., but at certain spots J. foetidissima forms pure

stands, sometimes intermixed with small stands of J. excelsa M. Bieb.

For identification we used the descriptions of Ryvarden & Gilbertson (1993: 376,

as Lenzitella malenconii) and Bernicchia (2005). The nomenclature follows Index

Fungorum (http://www.indexfungorum.org/names/Names.asp 2012) and Mycobank

(http://www.mycobank.org/Biolomics.aspx? Table=Mycobank 2012). The samples were

examined using classical methods. Sections were mounted in Melzer’s reagent and in

5% aqueous solution of potassium hydroxide (KOH). Measurements and photographs

were taken from sections mounted in KOH (5%) and examined at magnifications up to

1000x with an LW scientific microscope. Specimens are deposited in the Macedonian

Collection of Fungi (MCF), within the Mycological Laboratory (Ss Cyril and Methodius

University, Institute of Biology, Faculty of Natural Sciences and Mathematics).

Lenzitopsis oxycedri new for the Balkans ... 371

FiGuRES 2-4. Lenzitopsis oxycedri (MCF 10/11631). 2. Basidiocarp (left). 3. Spores (above right).

4, Generative hyphae with clamps (below right).

Taxonomy

Lenzitopsis oxycedri Malencon & Bertault, Bull. Soc. Mycol. Fr. 79: 82 (1963)

Fics 2-4

= Lenzitella malenconii Ryvarden, Syn. Fung. 5: 174 (1991), nom. illegit.

Type: Spain, Guadalajara, Tamajon, on living Juniperus thurifera, 26.4.1991, G. Moreno

& L. Gonzaga (neotype, AH; isoneotype, O; Ryvarden 1991).

BASIDIOCARP perennial, effuse reflexed to pileate, projecting up to 2-3 cm

wide, and 2-8 mm cm thick at base, corky to fragile when dry. PILEAL SURFACE

dark brown, velutinate, azonate, later becoming glabrous and almost black.

HyYMENOPHORE lenzitoid to lamellate, lamella 1-3 per mm, sinuous to raduloid

or hydnoid with flat teeth, approximately 0.5 to 1 mm between the lamellae or

spines, at first whitish, then ochraceous to pale brown. CoNnTEXT pinkish buff,

corky, up to 2 mm thick, lamella concolorous with context, corky, up to 6 mm

deep.

HyPHAL SYSTEM monomitic, generative hyphae with clamps, hyaline in

trama and context, pale brown in tomentum, 3-5 um wide, slightly thick-

walled, sparingly branched, some irregular crystals present on hyphae.

CysTipIA absent.

BASIDIA with 4 sterigmata, at first clavate but elongating considerably to

sinuous tube-like organs, 40-80 um long, with a basal clamp. BASIDIOSPORES

globose to subglobose, pale brownish, thick-walled, echinulate, often with small

inner crystals of pigment, negative in Melzer’s reagent, 5-6.9 x 4.9-5.9(-6.9)

um, L = 5.96 um, W = 5.52 um, Q = 1.08.

372 ... Karadelev, Rusevska & Avramovski

SPECIMENS EXAMINED: MACEDONIA, Galichica National Park: Bitolski Pat, 1200 m

a.s.l., Juniperus foetidissima forest with Quercus spp., at the base of branches of living

J. foetidissima, 01.05.2010, Mitko Karadelev (MCF 10/11634); Zli Dol, 1400 m a.s.L,

J. foetidissima forest with Quercus spp., at the base of branches of living J. foetidissima,

01.05.2010, Mitko Karadelev (MCF 10/11633); Velgoshti vill, 800 m a.sl., pure

J. foetidissima forest, at the base of branches of living J. foetidissima, 01.05.2010, Mitko

Karadelev (MCF 10/11631).

Discussion

Macroscopically Lenzitopsis oxycedri resembles Gloeophyllum sepiarium

(Wulfen) P. Karst. and Daedaleopsis tricolor (Bull.) Bondartsev & Singer in

having a lamellate hymenophore. Gloeophyllum sepiarium most commonly

grows on conifers and is yellowish to rusty brown in color, whereas D. tricolor

develops on broadleaved trees, especially Salix. Both species have a dimitic

hyphal structure and smooth basidiospores.

Since L. oxycedri is known from only a few localities in five Mediterranean

countries and China (FiGurE 1), it is considered a very rare white rot saprobe

or parasite that is apparently restricted to junipers. The Chinese specimens

were collected from living junipers in Beijing and Sichuan Provinces (Dai et al.

2007), while elsewhere it occurs on J. oxycedrus in Morocco and Italy (Malen¢gon

& Bertault 1963, Bernicchia 2000, 2005), J. oxycedrus and J. thurifera in Spain

(Garcia-Manjon & Moreno 1981), and J. foetidissima in Turkey (Dogan et al.

2007). Our specimens were also collected from J. foetidissima at three different

sites within Galichica National Park in southwestern Macedonia. Lenzitopsis

oxycedri may well prefer J. foetidissima in the eastern Mediterranean (Macedonia

and Turkey), for it is found exclusively on J. foetidissima even in mixed juniper

forests containing both J. excelsa and J. foetidissima.

Acknowledgments

The authors gratefully thank Drs. H.H. Dogan (Selcuk University, Turkey), H.

Kotiranta (Finnish Environment Institute, Helsinki), and Nomenclature Editor S.R.

Pennycook for helpful presubmission reviews.

Literature cited

Bernicchia A. 2000. Wood-inhabiting aphyllophoraceous fungi on Juniperus spp. in Italy.

Mycotaxon 75: 241-256.

Bernicchia A. 2005. Fungi Europaei, Polyporaceae Vol 10. Italy: Edizioni Candusso.

Dai Y-C, Yu C-J, He W. 2007. Lenzitella—a polypore genus new to China. Fung. Sci. 22(1, 2):

47-50.

Dogan HH, Karadelev M, Isiloglu M, Oztiirk C. 2007. Lenzitopsis oxycedri Malencon & Bertault

(Thelephoraceae, Basidiomycota), a very rare wood-decay fungus collected in Turkey. Turk. J.

Bot. 31: 349-352.

Garcia-Manjon JL, Moreno G. 1981. Estudios sobre Aphyllophorales. I. Fructificationes sobre

Juniperus. Anales Jard. Bot. Madrid 37: 407-416.

Lenzitopsis oxycedri new for the Balkans ... 373

Malencon G, Bertault R. 1963. Lenzitopsis oxycedri Malencon et Bertault, genre nouveau et espece

nouvelle daphyllophorale a spores colorees. Bull. Soc. Mycol. France 79: 75-82.

Ryvarden L. 1991. Genera of polypores, nomenclature and taxonomy. Synopsis fungorum 5. Oslo,

Norway: Fungiflora.

Ryvarden L, Gilbertson RL. 1993. European polypores. Synopsis fungorum 6. Oslo, Norway:

Fungiflora.

Zhou L-W, Koljalg U. 2013. A new species of Lenzitopsis (Thelephorales, Basidiomycota) and its

phylogenetic placement. Mycoscience 54:87-92. http://dx.doi.org/10.1016/j.myc.2012.06.002

MYCOTAXON

http://dx.doi.org/10.5248/123.375

Volume 123, pp. 375-402 January-March 2013

Cortinarius of California: eight new species in subg. Telamonia

DIMITAR BOJANTCHEV

MushroomHobby.com, 345 Shipwatch Lane, Hercules, CA 94547, USA

*CORRESPONDENCE TO: dimitar@pontix.com

ABSTRACT — Described are eight new species of Cortinarius subg. Telamonia from California

and the Pacific Northwest of North America. Four species fruit in the fall and the other four

are vernal. Cortinarius cisqhale is a tanoak associate that fruits abundantly in the fall and

winter. Cortinarius athabascus is an autumnal fruiting species and is broadly distributed

throughout the region in conifer and mixed woods. Cortinarius tuolumnensis, C. miwok,

C. gualalaensis, and C. ohlone belong to the /decipiens clade — the first two fruit vernally

in boreal and montane habitat while the other two are autumnal at low elevations under

coastal pine and live oak, respectively. Cortinarius eldoradoensis is a vernal fruiting species

abundant in the higher elevations near melting snow. Cortinarius truckeensis is a rare, but

locally abundant, species known only from its type location near Truckee, California.

Key Worps — Cortinariaceae, fungal taxonomy, nrITS data

Introduction

Cortinarius subgenus Telamonia (Fr.) Trog is well represented in the state

of California and the broader North American west. This geographical region

features two distinct fruiting periods for Cortinarius — autumn, which is

typical for the genus worldwide, followed by a rich vernal crop, predominantly

in the boreal and montane areas during or shortly after snowmelt. The species

composition is quite different between these fruiting periods with almost no

overlap; only a few species are known to fruit in both seasons. This suggests

that a long evolutionary path of adaptation to seasonal ecological factors has

influenced the time of fruiting. While the vernal fruiting phenomenon for

Cortinarius is not uncommon in Europe, it occurs far more frequently in the

arid mountains of western North America.

Subgenus Telamonia is poorly studied in California, with only a handful of

species reported or described. I am involved in a multi-year study of Cortinarius

in California and the adjacent subcontinent, supported by extensive molecular

376 ... Bojantchev

data analyses. Among the number of undescribed species that have emerged

are eight common, well-understood taxa in subg. Telamonia described here.

Materials & methods

Methods for morphological studies and DNA extraction, PCR conditions and

primers, PCR product clean-up, and sequencing follow Bojantchev & Davis (2011).

Collections are stored in the author’s private herbarium or at the University of California

herbarium in Berkeley (UC) where noted. Extensive iconography of all newly described

species is available on http://www.mushroomhobby.com. Tree genera are abbreviated

with their initial letter, with the exception of Pi. = Picea; P. = Pinus; Ps. = Pseudotsuga.

PHYLOGENETIC ANALYSIS—AIl Cortinarius nrDNA sequences from the public

databases GenBank (http://www.ncbi.nlm.nih.gov) and UNITE (http://unite.ut.ee/)

were downloaded and reviewed. A representative sample of 205 Telamonia sequences

from the northern hemisphere (including 72 sequences from the author's collections)

were selected for phylogenetic analysis based on whether they [1] provided good clade

delineation throughout the entire subgenus or [2] represented taxa closest to the new

species described here.

Sequence alignments were generated with MAFFT v6.821b (Katoh et al. 2002) with

the G-INS-i global alignment iterative refinement strategy. Minimal gap opening and

extension penalties were set for better resolution of the more variable sectors within the

nrITS. The alignments were visually inspected and corrected where needed.

Phylogenetic reconstruction utilized Bayesian inference (BI) running on Mr.Bayes

v.3.1.1 (Huelsenbeck & Ronquist 2003) with the General Time Reversible substitution

model plus gamma distribution (GTR + I’) as the best fit recommended by MrModeltest

v.2.3 (Posada & Crandall 1998). The BI ran two independent analyses with four chains for

16,000,000 generations with sampling frequency for every 100th generation and a burnin

value set at 400,000 (25%). The lowest average standard deviation of split frequencies

achieved was 0.008821. A 50% majority rule consensus tree (Fic. 1) was generated with

the posterior probability (PP) values showing above or below the branches.

Additional phylogenetic analysis of 38 sequences in the /decipiens clade (with

Cortinarius athabascus as an outgroup), including all in GenBank that have 298%

BLAST similarity index to C. tuolumnensis, C. miwok, C. gualalaensis and C. ohlone, was

inferred using the Maximum Likelihood method based on the Tamura-Nei model as

implemented by MEGAS (Tamura et al. 2007). The resulting tree (Fic. 20) offers a much

more detailed view of the /decipiens clade within Western North America, confirming

the phylogenetic position of the newly described species as well as illuminating some

commonly collected undescribed members of the clade.

Fic. 1. Phylogram generated by Bayesian inference of 205 Cortinarius subg. Telamonia nrITS

sequences from the northern hemisphere. Two independent analyses with four chains for 16,000,000

generations were run with sampling frequency for every 100th generation and a burnin value set at

40,000 (25%). The posterior probabilities (PP) of the 50% majority rule consensus tree are showing

above or below the branches. The branches of the new species described here are colored in red.

The GenBank accession numbers are presented after the species names and the author’s collection

numbers are enclosed in parentheses. Holotype collections are marked by an asterisk. Misapplied

species names on GenBank collections are enclosed in quotes.

New telamonias from California ... 377

82 /brunneus

/colymbadinus, ahsii

96 100 f

/alboviolaceus

/turgidus

95 99 : P

/biformis

100 100 :

/bovinus

8 C. eldoradoensis JQ906746 (DBB04565*)

C. eldoradoensis JQ906747 (DBB33888)

= C. "limonius" GQ159869

90 : /anisatus, furvolaesus

u /armillatus, paragaudis

= /betuletorum

od /balaustinus /subbalaustinus

57 ;

/bivelus

80 rv C. athabascus JN133295 (DBB27618*)

C. athabascus JN133294 (DBB27010)

/cagei

C. candelaris AY669675

100 C. duracinus AY669674

C. cisqhale JF795387 (DBB26828*)

C. cisqhale JF795388 (DBB26773)

C. fuscoperonatus EU433390

C. humidicola AF388785

C. laniger AF325591

100 C. solis-occasus AY669696

C. malachius AY669681

C. testaceofolius EU693242

C. truckeensis JQ937284 (DBB33100*)

100 ! C. truckeensis JQ937285 (DBB43920)

C. traganus HM240523

/torvus /agathosmus

C. decipiens AY083180

100 C. atrocoerulaeus AY083178

97 + C. decipiens FN428992

59 C. decipiens FN428994

100 C. tuolumnensis JQ906762 (DBB15027*)

C. tuolumnensis JQ906763 (DBB43454)

C. "parevernius" HQ604722 /decipiens

55 99 C. ohlone JQ906757 (DBB42193*)

C. ohlone JQ906759 (MD101205)

C. ohlone JQ906758 (DBB41040)

C. miwok JQ906753 (DBB43810"*)

C. miwok JQ906751 (DBB16312)

C. "sertipes" FJ039540

/saniosus

93 100

100

/comptulus

/casimiri

/hinnuleus

/flexipes

/vernus

378 ... Bojantchev

Fic. 2. Cortinarius cisqhale (UC 1860822, holotype).

Taxonomy

Cortinarius cisqhale Bojantchev, sp. nov. FIGS 2-7

MycoBAnk MB 800343

Differs from other Telamonia species by its distinct coloration, small spores, and

association with Tanoak.

Type: USA. California: Sonoma County, Salt Point State Park, 0.5 mi. North of Hwy 1

(38°33'48"N 123°18'27"W), elev. 300 ft, under Notholithocarpus densiflorus, P. muricata,

Ps. menziesii, 20 Nov 2009, Bojantchev DBB26828 (Holotype UC 1860822; Genbank

nrITS JF795387, RPB2 JQ906739).

EryMo.oey: Cigghale (= beautiful tree) is the word for tanoak in the language of the

Kashaya/Pomo Native American people.

Piteus 40-120 mm diam., very variable in shape, hemispherical to convex

when young, typically with a broad flattened umbo; margin involute then

straight to flanging; colors diverse, mostly in the light to dark brown spectrum

with grayish to olive tinges when wet, pale brownish when dry, often with

bluish tinges outside of the disc, with the margins remaining whitish due to

persistent velar remnants; surface smooth to shiny when wet, dull fibrillose

when dry, hygrophanous in radial streaks, leaving irregular shapes, drying

from the center out. LAMELLAE moderately crowded to subdistant, 5-18 mm

broad, light or dark clay brown, occasionally lilac when young, turning rusty

brown as the spores mature; edges even, pale; attachment sinuate; lamellulae

abundant in series of 3-7. Stipe 50-120 mm long, 12-40 mm wide, cylindrical

to subclavate, occasionally bulged at the base, frequently laterally subradicating,

New telamonias from California ... 379

Fic. 3. Cortinarius cisqhale a) Basidiospores (UC 1860820, holotype) b) DBB27382

light to dark brown beneath the cottony white universal veil, with bluish tinges

near the top on young basidiomata; mycelial strands occasionally present in

mature basidiomata. UNIVERSAL VEIL gray white, covering the stipe over the

entire length, often leaving an annular zone, easily bruised off. CoRTINA gray

white. CONTEXT mottled, with grayish white zones mixed with brown in the

lower stipe and with predominantly bluish tinges in the upper stipe; colors less

contrasting at age, settling to grayish brown; bluish tinges stronger near the

outer surface. ExsiccaTa dark gray to charcoal. HaBir scattered to frequently

caespitose of up to a dozen basidiomata. Opor earthy. Taste mild earthy.

UV Licurt no fluorescence detected. MACROCHEMICAL REACTIONS 5% KOH

negative. SPORE DEPOSIT rusty brown.

BASIDIOSPORES (6.5—)7.0-8.0(-9.0) x (3.8-)4.0-4.6(-5.0) um (mean 7.5 x

4.4 um), Q = 1.59-1.86, Q. = 1.73 (N = 198, 8 basidiomata, four collections),

amygdaliform to narrowly ellipsoid, slightly ovate, moderately finely verrucose,

non-dextrinoid. Basip1A 28-36 x 5-7 um, 4-spored, cylindro-clavate, clamped.

LAMELLA EDGE sparsely fertile. Cystrp1a not observed. LAMELLAR TRAMA

composed of filamentous hyphae, 4-8 um wide, with an olivaceous pigment.

PILEIPELLIS a cutis; upper layer composed of parallel hyphae, 4-12 um wide,

smooth, with olivaceous-brown pigment; lower layer hyphae 8-20 um wide,

thin walled. No distinct reaction with Melzer’s reagent. HyYPODERMIUM not

observed. OLEIFEROUS HYPHAE common in all parts. CLAMP CONNECTIONS

common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius cisqhale appears to be associated

solely with tanoak (Notholithocarpus densiflorus) and broadly distributed

throughout its range. It has been collected along the California coast, in

southern Oregon and the foothills of Sierra Nevada. At some locations in the

fall it can be the most prevalent Telamonia. The only matching sequences in

GenBank are from environmental studies on tanoak root tips (Bergemann &

Garbelotto 2006).

380 ... Bojantchev

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: MENDOCINO COUNTY,

Jackson State Demonstration Forest, off Little Lake Rd. (39°18'27"N 123°42'47”W), elev.

1800 ft, under N. densiflorus, Ps. menziesii, Tsuga heterophylla, Sequoia sempervirens, 14

Nov 2009, DBB26186; Feb 13, 2010, DBB27382, DBB27520; Jackson State Demonstration

Forest, off Hwy 20. (39°21'29"N 123°38'11"W), elev. 1700 ft, under N. densiflorus,

Ps. menziesii, T: heterophylla, S. sempervirens, Nov 24, 2007, DBB01833 (Genbank nrITS

JF795390, RPB2 JQ906737); DBB01834; Nov 23, 2008, DBB09828, DBB10324; SONOMA

County, Salt Point State Park, 0.5 mi. North of Hwy 1 (38°33'48"N 123°18'27"W), elev.

450 ft, under N. densiflorus, P. muricata, Ps. menziesii, 23 Nov 2007, DBB01378; 20 Nov

2009, DBB26773 (Genbank nrITS JF795388, RPB2 JQ906738), DBB26876, DBB27028;

YuBA County, New Bullard’s Bar Reservoir (39°25'23"N 121°05'27"W), elev. 2200 ft,

under N. densiflorus, P. ponderosa, 24 Nov 2010, DBB39790 (Genbank nrITS JQ906745

RPB2 JQ906740); DBB40759; OREGON: CuRRY COUNTY, Samuel Boardman State Park,

elev. 100 ft, under N. densiflorus, 11 Nov 2009, Bojantchev DBB27721.

Discussion — Cortinarius cisqhale is a very variable species with several distinct

forms that could easily be interpreted as different species. Careful collecting

and observation of basidiomata in all stages of development is required for the

synthesis of a refined species concept. In some locales like the Sierra Nevada

foothills and Santa Cruz this species tends to form clusters of long-stemmed

fruitbodies in a narrowly caespitose habit (Fic. 6a). In wet conditions the

coloration becomes distinctly olive-gray (Fic. 3b), while in dry conditions the

collections look more like Fic. 4b. The material sometimes can be fairly dark

(Fic. 1), while in age it fades to develop a strongly hygrophanous appearance

(Fic. 7a,b). The exsiccata have a distinctly blackish-charcoal appearance, similar

to C. brunneus (Pers.) Fr. and can easily be confused for a member of that clade.

The distinctly mottled stem and persistent bluish-grey context coloration in the

upper stem context are good taxonomic characters for identifying C. cisqhale in

the field. A key factor is also the presence of tanoak.

While the nrITS gene region is the primary fungal barcode marker in

the Cortinarius study used throughout this paper, the diversity of forms in

C. cisqhale prompted using nrRPB2, an additional gene region that previous

analysis of multiple collections (not shown) demonstrated a good species level

resolution. The sequences of all examined C. cisqhale collections matched 100%

in both gene regions.

Cortinarius cisqhale is quite isolated from any other sequences in GenBank,

with the BLAST max identity index to the nearest neighbor 94% for nrITS and

92% for RPB2. The position of C. cisqghale within subg. Telamonia was not easily

resolved as different methods and packages for phylogenetic reconstruction

generated somewhat conflicting results (data not shown). The Bayesian

analysis generated the most likely ancestral relationship scenario showing

C. cisghale as fairly isolated from most other known species. Future research

will undoubtedly shed additional light on its ancestral hierarchy as more closely

related species are found.

New telamonias from California ... 381

Fic. 7 Cortinarius ate ae dry basidiomata a) DBB01833 b) DBBO1834

382 ... Bojantchev

Fia. 8. Cortinarius athabascus DBB27073.

Cortinarius athabascus Bojantchey, sp. nov. Figs 8-11

MycoBank MB 800344

MISAPPLIED NAME - “Cortinarius brunneus” sensu Trudell & Ammirati (2009:151).

Differs from other Telamonia species by its distinct coloration, conifer association,

autumnal fruiting, and western American distribution.

Type: USA. California: Mendocino County, Mendocino, off Crestwood Dr. (39°18'18"N

123°47'26"W), elev. 400 ft, under P muricata, 21 Nov 2009, Bojantchev DBB27618

(Holotype UC 1860905; Genbank nrITS JN133295).

Erymo.oey: In honor of the Athabascan family of Native American tribes — this

species can be found throughout its range along the Pacific coast of North America.

Piteus 30-150 mm diam., campanulate to convex with 1-3 concentrically

raised and depressed zones, typically with a broad flattened umbo; margin

involute then straight, often flanging; dark brown with bluish-grey tinges when

moist, drying to brown, or pale brown at age, often hygrophanous in concentric

zones or radial streaks; veil remnants present at the extreme margin; surface

smooth, innately fibrillose, dull when moist, often shiny when dry. LAMELLAE

moderately crowded to subdistant, 6-22 mm broad, clay brown, turning rusty

brown as the spores mature; edges even to slightly uneven; attachment sinuate;

lamellulae abundant in series of 3-5. StrpE 50-180 mm long, 15-50 mm wide,

cylindrical to slightly subclavate, often twisted, pale brown beneath the white

veil remnants, bluish tinges at the extreme apex often preserved into maturity.

UNIVERSAL VEIL white, abundant, covering the stipe over the entire length,

often leaving a distinct annular zone, occasionally shedding white floccose

scales on the pileus that disappear at maturity. CorRTINA white. CONTEXT

mottled with white to brown vertical streaks in stipe, with predominantly

New telamonias from California ... 383

Fic. 9. Cortinarius athabascus (UC 1860905, holotype) a) Basidiospores b) Basidiomata

bluish tinges in the upper stipe and pronounced orange brown spots at the

base. ExstccatTa dark brown, the stipe brown. Haxit mostly scattered,

but frequently caespitose. OpoR earthy. Taste mild earthy. UV LicHT no

fluorescence detected. MACROCHEMICAL REACTIONS 5% KOH negative. SPORE

DEPOSIT rusty brown.

BASIDIOSPORES (8.0—)8.5-10.0(-10.5) x (5.0—)5.4-5.8(-6.3) um (mean 9.2

x 5.6 um), Q = 1.51-1.76, Q. = 1.65 (N = 138, 5 basidiomata, four collections),

amygdaliform to narrow ellipsoid, finely verrucose, non-dextrinoid. BAsIDIA

27-35 x 5-8 um, 4-spored, cylindro-clavate, clamped. LAMELLA EDGE

infertile. CysTIpIA not observed. LAMELLAR TRAMA composed of filamentous

hyphae, 3-8 um wide. PILEIPELLIS a cutis, upper layer composed of thin

parallel hyphae, 3-6 um wide, brownish pigmented in KOH, walls smooth to

very finely encrusted. Lower layer hyphae 5-18 um wide, with thick hyaline

walls. No distinct reaction to Melzer’s reagent. HYPODERMIUM not observed.

OLEIFEROUS HYPHAE sparse. CLAMP CONNECTIONS common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius athabascus is widely distributed

in northern coastal California and the Pacific Northwest in conifer and mixed

woods. It has often been collected in the woods along the northern California

coast, Oregon, and the Olympic Peninsula of Washington. There are several

matching collections in GenBank, one of which (FJ717542) from Washington

state is featured in Trudell & Ammirati (2007) as C. brunneus. This species

has not been reported yet from the inner mountain ranges of western North

America.

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: MENDOCINO COUNTY,

Mendocino, Jackson State Demonstration Forest, off Little Lake Rd. (39°18'27"N

123°42'47"W), elev. 100 ft, under Ps. menziesii, N. densiflorus, T: heterophylla,

S. sempervirens, 21 Nov 2009, Bojantchev DBB27073; Gualala, off Old State Hwy

(38°46'07"N 123°31'29" W), elev. 300 ft, under Ps. menziesii, N. densiflorus, S. sempervirens,

26 Nov 2010, Bojantchev DBB40030 (Genbank nrITS JN133296); Sonoma Counry,

Salt Point State Park, 0.5 mi. North of Hwy 1 (38°33'48"N 123°18'27"W), elev. 300 ft,

under N. densiflorus, P. muricata, Ps. menziesii, 23 Nov 2007, DBB01290 (Genbank

nrITS JN133298); 20 Nov 2009, DBB26792 (Genbank nrITS JN133293), DBB27010

(Genbank nrITS JN133294); HUMBOLDT County, Eureka (40°49'48"N 124°10'45"W),

384 ... Bojantchev

Fic. 10. Cortinarius athabascus a) DBB40030 b) DBB40030 c) DBB01290

Sade ty i Hh

elev. 100 ft, under Pi. sitchensis, 20 Nov 2010, DBB39388; OREGON: CURRY COUNTY,

Samuel Boardman State Park, elev. 100 ft, under Pi. sitchensis, 11 Nov 2009, Bojantchev

DBB37325; WASHINGTON: CLALLAM County, Olympic Peninsula, off Hot Spring

Rd. (48°04'29"N_ 123°57'18"W), elev. 1000 ft, under Pi. sitchensis, Pi. engelmannii,

Ps. menziesii, 21 Oct 2009, DBB24697;

Discussion — Cortinarius athabascus is one of the most common Telamonia

species in the coastal woods of the Pacific rim of North America. It is also one

of the larger Telamonia in the area, as it can achieve an impressive size. This

species is quite variable in appearance with diverse hygrophanous patterns on

the cap, while some forms remain quite pale even when moist. One of the key

taxonomic differentials against other local Telamonia taxa is the long lasting

and very distinct bluish coloration of the stem context in its upper part, even

in mature basidiomata. The stepped/depressed multiple rings on the cap with

smooth to innately fibrillose surface are good field characteristics too. Based on

GenBank records this species is very commonly misidentified in the PNW as

various European taxa. Careful analysis of the images presented in this study

will help researchers to better comprehend the range of coloration and stature

of this common western North American species.

Cortinarius eldoradoensis Bojantchey, sp. nov. FIGS 12-15

MycoBank MB 800346

Differs from other Telamonia species by the totality of its coloration, montane vernal

fruiting habit, and absence of fluorescent universal veil remnants.

New telamonias from California ... 385

di WA |

AH 77

‘A

4 -\ : Y E

ea eA ~~

Fic. 12. Cortinarius eldoradoensis DBB15611. Note the distinct cavities in the stipe.

Type: USA. California: El Dorado County, El Dorado National Forest, 0.1 mi. east of

Wrights Lake Road (38°48'20"N 120°14'18"W), elev. 6100 ft, under P. ponderosa, Abies

concolor, A. magnifica, Ps. menziesii, 26 May 2007, Bojantchev DBB04565 (Holotype UC

1861350; Genbank nrITS JQ906746).

EtyMoLoGy: after the Eldorado National Forest in the Central Sierra Nevada.

PitEus 50-120 mm diam., convex to plano-convex without umbo; margin

involute, frequently flanging at age; brown with bluish grey tints towards the disk;

the extreme margin frequently whitish due to velar remnants; surface smooth

to glossy, innately fibrillose, inconspicuously hygrophanous with occasional

radial streaks. LAMELLAE moderately crowded, 7-22 mm broad, latte brown at

first, turning rusty brown as the spores mature; edges even, attachment sinuate;

lamellulae abundant. Stipe 40-80 mm long, 15-30 mm wide, cylindrical to

subclavate, white to beige, sometimes shiny. UNIVERSAL VEIL white, partially

covering the lower stipe, often leaving an evanescent annular zone. CORTINA

white. CONTEXT white to beige, developing cavities along the length of the

stipe. ExsiccatTa dark brown. Hair scattered to sub-caespitose. ODOR earthy.

TasTE mild earthy. UV LicuT no fluorescence detected. MACROCHEMICAL

REACTIONS 5% KOH negative. SPORE DEPOSIT rusty brown.

BASIDIOSPORES (8.0-)8.5-10.5(-11.3) x (4.8-)5.0-6.0(—6.5) um (mean 9.5 x

5.5 um), Q = 1.34-2.08, Q. = 1.73 (N = 203, 7 basidiomata, five collections),

variable in shape, mainly amygdaliform, but varying from ovoid to cylindro-

amygdaliform to oblong-ellipsoid, moderately verrucose, non-dextrinoid.

386 ... Bojantchev

: Ths as Tinea d ;

Fia. 14. Cortinarius eldoradoensis a) DBB33888 b) DBB15626

BASIDIA 26-36 x 6-9 um, 4-spored, cylindro-clavate, clamped. LAMELLA EDGE

sparsely fertile. Cystrp1a not observed. LAMELLAR TRAMA regular, composed

of more or less parallel, cylindrical hyphae, 2-8 um wide. PILEIPELLIs a cutis,

upper layer composed of thin parallel hyphae, 3-5um wide with pale olivaceous

pigment in KOH, walls smooth, not encrusted. Lower layer hyphae 6-22 um

wide, with thin walls. No distinct reaction with Melzer’s reagent. HYPODERMIUM

\ ~ » : . { a

~ mn AL ‘ ‘G . ‘ lal 10um QO

\th ins ‘ . |

a”

a) Oleiferous hypha in the pileus trama (UC 1861350, holotype)

b) Basidiospores (UC 1861350, holotype)

New telamonias from California ... 387

not developed. OLEIFEROUS HYPHAE abundant in all parts, with thick refractive

walls and olivaceous pigment. CLAMP CONNECTIONS common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius eldoradoensis fruits abundantly

in late spring and early summer in the mountains of California during, or soon

after snow melt. Apparently it is broadly distributed in the Pacific Northwest,

yet probably uncommon, as there is only one matching sequence in GenBank

(GQ159869) from Mt. Washington, Vancouver Island, British Columbia (also

vernally fruiting). In California this species is very common in both the foothills

of the Sierra Nevada and the higher elevations of the Sierra—Cascade range

where it can frequently be observed right along the banks of melting snow. The

typical surrounding trees are conifers like ponderosa pine (P. ponderosa), white

fir (A. concolor), red fir (A. magnifica) and Douglas-fir (Ps. menziesii).

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: EL DoRADO County, El

Dorado National Forest, 0.1 mi. East of Wrights Lake Road (38°48'20"N 120°14'18"W),

elev. 6100 ft, under P ponderosa, A. concolor, A. magnifica, Ps. menziesii, 22 May 2009,

DBB15611 (Genbank nrITS JQ906747); DBB15626; DBB15638; TuoLUMNE County,

Yosemite National Park, off Hodgdon Meadow Access Rd (39°18'25"N 123°42'53"W),

elev. 5100 ft, under P ponderosa, A. concolor, A. magnifica, Ps. menziesii, 25 Jun 2010,

DBB33888 (Genbank nrITS JQ906748); DBB34062 (Genbank nrITS JQ906749); SERRA

County, Chapman Creek Campground (39°37'54"N 120°32'37"W), elev. 5900 ft, under

P. ponderosa, A. concolor, A. magnifica, Ps. menziesii, 14 May 2010, DBB43454 (Genbank

nrITS JQ906750);

Discussion — Cortinarius eldoradoensis is a typical member of the rich vernal

snowbank Telamonia biota of the Sierra—-Cascade range. Most of these species

share similar brownish coloration and overall stature, which makes them

difficult to separate in the field. The problem is exacerbated by the fact that

these species fruit in moist spots near melting snow where the soil is very soft

and obtaining clean material is difficult. Several years of field observation of

multiple collections were needed in order to understand this species better.

There is a cluster of several very similar, undescribed, vernal species, which can

be separated based on their positive UV fluorescence. The less similar vernal

species in the /decipiens clade, like C. miwok and C. tuolumnensis, develop

purplish coloration in the stipe context. In addition, most C. eldoradoensis

basidiomata develop distinct cavities in the stipe, which can be diagnostic for

this species.

Cortinarius truckeensis Bojantchev, sp. nov. Fics 16-19

MycoBank MB 800349

Differs from other Telamonia species by the distinct white coloration, montane vernal

fruiting habit, and large spores.

Type: USA. California: Nevada County, North of Truckee, off Hwy. 89 (39°24'05"N

120°11'19"W), elev. 6200 ft, under P jeffreyi, 12 Jun 2010, DBB33100 (Holotype UC

1861353; Genbank nrITS JQ937284).

388 ... Bojantchev

<i > ; “ aS

Fic. 16. Cortinarius truckeensis (UC 1861353, holotype).

EryMo_oey: after the town of Truckee, California in the vicinity of which this species

is abundant.

PiLEus 40-100 mm diam., convex to plano-convex to depressed; margin involute

into maturity, often wavy; white to light gray-white, remaining so into maturity;

surface fibrillose, smooth to shiny, inconspicuously hygrophanous. LAMELLAE

moderately crowded to subdistant, 8-26 mm broad, tan to latte brown, without

bluish tinges, turning rusty brown as the spores mature; edges even, paler than

the sides; attachment sinuate; lamellulae abundant in series of 3-7. STIPE 30-90

mm long, 20-44 mm wide, cylindrical, white, occasionally with a silky shiny

surface, solid well into maturity. UNIVERSAL VEIL white, occasionally leaving

an evanescent annular zone at the base of the stipe. CORTINA white. CONTEXT

white to pale beige, sometimes mottled with tan to brown longitudinal streaks.

ExsiccaTta brownish with silvery sheen, particularly on the stipes. HABIT

subcaespitose to scattered. OpoR earthy. Taste mild earthy. UV LicuT no

fluorescence detected. MACROCHEMICAL REACTIONS 5% KOH negative. SPORE

DEPOSIT rusty brown.

BASIDIOSPORES (8.5-)9.0-11.2(-12.0) x (4.8-)5.0-5.5(-6.0) um (mean 10.1

x 5.3 um), Q = 1.70-2.13, Q. = 1.91 (N = 146, 4 basidiomata, four collections),

inequilaterally amygdaliform to broadly fusiform, some strangulated near

the apex, very finely verrucose, non-dextrinoid. Basip1A 32-44 x 7-10 um,

4-spored (some 2-spored), cylindro-clavate, clamped. LAMELLA EDGE sterile.

CysTIDIA not observed. LAMELLAR TRAMA regular, composed of more or less

parallel, cylindrical hyphae, 4-10 um wide, with brown intracellular pigment

when viewed in KOH. PILEIPELLIS a cutis, upper layer composed of thin parallel

hyphae, 3-8 um wide with brown pigment in KOH, walls smooth. Lower layer

hyphae 9-25 um wide, with thin walls. No distinct reaction with Melzer’s

reagent. HYPODERMIUM well developed. OLEIFEROUS HYPHAE infrequent.

New telamonias from California ... 389

Fic. 17. Cortinarius truckeensis

a) Basidiospores (UC 1861353, holotype) b) DBB16054, young basidiomata.

CLAMP CONNECTIONS common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius truckeensis is known only from

its type location where it has fruited abundantly for several consecutive years.

The habitat is composed almost solely of Jeffrey’s Pine (P. jeffreyi). Despite

the extensive field research and many organized forays in the Sierra Nevada,

this species has never been collected outside of the type location. Typically,

C. truckeensis fruits under a very thick conifer duff and rarely breaks above the

surface of the forest floor. Most commonly the location of fruiting is marked by

a hump that may hide a dozen or more clustered basidiomata.

Fic. 18. Cortinarius truckeensis a) DBB33200 b) DBB43920

ar SE ie | lo Ae

) ; on a Mi

By SX ny, N 4 \

i f/

SS — = ee

nsis mature basidiomata DBB43920 a) DBB43920 b) DBB33200

Fie. 19.

Cortinarius truckee

390 ... Bojantchev

C. tuolumnensis JQ906762

C. tuolumnensis JQ906763

C. tuolumnensis JQ906761

29] | C. “saturninus" GQ159813

11! C. "parevernius" HQ604722

Cc. “vernus" GQ159806

of C. sp. FJ039536

5 C cf. "evernius" HQ604723

C. "pulchripes" FJ039537

Cc. "vernus" GQ159805

43 - C. gualalaensis JX501775

2 Wc. gualalaensis JX501776

C. gualalaensis JX501774

“ C. ohlone JQ906757

? =| C. ohlone JQ906760

C. ohlone JQ906758

Uncultured C. sp. FR851994

83) C. decipiens FN428992

= 34 1! C. decipiens FN428996 C. decipiens

' 7} 1 C. decipiens FN429002 _~—*| from Europe

Cc. decipiens FN428994 IB 2005/0010

Cc. "leucopus" HQ604720 Undescribed

C. "leucopus" GQ159824 Telamonia

= : C. "leucopus" GQ159855 from the PNW

C. sp. FJ039538

49 C. "vernus" GQ159840

1 Cc. "leucopus" HQ604721

: C. cf. "vernus" GQ159821 | Undescribed

C. "saturninus" GQ159841 | Telamonia

C. "vernus” FJ039539 from the PNW

84 C. "vernus" GQ159816

4 C. "leucopus" GQ159843

C. cf. "vernus" GQ159825

C. sp. DQ822800

: : C. miwok JQ906753

C. miwok JQ906751

C. "sertipes" FJ039540

C. "hinnuleoarmillatus" DQ499464

C. athabascus JN133295 | outgroup

Fic. 20. Phylogenetic tree inferred by Maximum Likelihood analysis of 38 nrITS Cortinarius

sequences in the /decipiens clade (with C. athabascus as an outgroup). The tree shows all GenBank

sequences that are within 2% BLAST similarity index to C. tuolumnensis, C. miwok, C. gualalaensis

and C. ohlone. The percentage of replicate trees in which the associated taxa clustered together in

the bootstrap test (1000 replicates) are shown above the branches and the branch length values are

shown below the branches. GenBank accession numbers are shown after the binomial. Misapplied

species names on GenBank collections are enclosed in quotes.

New telamonias from California ... 391

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: NEVADA County, North

of Truckee, off Hwy. 89 (39°24'05"N 120°11'19"W), elev. 6200 ft, under P jeffreyi, 15

Jun 2006, DBB00054; 18 Jun 2009, DBB16054; 12 Jun 2010, DBB33200; 9 Jun 2011,

DBB43920 (Genbank nrITS JQ937285);

Discussion — Cortinarius truckeensis is one of the most distinctive species

in subg. Telamonia, and is one of the easiest to identify in the field as the

combination of predominantly white coloration, habitat, and season of fruiting

sets it apart. One of the unusual features of this species is the spore shape

(Fic. 17a) and size that resemble those of the phylogenetically distant species in

subg. Phlegmacium sect. Variecolores Brandrud & Melot.

This species seems somewhat isolated from any of the Cortinarius collections

represented by sequence data in GenBank and even more so from those in

the European public database UNITE. As with C. cisqhale, establishing the

phylogenetic position of C. truckeensis presented difficulties as various packages

and methods produced contradictory results. Neighbor-joining and maximum

parsimony analyses suggested affinity towards either the /alboviolaceus or the

/evernius clades. The Bayesian analysis suggests that this species is somewhat

isolated on its own evolutionary path without known close relatives.

Species in the /decipiens clade

Cortinarius tuolumnensis Bojantchey, sp. nov. Fics 21-24

MycoBAnkK MB 800345

Differs from other Telamonia species by the distinct context coloration, montane vernal

fruiting habit, and spore shape.

Type: USA. California: Tuolumne County, Yosemite National Park, off Hodgdon

Meadow Access Rd. (37°48'28"N 119°51'19"W), elev. 4600 ft, under P ponderosa,

A. concolor, A. magnifica, Ps. menziesii, 14 May 2009, DBB15027 (Holotype UC 1861347;

Genbank nrITS JQ906762).

EryMo_oey: after the Tuolumne river and county in the Central Sierra Nevada.

PiLEus 20-90 mm diam., hemispherical to convex when young, plano-convex

at age, often with a broad rounded umbo; margin involute then straight;

blackish-brown when young and/or moist, lighter reddish-brown to chestnut-

brown to cinnamon-brown at maturity or when dry; the disk and umbo

remain darker, fading at age; bruising purplish-black, hygrophanous in radial

streaks. LAMELLAE moderately crowded to subdistant, 5-12 mm broad, latte

brown at first, turning rusty brown as the spores mature; edges slightly uneven,

conspicuously lighter in color; attachment sinuate; lamellulae abundant in series

of 3-5. Stipe 40-130 mm long, 6-15 mm wide, cylindrical, straight or curved

due to caespitose habit, light brown beneath the universal veil, sometimes with

grayish-blue tinges near the top; with longitudinal cavity. UNIVERSAL VEIL

white, covering the stipe over the entire length, often leaving an annular zone,

392 ... Bojantchev

A 2S —

NOR OS RNa Ver

Fic. 21. Cortinarius tuolumnensis (UC 1861347, holotype).

fibrous at first, sometimes glossy at age. CORTINA white. CONTEXT beige with

conspicuous purple-brown bruising in young material, often completely absent

with age. ExstccaTa dark brown, stipes brown. HasitT often caespitose of up

to a dozen basidiomata, but frequently scattered. OpoR distinctly spicy with

a hint of fruit (especially in young basidiomata). Taste mild. UV LicuT no

fluorescence detected. MACROCHEMICAL REACTIONS 5% KOH negative. SPORE

DEPOSIT rusty brown.

BASIDIOSPORES (8.5—)9.5-11.2(-12.5) x (5.0-)5.7-6.5(-7.0) um (mean 10.2

x 6.1 um), Q = 1.52-1.88, Q. = 1.67 (N = 153, 6 basidiomata, five collections),

amygdaliform to narrow ellipsoid, slightly ovate, strongly verrucose, more

so towards the apex, non-dextrinoid. Basrp1a 34-40 x 8-10 um, 4-spored

(frequently 2-spored), cylindro-clavate, clamped. LAMELLA EDGE sparsely

; - a ss ——— 7 La, oy

g oe =, - Zs

w “a = mei “ we Pe SP

4G » Nd — £ a a eee’

Fic. 22. Cortinarius tuolumnensis a) Basidiospores (UC 1861347, holotype) b) DBB00114

New telamonias from California ... 393

in, ’ 4 ‘ / -

rd t

+ ; ™ 5

SS : &

4 i 2 a yi : : “s

ae : _~ . a

Fic. 23. Cortinarius tuolumnensis a) DBB09856 b) DBB43454 c) DBB00112

Fic. 24. Cortinarius tuolumnensis DBB00112

a) context discoloration in young basidiomata b) cortina c) clustered/caespitose habit

fertile. CysTip1A not observed. LAMELLAR TRAMA regular, composed of more

or less parallel, cylindrical hyphae, 2-10 um wide. PILEIPELLIS a cutis, upper

layer composed of thin parallel hyphae, 2-5 um wide with red-brown pigment

in KOH, walls smooth to finely zebra-stripe encrusted. Lower layer hyphae

6-22 um wide, with thin walls. No distinct reaction with Melzer’s reagent.

HYPODERMIUM well developed. OLEIFEROUS HYPHAE none observed. CLAMP

CONNECTIONS common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius tuolumnensis fruits abundantly

in late spring and early summer in the mountains of California and in the

broader Pacific Northwest. There are two matching vernally fruiting collections

represented in GenBank (HQ604722, HQ604723) from Victoria, British

Columbia. It appears that the preferred habitat of this species in California is

the wet montane meadows (soon after snow melt), surrounded by conifers,

mostly pines like Ponderosa pine (P. ponderosa).

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: EL DoRADO COUNTY,

El Dorado National Forest, 0.1 mi. SE of Icehouse Road (38°51'24"N 120°22'32”W),

elev. 5100 ft, under P ponderosa, A. concolor, A. magnifica, Ps. menziesii, 14 May 2011,

DBB43454 (Genbank nrITS JQ906763); TUoLUMNE CouNTY, Stanislaus National

Forest, off Evergreen Rd. (37°49'55"N 119°51'04"W), elev. 4500 ft, under P ponderosa,

A. concolor, Ps. menziesii, 13 May 2007, DBB00112 (Genbank nrITS JQ906761) and

DBBO00114; 22 May 2008, DBB09856.

Discussion — When first observed in the field, C. tuolumnensis was easily

placed in the /decipiens clade (Fic. 20) due to the purplish discoloration on

394 ... Bojantchev

the stipe context and distinct spicy odor. That position was confirmed later by

the nrITS generated phylogeny. Cortinarius tuolumnensis stands out from other

clade representatives (such as C. miwok) by its caespitose habit and typically

longer stems.

Fic. 25. Cortinarius gualalaensis (UC 1998883, holotype).

Cortinarius gualalaensis Bojantchey, sp. nov. FIGS:25=27

MycoBank MB 801217

Differs from C. tuolumnensis by the autumnal fruiting habit and slightly smaller spores.

Type: USA. California: Sonoma County, Jenner, Salt Point Lodge, off Hwy 1. (38°33'24"N

123°18'10"W), elev. 120 ft, under P muricata, 21 November 2010, DBB39900 (Holotype

UC 1998883; Genbank nrITS JX501775).

EryMo_oey: after the coastal city of Gualala in northern California — the name means

“where the river meets the sea” in the language of the Pomo tribe.

PiLEus 20-70 mm diam., hemispherical to convex when young, plano-convex

at age, often with a broad rounded umbo; margin straight then flanging; dark-

brown when young and/or moist, lighter reddish-brown to chestnut-brown

to dull cinnamon-brown at maturity or when dry; the disk and umbo remain

darker, fading at age; bruising purplish-black, strongly hygrophanous with

fading radial streaks. LAMELLAE moderately crowded to subdistant, 4-15 mm

broad, latte brown at first, turning rusty brown as the spores mature; edges

slightly uneven, conspicuously lighter in color; attachment sinuate; lamellulae

abundant in series of 3-5. Stipe 30-120 mm long, 5-14 mm wide, cylindrical,

straight or curved due to caespitose habit, light brown beneath the universal veil

remnants, sometimes with grayish-blue tinges near the top; with longitudinal

cavity. UNIVERSAL VEIL white, covering the entire stipe, but more prominent in

the lower portion, often leaving a faint annular zone, sometimes glossy in age.

New telamonias from California ... 395

CorTINA white. CONTEXT beige with conspicuous purple-brown bruising in

young material, often completely absent at age. Exstccata dark brown, stipes

lighter in color. Hasit mostly caespitose of up to a dozen basidiomata, but

frequently scattered. ODoR somewhat spicy (especially in young basidiomata).

TasTE mild. UV LicuT no fluorescence detected. MACROCHEMICAL REACTIONS

5% KOH negative. SPORE DEPOSIT rusty brown.

BASIDIOSPORES (8.5—)9.0-11.0(-11.5) x (5.8-)6.0-6.8(-7.3) um (mean

9.9 x 6.5 um), Q = 1.42-1.75, Q. = 1.54 (N = 143, 5 basidiomata, three

collections), amygdaliform to ellipsoid, distinctly verrucose, more so towards

the apex, non-dextrinoid. Basip1a 32-42 x 7-11 um, 4-spored (frequently

2-spored), cylindro-clavate, clamped. LAMELLA EDGE sparsely fertile. CysTIDIA

not observed. LAMELLAR TRAMA regular, composed of more or less parallel,

cylindrical hyphae, 3-12 um wide. PILEIPELLIs a cutis, upper layer composed

of thin parallel hyphae, 2-6 um wide with red-brown pigment in KOH, walls

smooth to finely zebra-stripe encrusted. Lower layer hyphae 5-25 um wide,

with thin walls. No distinct reaction with Melzer’s reagent. HyPODERMIUM

well developed. OLEIFEROUS HYPHAE none observed. CLAMP CONNECTIONS

common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius gualalaensis fruits in the fall and

is known only from the coastal areas of California and Oregon. In California,

C. gualalaensis was collected in dense stands of Bishop pine (P. muricata) while

in the coastal sand dunes of Oregon this species fruited under Shore pine

(P. contorta var. contorta).

Fic. a7. Gunna ee RE x DBB43121 b) DBB43121 5 DBB23436

396 ... Bojantchev

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: SONOMA COUNTY, Salt

Point State Park, 0.5 mi. North of Hwy 1 (38°33'48"N 123°18'27"W), elev. 300 ft, under

P. muricata, 27 Nov 2011, DBB43121 (Genbank nrITS JX501776); OREGON: LANE

Counry, Jessie M. Honeyman Memorial State Park (43°55'48"N 124°06'26"W), elev. 80

ft, under P. contorta var. contorta, 9 Nov 2009, DBB23436 (Genbank nrITS JX501774);

Discussion — Cortinarius gualalaensis is very closely related to C. tuolumnensis

and appears very similar in stature, colors, and the clustered habit. The major

differentiating character is the time of fruiting — C. tuolumnensis fruits in

the spring and C. gualalaensis in the autumn. Microscopically, C. gualalaensis

spores are a bit shorter and wider, with a smaller Q

—--

Fic. 28. Cortinarius ohlone DBB41040.

Cortinarius ohlone Bojantchey, sp. nov. FIGs 28-31

MycoBAnk MB 800348

Differs from other Telamonia by the unique coloration and association to oaks in

California and adjacent regions.

Type: USA. California: Contra Costa County, Kennedy Grove (37°56'47"N

122°15'58"W), elev. 100 ft, under Quercus agrifolia, 23 Jan 2011, DBB42193 (Holotype

UC 1861349; Genbank nrITS JQ906757).

Erymo toey: In honor of the Ohlone Native American people — this species can be

found in the oak woods of northern California that they inhabited.

PILEus 30-80 mm diam., convex to plano-convex, rarely with a broad rounded

umbo; margin more or less involute into maturity; dark brown to grey-

brown. When young with dark bluish-gray cast, turning cinnamon brown at

New telamonias from California ... 397

Fic. 29. Cortinarius ohlone

a) Basidiospores (UC 1861349, holotype)

b) DBB05460, young basidiomata with dark bluish-gray cast on the pileus

maturity; surface fibrillose, smooth to shiny, inconspicuously hygrophanous

with radial streaks. LAMELLAE moderately crowded to subdistant, 5-18 mm

broad, dark clay brown, without bluish tinges, turning rusty brown as the

spores mature; edges slightly uneven, much paler than the sides; attachment

sinuate; lamellulae abundant in series of 3-7. Stipe 30-120 mm long, 12-24

mm wide, cylindrical, white to pale beige. UNIVERSAL VEIL white, partially

covering the lower stipe, leaving an evanescent annular zone. CoRTINA white.

CONTEXT beige to white, bruising purplish-brown slightly, stipe frequently

hollow at maturity. Exstccata dark brown. Hasir scattered to subcaespitose.

Opor distinctly spicy somewhat like cedar wood. Taste mild. UV LicuT no

fluorescence detected. MACROCHEMICAL REACTIONS 5% KOH negative. SPORE

DEPOSIT rusty brown.

BASIDIOSPORES (6.8—)7.2-8.2(-9.0) x (4.8-)5.0-5.6(-6.0) um (mean 7.8 x

5.2 um), Q = 1.42-1.60, Q. = 1.50 (N = 134, 5 basidiomata, four collections),

amygdaliform to ellipsoid, ovoid, or broadly ellipsoid, finely verrucose, non-

dextrinoid. BAsip1A 26-40 x 6-8 um, 4-spored (rarely 2-spored), cylindro-

clavate, clamped. LAMELLA EDGE sterile. CysTipIA not observed. LAMELLAR

TRAMA regular, composed of more or less parallel, cylindrical hyphae, 3-7 um

wide. PILEIPELLIS a cutis; epicutis composed of thin parallel hyphae, 2-7 um

wide with brown pigment in KOH; walls smooth to very finely zebra-stripe

encrusted. Lower layer hyphae 8-22 um wide, with thin walls. No distinct

reaction with Melzer’s reagent. HYPODERMIUM well developed. OLEIFEROUS

HYPHAE infrequent. CLAMP CONNECTIONS common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius ohlone is fairly common under

evergreen oaks throughout California and its distribution appears limited to

the state. The primary mycorrhizal associates are coast live oak (Q. agrifolia)

and interior live oak (Q. wislizenii).

398 ... Bojantchev

Fic. 30. Cortinarius ohlone

a) DBB41040, an annular zone on the stipe b) DBB42193 (UC 1861349, holotype)

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: CONTRA COSTA COUNTY,

San Pablo Dam (37°54'16"N 122°12'55"W), elev. 150 ft, under Q. agrifolia, 2 Jan 2008,

DBB05460 (Genbank nrITS JQ906760); SANTA Cruz County, Santa Cruz, UCSC

Campus (36°59'49"N 122°04'02"W), elev. 600 ft, under Q. agrifolia, 22 Dec 2010,

DBB41040 (Genbank nrITS JQ906758); RIVERSIDE CouNTy, Cleveland National

Forest, off Forest Road 6805 (33°38'18"N 117°25'21"W), elev. 1300 ft, under Q. agrifolia,

Q. wislizenii, 12 Feb 2011, DBB42536; YoLo County, Putah Creek State Wildlife

Reserve, Franklin Canyon, (38°30'23"N 122°05'51"W), elev. 1000 ft, under Q. agrifolia,

Q. wislizenii, 6 Jan 2011, R. M. Davis RMD101205 (Genbank nrITS JQ906759);

Discuss1on — Cortinarius ohlone is the most common of several small to

medium-sized brownish Telamonia species in the oak woods of northern

California. The diversity of this group seems to increase in the warmer oak

habitats of southern California. A key diagnostic feature for differentiating

C. ohlone from other similar cortinarii in the same habitat is the slight to

pronounced spicy-fragrant odor (reminiscent of cedar wood), which is typical

for the members of the /decipiens clade.

phage.

ip SE

Fic. 31. Cortinarius ohlone a) DBB42536, older basidiomata b) RMD101205

New telamonias from California ... 399

Fic. 32. Cortinarius miwok (UC 1861351, holotype).

Cortinarius miwok Bojantchey, sp. nov. FIGS 32-34

MycoBANK MB 800347

Differs from C. ohlone by its coniferous association and montane vernal fruiting habit.

Type: USA. California: El Dorado County, El Dorado National Forest, 0.1 mi. SE of

Icehouse Road (38°51'24"N 120°22'32"W), elev. 5100 ft, under P ponderosa, A. concolor,

A. magnifica, Ps. menziesii, 5 Jun 2011, DBB43810 (Holotype UC 1861351; Genbank

nrITS JQ906753).

Erymotoey: In honor of the Miwok Native American people — this species can

frequently be found in the Sierra Nevada areas that they inhabited.

PILEus 30-70 mm diam., convex to plano-convex, often with a broad flattened

umbo; margin involute then straight to flanging; dark to light grey-brown,

turning brown at maturity, the extreme margin whitish due to persistent veil

remnants; surface fibrillose, smooth to shiny, inconspicuously hygrophanous.

LAMELLAE moderately crowded to subdistant, 6-20 mm broad, clay brown, no

bluish tinges, turning rusty brown as the spores mature; edges slightly uneven,

pale; attachment sinuate; lamellulae abundant in series of 3-7. StrPE 30-80 mm

long, 15-30 mm wide, cylindrical to subclavate, white to light brown beneath

the universal veil remnants, bruising purplish-brown, rarely with bluish tinges

400 ... Bojantchev

Fic. 33. Cortinarius miwok a) Basidiospores (UC 1861351, holotype) b) DBB43988

near the top on young basidiomata. UNIVERSAL VEIL white, partially covering

the stipe in the lower half, leaving an evanescent annular zone. CorTINA white.

CONTEXT beige to white, bruising purplish-brown slightly, stipe frequently

hollow at maturity. Exstccata dark brown. Hasir scattered to subcaespitose.

Opor slightly spicy, or moldy if surrounded by wet soil. Taste mild. UV LIGHT

no fluorescence detected. MACROCHEMICAL REACTIONS 5% KOH negative.

SPORE DEPOSIT rusty brown.

BASIDIOSPORES (6.8-)7.2-9.0(-9.3) x (3.8-)4.0-5.0(-5.3) um (mean 8.2 x

4.5 um), Q = 1.60-2.14, Q. = 1.82 (N = 177, 7 basidiomata, four collections),

amygdaliform to cylindro-amygdaliform, finely verrucose, non-dextrinoid.

BASIDIA 28-38 x 6-9 um, 4-spored (occasionally 2-spored), cylindro-clavate,

clamped. LAMELLA EDGE sparsely fertile. Cystip1A not observed. LAMELLAR

TRAMA regular, composed of more or less parallel, cylindrical hyphae, 2-8 um

wide. PILEIPELLIS a cutis; upper layer composed of thin parallel hyphae, 2-6

um wide with olivaceous-brown pigment in KOH; walls smooth to finely zebra-

stripe encrusted. Lower layer hyphae 7-20 um wide, with thin walls. No distinct

reaction with Melzer’s reagent. HYPODERMIUM well developed. OLEIFEROUS

HYPHAE infrequent. CLAMP CONNECTIONS common on all septa.

HABITAT AND DISTRIBUTION — Cortinarius miwok fruits in late spring and

early summer in the California mountains during, or soon after, snow melt.

It is one of the less common species amongst the vernal fruiting cortinarii.

Apparently it occurs in the Pacific Northwest, where it may be uncommon,

as there is only one matching sequence in GenBank (FJ039540; also vernally

fruiting but incorrectly designated as C. sertipes from a collection made at

Observatory Hill, Vancouver Island, British Columbia]. In California C. miwok

occurs in the foothills and higher elevations of the Sierra—Cascade range,

where it can frequently be observed near melting snow. The surrounding

conifer habitat consists primarily of ponderosa pine (P ponderosa), white fir

(A. concolor), red fir (A. magnifica), lodgepole pine (P. contorta var. latifolia)

and Douglas-fir (Ps. menziesii).

New telamonias from California ... 401

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: EL Dorapo County, El

Dorado National Forest, 0.1 mi. SE of Icehouse Road (38°51'24"N 120°22'32"W), elev.

5100 ft, under P. ponderosa, A. concolor, A. magnifica, Ps. menziesii, 5 Jun 2011, DBB43976

(Genbank nrITS JQ906754); TUOLUMNE County, Yosemite National Park, off Tioga

Pass Rd. (37°50'12"N 119°40'48"W), elev. 7300 ft, under P. ponderosa, A. concolor, A.

magnifica, Ps. menziesii, 20 Jun 2009, DBB16312 (Genbank nrITS JQ906751); StERRA

County, SF State Sierra Nevada Field campus (39°37'23"N 120°34'48"W), elev. 5700

ft, under P ponderosa, A. concolor, A. magnifica, Ps. menziesii, 9 Jun 2011, DBB43988

(Genbank nrITS JQ906755); Chapman Creek Campground (39°37'54"N 120°32'37"W),

elev. 5900 ft, under P ponderosa, A. concolor, A. magnifica, Ps. menziesii, 22 Jun 2010,

DBB33741 (Genbank nrITS JQ906752);

Discussion — Cortinarius miwok stands out from the other vernal cortinarii

with similar stature in the same surroundings by the more bluish-grey cap

and slight purplish discoloration of the context. This species also exhibits the

slightly spicy odor typical of the /decipiens clade. Its closest relative in the same

habitat is C. tuolumnensis, which can be distinguished by its predominantly

caespitose habit and longer stems. Microscopically, C. miwok can be separated

from C. tuolumnensis by the smaller spores.

Fic. 34. Cortinarius miwok a) DBB16312 b) DBB43976

Acknowledgements

Gratitude goes to Dr. Else C. Vellinga and Nhu Nguyen for their presubmission

reviews and comments. Prof. R. Michael Davis contributed collections of C. ohlone and

most molecular data. Dr. Shaun Pennycook was very helpful in answering nomenclatural

questions during the preparation of this manuscript. Dr. John David helped with

inquiries on Greek language.

Literature cited

Bergemann, SE, Garbelotto M. 2006. High diversity of fungi recovered from the roots of

mature tanoak (Lithocarpus densiflorus) in northern California. Can J Bot 84: 1380-1394.

http://dx.doi.org/10.1139/B06-097

Bojantchev D, Davis RM. 2011. Cortinarius xanthodryophilus sp. nov. — a common Phlegmacium

under oaks in California. Mycotaxon 116: 317-328. http://dx.doi.org/10.5248/116.317

402 ... Bojantchev

Huelsenbeck JP, Ronquist F. 2001. MRBAYES: Bayesian inference of phylogenetic trees.

Bioinformatics 17: 754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

Katoh K, Misawa K, Kuma KI, Miyata T. 2002. MAFFT: a novel method for rapid multiple

sequence alignment based on fast Fourier transform. Nucleic Acids Res 30: 3059-3066.

http://dx.doi.org/10.1093/nar/gkf436

Posada D, Crandall KA. 1998. Modeltest: testing the model of DNA substitution. Bioinformatics

14: 817-818.

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

Trudell $, Ammirati JE. 2009. Mushrooms of the Pacific Northwest. Timber Press. Portland.

MY COTAXON

http://dx.doi.org/10.5248/123.403

Volume 123, pp. 403-408 January-March 2013

Acaulospora endographis (Glomeromycetes),

a new fungus with a complex spore wall

BRUNO T. GOTO!’", CAMILLA M.R. PEREIRA”, CAMILA P. NOBRE?,

NATALIA P. ZATORRE?, FERNANDA COVACEVICH4,

RICARDO L.L. BERBARA?& LEONOR C. MAIA?

"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

*Departamento de Ciéncias do Solo, Universidade Federal Rural do Rio de Janeiro,

Seropédica, Rio de Janeiro, Brazil

‘CONICET, Laboratorio de Microbiologia de Suelo Estacién Experimental Agropecuaria INTA,

Balcarce, Argentina

‘CORRESPONDENCE TO: brunogoto@hotmail.com

ABSTRACT — A new acaulosporoid fungal species characterized by glomerospores with a

complex ten-layered wall structure and uniquely ornamented fourth layer (outer wall layer

OWL4) found in impacted areas of the Amazon Forest and cultivated areas of the Atlantic

Forest in Brazil is described as Acaulospora endographis.

Key worps — Glomeromycota, Diversisporales, Acaulosporaceae, morphology

Introduction

Several arbuscular mycorrhizal (AM) fungal species with a generally

scutellosporoid spore development have been described from tropical areas

(Cuenca & Herrera-Peraza 2008; Silva et al. 2008; Oehl et al. 2008, Goto et

al. 2009, 2010, 2011, 2012; Tchabi et al. 2009). In contrast, only a few species

with an acaulosporoid development have been described during the last 10

years from the tropics [Ambispora brasiliensis (Goto et al. 2008), Acaulospora

punctata (Oehl et al. 201la), Ac. sieverdingii (Oehl et al. 2011b), and

Ac. soloidea (Vaingankar & Rodrigues 2011)], with the first two described from

South America.

An undescribed ornamented acaulosporoid fungus frequently found during

diversity studies of arbuscular mycorrhizal fungi in Brazilian biomes is here

described and illustrated as Acaulospora endographis.

A404 ... Goto & al.

Material & methods

Study area and sites

Soilsamples were collected in the Seringal Cachoeira (7°07'-11°08'S 66°30'-74°00'W),

located in the municipality of Xapuri, Acre State, north Brazil. Soils were characterized

as red-yellow Argissols (Ultisols). The climate in the region is Am’ Equatorial (Képpen

classification): hot and humid with 24.5-32°C average temperatures with two distinct

seasons: dry (300-375 mm precipitation) and rainy (1200-1360 mm). The study areas

consisted of an open rainforest dominated by palms, a 20 year-old secondary forest, and

a 30 year-old regenerated pasture with Brachiaria decumbens Stapf, Bertholletia excelsa

Bonpl., and Zea mays L. The soil contained 4.77-3.44 mg.dm”’ organic phosphorus,

0.31-0.21 cmol_.kg” potassium, 6.04-3.78 cmol..kg™ calcium, 2.88-2.62 cmol_.kg”

magnesium, 2.62-2.44 cmol_.kg' H+Al, and 41-26 g.kg™ organic matter with a pH

of 5.60-5.35. Other soil samples, also characterized as red-yellow Argissols (Ultisols),

were collected from the Experimental Station of Itapirema (Instituto Agronémico de

Pernambuco) in the municipality of Goiana (7°38'20"S 34°57'10"W), Pernambuco

State, northeast Brazil, where the climate is Ams’ (K6ppen classification): tropical rainy

with 21.1-31.3°C average annual temperature and 2000 mm average annual rainfall.

The soil was sampled during the rainy season (June 2011) in an area cultivated for nine

months with cassava (Manihot esculenta Crantz); this area was fertilized with NPK and

the pH corrected with calcitic limestone at planting time. The soil composition was 3.8

mg.dm~ organic phosphorus, 0.006 cmol_.kg’ potassium, 0.005 cmol..kg™ calcium,

0.003 cmol_.kg™' magnesium, and 31.0 g.kg"' organic matter with a pH of 5.3.

Morphological analyses

Acaulosporoid spores were extracted from field soils and from trap cultures by wet

sieving and sucrose centrifugation (Sieverding 1991). The spores were isolated in a

stereomicroscope and mounted in polyvinyl alcohol-lacto-glycerin (PVLG) and PVLG

+ Melzer’s reagent (Brundrett et al. 1994). Slide mounts were dried (3-5 days at 60°C)

to clarify oil drops and observed with a Zeiss Axioskop compound microscope. Images of

glomerospores were taken using a Canon digital camera.

General terminology follows Goto et al. (2008) and Oehl et al. (2011a), and spore

terminology follows Goto & Maia (2006). Holotype and isotype specimens are conserved

at the Herbarium of Departamento de Micologia, Universidade Federal de Pernambuco

(URM), and other specimens at the Herbarium of Departamento de Botanica, Ecologia

e Zoologia, Universidade Federal do Rio Grande do Norte (UFRN).

Arbuscular mycorrhizal cultures

Cultures of the new species were tentatively established in three institutions, two

in Brazil and one in Argentina. For culturing the native AM fungal communities

400 g of field soil were placed into 500 ml pots at the Departamento de Ciéncia do

Solo, Universidade Federal Rural do Rio de Janeiro (Seropédica, Rio de Janeiro, BR).

Brachiaria decumbens was used as bait plant and the culture pots were maintained in the

greenhouse. At the Departamento de Micologia, Universidade Federal de Pernambuco

(Recife, Pernambuco, BR), cultures were set up using 800 g of field soil collected from

Pernambuco into 1000 mL pots. Maize (Zea mays), sunflower (Helianthus annuus L.),

and sorghum (Sorghum bicolor (L.) Moench) were used as bait plants and the pots were

kept in the greenhouse. At the Microbiology Laboratory, EEA INTA Balcarce, Argentina,

Acaulospora endographis sp. nov. (Brazil) ... 405

for single species cultures multiple glomerospores were separated from field samples

and used as infective propagules using 800 g of field soil collected from Acre into 1000

mL pots, with Secale cereale L. as host plant. The cultures were maintained (one year)

in the greenhouse. So far, all trials failed to establish a successful symbiosis of the new

AMF species either in trap cultures or in the single species cultures.

Taxonomy

Acaulospora endographis B.T. Goto, sp. nov. FIGs 1-9

MycoBank MB 564888

Differs from other Acaulospora species by its complex ten-layered spore wall structure

with spinose ornamentation in the fourth outer wall layer.

Type: Brazil, Pernambuco State, Goiana, Experimental Station of Itapirema (Instituto

Agronémico de Pernambuco), from red-yellow Argissols under Cassava cultivation

(Holotype, URM83586; isotypes, URM83586a, URM83586b).

EryMmo ocy: endographis refers to ornamentation similar to pencil imprints in an inner

layer of the spore wall.

SPOROCARPS unknown. Acaulosporoid spores form singly in soils upon a short

pedicel arising laterally on the tapering hyphal neck of a sporiferous saccule.

The glomerospore forms three spore walls — outer wall (OW), middle wall

(MW), and inner wall (IW) — with ten layers total.

SPORIFEROUS SACCULE hyaline to white, globose to subglobose and formed

terminally on a hypha, generally absent and detached from the mature spore

while the pedicel or collar often persists on the glomerospore. ‘The saccule wall

is generally bi-layered with a rapidly degrading evanescent hyaline to light

yellow outer layer (0.5-1.5(-2.0) um) and semi-persistent hyaline to subhyaline

inner layer 0.8-1.5(-2.2) um.

GLOMEROSPORES form singly on a short hyphal pedicel. The spores are

globose (74-128 um diam.) to subglobose to oval (68-110 x 69-100(-180)),

yellow brown to orange brown in water (generally dark orange to dark orange

brown in PVLG-based mountants). The spore wall is complex with ten layers

distributed in the three walls (OW, MW, IW).

OUTER WALL consists of five layers, with the outermost layer (OWL1)

evanescent, 0.5-1.5 um thick, hyaline to sub-hyaline, and often difficult to

detect, as it tightly adheres to OWL2. OWL2 is laminated, 2.2-5.4 um thick,

and yellow to orange brown. OWL3 is thin (< 0.5 um), hyaline, and tightly

attached to OWL2, making it difficult to detect. In older spores, the outer wall

layer might degrade completely, but usually some fragments remain on the

OWL12 surface. A fourth layer (OWL4) is 2.5-4.5 um thick, yellow to yellow

brown and ornamented with spines, generally 0.5-1.0(-1.5) um wide and

1.5-2.0(-2.5) um high. OWLS tightly adheres to OWL4 and is flexible, thin

(< 1.0 um), hyaline and difficult to see. No Melzer’s reaction is evident in any

outer wall layer.

406 ... Goto & al.

Fics 1-9. Acaulospora endographis. 1. Young spores with hyaline to sub-hyaline sporiferous saccule

attached. 2. Intact glomerospores with distinctive outer wall (ow) and inner wall (1w). 3. Cicatrix

globose to subglobose commonly observed in healthy spores; note the ornamentation (Orn) in a

planar view. 4. Insertion point of hypha in spore with a short pedicel (Pdcel). In this case the pedicel

wall is continuous with owL1. 5. Ornamentation in a planar view. 6. Spore wall with distinctive

outer wall (OW) and inner wall (IW). 7-9. Spore wall structure with OW, middle wall (MW) and

IW. Note the outer wall with five layers (ow11,2,3,4,5) and ornamentation in OwL4. Inner wall

presents a beaded layer (IWL1) and a strong Melzer’s reaction.

MIDDLE WALL is bi-layered and 2.0-4.0 um thick. M“WLI is semi-rigid to

flexible, 1.0-1.5 um thick, hyaline, and tightly adheres to MWL2, also semi-

rigid to flexible, 1.0-2.0 um, and hyaline. The bi-layered wall component

appears laminated in some spores. No Melzer’s reaction is evident in either

middle wall layer.

INNER WALL is three-layered, with all three layers hyaline. The outer layer

IWLI is 0.5-1.0 um and easy to detect, ornamented with granular (beaded)

projections, and tightly adherent to IWL2 which is amorphous and generally

1.5-3.5 um thick. [WL3 adheres to IWL2 and is thin (0.5-1.0 um thick) and

difficult to see. The Melzer’s reaction is dark red purple in IWL2.

PEDICEL short, 2-4 um long, and 4-6(-8) um wide at the glomerospore

base. The wall layer of the pedicel is 3-4 um thick and sub-hyaline to light

yellow. The pedicel wall layer is always continuous with the outer spore wall

Acaulospora endographis sp. nov. (Brazil) ... 407

layer (OWL1), and the OWL2 sometimes forms a collar on the glomerospore

(2.5-4 um diam.)..

GERMINATION was not observed. The germination structure is believed to

resemble those observed in other Acaulospora species (see Spain 1992).

ARBUSCULAR MYCORRHIZA FORMATION is unknown.

DISTRIBUTION — So far, known only from Brazil in soils from disturbed

areas of the Amazon Forest (North Brazil) and in agricultural systems from the

Atlantic rainforest (Northeast Brazil).

ADDITIONAL MATERIAL EXAMINED: BRAZIL, ACRE STATE, Xapuri, Seringal Cachoeira,

from red-yellow Argissols (UFRN1742 [permanent slides with PVLG]).

Discussion

The low number of glomerospores (2-6) in 100g of the field soil samples

from both sites explains the difficulty in obtaining trap and pure cultures of the

new fungus during this study.

Its complex distinct spore wall and ornamentation easily separate Acaulospora

endographis from other acaulosporoid species. A similar ornamentation is

found only in Entrophospora infrequens (I.R. Hall) R.N. Ames & R.W. Schneid.,

but the entrophosporoid spore development and two-walled structure easily

distinguish E. infrequens from A. endographis. Furthermore, the evanescent or

semi-persistent layers covering the ornamentation in E. infrequens (Sieverding

& Oehl 2006, Oehl et al. 2011c) is quite distinct from the permanent rigid

laminated layer covering the ornamentation in A. endographis.

The complex 10-layered spore wall structure with one spiny ornamented

layer (OWL4) easily distinguishes A. endographis from all other ornamented

Acaulospora species.

Acknowledgements

The authors especially acknowledge Dr. Fabien Hountondji (University of Parakou,

Benin) and Dr. Eduardo Furrazola (Instituto de Ecologia e Sistematica, IES-CITMA,

de la Habana, Cuba) 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. This work was supported by:

Protax (Programa Capacitagao em Taxonomia), and INCT Herbario Virtual da Flora

e dos Fungos, both from the Conselho Nacional de Desenvolvimento Cientifico e

Tecnoldgico (CNPq) that provided research grants to Ricardo L.L. Berbara and Leonor

C. Maia; aid of a grant from the Inter-American Institute for Global Change Research

(IAI) CRN 2014 which is supported by the US National Science Foundation (Grant

GEO-04523250).

Literature cited

Brundrett M, Melville L, Peterson L. 1994. Practical methods in mycorrhizal research. University of

Guelph, Mycologue Publications, Guelph, Ontario.

A408 ... Goto & al.

Cuenca G, Herrera-Peraza R. 2008. Scutellospora striata sp. nov., a newly described glomeromycotan

fungus from La Gran Sabana, Venezuela. Mycotaxon 105: 79-87.

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

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

Goto BT, Maia LC, Oehl E. 2008. Ambispora brasiliensis, a new ornamented species in the arbuscular

mycorrhiza-forming Glomeromycetes. Mycotaxon 105: 11-18.

Goto BT, Silva GA, Maia LC, Oehl F. 2009. Racocetra intraornata, a new species in

the Glomeromycetes with a unique spore wall structure. Mycotaxon 109: 483-491.

http://dx.doi.org/10.5248/109.483

Goto BT, Silva GA, Maia LC, Oehl E 2010. Dentiscutata colliculosa, a new species in the

Glomeromycetes from Northeastern Brazil with colliculate spore ornamentation. Nova Hedwigia

90: 383-393. http://dx.doi.org/10.1127/0029-5035/2010/0090-0383

Goto BT, Silva GA, Maia LC, Souza RG, Coyne D, Tchabi A, Lawouin L, Hountondji F Oehl F. 2011.

Racocetra tropicana, a new species in the Glomeromycetes from tropical areas. Nova Hedwigia

92: 69-82. http://dx.doi.org/10.1127/0029-5035/2011/0092-0069

Goto BT, Silva GA, Assis DMA, Silva DKA, Souza RG, Ferreira ACA, Jobim K, Mello CMA,

Evangelista-Vieira HE, Maia LC, Oehl FE 2012. Intraornatosporaceae (Gigasporales),

a new family with two new genera and two new species. Mycotaxon 119: 117-132.

http://dx.doi.org/10.5248/119.117

Oehl F, de Souza FA, Sieverding E. 2008. Revision of Scutellospora and description of five new genera

and three new families in the arbuscular mycorrhiza forming Glomeromycetes. Mycotaxon 106:

311-360.

Oehl F, Silva GA, Palenzuela J, Sanchez-Castro I, Castillo C, Sieverding E. 201la. Acaulospora

punctata, a new fungal species in the Glomeromycetes from mountainous altitudes of the Swiss

Alps and Chilean Andes. Nova Hedwigia 93:353-362.

http://dx.doi.org/10.1127/0029-5035/2011/0093-0353

Oehl F, Sykorova Z, Blaszkowski J, Sanchez-Castro I, Coyne D, Tchabi A, Lawouin L, Hountondji

FCC, Silva GA. 2011b. Acaulospora sieverdingii, an ecologically diverse new fungus in the

Glomeromycota, described from lowland temperate Europe and tropical West Africa. J. Appl.

Bot. Food Qual. 84: 47-53.

Oehl F, Silva GA, Sanchez-Castro I, Goto BT, Maia LC, Evangelista- Vieira HE, Barea JM, Sieverding

E, Palenzuela J. 2011c. Revision of Glomeromycetes with entrophosporoid and glomoid spore

formation with three new genera. Mycotaxon 117: 297-316. http://dx.doi.org/10.5248/117.297

Sieverding E. 1991. Vesicular-arbuscular mycorrhiza management in tropical agrosystems.

Deutsche Gesellschaft fiir Technische Zusammenarbeit (GTZ) GmbH, Eschborn, Germany.

Sieverding E, Oehl F. 2006. Revision of Entrophospora and description of Kuklospora and Intraspora,

two new genera in the arbuscular mycorrhizal Glomeromycota. J. Appl. Bot. Food Qual. 80:

69-81.

Silva DKA, Freitas NO, Cuenca G, Maia LC, Oehl FE. 2008. Scutellospora pernambucana, a new

fungal species in the Glomeromycetes with a diagnostic germination orb. Mycotaxon 106:

361-370.

Spain JL. 1992. Patency of shields in water mounted spores of four species in Acaulosporaceae

(Glomales). Mycotaxon 43: 331-339.

Tchabi A, Hountoudji F, Laouwin L, Coyne D, Oehl F. 2009. Racocetra beninensis, from sub-saharan

savannas: a new species in the Glomeromycetes with ornamented spores. Mycotaxon 110:

199-209. http://dx.doi.org/10.5248/110.199

Vaingankar JD, Rodrigues BE. 2011. Acaulospora soloidea, a new arbuscular mycorrhizal

fungus from rhizosphere soils of Murraya paniculata. Mycotaxon 115: 323-326.

http://dx.doi.org/10.5248/115.323

MYCOTAXON

http://dx.doi.org/10.5248/123.409

Volume 123, pp. 409-417 January-March 2013

Basidiospore ultrastructure of some Dacrymycetales from Mexico

SIGFRIDO SIERRA? , JOAQUIN CIFUENTES””,

OLGA M. ECHEVERRIA-MARTINEZ‘* & SANDRA CASTRO-SANTIUSTE!

‘Lab. Taxonomia de Hongos Tremeloides (Heterobasidiomycetes)

’Lab. Biodiversidad y Biogeografia Ecologica de Hongos

°Secc. de Micologia Herbario FCME

‘Lab. de Microscopia Electronica. Fac. de Ciencias,

UNAM. A.P. 70 -181, Coyoacan, C.P. 04510 México, D.F., México.

* CORRESPONDENCE TO: sigfridosg@ciencias.unam.mx

AsstTract — An ultrastructural study was made of basidiospores from nine species of

Dacrymycetales: Calocera macrospora, C. viscosa, Dacrymyces dictyosporus, D. chrysospermus,

Dacryopinax elegans, Da. spathularia, Da. lowyi, Guepiniopsis alpina, and G. buccina. Spore

walls and septa analyzed with transmission electron microscopy revealed the absence or

presence of discrete lateral wall layers as well as septal pores that appear diagnostic at the

species level.

Key worps —jelly fungi, morphology, systematics, taxonomic novelty

Introduction

Septum ultrastructure has been shown to serve as a highly significant

phylogenetic marker of the order Dacrymycetales (Grand & Moore 1971, Khan

& Talbot 1976, Moore 1978, 1994; Khan & Kimbrough 1982, Berbee & Wells

1988, 1989; Boehm & McLaughlin 1989, Lu & McLaughlin 1991, Wells 1994).

Even when teleomorphic characters are lacking, the septal pore ultrastructure

together with some other character (e.g. pigmentation) facilitates placement of

certain taxa within the order (Kirschner & Yang 2005). It is, however, not clear

whether there are other ultrastructural characters that permit differentiation

at the specific, generic, or family level within the order. Here we describe and

compare the ultrastructure of basidiospore walls and septa in several taxa of

Dacrymycetales from Mexico.

Materials & methods

Seventy-five specimens representing nine dacrymycete species from México were

collected. Species represented were Calocera macrospora, C. viscosa, Dacrymyces

410 ... Sierra & al.

chrysospermus, D. dictyosporus, Dacryopinax elegans, Da. lowyi, Da. spathularia,

Guepiniopsis alpina, and G. buccina. The specimens were examined using standard

mycological techniques (Martin 1952, Lowy 1971, Cifuentes et al. 1986, Sierra 1992,

1995, 2000; Sierra & Cifuentes 1993) and deposited in FCME and TLXM herbaria

(Holmgren & Holmgren 1990).

Ten specimens with a large number of spores were selected for processing with

transmission electron microscopy (TEM). Small (2 mm’) basidiome pieces were

rehydrated for 1 day in distilled water. Samples were fixed with 2.5% glutaraldehyde in

0.05 M sodium cacodylate buffer, pH 7.4 for 2 hours. A post-fixation with 1% osmium

tetroxide in 0.05 M sodium cacodylate buffer was carried out for 2 hours. The samples

were dehydrated via a graded ethanol series (70-100%), prepared with a mixture of

propylene oxide and epoxy resin (in 2:1, 1:1, and 1:2 proportions) for 24 hours, and

embedded in pure epoxy resin. The block face was then shaped to a 0.1 mm/’ trapezoid.

Sections were cut with glass cleavers in a Sorvall Ultramicrotome MT2. No serial-sections

were made due to the scarcity of herbarium materials in good condition. Sections were

placed on copper grids and stained in uranyl acetate (10 minutes) followed by lead

citrate (5 minutes). The grids were examined with a Zeiss EM-10 TEM.

Species & collections

Calocera macrospora Brasf., Lloydia 1: 156. 1938. Fics la,b

SPECIMEN EXAMINED: MEXICO, TLaxcata, Mpio. de Tlaxco, Parque recreativo El

Rodeo, 19°39'54"N 98°08'17"W, alt. 2900 m, on rotten wood, 14 VIII 1992, Pérez-

Ramirez 1604 (FCME 4856).

Calocera viscosa (Pers.) Fr., Syst. Mycol. 1: 486. 1821. Figs Ic,d

SPECIMEN EXAMINED: MEXICO, Micuoacan, Mpio. de Zinapecuaro, Desviacién

al Balneario Erendira, km 5.5 carr. San Pedro Jacuaro—Los Azufres, 19°45'36"N

100°41'21"W, alt. 2600 m, on soil, 4 IX 1987, Ojeda y Villegas-Rios 952 (FCME 14002).

Dacrymyces chrysospermus Berk. & M.A. Curtis, Grevillea 2: 20. 1873. Figs 3a,b

SPECIMEN EXAMINED: MEXICO, GUERRERO, Mpio. de Chilpancingo de los Bravo,

Canada de Agua Fria, 17°32'30"N 99°41'30"W, alt. 2500 m, on rotten wood, 8 VII 1984,

Lopez L. s/n (FCME 13012).

Dacrymyces dictyosporus G.W. Martin, Mycologia 50: 939. 1959. Figs. 2

SPECIMEN EXAMINED: MEXICO, MicHoacan, Mpio. de Zinapecuaro, Desviacioén

al Balneario Erendira, km 5.5 carr. San Pedro Jacuaro-Los Azufres, 19°45'36"N

100°41'21"W, alt. 2600 m, on rotten wood, 4 IX 1987, Molina y Pérez-Ramirez 805

(FCME 14137).

Dacryopinax elegans (Berk. & M.A. Curtis) G.W. Martin, Lloydia 11: 116. 1948. Fic. 3c

SPECIMEN EXAMINED: MEXICO, TaBasco, Mpio. de Teapa, Centro Regional

Universitario del Sureste (CRUSE) Puyacatengo, 17°31'39"N 92°55'51"W, alt. 80 m, on

rotten wood, 20 X 1991, Sierra 46 (FCME 4523).

Dacryopinax lowyi S. Sierra & Cifuentes, Mycotaxon 92: 244. 2005. Fics 3f,g

SPECIMENS EXAMINED: MEXICO, Micuoacan, Mpio. de Zinapecuaro, Laguna Larga,

zona de proteccién forestal Los Azufres, 19°48'05"N 100°41'02"W, alt. 2800 m, on

Ultrastructure in Dacrymycetales (Mexico) ... 411

FIGURE 1. Calocera macrospora: a. Biseptate basidiospore. b. Detail of spore wall and septum

[coriotunic a = CTa, coriotunic b = CTb, endosporium = EnS, episporium= EpS, septum = S, pore

= P, pore aperture = Pa]. Calocera viscosa: c. Uniseptate basidiospore. d. Detail of septal wall [S].

Bars: a,c = 5 um; b, d= 1 um.

412 ... Sierra & al.

FIGURE 2. Dacrymyces dictyosporus: a. Multiseptate basidiospore (transverse and longitudinal

septa). b. Detail of spore wall and septum [coriotunic a = CTa, coriotunic b = CTb, endosporium

= EnS, episporium = EpS, transverse septum = TS, longitudinal septum = LS]. c. Detail of a

multiseptate basidiospore; only transverse septum [TS] and spore wall with 4 layers [coriotunic

a = CTa, coriotunic b = CTb, endosporium = EnS, episporium = EpS, pore = P]. d. Uniseptate

basidiospore. e. Detail of septal wall [pore = P, pore aperture = Pa]. Bars: a = 5 um; b-d = 2 um;

e=1 um.

Ultrastructure in Dacrymycetales (Mexico) ... 413

FIGURE 3. Dacrymyces chrysospermus: a. Multiseptate basidiospore with transverse septa [TS]

only. b. Detail of spore wall and septa. Dacryopinax elegans: c. Spore walls [W] and transverse

septa [TS]. Dacryopinax spathularia: d. Uniseptate basidiospore with hilar appendix [HA].

e. Detail of spore wall [W]. Dacryopinax lowyi: f. Biseptate basidiospore with hilar appendix [HA].

g. Detail of transverse septum [TS] and pore [P]. Guepiniopsis buccina: h. Detail of spore wall [W],

transverse septum [S], organelle, and pore [P]. Guepiniopsis alpina: i. Detail of transverse septum

[TS] and pore [P?]. Bar: a, b = 5 um.

414 ... Sierra & al.

rotten wood, 3 VII 1987, Nufiez Mariel s/n (FCME 13863); ESTADO DE MEXxIco, Mpio.

de Amecameca, Km 14 carr. Amecameca—Tlamacas, 19°05'14"N 98°40'43”"W, alt. 3300

m, on rotten wood, 15 VII 1993, Sierra 165 (FCME 20406).

Dacryopinax spathularia (Schwein.) G.W. Martin, Lloydia 11: 116. 1948. Fics 3d,e

SPECIMEN EXAMINED: MEXICO, Nayarit, Mpio. de Tepic, La Capilla, Reserva

Ecoldgica del Cerro San Juan, 21°28'11"N 105°00'31"W, alt. 1371 m, on rotten wood, 29

VIII 1991, Rodriguez Castafieda y Pérez-Ramirez 1467 (FCME 3903).

Guepiniopsis alpina (Tracy & Earle) Brasf., Amer. Midl. Nat. 20: 225. 1938. Fia. 3i

SPECIMEN EXAMINED: MEXICO, HipatGo, Mpio. de Tenango de Doria, Ejido

Muridores a 2 km de Apulco via Agua Blanca, 20°20'05"N 98°20'52"W, alt. 2200 m, on

rotten wood, 3 IX 1980, Cifuentes 827 (FCME 10620).

Guepiniopsis buccina (Pers.) L.L. Kenn., Mycologia 50(6): 888. 1959. Fig. 3h

SPECIMEN EXAMINED: MEXICO, HipAtco, Mpio. de Calnali, a 8 km de Ahuacatlan,

20°49'N 99°03'W, alt. 1300 m, on rotten wood, 8 VII 1980, Cifuentes 480 (FCME

10159).

Results

Calocera macrospora and C. viscosa have uniformly thick walls. In C. viscosa

the wall layers are not defined, but they are in septa, while in C. macrospora

the layers are clearly defined. A septal pore-like structure was observed only in

C. macrospora.

Dacrymyces dictyosporus has both transverse and longitudinal septa, while

only longitudinal septa are present in D. chrysospermus. ‘The spore walls of

these two species are thick, with four well-differentiated layers, and 3-layered

septa are visible in each one. The D. dictyosporus septa have a wall with a central

swelling (pore-like structure) that is not observed in D. chrysospermus.

Spore walls are not uniformly thick within Dacryopinax. In Da. elegans

spore walls are thick but with undefined layers; two septal wall layers can be

seen, but no septal pores are distinguished. Spore walls in Da. spathularia are

thick (but not to the same degree as in Da. elegans) and the walls are 3-layered.

In Da. lowyi (Sierra & Cifuentes 2005), a hilar appendix, broken hilum wall,

and septal pores with a dolipore-like structure can be discerned.

Spore wall layers are visible in Guepiniopsis buccina and there is a pore-like

structure in the septum. In G. alpina there is a swollen part in the transverse

septal wall that is not dolipore-like. The results are summarized in TABLE 1.

Discussion

All our conclusions are based on well-fixed material, but the material

may have been damaged during drying before fixing, and we could not make

serial sections. Nonetheless, the presence or absence of spore septal pores

Ultrastructure in Dacrymycetales (Mexico) ...415

TABLE 1. Presence or absence of spore wall layers and septa with dolipore.

LATERAL SPORE WALL SPORE SEPTA

SPECIES

A DISTINCT LAYER WITH DOLIPORE

[Giiocramacopow ||

| Cviscosa TC

Dacrymyces chrysospermus +

D. dictyosporus + +

Dacryopinax elegans

Da. lowyi

Da. spathularia

and distinguishable lateral wall layers appear species specific in Calocera,

Dacrymyces, Dacryopinax, and Guepiniopsis. The differences observed are

unlikely to be based on fixation artifacts. We could not distinguish lateral wall

layers in the well-preserved spores of either Da. elegans or C. macrospora, so

that the absence of distinguishable lateral wall layers in the less well preserved

spores of C. viscosa is, therefore, not likely to result from poor fixation. On the

other hand, the septal swelling seen in G. alpina might well be an artifact that

requires further study for verification. Pore-like structures that can be detected

in even poorly fixed material can easily be overlooked in single sections. In

Basidiomycota, the absence of septal pores has been indicated as an important

ultrastructural marker at the family and ordinal levels for of smut fungi (e.g.,

Microbotryaceae, Georgefischeriales, Ustilaginales; Bauer et al. 1997). Keller

(1986, 1992, 1997), Keller & Job (1992), and Garnica et al. (2007) have all

studied spore walls, yet none of them have commented on pores in the spore

septa.

The morphology and DNA analysis of taxa within the Dacrymycetales

does not appear to be consistent (Shirouzu et al. 2012). Perhaps some of the

ultrastructural differences described here might be useful for characterizing

taxa in a new classification more congruent with the DNA analysis. The first

step might be in producing good serial sections for a better morphological

reconstruction of septal pores in spores.

Acknowledgments

We wish to thank Dr. Roland Kirschner and Dr. Charles Chee-Jen Chen for

reviewing the manuscript and for helpful comments; Biol. Lilia Pérez-Ramirez from

FCME and MC. Alejandro Kong from TLXM Herbaria for curatorial support. This work

was financed by DGAPA/UNAM Projects numbers IN-206901, IN-209605, IN-218008,

IN-207311.

416... Sierra & al.

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http://dx.doi.org/10.2307/3760717

MY COTAXON

http://dx.doi.org/10.5248/123.419

Volume 123, pp. 419-429 January-March 2013

Studies of two Corner types (Marasmius nigroimplicatus

and M. subrigidichorda) and new Gymnopus combinations

ZDENKO TKALCEC & ARMIN MESIC

Ruder Boskovic Institute, Bijenicka cesta 54, HR-10000 Zagreb, Croatia

* CORRESPONDENCE TO: Ztkalcec@irb.hr & *amesic@irb.hr

ABSTRACT — Type specimens of Marasmius nigroimplicatus and M. subrigidichorda were

studied. Based on morphological studies, both species belong to Gymnopus, sect. Androsacei.

Color photographs of dried basidiomata and microscopic characters accompany the detailed

descriptions. New combinations are proposed for these two taxa. Nine related marasmioid

species are transferred to Gymnopus.

Key worps — Agaricales, Basidiomycota, Omphalotaceae, taxonomy

Introduction

Corner (1996) described 99 new white-spored agaric species from Malesian

region, of which 87 were placed in the genus Marasmius. However, he used

a very broad generic concept of Marasmius that includes several genera

(Gloiocephala, Gymnopus, Marasmiellus, Rhodocollybia, etc.) which have mostly

been accepted as good genera and whose concepts have also been supported by

phylogenetic analyses (e.g., Wilson & Desjardin 2005, Tan et al. 2009, Antonin

& Noordeloos 2010). Consequently, most of the Marasmius species described

by Corner (1996) should be transferred into the new genera.

During the process of describing the new species, Gymnopus fuscotramus

MeSic¢ et al. (MeSi¢ et al. 2011), we studied types of two similar species described

by Corner (1996), Marasmius nigroimplicatus and M. subrigidichorda, known

only from their type localities in Singapore. In this paper we give detailed

species descriptions, as well as color photographs of their dried basidiomata

and microscopic characters. The two species are characterized by small

marasmioid basidiomata, thin insititious dark colored stipes, well-developed

black rhizomorphs, dextrinoid trama of stipe, and non-hymeniform pileipellis.

According to these characters, neither species belongs to Marasmius in the

modern sense, but to Gymnopus, sect. Androsacei (Kithner) Antonin & Noordel.

420 ... Tkaléec & Medi¢

(Wilson & Desjardin 2005, Noordeloos & Antonin 2008, Antonin & Noordeloos

2010). Therefore, we propose new combinations in Gymnopus (Pers.) Roussel

for the two studied species, as well as for nine related marasmioid species in

sect. Androsacei based on their morphological characters.

Materials & methods

The descriptions of Gymnopus nigroimplicatus and G. subrigidichorda are completely

based on the type collections deposited at the Royal Botanic Garden Edinburgh (E).

The following abbreviations are used in macroscopic description: L for number of

lamellae and | for number of lamellulae between two lamellae. Microscopic features

were observed with a light microscope (brightfield and phase contrast (PhC)) under

magnification up to 1500x and photographed with a digital camera. Descriptions and

images of microscopic characters were made from rehydrated dried specimens mounted

in 2.5% potassium hydroxide (KOH) solution. Amyloidity and dextrinoidity were tested

in Melzer’s reagent (Erb & Matheis 1983). Basidiospore measurements were made from

mounts of lamellae and based on calibrated digital images. For G. nigroimplicatus, three

mature basidiomata were chosen and 20 basidiospores were measured in total (spores

were scarce), while for G. subrigidichorda, two mature basidiomata were chosen and 50

randomly selected basidiospores were measured in total. Spore measurements (length,

width) include apiculus and are given as: (min.) stat. min. - av. - stat. max. (max),

where “min.” = minimum (lowest measured value), “stat. min.” = statistical minimum

(arithmetic average minus 2x standard deviation), “av.” = arithmetic average, “stat.

max.’ = statistical maximum (arithmetic average plus 2x standard deviation), “max.”

= maximum (highest measured value). Standard deviation (SD) of spore length and

width is also given. The length/width ratio of spores is given as the “Q” value (min. — av.

— max.). Comparison of G. nigroimplicatus and G. subrigidichorda with similar species

is based on descriptions and illustrations in the following literature: Petch 1948, Singer

1965, 1969, 1976, 1989, Pegler 1986, Desjardin 1987, Corner 1996, Antonin 2007, Mesi¢

et al. 2011.

Taxonomy

Type studies

Gymnopus nigroimplicatus (Corner) Mesi¢é & Tkaléec, comb. nov. — PLATES 1, 3-7

MycoBank MB 564875

= Marasmius nigroimplicatus Corner, Beih. Nova Hedwigia 111: 75. 1996. TyPE:

SINGAPORE: Botanic Garden, 14 Aug 1940, leg. E. J. H. Corner, (E 206719,

holotype).

The following description is compiled from the dried holotype collection,

which consists of approximately 12 basidiomata, partially fragmented or

badly preserved, accompanied with abundant rhizomorphs. Basidiomata

and rhizomorphs arising from dead leaves. Basidiomata are not connected

with rhizomorphs. PILEus 3-8.5 mm broad, convex or conico-convex at first,

later applanate, slightly striate-sulcate, matt, homogeneous brown. LAMELLAE

Two Marasmius types + new Gymnopus combinations ... 421

PLatTEs 1-2. Dried basidiomata (holotypes).

1. Marasmius nigroimplicatus. 2. Marasmius subrigidichorda. Bars: 1 = 5 mm; 2 = 10 mm.

adnate or attached to a pseudocollarium, distant, L = 10-14, 1 = 1-3, <1 mm

broad, sometimes furcate, brown (concolorous with pileus), with an entire,

concolorous edge. STIPE 7-23 x 0.15-0.40 mm, subcylindrical, dark brown,

surface densely minutely dark brown pubescent, insititious. CONTEXT brown.

422 ... Tkaléec & MeSié

RuIzomorpHs filiform, mostly gradually tapering towards the apex, <70 mm

long, 0.05-0.20 mm thick near the base, rarely branched, mostly minutely

pubescent (especially in the lower half), dark brown to black.

BASIDIOSPORES [20/3/1] (7.1—)6.8-8.7-10.5(-11.0) x (2.3-)2.3-3.0-3.7

(-3.6) um, SD = 0.93 x 0.36, Q = 2.35-2.95-3.87, cylindrical to bacilliform,

mostly with conical base and rounded apex in frontal view, (sub)amygdaliform

or lacrymoid in side view, smooth, hyaline, thin-walled, non-amyloid, non-

dextrinoid. Bastp1A 20-25 x 6.5-7 um, clavate, 4-spored, thin-walled, pale

brownish to pale brown, light brown in mass. BAsIDIOLEs mostly fusoid, but

also subcylindrical, narrowly utriform or narrowly clavate, often mucronate

to rostrate (and then sometimes with subcapitate apex). LAMELLAR EDGE

sterile. CHEILOcysTiIpIA abundant, 21-50 x 8-27 um, coralloid, with

clavate to irregularly shaped body (10-30 x 3-9 um) and mostly abundant,

strongly branched projections, thin-walled, hyaline to pale yellowish brown.

PLEUROCYSTIDIA absent. HYMENOPHORAL TRAMA rather irregular, composed

of pale brownish to pale brown (light brown in mass), often branched, 1-6

uum broad, thin-walled hyphae. PILEIPELLIs an irregular cutis, hyphae 2-10

um broad, thin-walled, smooth to diverticulate, terminal elements sometimes

subcoralloid, subhyaline to brown, pigment often minutely to coarsely

encrusted. STIPITIPELLIS a cutis of parallel, cylindrical, 3-10 um broad, brown-

gray hyphae with encrusted pigment. CauLocystip14 abundant, formed as

lateral or terminal elevations of stipitipellis cells, 5-93 x 4-12 um, cylindrical,

subcylindrical, less frequently somewhat flexuose, rarely branched, always with

obtuse apex, thick-walled (1.8-5 um thick), brown-gray, pigment parietal.

STIPE TRAMA composed of (sub-)hyaline, parallel, thin- to thick-walled (<4 um

thick), 2-9 um broad hyphae. CLAMP CONNECTIONS present in all tissues.

CHEMICAL REACTIONS — All parts of basidioma non-amyloid and non-

dextrinoid except hyphae in cortical layer of stipe and caulocystidia, which are

strongly dextrinoid.

Hasitat — On dead leaves in botanic garden.

DISTRIBUTION — Known only from the type locality in Singapore, Asia.

REMARKS — Gymnopus nigroimplicatus is characterized by purplish brown to

cinnamon brown colors in pileus, lamellae, and stipe; slightly striate-sulcate

pileus; lamellae adnate or attached to a pseudocollarium; an insititious, dark

brown, and minutely pubescent stipe; well-developed black rhizomorphs;

basidiomata growing only on dead leaves (not arising from rhizomorphs); non-

hymeniform pileipellis (irregular cutis); subcylindrical, cylindrical or lacrymoid

spores; presence of clamp connections in all tissues; coralloid cheilocystidia;

abundant thick-walled caulocystidia with obtuse apex; and dextrinoid hyphae

in cortex of stipe. We consider it a good species.

Two Marasmius types + new Gymnopus combinations ... 423

PLaTEs 3-7. Marasmius nigroimplicatus. 3. Spores (PhC). 4. Cheilocystidia (PhC). 5. Hymenophoral

trama. 6. Caulocystidia. 7. Pileipellis. Bars: 3 = 5 um; 4 = 10 um; 5-7 = 30 um.

424 ... Tkaléec & MeSi¢

Among other species in sect. Androsacei with clamp connections, black

rhizomorphs, non-glabrous stipe, and colored (not white or cream) pileus and

lamellae, G. bactrosporus has a smaller pileus (2.5-3.5 mm broad), whitish to

light sordid gray lamellae, longer spores (10-12 x 2.7-3.7 um), well-developed

pleurocystidia, and different cheilocystidia. Gymnopus brevistipitatus has a

smaller pileus (2-4 mm broad), shorter stipe (2-3 mm long), fewer lamellae

(L = 7-9), and smaller caulocystidia (20-40 um long) with apical projections

or coralloid. Gymnopus campinaranae has a smaller pileus (2-3 mm broad),

thinner stipe (<0.1 mm wide), and larger spores (9-13 x 3.2-4.5 um).

Gymnopus fuscotramus has free lamellae, glabrous and thicker rhizomorphs

(<0.5 mm wide), less elongated spores, and caulocystidia with thinner walls

[0.5-2(-2.5) um thick]. Gymnopus rhizomorphicola has a shorter (+3 mm

long) eccentric curved stipe, fewer lamellae (L = 7-8), and basidiomata arising

from rhizomorphs. Gymnopus rigidichorda has mature creamy white lamellae,

thicker (0.4-0.6 mm wide) glabrous rhizomorphs, and differently shaped non-

coralloid cheilocystidia. Gymnopus subrigidichorda has an umbilicate pileus,

broader lamellae, longer and thicker rhizomorphs, broader spores (4-5 um

wide), and rare clamp connections (seen only in stipe trama). Gymnopus thiersii

has rare or absent rhizomorphs, crowded lamellae, and lacks cheilocystidia.

Gymnopus subrigidichorda (Corner) Tkaléec & Megi¢, comb. nov. PLATES 2, 8-12

MycoBank MB 564876

= Marasmius subrigidichorda Corner, Beih. Nova Hedwigia 111: 99, 1996. TyPE:

SINGAPORE: Bukit Timah, 11 Nov 1929, leg. E. J. H. Corner, (E 206861, holotype).

The following description is compiled from the dried holotype collection,

which consists of 20 basidiomata in good condition accompanied with

abundant rhizomorphs. Basidiomata arising from 2-8 mm wide twigs as well

as from rhizomorphs. PILEus 6.5-19 mm broad, convex to plano-convex with

an umbilicate center, sometimes with a small papilla in the central depression,

striate-sulcate up to the center, matt, homogeneous brown to dark brown.

LAMELLAE attached to a pseudocollarium, distant, L = 12-14, 1 = 1-3, <2.5mm

broad, often intervenose, yellowish brown to brown, with an entire, concolorous

edge. STIPE 4-17 x 0.6-0.9 mm, subcylindrical (slightly broadened at the base

and the apex), brown to dark brown, surface moderately to densely minutely

brown pubescent (under a hand-lens), sub-insititious with small pale brown

fibrils at the base. CONTEXT pale brown. RHIZOMORPHSs robust, <410 mm long

and <0.9 mm thick, occasionally branched, glabrous, shiny, stiff, black with a

pale brown apex, medulla pale brown.

BASIDIOSPORES [50/2/1] (6.2—)6.4-8.3-10.2(-10.2) x (4.0-)4.0-4.5-5.1

(-5.1) um, SD =0.95 x 0.27, Q = 1.43-1.82-2.18, ellipsoid to elongate (sometimes

cylindrical), mostly with conical base and rounded apex in frontal view, mostly

Two Marasmius types + new Gymnopus combinations ... 425

Piates 8-12. Marasmius subrigidichorda. 8. Spores (PhC). 9. Cheilocystidia. 10. Hymenophoral

trama. 11. Caulocystidia. 12. Pileipellis. Bars: 8 = 5 um; 9 = 10 um; 10-12 = 30 um.

426 ... Tkaléec & Medi¢

(sub)amygdaliform or lacrymoid in side view, smooth, hyaline, thin-walled,

non-amyloid, non-dextrinoid. Basrp1a 26-38 x 6.5-10 um, clavate, 4-spored,

thin-walled, subhyaline to pale brown, light brown in mass, pigment intracellular

and granular. BASIDIOLES narrowly clavate, cylindrical or fusoid, sometimes

mucronate to rostrate. LAMELLAR EDGE heterogeneous. CHEILOCYSTIDIA

scattered to locally abundant, 13-32 x 7.5-20 um, + clavate with irregular

apical projections of different types (nodose, digitate or branched), thin-walled,

subhyaline to pale brown in KOH. PLeurocystip14 absent. HYMENOPHORAL

TRAMA irregular, composed of hyaline, sometimes slightly gelatinized, often

branched hyphae, 3-14 um broad, thin- to moderately thick-walled (<0.8 um

thick). PILEIPELLIs an irregular cutis of mostly nodose to diverticulate + repent

hyphae (3-10 um broad) and abundant + erect, lobed to irregularly coralloid

terminal elements, projections knob-like, digitate or clavate, 2-10(-20) x 2-5

(-9) um, hyphae thin-walled, subhyaline to light brown in KOH. PILEUS TRAMA

composed of hyaline, interwoven, partly gelatinized, thin- to moderately thick-

walled (<1 um thick), 1.5-11 um broad hyphae. ST1PITIPELLIs a cutis of parallel,

cylindrical, <13 um broad, thick-walled hyphae (<6 um thick), mostly brown

in KOH, pigment parietal. CAuLocystTrip1A abundant in upper part of stipe,

18-50(60) x 7-12 um, subcylindrical to narrowly clavate with an obtuse apex,

sometimes branched, thick-walled (2-5 um thick), subhyaline to light brown in

KOH. STIPE TRAMA composed of (sub)hyaline, parallel, thin- to thick-walled

(<4 um thick), 2-8 um broad hyphae. CLAMP CONNECTIONS rare, seen only in

stipe trama.

CHEMICAL REACTIONS — all parts of basidioma non-amyloid and non-

dextrinoid except hyphae in cortical layer of stipe and caulocystidia, which are

strongly dextrinoid.

Habitat — In forest, on dead twigs.

DISTRIBUTION — Known only from the type locality in Singapore, Asia.

REMARKS — Gymnopus subrigidichorda is characterized by a rufous brown

striate-sulcate pileus with umbilicate center; incarnate to rufous brown lamellae

attached to the pseudocollarium; sub-insititious minutely pubescent, brown

stipe; abundant robust black rhizomorphs; basidiomata both growing on twigs

and arising from rhizomorphs; non-hymeniform pileipellis (irregular cutis with

lobed to coralloid terminal elements); ellipsoid, oblong, or lacrymoid spores;

rare clamp connections (present only in stipe trama); clavate cheilocystidia

with irregular apical projections; abundant thick-walled caulocystidia with

obtuse apex; and dextrinoid hyphae in the stipe cortex. We consider it a good

species. Our description pretty well agrees with the original description, except

in spore size, which is significantly smaller (4.5-6.0 x 3.5-4.0 um) in original

description (Corner 1996).

Two Marasmius types + new Gymnopus combinations ... 427

Among other species in sect. Androsacei with clamp connections, black

rhizomorphs, non-glabrous stipe, and colored (not white or cream) pileus and

lamellae, G. bactrosporus has a smaller pileus (2.5-3.5 mm broad), hyaline-

pilose stipe (under a hand-lens), longer (10-12 x 2.7-3.7 um) and more

elongated spores, well-developed pleurocystidia, and thin-walled caulocystidia.

Gymnopus brevistipitatus has a smaller pileus (2-4 mm broad), shorter stipe

(2-3 mm long), fewer lamellae (L = 7-9), and more elongated, narrower spores

(2.5-3.7 um wide). Gymnopus campinaranae has a smaller pileus (2-3 mm

broad), thinner stipe (<0.1 mm wide), and larger spores (9-13 x 3.2-4.5 um).

Gymnopus fuscotramus has free lamellae, shorter (<135 mm long) and thinner

rhizomorphs (<0.5 mm wide), more elongated spores, gray-brown pileal and

hymenophoral trama, and caulocystidia with thinner walls (0.5-2(-2.5) um

thick). Gymnopus nigroimplicatus has a pileus without central depression,

narrower lamellae, shorter and thinner rhizomorphs, narrower spores

(2.3-3.7 um wide) and clamp connections present in all tissues (not only in

stipe trama). Gymnopus rhizomorphicola has a shorter (+3 mm long), eccentric

and curved stipe, fewer lamellae (L = 7-8), more elongated spores (Q = 2.1-2.9),

and larger caulocystidia (45-190 x 6-26 um). Gymnopus rigidichorda has

creamy white mature lamellae and very narrow thinner (0.4-0.6 mm wide)

rhizomorphs, differently shaped non-coralloid cheilocystidia, and narrower

spores (3-3.5 um wide). Gymnopus thiersii has rare or absent rhizomorphs,

crowded lamellae, and lacks cheilocystidia. Because G. subrigidichorda has

rare clamp connections (seen only in stipe trama) it could also be confused

with a few species that lack clamp connections. Gymnopus aporpohyphes differs

by a smaller (<5 mm broad) and convex pileus with small umbo, whitish to

pale avellaneous lamellae, narrower cheilocystidia (4-6.5 um wide), and more

elongated narrower (2.8-3.2 um wide) spores. Gymnopus pacificus differs by a

smaller pileus (5-6.5 mm broad), narrower stipe (0.2-0.3 mm wide), narrower

spores (3.5-4.1 um wide), and caulocystidia with thinner walls (0.5-1.5 um

wide). Gymnopus tomentellus differs by a smaller pileus (1-3 mm broad)

without a central depression, smaller stipe (1-4 x 0.2-0.6 mm), larger and

more elongated spores (11-11.5 x 4-5 um), and caulocystidia with thinner

walls (ca. 1 um wide).

Additional new combinations in Gymnopus

Gymnopus aporpohyphes (Singer) Tkaléec & MeSié, comb. nov.

MycoBank MB 800668

= Marasmius aporpohyphes Singer, Sydowia 18: 333, 1965 [“1964”].

Gymnopus bactrosporus (Singer) Mesi¢ & Tkaléec, comb. nov.

MycoBank MB 800669

= Marasmius bactrosporus Singer, Sydowia 18: 334, 1965 [“1964”].

428 ... Tkaléec & Medi¢

Gymnopus brevistipitatus (Antonin) Tkaléec & Mesi¢, comb. nov.

MycoBank MB 800670

= Setulipes brevistipitatus Antonin, Mycotaxon 88: 71, 2003.

Gymnopus campinaranae (Singer) Mesi¢ & Tkaléec, comb. nov.

MycoBank MB 800671

= Marasmius campinaranae Singer, Fieldiana, Bot. 21: 57, 1989.

Gymnopus pacificus (Singer) Tkaléec & MeSi¢, comb. nov.

MycoBank MB 800672

= Marasmius pacificus Singer, Beih. Nova Hedwigia 29: 95, 1969.

Gymnopus rhizomorphicola (Antonin) Mesi¢ & Tkaléec, comb. nov.

MycoBank MB 800673

= Setulipes rhizomorphicola Antonin, Mycotaxon 88: 75, 2003.

Gymnopus rigidichorda (Petch) Tkaléec & MeSi¢, comb. nov.

MycoBank MB 800674

= Marasmius rigidichorda Petch, Trans. Brit. Mycol. Soc. 27(3-4): 138, 1945 [“1944”].

Gymnopus thiersii (Desjardin) Mesi¢ & Tkaléec, comb. nov.

MycoBank MB 800675

= Marasmius thiersii Desjardin, Mycologia 79(1): 123, 1987.

Gymnopus tomentellus (Berk. & M.A. Curtis) Tkaléec & MeSi¢, comb. nov.

MycoBank MB 800676

= Marasmius tomentellus Berk. & M.A. Curtis, J. Linn.

Soc., Bot. 10(45): 298, 1868 [“1869”].

Acknowledgments

We are grateful to the Royal Botanic Garden Edinburgh (E) for the loan of Corner’s

holotype collections. We would also like to thank Vladimir Antonin (Brno, Czech

Republic) and Clark L. Ovrebo (Edmond, OK, USA) for their critical reviews of the

manuscript.

Literature cited

Antonin V. 2007. Monograph of Marasmius, Gloiocephala, Palaeocephala and Setulipes in tropical

Africa. Fungus flora of tropical Africa 1. National Botanic Garden: Meise (Belgium).

Antonin V, Noordeloos ME. 2010. A monograph of marasmioid and collybioid fungi in Europe.

IHW- Verlag: Eching (Germany).

Corner EJH. 1996. The agaric genera Marasmius, Chaetocalathus, Crinipellis, Heimiomyces,

Resupinatus, Xerula and Xerulina in Malesia. Beih. Nova Hedwig. 111: 1-175.

Desjardin DE. 1987. New and noteworthy marasmioid fungi from California. Mycologia 79(1):

123-134. http://dx.doi.org/10.2307/3807751

Erb B, Matheis W. 1983. Pilzmikroskopie. Kosmos: Stuttgart (Germany).

Me’i¢ A, Tkaléec Z, Deng C-Y, Li T-H, PleSe B, Cetkovié H. 2011. Gymnopus fuscotramus

(Agaricales), a new species from southern China. Mycotaxon 117: 321-330.

http://dx.doi.org/10.5248/117.321

Two Marasmius types + new Gymnopus combinations ... 429

Noordeloos ME, Antonin V. 2008. Contribution to a monograph of marasmioid and collybioid

fungi in Europe. Czech Mycol. 60(1): 21-27.

Pegler DN. 1986. Agaric flora of Sri Lanka. Kew Bulletin Additional Series 12. HMSO, London.

Petch T. 1948. A revision of Ceylon Marasmii. Trans. Brit. Mycol. Soc. 18(2): 19-44 + pl. II-IV.

Singer R. 1965 [“1964”]. Monographic studies on South American Basidiomycetes, especially

those of the East Slope of the Andes and Brazil. 2.) The genus Marasmius in South America.

Sydowia 18: 106-358.

Singer R. 1969. Mycoflora australis. Beih. Nova Hedwigia 29. J. Cramer, Lehre.

Singer R. 1976. Marasmieae (Basidiomycetes - Tricholomataceae). Flora Neotropica 17. The New

York Botanical Garden, New York.

Singer R. 1989. New taxa and new combinations of Agaricales (Diagnoses fungorum novorum

agaricalium IV). Fieldiana, Bot. 21: 1-133.

Tan Y-S, Desjardin DE, Perry BA, Vikineswary S, Noorlidah A. 2009. Marasmius sensu stricto in

Peninsular Malaysia. Fungal Diversity 37: 9-100.

Wilson AW, Desjardin DE. 2005. Phylogenetic relationships in the gymnopoid and marasmioid

fungi (Basidiomycetes, euagarics clade). Mycologia 97(3): 667-679.

http://dx.doi.org/10.3852/mycologia.97.3.667

MY COTAXON

http://dx.doi.org/10.5248/123.431

Volume 123, pp. 431-437 January-March 2013

An emendation of Fusticeps and

two new species from the Brazilian Amazon Forest

JOSIANE SANTANA MONTEIRO'?& Luis FERNANDO PASCHOLATI GUSMAO”

"Departamento de Micologia, Universidade Federal de Pernambuco,

Pernambuco, 50670-420, Brazil

*Departamento de Ciéncias Biologicas, Universidade Estadual de Feira de Santana,

Bahia, 44036-900, Brazil

* CORRESPONDENCE TO: lgusmao@uefs.br

ABSTRACT — Two new species, Fusticeps lampadiformis and F. papillatus, are described and

illustrated. The description of Fusticeps is emended to accommodate smooth-spored species,

and a key to the four Fusticeps species is provided.

Key worps—freshwater fungi, anamorphic fungi, taxonomy

Introduction

Webster & Davey (1980) established Fusticeps for the single species F. bullatus,

collected on submerged leaves in Malaysia. The most conspicuous feature of the

surface ornamentation of the conidia isa projection that expands to a thin-walled

domed cap; according to Webster & Davey (1980), the air entrapped between

the projections may possibly aid conidia flotation. This character was used to

include this genus in the aero-aquatic hyphomycetes (Fisher 1977, Goh & Hyde

1996, Hyde & Goh 1998). The genus remained monotypic until Matsushima

(1993) described F. laevisporus collected on dead leaves in Ecuador. The original

generic description — “Mycelium septate, dark. Conidiophores erect, septate,

unbranched or occasionally branched. Conidia holoblastic, integrated, solitary,

dark, transversely septate, epispore fractured into irregular scales which entrap

air” — does not accommodate F. laevisporus, which has smooth conidia; the

rhexolytic conidial secession is not mentioned and only the broad basal scar is

described. However, several characters shared by F. bullatus and F. laevisporus

(e.g., conidial ontogeny, secession, morphology) justify emending the generic

description of Fusticeps to accommodate species with smooth conidia.

There are few records of this genus in the literature. It has been collected

in Malaysia and Ecuador in lentic environments (Webster & Davey 1980;

432 ... Monteiro & Gusmao

Matsushima 1993) and in Poland in lentic and lotic environments (Orlowska et

al. 2004, Orlowska et al. 2006, Czeczuga et al. 2007).

During study of anamorphic fungi associated with submerged litter in the

Brazilian Amazon forest, Fusticeps was very well represented by collections of

the two previously described species and two new species, described here as

F. lampadiformis (with ornamented conidia) and F papillatus (with smooth

conidia).

Materials& methods

Collecting expeditions were made in two different sites in Amazon forest, near

Belém County, Para State: Parque Estadual de Utinga and Area de Protecéo Ambiental

Ilha do Combu. Submerged litter was collected from streams and placed in plastic bags

containing humid paper towels. In the laboratory, the samples were removed from

the plastic bags and incubated in Petri dishes with moistened paper towels at ambient

temperature (about 24°C). Samples were examined within two weeks after collection,

and then periodically, using a dissecting microscope. For microscopic identification the

specimens were placed on glass slides containing PVL resin (polyvinyl alcohol, lactic

acid, and phenol). Measurements and digital images were made using an Olympus

microscope BX51 equipped with bright field and Nomarski interference optics and a

microscopy image acquisition DP25 digital color camera. Specimens in glass slides and

dry material were deposited in the Herbarium of Universidade Estadual de Feira de

Santana, Bahia, Brazil (HUEFS).

Taxonomy

Fusticeps J. Webster & R.A. Davey, Trans. Br. Mycol. Soc. 75: 341, 1980; emend. J.S.

Monteiro & Gusmao

COLONIES on natural substratum effuse, olivaceous to olivaceous brown,

dark to very dark brown. Mycet1um mostly immersed in the substratum,

composed of septate, brown to pale brown hyphae. CONIDIOPHORES macro-

or micronematous, erect, septate, unbranched or occasionally branched,

pale brown. CONIDIOGENOUS CELLS holoblastic, integrated, determinate,

terminal, cylindrical, pale brown. CONIDIAL SECESSION rhexolytic. CONIDIA

solitary, clavate to pyriform, transversally septate, smooth or with conspicuous

ornamentation, pale brown. Teleomorph unknown.

TYPE SPECIES: Fusticeps bullatus J. Webster & R.A. Davey

Fusticeps lampadiformis J.S. Monteiro & Gusmao, sp. nov. PLATE 1a-j

MycoBank MB 800031

Differs from Fusticeps bullatus in producing pyriform to lampadiform, 2-3-septate

conidia.

Type: Brazil. Para: Parque Estadual do Utinga, 1°25'S 48°27'W, on unidentified

submerged leaves, 18 Jan 2012, J. S. Monteiro (holotype, HUEFS 42783).

Erymo oey: lampas (Latin), referring to the conidial shape.

Fusticeps spp. nov. (Brazil) ... 433

PLATE 1.

Fusticeps lampadiformis: a. Immature conidia. b, c. Conidiogenous cell. d—j. Mature conidia.

Fusticeps papillatus: k, |. Conidiogenous cell. m-t. Mature conidia.

Scale bar = 10 um.

434 ... Monteiro & Gusmao

COLONIES on natural substratum effuse, olivaceous to olivaceous brown.

MyceELium mostly immersed in the substratum, composed of septate, smooth,

pale brown hyphae. CoNIDIOPHORES micronematous, mononematous,

unbranched or occasionally branched, straight, erect or slightly flexuous,

septate, smooth, pale-brown, 22.5-62.5 x 2.5-5 um. CONIDIOGENOUS CELLS

monoblastic or rare polyblastic, integrate, determinate, terminal, cylindric,

pale brown. CONIDIAL SECESSION rhexolytic. Conrp1A solitary, pyriform to

lampadiform, 2-3 transversally septate, with cup-shape projections (3-5 x

1.5-2.5 um) arising from the conidial wall, distal cell and intermediate cell

brown, proximal cell pale brown, 21.5-35.5 x 12.5-18 um; basal scar 2-3 um.

CoMMENTS — Fusticeps lampadiformis is close to F. bullatus, which differs

by conidia that are clavate, 3-4 septate, and have predominantly mushroom-

shaped projections (Webster & Davey 1980).

Fusticeps papillatus J.S. Monteiro & Gusmao, sp. nov. PLATE 1k-t

MycoBank MB 800032

Differs from Fusticeps laevisporus in producing clavate, 4—6 septate conidia with papillate

apices.

Type: Brazil. Para: Parque Estadual do Utinga, 1°25’S 48°27'W, on unidentified

submerged leaves, 18 Jan 2012, J. S. Monteiro (holotype, HUEFS 42784).

Erymo oey: diminutive of papula (Latin), referring to the papilla at the conidial distal

cell.

Co.Lonigs on the natural substratum effuse, dark brown. Mycetrum mostly

immersed in the substratum, composed of septate, smooth, brown to pale

brown hyphae. CONIDIOPHORES micronematous, mononematous, unbranched

or occasionally branched, grouped into 3-4, straight, erect or slightly flexuous,

septate, smooth, pale-brown, 17.5-67.5 x 2.5-3.8 um. CONIDIOGENOUS CELLS

monoblastic, integrate, determinate, terminal, cylindric, brown to pale brown.

CONIDIAL SECESSION rhexolytic. Conip1a solitary, clavate, 4-6 transversally

septate, smooth, distal cell thicker wall 1.5-2 um, with a papilla (2-4 um

longer), distal cell and intermediate cells brown to pale brown, proximal cell

pale brown, 35-45.5 x 11.5-15 um; basal scar 2-3 um.

CoMMENTS— Fusticeps papillatus is characterized by clavate, smooth, 4-6

septate conidia with a papilla at the apex of the distal cells. Fusticeps laevisporus

is a comparable species but is distinguished by absence of a papilla and fewer

conidial septa (Matsushima 1993).

Fusticeps bullatus J. Webster & R.A. Davey, Trans. Br. Mycol. Soc. 75: 342, 1980.

PLATE 2a-e

Cotoniges on the natural substratum effuse, dark. MycEeL1um mostly

immersed in the substratum, composed of septate, smooth, pale brown hyphae.

Fusticeps spp. nov. (Brazil) ... 435

PLATE 2. Fusticeps bullatus: a-e. Mature conidia. Fusticeps laevisporus: f-j. Mature conidia.

Scale bar = 10 um.

CONIDIOPHORES micronematous, mononematous, unbranched, septate,

smooth, brown to pale brown 20-22.5 x 2.5-3.75 um. CONIDIOGENOUS CELLS

holoblastic, integrate, determinate, terminal, cylindric. CONIDIAL SECESSION

rhexolytic. Conrp1A solitary, clavate, 2-3 transversally septate, with mushroom-

shaped to cup-shaped projections (2.5 um long) arising from the conidial wall,

pale brown 23.5-34 x 10-13 um; basal scar 2-3 um.

SPECIMEN EXAMINED: BRAZIL, ParA, Area de Protecéo Ambiental Ilha do Combu,

1°29'S 48°25'W, on unidentified submerged leaves, 17 Jan 2012, J.S. Monteiro (HUEFS

42785).

ComMENTS — Fusticeps bullatus is characterized by clavate, 3-4 septate conidia

with mushroom-shaped to cup-shaped projections (Webster & Davey 1980).

Our Brazilian specimens and Matsushima’s (1993) Ecuadorian material, both

from Amazon forest, have similar conidial dimensions and septation but differ

slightly from the original Malaysian material (TABLE 1). Previously known from

Malaysia, Ecuador, and Poland (Webster & Davey 1980, Matsushima 1993,

Orlowska et al. 2004, Czeczuga et al. 2007); this is the first record for Brazil.

Fusticeps laevisporus Matsush., Matsush. Mycol. Mem. 7: 52, 1993. PLATE 2f-j

COLONIES on natural substratum effuse, dark. MycELIuM mostly immersed

in the substratum, composed of septate, smooth, pale brown hyphae.

436 ... Monteiro & Gusmao

TABLE 1. Synopsis of Fusticeps conidia

Pe, CONIDIA

SIZE (um) SHAPE SEPTA REFERENCES

EF. bullatus 28-34 x 9-12 Clavate 3-4 Webster & Davey (1980)

23.5-35 x 8-12.5 Clavate 2-3 Matsushima (1993)

23.5-34 x 10-13 Clavate 2-3 This paper

EF laevisporus 30-44 x 9.5-13 Clavate 2-3 Matsushima (1993)

25-40 x 10-20 Clavate 3-4 This paper

EF. lampadiformis 21.5-35.5 x 12.5-18 Pyriform 2-3 This paper

E papillatus 35.5-45.5 x 11.5-15 Clavate 4-6 This paper

CONIDIOPHORES micronematous, mononematous, unbranched, septate,

smooth, brown to pale brown 20-47.5 x 2.5-3.75 um. CONIDIOGENOUS CELLS

monoblastic, integrate, determinate, terminal, cylindrical. CONIDIAL SECESSION

rhexolytic. Conip1A solitary, clavate, 3-4 transversally septate, smooth, distal

cell with rounded apex and intermediate cells brown, proximal cell pale brown,

25-40 x 10-20 um; basal scar 2-3 um.

SPECIMEN EXAMINED: BRAZIL, ParA, Parque Estadual do Utinga, 1°25'S 48°27'W, on

unidentified submerged leaves, 18 Jan 2012, coll. J.S. Monteiro (HUEFS 42786).

CoMMENTS— Fusticeps laevisporus has smooth conidia and is comparable with

EF. papillatus, but differs by the rounded apex of the distal cell. Matsushima

(1993) based his description of FE laevisporus on pure culture, and his conidia are

narrower (9.5-13 um) and have fewer septa (2-3) than our Brazilian material,

but we consider the Ecuadorian and Brazilian collections to be conspecific.

This species is a new record for Brazil; previously known from Ecuador and

Poland (Matsushima 1993, Orlowska et al. 2006).

Key to Fusticeps species

Conidial measurements and morphologies are summarized in TABLE 1.

REC OnIG aISIMOOUE fat Bits’ Bid. oe Bowl ne Bie OEE we O8E Wh OE EE ae NOT EF ot RR es 2

la. Conidia with conspicuous ornamentation ........... 0... e eee cece eee tees =)

2; Conidia 2-4-septate; apex.roundeéd 5......2hs0e2gbibaahseaaiea rages eed EF. laevisporus

2a Conidia A=6-septate; apex papillate sc. <ep-2 ss ced oca eee ota, deed ote as Bs E papillatus

3. Conidia pyriform, 2-3-septate ........ 0... eee eee eee eae E lampadiformis

Sa. onidiarclavatey s=4-sep balers, eha82 Feats FON al UBB WB Bel UA al LAI. E bullatus

Acknowledgments

The authors thank Dr. Marc Stadler (Department Microbial Drugs, Helmholtz

Centre for Infection Research - Germany) and Dr. R.E Castafieda Ruiz (INIFAT -

Cuba) for their great contribution in improving the manuscript. The authors extend

thank to Coordination for the Improvement of Higher Education Personnel (CAPES)

for financial support and the “Programa de Pés-graduacao em Biologia de Fungos -

Fusticeps spp. nov. (Brazil) ... 437

PPGBE/UFPE”. This study was supported by The National Council for Scientific and

Technological Development (CNPq) (Grant for LFP Gusmao, Proc. 303924/2008-0).

Literature cited

Czeczuga B, Muszynska E, Godlewska A, Mazalska B. 2007. Aquatic fungi and straminipilous

organisms on decomposing fragments of wetland plants. Mycologia Balcanica 4: 31-44.

Fisher PJ. 1977. New methods of detecting and studying saprophytic behavior of aero-aquatic

hyphomycetes from stagnant water. Trans. Br. Mycol. Soc. 68: 407-411.

http://dx.doi.org/10.1016/S0007-1536(77)80194-0

Goh TK, Hyde KD. 1996. Biodiversity of freshwater fungi. J. Ind. Microbiol. Biotechnol. 17:

328-345. http://dx.doi.org/10.1007/BF01574764

Hyde KD, Goh TK. 1998. Acanthophysis-like structures from wood submerged in freshwater

streams in the tropics. Mycoscience 39: 199-203. http://dx.doi.org/10.1007/BF02464060

Matsushima T. 1993. Matsushima Mycological Memoirs, vol. 7. Published by the author, Kobe.

Orlowska M, Lengiewicz I, Suszycka M. 2004. Hyphomycetes developing on water plants and

bulrushes in fish ponds. Pol. J. Environ. Stud. 13: 703-707.

Orlowska M, Lengiewicz I, Ostrowska H. 2006. Conidial fungi on plants in the Biebrzanski National

Park. Acta Hydrochim. Hydrobiol. 34: 53-57.

Webster J, Davey RA. 1980. Two aero-aquatic hyphomycetes from Malaysia. Trans. Br. Mycol. Soc.

75: 341-345. http://dx.doi.org/10.1016/S0007-1536(80)80105-7

MY COTAXON

http://dx.doi.org/10.5248/123.439

Volume 123, pp. 439-444 January-March 2013

Two Ropalospora lichens new to mainland China

Linc Hu, Ha1-YING WANG, JING LIU & ZUN-TIAN ZHAO

Key Laboratory of Plant Stress Research, College of Life Sciences,

Shandong Normal University, Jinan, 250014, P. R. China;

“CORRESPONDENCE TO: endolichen@gmail.com

ABSTRACT —'The genus Ropalospora is reported from mainland China for the first time;

R. chlorantha is new to China, while R. phaeoplaca is new to mainland China.

Key Worps —taxonomy, corticolous, fungi, Ropalosporaceae

Introduction

Ropalospora A. Massal., first described in 1860, belongs to the Ropalosporaceae

(Kirk et al. 2008). The genus is characterized by black prothallus, dark brown

to black lecideine apothecia, Fuscidea-type ascus, bacilliform conidia, and

multiseptate ascospores (Ekman 1993, Brodo et al. 2001, Kantvilas 2004).

Ropalospora includes six species worldwide: R. chlorantha, R. hibernica

(P. James & Poelt) Tonsberg, R. Iugubris (Sommerf.) Poelt, R. phaeoplaca,

R. rossii Ovstedal, and R. viridis (Tonsberg) Tonsberg (Ekman 1993, 1996;

Tonsberg 1993; Brodo et al. 2001; Ovstedal et al. 2001; Smith et al. 2009; Fryday

& Coppins 2012).

In China, Ropalospora phaeoplaca has been reported only from Taiwan

(Aptroot & Sparrius 2003). Here we present records of R. phaeoplaca and

R. chlorantha from mainland China.

Materials & methods

The specimens studied are preserved in SDNU (Lichen Section of Botanical

Herbarium, Shandong Normal University) and KUN-L (Lichen Section, Cryptogamic

Herbarium of Kunming Institute of Botany, Academia Sinica). Morphological and

anatomical characters were examined under a stereomicroscope (COIC XTL7045B2)

and polarizing microscope (OLympus CX41). The lichen substances were identified

using standardized thin layer chromatography techniques (TLC) with system C and

microcrystal test (MCT) with GE solution (Orange et al. 2010). Photos of these lichens

were taken under OLymPus SZX16 and BX61 with DP72.

440 ... Hu & al.

Taxonomic descriptions

Ropalospora chlorantha (Tuck.) S. Ekman, Bryologist 96: 586 (1993) Figs 1-2

MorpHoLocy — Thallus corticolous, crustose, pale brown, discrete or

continuously verrucose areolate, thin, esorediate; prothallus black; medulla

with obvious crystals that are soluble in K and C. Apothecia lecideine, sessile,

0.4-0.8 mm diam.; disc black, concave; margin black, often flexuous. Exciple

externally brown, internally hyaline to straw-colored, without crystals;

epihymenium brown to black; hymenium hyaline; hypothecium hyaline, with

oil droplets. Asci clavate, 30-50-spored, Fuscidea-type; ascospores hyaline,

needle-shaped, 3-5-septate, with one attenuate end, curved, 26-35 x 1-2 um;

s : ; EO. % an ;

= a . oS he ee

Fic. 1 Ropalospora chlorantha (20100315a, SDNU). A: Thallus; B: Crystals in thallus; C: Prothallus;

D: Apothecia.

Ropalospora spp. new to China... 441

section, showing oil droplets in hypothecium; C: Apothecium section, showing exciple without

crystals; D: Apothecium section, showing K reaction (turning green) of epihymenium and outer

exciple; E: Ascus and ascospores; F: Amyloid reaction of ascus; G: Ascospores; H: Paraphyses.

paraphysis 1.5-2.0 um diam., branched in upper part and slightly thickened at

top. Pycnidia not observed.

CHEMISTRY — Thallus and medulla K-, C-, KC-, P-. Epihymenium and

exciple K+ green, C-. No lichen substances detected by TLC or MCT.

DISTRIBUTION — Ropalospora chlorantha has been reported from Canada,

USA, and Bhutan (Ekman 1993, Aptroot & Feijen 2002). New to China.

SPECIMEN EXAMINED: CHINA. Yunnan, Lijiang, Mt. Laojun, alt. 4000 m, on bark, 5 Nov

2009, Q. Tian 20100315a (SDNU).

442 ... Hu &al.

ComMENTs —Ropalospora chlorantha is characterized by the corticolous and

esorediate thalli, the exciple without crystals, the >30-spored asci, and perlatolic

acid. However, we detected no lichen substance in this specimen from China.

Ropalospora phaeoplaca (Zahlbr.) S. Ekman, Op. Bot. 127: 128 (1996) Figs 3-4

MorpHo.ocy — ‘Thallus corticolous, crustose, grayish-white, smooth to

verrucose areolate, without soredia; prothallus not seen; medulla with obvious

a ite eke g “a hey ~—

Fic. 3 Ropalospora phaeoplaca(20105161,SDNU). A: Thallus; B: Crystals in thallus; C: Apothecium

section, showing oil droplets in apothecium; D: Apothecium section, showing exciple with crystals;

E: Apothecium section, showing K reaction (turning green) of epihymenium and outer exciple;

F: Apothecium section, showing C reaction (turning green, then purple) of epihymenium and

outer exciple.

Ropalospora spp. new to China... 443

Fic. 4 Ropalospora phaeoplaca (20105161, SDNU). A: Amyloid reaction of asci; B: Ascus

and ascospores; C: Paraphyses; D: Ascospores; E: MCT result (GE solution); F: TLC result

(R = Reference).

crystals that are soluble in K and C. Apothecia lecideine, sessile, 0.4-0.9 mm

diam.; disk black, slightly convex; margin black, smooth or flexuose. Exciple

brown externally, inside pale brown to hyaline, with abundant crystals that are

soluble in K and C; epihymenium brown; hymenium hyaline; hypothecium

hyaline, with oil droplets. Asci clavate, 10-16-spored, Fuscidea-type; paraphysis

simple, slightly branched at top; ascospores hyaline, needle-shaped, 5-6

(-8)-septate, with one attenuate end, curved, 30-47 x 2-3 um. Pycnidia not

observed.

444 ... Hu &al.

CHEMISTRY —Thallus and medulla K-, C-, KC-, P-. Epihymenium and

outer exciple K+ green, C+ green then turn to purple. An unknown lichen

substance was detected by TLC and MCT.

DISTRIBUTION — Ropalospora phaeoplaca has been reported from Japan

and Taiwan (Ekman 1996, Aptroot & Sparrius 2003). New to mainland China.

SPECIMENS EXAMINED: CHINA. Fujian, Longyan, Mt. Huanglianyu, alt. 1400 m, on

bark, 29 Oct, 2010, H.Y. Wang 20105161, X.R. Kou 20105920 (SDNU). GuizHou,

Kaili, Mt. Leigong, alt. 1600 m, on bark, 23 Aug 2010, D.F Jiang 20103297, Z.L. Huang

20103179, 20103155, 20103128, L.L. Zhang 20103148a, 20103148b (SDNU); Tongren,

Mt. Fanjing, alt. 2000 m, on bark, 2 Nov 2009, Z.J. Ren 20128007 (SDNU). YUNNAN,

Chuxiong, Mt. Zixi, alt. 2100 m, on bark, 28 Aug 1994, L.S. Wang 94-15646 (HKAS).

ComMENTsS — Ropalospora phaeoplaca is closely related to R. chlorantha, but

differs in having minute K-soluble crystals in the exciple and usually eight

(occasionally <16) spores per ascus (Ekman 1996). The Chinese specimens

have abundant K-soluble crystals in the exciple, 10-16-spored asci, and an

unknown lichen substance.

Acknowledgements

The authors thank Dr. A. Aptroot (ABL Herbarium, Soest, the Netherlands) and

Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing, China) for presubmission reviews. This study

was supported by Program for Scientific Research Innovation Team in Colleges and

Universities of Shandong Province, and the National Natural Science Foundation of

China (31170187 & 31270059).

Literature cited

Aptroot A, Sparrius LB. 2003. New microlichens from Taiwan. Fungal Diversity 14: 1-50.

Brodo IM, Sharnoff DS, Sharnoff S. 2001. Lichens of North America. Yale University Press, New

Haven and London. 795 p.

Ekman S. 1993. A taxonomic study of Ropalospora chlorantha, and a comparison between

Ropalospora and Fuscidea. Bryologist 96: 582-591.

Ekman S. 1996. The corticolous and lignicolous species of Bacidia and Bacidina in North America.

Opera Botanica 127: 1-148.

Fryday AM, Coppins BJ. 2012. New taxa, reports, and names of lichenized and lichenicolous fungi,

mainly from the Scottish Highlands. Lichenologist 44: 723-737.

http://dx.doi.org/10.1017/S0024282912000369

Kantvilas G. 2004. Ropalospora. 186-187, in: PM McCarthy, K Mallett (eds). Flora of Australia,

volume 56 A, Lichens 4. ABRS, CSIRO Australia, Melbourne.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th Edition. CABI

Publishing, UK. 771 p.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens.

2nd edition. London: British Lichen Society.

@Ovstedal DO, Lewis Smith RI. 2001. Lichens of Antarctica and South Georgia: a guide to their

identification and ecology. Cambridge University Press, Cambridge, England. 411 p.

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA (eds). 2009.

The lichens of Great Britain and Ireland. Natural History Museum Publications, in association

with The British Lichen Society. 1046 p.

Tonsberg T. 1993. Additions to the lichen flora of North America II. Bryologist 96: 629-630.

MYCOTAXON

http://dx.doi.org/10.5248/123.445

Volume 123, pp. 445-450 January-March 2013

South Florida microfungi: a new species of Ellisembia

(hyphomycetes) with new records from the U.S.A.

GREGORIO DELGADO*

EMLab Pe&K North Phoenix, 1501 West Knudsen Drive, Phoenix, AZ 85027, U.S.A.

* CORRESPONDENCE TO: gdelgado@emlabpk.com

ABSTRACT — Ellisembia mercadoi sp. nov., collected on rachides of dead leaves of Sabal

palmetto in southwestern Florida, U.S.A., is described and illustrated. The fungus is

characterized by cylindrical, determinate conidiogenous cells and verruculose, subfusiform

to narrowly obclavate or obclavate-rostrate, 7-16 distoseptate conidia often with remnants

of an apical mucilaginous sheath. Similarities and differences with morphologically related

species within the genus are discussed. Berkleasmium leonense, Ceratosporella basibicellularia,

Hermatomyces tucumanensis, Monodictys capensis, and Selenosporella perramosa are newly

recorded from the U.S.A.

KEY worpDs — anamorphic fungi, palm fungi, Sporidesmium, taxonomy

Introduction

The southern tip of the Florida peninsula, with its humid subtropical

climate, extensive habitat diversity, and distinctive vegetation that includes

a high percentage of tropical plant species, is likely to harbor a rich but still

under-explored mycobiota. In the course of our continuing studies on saprobic

microfungi occurring in the area, particularly those hyphomycetous anamorphs

associated with dead plant debris (Delgado 2008a, b, 2009, 2010), a distinct

and unusual species of Ellisembia Subram. was found. The fungus agrees well

with the generic concept originally introduced by Subramanian (1992) for

Sporidesmium-like taxa with distoseptate conidia and conidiophores with or

without proliferating conidiogenous cells. However, it differs morphologically

from previously described Ellisembia species and therefore is proposed here

as new. Five other hyphomycetes are also recorded for the first time from the

U.S As

Materials & methods

Samples of dead leaves belonging to two palm tree species commonly found in

south Florida, the cabbage palm (Sabal palmetto (Walter) Lodd. ex Schult. & Schult.

446 ... Delgado

f.,) and the Everglades or paurotis palm (Acoelorrhaphe wrightii (Griseb. & H. WendL.)

H. Wendl. ex Becc.) were collected in forested areas of Broward and Collier counties

during 2007-2010. They were cut in smaller pieces, air-dried, and placed in plastic

bags for further processing and examination following Cannon & Sutton (2004). Slides

were prepped using lacto-cotton blue as mounting medium. Fungal structures were

examined, measured at 1000x, and photographed using an Olympus BX-45 microscope.

All specimens examined, including the type specimen of Ellisembia mercadoi as well

as semi-permanent slides, are deposited in the U.S. National Fungus Collections

Herbarium (BPI).

Taxonomy

Ellisembia mercadoi G. Delgado, sp. nov. FIGs 1-9

MycoBank MB# 804032

Differs from Ellisembia fusiformis in having cylindrical conidiogenous cells and longer

verruculose, brown, obclavate-rostrate, conidia with an apical mucilaginous sheath,

and from E. crassispora in having smaller conidia, determinate conidiogenous cells, and

shorter conidiophores.

Type — U.S.A. Florida, Collier Co., Naples, 26°3'35"N 81°41'43"W, on rachis of dead

leaf of Sabal palmetto, 23.X1.2007, coll. G. Delgado (Holotype: BPI 880515F).

EryMoLocy — Latin, mercadoi, in honor of the Cuban mycologist Dr. Angel Mercado-

Sierra (1937-2008) for his many contributions to the study of tropical and worldwide

hyphomycetes.

COLONIES on natural substrate effuse, hairy, inconspicuous. MycELIUM

predominantly immersed in the substrate, composed of branched, septate,

smooth-walled, pale brown to brown hyphae, 1-2.5 um wide. STROMATA none or

rudimentary, dark brown to blackish brown. CONIDIOPHORES macronematous,

mononematous, single or aggregated in small groups, simple, erect, straight or

flexuous, cylindrical, 2-5-septate, smooth, dark brown to dark reddish brown,

33-76 x 6-9 um, 10-14 um wide at the swollen base. CONIDIOGENOUS CELLS

monoblastic, integrated, terminal, cylindrical, determinate, dark brown to dark

reddish brown; apex truncate, often darkened. CONIDIAL SECESSION schizolytic.

Conip1A holoblastic, solitary, dry, acrogenous, straight or slightly curved,

subfusiform to narrowly obclavate, older conidia often obclavate-rostrate,

7-16-distoseptate, sometimes abruptly constricted at one distoseptum, brown,

paler toward the apex, verruculose, 63-168 x 12-15 um; apex usually rounded

or slightly acute and often with remnants of an apical mucilaginous sheath;

basal cell conico-truncate, dark brown to black. TELEOMORPH unknown.

ADDITIONAL SPECIMEN EXAMINED — U.S.A. FLORIDA, COLLIER Co., Naples, 26°3'35"N

81°41'43"W, on rachides of dead leaves of Sabal palmetto, 23.X1.2007, coll. G. Delgado

(BPI 880521L).

Ellisembia mercadoi sp. nov. (U.S.A.) ... 447

ca ee V9 ERS ~ &

Fics 1-14. Ellisembia mercadoi (holotype, BPI 880515F): 1-5. Conidia. 6. Conidium initials.

7. Young conidia. 8-9. Conidiophores and conidia. Ceratosporella basibicellularia (BPI 884153A):

10. Conidiophore and conidium. Monodictys capensis (BPI 880518D): 11. Conidiophores and

conidia. Berkleasmium leonense (BPI 884154G): 12. Sporodochium with conidia. Selenosporella

perramosa (BPI 884156D): 13. Conidiophore, conidiogenous cells and conidia. Hermatomyces

tucumanensis (BPI 884154D): 14. Conidia. Scale bars: 1-8 = 20 um; 9 = 15 um; 10-12; 13 = 30 um;

14 = 10 um.

448 ... Delgado

Discussion

The genus Ellisembia has recently undergone numerous additions (Ma et

al. 2008, 2010, 2011, McKenzie 2010, Rajeshkumar et al. 2012, Ren et al. 2012,

Santa Izabel et al. 2013) totaling 59 entries in the Mycobank Database (www.

mycobank.org). Wu & Zhuang (2005), however, had previously enlarged the

original concept of Subramanian (1992) after placing Imicles Shoemaker &

Hambl. (= Imimyces A. Hern. Gut. & B. Sutton) (Shoemaker & Hambleton

2001) into synonymy with Ellisembia. Consequently, a small group of additional

Sporidesmium-like taxa producing lageniform, doliiform, or ovoid percurrent

proliferations and distoseptate conidia could be transferred to Ellisembia

following this broad concept, but only three combinations were formally made:

E. bambusae (M.B. Ellis) W.P. Wu, E. carrii (Morgan-Jones) W.P. Wu, and E.

leptospora (Sacc. & Roum.) W.P. Wu. This left three species in Imicles (I. aquatica

(Cabello etal.) Shoemaker & Hambl., I. heterocateniformis (Matsush.) Shoemaker

& Hambl., I. pseudobambusae (P.M. Kirk) Shoemaker & Hambl.) and one in

Imimyces (I. hollowayensis A. Hern. Gut. & B. Sutton (Hernandez-Gutierrez &

Sutton 1997) as possible members of Ellisembia. Seifert et al. (2011), however,

retained Imicles as a separate genus, and in the case of I. heterocateniformis

(= Sporidesmium heterocateniforme Matsush.) (Hernandez-Gutierrez & Sutton

1997, Matsushima 1987), a reexamination of the type specimen is certainly

necessary to clarify its correct taxonomic placement.

Ellisembia mercadoi is characterized by dark brown to dark reddish brown,

cylindrical conidiophores often arising from a rudimentary stroma and

producing determinate, not proliferating, conidiogenous cells. The conidium

initial is distinctly mitriform, pale brown, and delimited by an incipient black

basal cell with the outer wall apparently showing signs of breakage resembling

longitudinal splits from the pointed apex downwards. After elongation, the

young conidia become cylindrical or subcylindrical with a slightly acute apex,

and when fully developed they are usually subfusiform or narrowly obclavate

but also obclavate-rostrate with a well-defined conico-truncate basal cell.

Remnants of an apical mucilaginous sheath were often visible near the conidial

apex as traces of mucilage or as short, lateral and subapical, thread-like filaments.

A well defined cap as seen in many other Ellisembia species (McKenzie 1995,

Wu & Zhuang 2005) was not observed, although further collections may

provide evidence of its presence.

Among the accepted Ellisembia species, E. mercadoi is morphologically

similar to E. fusiformis (Nees & T. Nees) Subram. (Ellis 1958) in conidial shape

and width, number of septa, the presence of a black conico-truncate basal cell,

and determinate conidiogenous cells. However, E. fusiformis has dark reddish

brown smooth-walled non-rostrate shorter conidia (<136 um long) without an

apical mucilaginous sheath, and the conidiogenous cells are not darkened and

Ellisembia mercadoi sp. nov. (U.S.A.) ... 449

taper abruptly toward the apex. Ellisembia crassispora (M.B. Ellis) Subram. (Ellis

1958, Wu & Zhuang 2005) also has verruculose, obclavate and rostrate conidia

with a conico-truncate basal cell but clearly differs in longer conidiophores

(<300 um long) with up to 3 percurrent proliferations and dark brown to

blackish brown larger conidia (100-250 x 18-30 um) with up to 22 distosepta

and also lacking a mucilaginous sheath.

Additional new records from U.S.A.

Berkleasmium leonense M.B. Ellis, More Dematiaceous Hyphomycetes: 69, 1976.

Fic. 12

SPECIMEN EXAMINED: U.S.A.. FLORIDA, BROWARD Co., Plantation, Plantation Heritage

Park, Anne Kolb Memorial Trail, 26°6'25"N 80°13'19"W, on rachides of dead leaves of

Acoelorrhaphe wrightii, 30.V.2010, coll. G. Delgado (BPI 884154G).

Ceratosporella basibicellularia Matsush., Matsush. Mycol. Mem. 7: 45, 1993.F1G. 10

SPECIMEN EXAMINED: U.S.A.. FLORIDA, BROWARD Co., Plantation, Plantation Heritage

Park, Anne Kolb Memorial Trail, 26°6'25"N 80°13'19"W, on petiole of dead leaf of

Acoelorrhaphe wrightii and overgrowing the surface of old hysterothecia, 30.V.2010, coll.

G. Delgado (BPI 884153A).

Hermatomyces tucumanensis Speg., Anal. Mus. nac. Hist. nat. B. Aires 13: 446,

LOT) FIG. 14

SPECIMEN EXAMINED: U.S.A.. FLORIDA, BROWARD Co., Plantation, Plantation Heritage

Park, Anne Kolb Memorial Trail, 26°6'25"N 80°13'19"W, on rachides of dead leaves of

Acoelorrhaphe wrightii, 30.V.2010, coll. G. Delgado (BPI 884154D).

Monodictys capensis R.C. Sinclair, Boshoff & Eicker, Mycotaxon 59: 359, 1996. Fic. 11

SPECIMENS EXAMINED: U.S.A.. FLORIDA, COLLIER Co., Naples, 26°3'35"N 81°41'43"W,

on rachides of dead leaves of Sabal palmetto, 23.XI.2007, coll. G. Delgado (BPI 880518D,

880521E, 880515C); BRowARD Co., Plantation, Plantation Heritage Park, Anne Kolb

Memorial Trail, 26°6’25"N 80°13'19"W, on rachides and petioles of dead leaves of

Acoelorrhaphe wrightii, 30.V.2010, coll. G. Delgado (BPI 884154F, 884155A).

Selenosporella perramosa (WB. Kendr. & R.F. Castafieda) R.F. Castafieda,

Mycotaxon 109: 69, 2009. FIG. 13

SPECIMENS EXAMINED: U.S.A.. FLORIDA, BROWARD Co., Plantation, Plantation Heritage

Park, Anne Kolb Memorial Trail, 26°6'25"N 80°13'19"W, on rachides, petiole, petiole-

lamina junction and segments of dead leaves of Acoelorrhaphe wrightii, 30.V.2010, coll.

G. Delgado (BPI 884151C, 884152D, 884154B, 884156D).

Acknowledgments

I am grateful to Drs. Eric McKenzie (Landcare Research) and Vadim Melnik

(Komarov Botanical Institute) for critically reading the manuscript and serving as

pre-submission reviewers. Thanks are also due to Dr. Zhao Guo-Zhu (Beijing Forestry

University) for the translation of a Chinese description of Monodictys capensis, Dominick

Shannon (BPI, USDA) for depositing specimens in BPI, and Elsa Delgado for assistance

450 ... Delgado

in the field. Dr. Kamash Ramanathan (EMLab P&K) is acknowledged for provision of

laboratory facilities and financial support.

Literature cited

Cannon P, Sutton B. 2004. Microfungi on wood and plant debris. 217—239, in: G Mueller et al.

(eds). Biodiversity of fungi: inventory and monitoring methods. Academic Press, Burlington.

Delgado G. 2008a. South Florida microfungi: a new species of Stanjehughesia (hyphomycetes) from

Sabal palm. Mycotaxon 103: 229-234.

Delgado G. 2008b. South Florida microfungi: new records of saprophytic hyphomycetes on plant

debris. Florida Scientist 71: 76-89.

Delgado G. 2009. South Florida microfungi: Veramycella bispora, a new palmicolous anamorphic

genus and species, with some new records for the continental U.S.A.. Mycotaxon 107: 357-373.

http://dx.doi.org/10.5248/107.357

Delgado G. 2010. South Florida microfungi: Kalamarospora multiflagellata gen. et sp. nov.

(hyphomycetes), with additional new records from U.S.A.. Mycotaxon 114: 231-246.

http://dx.doi.org/10.5248/114.231

Ellis MB. 1958. Clasterosporium and some allied Dematiaceae-Phragmosporae I. Mycol. Pap. 70:

1-89.

Hernandez-Gutierrez A, Sutton BC. 1997. Imimyces and Linkosia, two new genera segregated

from Sporidesmium sensu lato, and redescription of Polydesmus. Mycol. Res. 101: 201-209.

http://dx.doi.org/10.1017/S0953756296002419

Ma J, Zhang K, Zhang XG. 2008. Two new Ellisembia species from Hainan, China. Mycotaxon 104:

141-145.

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, Zhang XG. 2011. Three new hyphomycetes from southern China.

Mycotaxon 117: 247-253. http://dx.doi.org/10.5248/117.247

Matsushima, T. 1987. Matsushima Mycological Memoirs No. 5: 1-100.

McKenzie EHC. 1995. Dematiaceous hyphomycetes on Pandanaceae. 5. Sporidesmium sensu lato.

Mycotaxon 56: 9-29.

McKenzie EHC. 2010. Three new phragmosporous hyphomycetes on Ripogonum from an ‘ecological

island’ in New Zealand. Mycotaxon 111: 183-196. http://dx.doi.org/10.5248/111.183

Rajeshkumar KC, Kajale S, Sutar S, Singh SK. 2012. Ellisembia karadkensis sp. nov. from southern

Western Ghats, India. Mycotaxon 121: 181-186. http://dx.doi.org/10.5248/121.181

Ren SC, Ma J, Zhang XG. 2012. Two new Ellisembia species from Hainan and Yunnan, China.

Mycotaxon 122: 83-87. http://dx.doi.org/10.5248/122.83

Santa Izabel TS, Cruz ACR, Gusmao LFP. 2013. Conidial fungi from the semi-arid Caatinga biome

of Brazil. Ellisembiopsis gen. nov., new variety of Sporidesmiella and some notes of Sporidesmium

complex. Mycosphere 4: 156-163.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS- KNAW

Fungal Biodiversity Centre, Utrecht.

Shoemaker RA, Hambleton S. 2001. “Helminthosporium” asterinum, Polydesmus elegans, Imimyces,

and allies. Can. J. Bot. 79: 592-599.

Subramanian CV. 1992. A reassessment of Sporidesmium (hyphomycetes) and some related taxa.

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Diversity Press, Hong Kong.

MY COTAXON

http://dx.doi.org/10.5248/123.451

Volume 123, pp. 451-456 January-March 2013

Aureoboletus zangii (Boletaceae), a new species from China

X1A0-FEI SHI *?& PEI-Gu1 Liu”

' Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany

Chinese Academy of Sciences, Kunming 650201, Yunnan, China

? University of Chinese Academy of Sciences, Beijing 100049, China

* CORRESPONDENCE TO: pgliu@mail.kib.ac.cn

ABSTRACT —Aureoboletus zangii sp. nov. is described from central China based on

morphological and molecular analysis. This species is found fruiting in association with

hardwood trees. It is similar to the European A. gentilis and Asian A. thibetanus but is

characterized by the yellowish brown or reddish golden basidiomata, glutinous pileus

with subtomentum, and viscid stipe. LSU sequence analysis supports the new species in

Aureoboletus (Boletaceae). Photographs, line drawings, and a phylogenetic tree showing

relationships with closely allied taxa are provided.

Key worps — Chinese fungal diversity, Boletales, taxonomy

Introduction

Two Aureoboletus species have previously been reported from China,

A. thibetanus (Pat.) Hongo & Nagas. (Ying & Zang 1994) and A. reticuloceps

M. Zang et al. (Zang et al. 1993), although the latter species has been transferred

to Boletus (Wang & Yao 2005). Worldwide only 10 Aureoboletus species are

recognized (Kirk et al. 2008). Although Pouzar named Aureoboletus with the

type A. gentilis (Quél.) Pouzar in 1957 (Corner 1972, Kirk et al. 2008), the

genus was not accepted for some time (Smith & Thiers 1971, Corner 1972,

Singer 1986, Sutara 2005). After recent molecular analyses by Binder (1999)

and Binder & Hibbett (2006) helped resolve the taxonomy of the genus, Klofac

(2010) presented a world monograph in which he transferred 13 additional taxa

to Aureoboletus and presented a species key (including some critical species in

other genera). The morphological characters differentiating this genus from

other bolete genera include: medium-small sized basidiomata, a pileus that is

both viscid and subtomentose, yellow-colored tubes and pores, a hymenophoral

surface more or less depressed around the stipe apex, a cylindrical to slightly

swollen glutinous stipe that is never reticulate nor glandulose, and (most

species) ectomycorrhizal with frondose trees (Klofac 2010).

452 ... Shi & Liu

Materials & methods

Macroscopic and microscopic descriptions were based on fresh and dried material

respectively. Color codes follow Kornerup & Wanscher (1961). For microscopical

observations, slides were revived in 5% KOH solution and Melzer’s reagent. The

observation of the pileipellis structure was performed following Wang & Verbeken

(2006). Micro-morphological features were examined under a Nikon E400 microscope

at 1000x magnification and drawings were made through a drawing tube (Y-IDT)

installed on the microscope. For further explanation of basidiospore data, see Yang

(2000). All specimens examined were deposited in the Herbarium of Cryptogams,

Kunming Institute of Botany, Chinese Academy of Sciences (KUN-HKAS).

Total DNA was extracted from silica-dried materials using a modified CTAB

procedure by Doyle & Doyle (1987). The polymerase chain reaction (PCR) was performed

using the primer pairs LROR/ LRS5 to amply the ribosome larger subunit (nrDNA-LSU)

region of the nuclear ribosomal DNA (White et al. 1990). Sequencing of both strands

was performed with an ABI 3730 DNA analyzer and an ABI BigDye 3.1 terminator cycle

sequencing kit (Shanghai Sangon Biological Engineering Technology & Services CO.,

Ltd, Shanghai). The sequences have been submitted to GenBank (accession numbers

JQ734420-734422),

Taxonomy

Aureoboletus zangii X.F. Shi & P.G. Liu, sp. nov. FIGs 1, 2

MycoBAnk 564683

Differs from Aureoboletus gentilis and A. thibetanus by its concolorous stipe and pileus

and its shorter basidiospores.

Type: China, Shaanxi Province, Baoji City, Meixian County, Haoping Town, Dali

Village, 34°05'N 107°42’E, alt. 1200 m, 4.1X.2010, X.F. Shi 654 (Holotype, HKAS63217;

GenBank JQ734420).

ETyMOLOoGy: zangii is in honor of Prof. Mu Zang (28 Dec. 1930-10 Nov. 2011), a

distinguished Chinese mycologist who founded the Cryptogamic Herbarium at

Kunming Institute of Botany, Academia Sinica (KUN-HKAS).

MACROCHARACTERS — BASIDIOMATA small to medium, scarcely fleshy. PILEUS

at first hemispherical then expand to convex, 3-4 cm in diam; surface glutinous

when fresh, grayish orange to yellowish brown or reddish golden to light brown

(6B3, 5D5—-5E5, 6C4-6D5); with concolorous subtomentum, without sterile

margin. STIPE 5.5-7 x 0.5-0.8 cm, cylindrical or clavate, usually tapering

towards the base, mostly concolorous with the pileus or more reddish, light

brown to brownish orange (7C6, 6D5), sometimes darkening towards the base,

violet-brown (11E8), sometimes changing to reddish-white (9A2) or violet-

brown (10F7) when bruised, distinctly viscid especially in wet weather, entirely

fistulose and showing white mycelium at the base. CONTEXT 0.2-0.3 cm thick,

pale yellow or orange-white (2A3, 6A2), always more or less pinkish under the

cuticle, unchanging when bruised or exposed to air. TuBEs 0.3-0.5 cm deep,

vivid yellow at first and then olive yellow, grayish yellow or vivid yellow (2C6,

Aureoboletus zangii sp. nov. (China) ... 453

Fic 1. Aureoboletus zangii (Holotype).

2B7, 2A8, 2A6), unchanging when exposed to air. POREs 1-2 per mm, bright

yellow or concolorous with tubes, unchanging, depressed around the stipe.

Ovor not distinctive. TasTE mild.

MICROCHARACTERS — BASIDIOSPORES [80/3/3] (10-)10.5-11.0 x (4.0-)

4.5-5.0 [m, Q = (2.1-)2.2-2.4(-2.5), Q. = 2.23 + 0.09, elongate elliptical,

yellowish-olivaceous in 5% KOH, thin-walled, sometimes with tiny oil drops;

yellow to yellowish-brown in Melzer’s. BAsip1a 30-38 x 11-14 um, clavate, with

4 sterigmata. PLEUROCYSTIDIA 35-65 x 11-18 um, narrowly clavate, yellow in

5% KOH and lacking any incrustations. CHEILOCYSTIDIA 35-42.5 x 11-17 um,

similar to pleurocystidia. CAULOCysTIDIA absent. HYMENOPHORAL TRAMA

divergent, with hyphae 4-5 um in diameter, thin-walled. PILEIPELLIs a layer

consisting of mostly inflated hyphae tending to repent under the gelatinous

layer, slightly brownish in 5% KOH, 7-10 um in diameter.

454 ... Shi & Liu

Fic 2. Aureoboletus zangii (Holotype). a. Basidiospores; b. Cystidia; c. Hymenium; d. Pileipellis.

ECOLOGY & DISTRIBUTION —Solitary, rarely gregarious, widespread in

central China, on the ground, in subtropical broad-leaved mixed forests

dominated by Cyclobalanopsis and Quercus, alt. 1200-1400 m.

ADDITIONAL SPECIMENS EXAMINED — CHINA. GuizHou, Daozhen: near Yangxi

Town, 28°50'N 107°36’E, alt. 1200 m, 28.V1I.2010, X.F. Shi 394 (HKAS 63220; GenBank

JQ734422); Yangxi Town, Dashahe Conservation Area, 28°50'N 107°36’E, alt. 1400 m,

27.VIL.2010, X.F. Shi 388 (HKAS 63219; GenBank JQ734421).

Aureoboletus zangii sp. nov. (China) ... 455

Pulveroboletus ravenelii DQ534662**

Chalciporus piperatus AF336244***

Boletellus ananas AY612799

Heimiella retispora AFO50650*

Aureoboletus thibetanus AY700189

Aureoboletus zangii

| apelD

Aureoboletus gentilis DQ534635**

Aureoboletus auriporus DQ534636**

Boletus edulis AFO050643*

10 Octaviania asterosperma DQ534619**

Fic 3. The MP tree from a phylogenetic analysis of LSU sequences of Boletaceae. Numbers above

branches are bootstrap values (>50%). Sequences obtained in this study are presented in bold front;

* = sequences by Binder & Fischer (1997); ** = sequences by Binder (2006); *** = sequences by

Wang et al.(2002); all other sequences were downloaded from Genbank.

Phylogenetic results

Three samples were used for phylogenetic analysis. Aureoboletus zangii

fell into the well-supported Clade I in Fic. 3. Phylogenetic analysis of nLSU

supports A. zangii should be a member of the genus Aureoboletus along with

the European species A. gentilis and the Asian species A. thibetanus.

Discussion

The phylogeny confirms the new taxonas sister to A. gentilisand A. thibetanus.

Aureoboletus zangii is similar to the European A. gentilis in its viscid reddish

orange cap and viscid stipe, but the stipe of the new species is concolorous

with its pileus, while in A. gentilis the stipe is bright yellow with just pinkish to

brownish tinges in places. Aureoboletus zangii might also be confused with the

Asian A. thibetanus, which can be distinguished by its reddish brown shrunken

cap and yellowish white stipe. Anatomically, the A. gentilis spores are with

largest (<15 um long) with those of A. thibetanus around 12um long and those of

A. zangii around 10 um long. In addition, the A. zangii cystidia are <65 um long

while those of A. gentilis and A. thibetanus are <70 um long (Klofac 2010; Yang

et al. 2003). Moreover, the outer surface of A. thibetanus is characteristically

covered by a 5-8 um thick layer of a strongly refractive yellow substance that is

quickly and completely dissolved in 5% KOH (Yang et al. 2003).

456 ... Shi & Liu

Acknowledgements

The authors are grateful to the two peer reviewers, Dr. Matteo Gelardi and Prof.

Taihui Li, who critically reviewed the manuscript and provided invaluable suggestions.

This study was financed by the National Science Foundation of China (No. 31270075),

the Joint Founds of the National Science Foundation of China and Yunnan Province

Government (No. 1202262), and Yunnan Program of Innovation to Strong Province by

Science & Technology (No. 2009AC013).

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Binder M, Hibbett DS. 2006. Molecular systematics and biological diversification of Boletales.

Mycologia 98: 971-981.

Corner EJH. 1972. Boletus in Malaysia. Singapore Publishing House.

Doyle JJ, Doyle JL. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue.

Phytochem. Bull. 19: 11-15.

Kirk PM, Cannon PF, Minter DW, Stalpers JA (eds). 2008. Dictionary of the fungi. 10th edition.

CABI Publishing, UK.

Klofac W. 2010. The genus Aureoboletus, a world-wide survey. A contribution to a monographic

treatment. Osterr. Z. Pilzk. 19: 133-174.

Kornerup A, Wanscher JH. 1961. Farver i Farver. Politikens Forlag. Kobenhavn.

Singer R. 1986. The Agaricales in modern taxonomy. 4th ed. Koenigstein, Koeltz Scientific Books.

Smith AH, Thiers HD. 1971. The Boletes of Michigan. Ann Arbor. Michigan, The University of

Michigan Press.

Sutara J. 2005. Central European genera of the Boletaceae and Suillaceae, with notes on their

anatomical characters. Ceska Mykol. 57(1-2): 1-20.

Wang QB, Yao YJ. 2005. Boletus reticuloceps, a new combination for Aureoboletus reticuloceps.

Sydowia 57(1): 131-136.

Wang XH, Verbeken A. 2006. Three new species of Lactarius subgenus Lactiflui (Russulaceae,

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http://dx.doi.org/10.1127/0029-5035/2006/0083-0167

White TJ, Bruns TD, Lee SB, 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 applications. San Diego: Academic Press.

Yang ZL. 2000. Species diversity of the genus Amanita (Basidiomycetes) in China. Acta Botanica

Yunnanica 22(2): 135-142.

Yang ZL, Wang XH, Binder M. 2003. A study of the type and additional materials of Boletus

thibetanus. Mycotaxon 86: 283-290.

Ying JZ, Zang M.1994. Economic macrofungi from southwestern China. Beijing, Science Press.

Zang M, Yuan MS, Gong MQ. 1993. Notes on and additions to Chinese members of the Boletales.

Acta Mycologica Sinica 12(4): 275-282.

MY COTAXON

http://dx.doi.org/10.5248/123.457

Volume 123, pp. 457-465 January-March 2013

Molecular characterization of Colletotrichum species

causing soybean anthracnose in Argentina

ARACELI M. RAmos', Luis FRANCO TapiIc’, ISABEL CINTO?’,

MARCELO CARMONA? & MARCELA GALLY?

' Departamento BBE, Lab. 9, FCEN (UBA), PROPLAME-PRHIDEB (CONICET) &

? Catedra de Fitopatologia, Facultad de Agronomia, UBA C.A.B.A. Argentina

* CORRESPONDENCE TO isa.cinto@gmail.com or icinto@bg.fcen.uba.ar

ABSTRACT —Twenty-six isolates obtained from soybean crops (Glycine max) with typical

anthracnose symptoms were identified as Colletotrichum truncatum (73 %) and C. destructivum

(26 %). Their genetic relationships were studied using the AFLP method. A UPGMA

phenogram divided the strains into two clusters corresponding with the two species. Genetic

distances based on association coefficient were 0.71-0.89 among the 18 C. truncatum strains

and 0.67-1 among the eight C. destructivum strains. Genetic variability within species,

measured in terms of percentage of polymorphic loci, was high (<90%). Only two isolates

showed 100% similarity, suggesting high intraspecific variability.

Key worps —Glomerella glycines, teleomorph, genetic diversity

Introduction

Soybean (Glycine max (L.) Merr.) is the most important crop in Argentina,

with late-season diseases caused by Septoria glycines, Cercospora kikuchii,

Phomopsis sojae, Colletotrichum destructivum, and C. truncatum causing major

losses (Carmona et al. 2009, 2011).

The two anthracnose pathogens most frequently associated with soybean are

Colletotrichum truncatum (Schwein.) Andrus & W.D. Moore and C. destructivum

O’Gara, [= Glomerella glycines Lehman & EA. Wolf]. The traditional

characterization and identification of Colletotrichum species has relied on

morphological differences (Sutton 1992). Molecular markers constitute

the current choice to obtain diagnostic characters as an aid for species-level

identification and to evaluate genetic diversity. The amplified fragment length

polymorphism (AFLP) method (Vos et al. 1995) is frequently used to monitor

the identity of an isolate or to determine the degree of similarity among isolates,

owing to certain advantages over other techniques (i.e. high level of identified

polymorphism, high reproducibility, relative technical simplicity; Briad et

458 ... Ramos & al.

al. 2000). The taxonomy and nomenclature of the anthracnose pathogens

(Colletotrichum spp.) is confusing (Hyde et al. 2009 a) and have recently been

summarised by Hyde et al. (2009b), incorporating data obtained by DNA

analysis and phylogenetic systematics.

Physiological studies have been performed on C. truncatum isolates obtained

from soybean plants in Argentina (Levin et al. 2007, Ramos et al. 2010), but the

genetic structure of anthracnose fungi associated with the Argentinean soybean

crop is not available. Our study aimed to evaluate genetic variability and

genetic relationships within Colletotrichum species obtained from symptomatic

soybean plants in Argentina.

Materials & methods

Fungal Isolation

Diseased soybean plants showing typical anthracnose symptoms were collected from

different geographic locations in Argentina. The isolates were obtained from lesions of

stems and pods, using the procedure of Levin et al. (2007).

All fungal strains used in this survey were deposited at the Herbarium and Culture

Collection of the Departamento de Biodiversidad, Facultad de Ciencias Exactas y

Naturales Universidad de Buenos Aires (BAFC). TABLE 1 shows the collection locality

and BAFC accession number of each strain used.

TABLE 1. Colletotrichum truncatum and C. destructivum fungal strains studied

SPECIES COLLECTION LOCALITY* STRAINS TESTED *

C. truncatum Godeken, SE, Arg. Cl (3093)

Cerro Azul, SE, Arg. C2 (3094)

Armstrong, SF, Arg. C4 (3096), C5 (3097)

Cruz Alta, CO, Arg. C6 (3098)

Fac. Agronomia CABA, Arg. C7 (3099)

Lobos, BA, Arg. C8 (3100), C9 (3101)

Saenz Pefia, CH, Arg. C10 (3102)

Salto, BA, Arg. C14 (3396)

Las Lajitas, SA, Arg. C15 (3397)

Piamonte, SF, Arg. C26 (3398)

Otamendi, BA, Arg. C24 (3399), C32 (3448)

Las Varillas, SE, Arg. C25 (3400)

Fac. Agronomia CABA, Arg. C29 (3443), C30 (3444)

Venado Tuerto, SE, Arg. C31 (3447)

C. destructivum Urdinarrain, ER, Arg. C16 (3434)

Chivilcoy, BA, Arg. C17 (3435)

Rio Cuarto, CO, Arg. C18 (3436)

9 de Julio, BA, Arg. C19 (3437)

Barranca Yaco, SF, Arg. C21 (3439)

Piamonte, SF, Arg. C23 (3441)

Chivilcoy, BA. Arg. C27 (3401), C28 (3442)

* Abbreviations: BA = Buenos Aires Province; CABA = Ciudad Auténoma de Buenos Aires;

CH = Chaco Province; CO = Cordoba Province; ER = Entre Rios Province; LR = La Rioja

Province; SA = Salta Province; SF = Santa Fé Province; Arg.: = Argentina.

*Numbers in parentheses correspond to the BAFC culture number (see Materials & methods).

Colletotrichum anthracnose on soybean (Argentina) ... 459

TABLE 2. AFLP primers used in this study

PRIMERS SEQUENCE 5-3

E+AC GAC TGC GTA CCA ATT CAC

E+AA GAC TGC GTA CCA ATT CAA

E+ACG GAC TGC GTA CCA ATT CACG

E+AAC GAC TGC GTA CCA ATT CAA C

E+AAG GAC TGC GTA CCA ATT CAA G

M+A GAT GAG TCC TGA GTA AA

M+G GAT GAG TCC TGA GTA AG

M+C GAT GAG TCC TGA GTA AC

M+T GAT GAG TCC TGA GTA AT

* E = EcoRI; M = Msel.

DNA extraction

Actively growing mycelia were obtained and then ground in liquid nitrogen (Ramos

et al. 2000). Genomic DNA was extracted following Gottlieb & Lichtwardt (2001).

Quality control and quantification of genomic DNA was carried out by agarose gel

(0.8% w/v) electrophoresis and by comparison with a DNA molecular-size standard

(Lambda EcoRI/HindIH, Promega Corp.). Gels, containing ethidium bromide, were

photographed under UV light.

AFLP

The AFLP methodology was carried out on 250 ng of genomic DNA using the

AFLP® Analysis System for Microorganisms Primer Kit (Invitrogen) as described in the

instructions manual with minor modifications (Gottlieb et al. 2005). Selective primers

used herein, containing one, two or three additional bases, were combined as in TABLE 2.

All PCR amplifications were performed in a TECHNE PROGENE thermal cycler.

Polyacrylamide gel electrophoresis conditions were conducted as in Gottlieb et al.

(2005). A 30-330-bp AFLP’ DNA Ladder (Invitrogen) size marker was included twice

in each electrophoresis run. Thus, the size of scored AFLP bands ranged from 90 to

330 bp. AFLP bands were visualized using the SILVER SEQUENCE ™ DNA Sequencing

System (Promega). Air-dried gels were digitalized and visually analyzed using the Adobe

Photoshop TM (Adobe Systems, Mountain View, CA, USA).

Data analysis

Each AFLP band, regardless of its relative intensity, was considered as a dominant

allele at only one locus. The data, as either present (1) or absent (0), were extracted

as a table. Monomorphic bands (bands present in all individuals of a species) were

discriminated within each species and across the entire data set. The binary matrix

was analyzed with the program NTSYS-PC version 2.02i (Rohlf 1993). The unweighted

pair-group method with arithmetic mean (UPGMA) cluster analysis was performed

based on the simple matching (SM), Jaccard (J), and Dice (D) association coefficients

(Sneath & Sokal 1973). Phenograms showing similarity relations were generated by the

same program, with the tree display option (TREE). Correlation cophenetic coefficients

(r) were calculated for each association coefficient (Sokal & Rohlf 1962). A cophenetic

correlation (r) of 0.8 was considered a good fit. A three-dimensional graphic was

obtained with the Principal Coordinates ordination method (Gower 1966).

The percentage of polymorphic loci (P = number of polymorphic loci/ number of

loci analyzed) for each primer combination was calculated.

460 ... Ramos & al.

Results

Twenty-six isolates obtained from soybean crops that showed typical

anthracnose symptoms were identified as Colletotrichum truncatum (73%) and

C. destructivum (26%), based on their morphological characteristics (Sutton

1992).

Characterization of AFLP markers

Six primers (TABLE 3) that produced high numbers of clear, reproducible

bands by AFLP amplification were selected. The analysis of the six primers

yielded a total of 812 bands that appeared consistently in all experiments among

the amplified fragments of the 26 isolates. The size of the amplified bands ranged

from 50 to 550 bp but we analyzed only bands from 90 to 330 bp.

The total number of AFLP bands (polymorphic and monomorphic) obtained

and the percentage of polymorphic loci for each selective primer combination

in two fungal species assayed are described in TABLE 3. All primers detected a

high number of polymorphic bands within species and a high percentage of

polymorphic loci was obtained.

TABLE 3. Total AFLP bands (polymorphic and monomorphic) detected for each

primer combination in each assayed fungal species with polymorphic loci

percentages (P%).

PRIMER SPECIES ToTAL N° POLYMORPHIC MONOMORPHIC P%

COMBINATION OF BANDS BANDS BANDS

M+T/E+ACG C. truncatum 73 65 8 89

C. destructivum 81 74 7 91

M+C/E+AAG C. truncatum 117 111 6 95

C. destructivum 111 109 2 98

M+A/E+AAG C. truncatum 95 93 2 98

C. destructivum 56 51 5 91

M+A/E+ACG C. truncatum 73 71 2 97

C. destructivum 100 96 4 96

M+G/E+ACG C. truncatum 82 80 2 97

C. destructivum 54 52 2 96

M+A/E+AAC C. truncatum 103 82 21 79

C. destructivum 56 48 8 86

Only 41 (7.6%) of the bands obtained in C. truncatum and 28 (6.1%) in

C. destructivum were monomorphic. Genetic variability within species,

measured in terms of percentage of polymorphic loci, was high: C. truncatum

(P = 92%) and C. destructivum (P = 94%).

The UPGMA phenogram (FiGuRE 1) grouped all strains into two defined

clusters with a SM coefficient of 0.6 between them. The main cluster included

Colletotrichum anthracnose on soybean (Argentina) ... 461

C1 )

ve eS)

24 >

$G1 < 8

26 =

G1 C2 ie

C3 10

SG2 14

‘o)

16 °

SG3 17 rm

G2 18 gS

C5 19 |

SG4 aes

C6 27 =

0.60 0.68 0.77 0.85 093

Coefficient

FiGuRE 1. UPGMA phenogram showing relationships of Colletotrichum truncatum and

C. destructivum isolates based on the simple matching (SM) association coefficient estimated from

AFLP loci.

the 18 isolates identified as C. truncatum (FiGuRE 1, G1), whereas the other

(FIGURE 1, G2) included the eight isolates identified as C. destructivum. There

was no direct correlation between molecular genotype and geographic origin.

Genetic distance and cluster analysis

The phenogram obtained using the UPGMA method was constructed

based on Simple Matching (SM) coefficients. Little distortion occurred while

constructing this phenogram, as implied by the value of the correlation

cophenetic coefficient (r = 0.91903).

The main group (FicurE 1, G1) comprised two subgroups (SG1 and SG2),

one with 14 isolates and two conjuncts (Cl and C2), and the other (SG2)

with two conjuncts (C3 and C4), each with two isolates. Group 2, formed by

C. destructivum isolates, showed two subgroups, SG3 formed by three isolates

(16, 17 and 18), and SG4, which included conjunct C5 and C6 formed by the

rest of isolates belonging to C. destructivum. The relatedness of the 18 isolates of

C. truncatum was 0.71-0.89, whereas that among the 8 isolates of C. truncatum

was 0.67-1. Only two isolates (16 and 17, SG3) showed 100% similarity.

The three-dimensional graphic generated by the ordination technique

distinguished two groups (FIGURE 2, GI and GII) and showed nearly the same

relations between isolates as the phenogram (FiGurRe 1). The first group included

the 18 isolates belonging to C. truncatum, which are slightly differentiated in

462 ... Ramos & al.

C. truncatum

GII

C. destructivum

FIGURE 2. Three-dimensional graphic obtained with

the Principal Coordinates ordination technique.

axis 1 and largely separated in the other two axes. The second group showed

the seven isolates of C. destructivum largely separated in axis 1 and moderately

differentiated in axes 2 and 3. Group GI included isolates of SGI and SGII of the

phenogram and group GII comprised the isolates of SGHI and SGIV.

Discussion

C. truncatum and C. destructivum are highly variable pathogens in Argentina.

Sharma (2009) also reported a high degree of variability among different

isolates of C. truncatum studied through Random Amplified Polymorphic

DNA (RAPD) analysis.

Perithecia of the C. destructivum teleomorph (the state referred to in dual

nomenclature as Glomerella glycines) are frequently found in the fields of

Argentina, indicating that the sexual mechanism is an important source of

variability in the populations of this species. However, sexual recombination in

most Colletotrichum species is rare in nature (Wharton & Diéguez-Uribeondo

2004; Hyde et al. 2009b).

Nevertheless, in our study, we found a high degree of variability in the

populations analyzed. Alternatives to sexual recombination for genetic

interaction among fungi are heterokaryosis and mitotic recombination (Esser

Colletotrichum anthracnose on soybean (Argentina) ... 463

& Kuenen 1967). In many Colletotrichum populations, the only means of

exchanging genetic material between two strains is mainly or exclusively

vegetative, by means of anastomosis and heterokaryosis (Correll et al. 2000).

These processes have been studied in some Colletotrichum species (Vaillancourt

1994; Correll et al. 1993; Elmer 2001; Nitzan et al. 2002; Rodriguez-Guerra et

al. 2003; Barcelos et al. 2011). Sant’Anna et al. (2010) related genetic diversity

analyzed by RAPD with the presence of vegetative compatibility groups in

C. truncatum, and concluded that the parasexual process might be an important

factor to increase the genetic variability in the pathogen.

Almost the entire soybean crop area of Argentina is cultivated under

no-tillage systems. The crop debris remaining on the soil surface provides

optimum conditions for the active development of reproductive structures of

C. truncatum and C. destructivum. Any event of mutation or sexual or vegetative

recombination is enhanced by the high reproductive rate occurring in these

conditions. Gene flow and migration forces seem to be relatively unimportant

due to the weak dispersal ability of the spores, as both conidia and ascospores

are immersed in a mucous matrix, which prevents dispersal, by wind. The only

possibility of migration to long distances is by means of seed transmission.

Nevertheless, both species are not frequent seed pathogens in the Pampean

Region.

The level of genetic variability within populations of plant pathogens provides

an indication of their adaptability of pathogen in overcoming plant resistance

and other measures used to control the pathogen, such as the application of

fungicides in agro-ecosystems (McDonald et al. 1989; McDonald & Linde

2002). In our study, the high genetic diversity indicates increasing fitness of the

population.

This paper constitutes the first report of genetic variability in C. truncatum

and C. destructivum isolates collected from different soybean growing areas in

Argentina.

Acknowledgments

This study was supported by a grant from Consejo Nacional de Investigaciones

Cientificas y Técnicas (CONICET) and University of Buenos Aires, Argentina. The

authors thank Dr. S. Lupo (Universidad de la Republica, Montevideo, Uruguay) and Dr.

E.M. Reis (University of Passo Fundo, Brazil) for expert presubmission review.

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MY COTAXON

http://dx.doi.org/10.5248/123.467

Volume 123, pp. 467-472 January-March 2013

A new species of Cantharomyces (Laboulbeniales, Ascomycota)

from the Netherlands

D. HAELEWATERS! & A. DE KESEL?”

‘Department of Organismic and Evolutionary Biology, Harvard University

22 Divinity Avenue, Cambridge, Massachusetts 02138, U.S.A.

*National Botanic Garden of Belgium, Domein van Bouchout, 1860 Meise, Belgium

CORRESPONDENCE TO: ' dhaelewaters@fas.harvard.edu & * adk@br.fgov.be

ABSTRACT — This paper describes and illustrates Cantharomyces elongatus sp. nov., a parasitic

fungus from Syntomium aeneum (Staphylinidae, Oxytelinae, Euphaniini). Syntomium is anew

host genus for Cantharomyces. Comments on its position among related taxa are given.

KEY worps — insect-associated fungi, morphology, taxonomy

Introduction

Laboulbeniales are obligate ectoparasitic ascomycetes that live associated

with arthropods, mostly insects. At present the order comprises about 2050

species in 140 genera (Rossi & Santamaria 2012). Systematic and taxonomic

contributions are available for several countries in western Europe and North

America.

The genus Cantharomyces Thaxt. includes 28 species (Tavares 1985, Huldén

1983, Majewski 1990, Rossi & Santamaria 2000), characterized by a receptacle

consisting of three superposed cells, a compound antheridium subtending a

simple or variably branched primary appendage, and a perithecium having

four to five cells in each vertical row of outer wall cells (Thaxter 1931, Majewski

1994). Detailed information on Cantharomyces and its morphology and

position among other Laboulbeniales is given in Thaxter (1890, 1896, 1908,

1931), Tavares (1985), Majewski (1990, 1994), and Santamaria (2003).

The host range of Laboulbeniales on Staphylinidae and other Coleoptera has

been studied by Frank (1982), who listed 17 Cantharomyces species. In addition

to Staphylinidae, Cantharomyces species have been reported from Dryopidae,

Limnichidae, and Hydrophilidae (Tavares 1985). Based on recent taxonomic

insights in Staphylinidae from Newton & Thayer (2005) and Bouchard et al.

(2011), four tribes of the Staphylinidae are host to Cantharomyces: Blediini

468 ... Haelewaters & De Kesel

and Oxyteliini (subfam. Oxytelinae), and Aleocharini and Oxypodini (subfam.

Aleocharinae) (Thaxter 1890, 1896, 1908, 1931, Sugiyama 1973, Frank 1982,

Huldén 1983, Tavares 1985, Majewski 1990, 1994, Rossi & Santamaria 2000,

Santamaria 2003).

This paper presents a new species of Cantharomyces, collected from a

staphylinid beetle belonging to a fifth and so far unreported host tribe.

Materials & methods

The host was found by sifting soil from an ash forest on clay. Infected material was

dried, pinned, and identified by O. Vorst (using Lohse 1964). Screening for infection

and removal of thalli was done at 50x magnification. Thalli were transferred with insect

pin 0 and embedded in Amann solution (Benjamin 1971). Cover slips were ringed with

transparent nail varnish. Both insect specimen and the examined microscope slides are

deposited at the National Herbarium of Belgium (BR). Drawings and measurements

were made using an Olympus BX51 light microscope with drawing tube, digital camera

and AnalySIS Five imaging software (Soft Imaging System GmbH). Differences in the

proportions of basal cell and suprabasal cell were analyzed using the ratios R, = length

basal cell / length suprabasal cell and R,, = width basal cell / width suprabasal cell.

Average ratios are given for adult thalli, including minimum and maximum values in

parenthesis.

Host taxonomy follows Newton & Thayer (2005) and Bouchard et al. (2011).

Taxonomy

Cantharomyces elongatus Haelew. & De Kesel, sp. nov. PLATE 1

MycoBank MB 800871

Differs from Cantharomyces orientalis by its single long, strictly unbranched, straight

primary appendage with the basal cell > 2.5x longer and wider than the suprabasal cell.

Type: The Netherlands, Prov. Gelderland, Rha, 52°02.9'N 6°08.5’E, on Syntomium

aeneum (Miller, 1821) (Coleoptera, Staphylinidae, Oxytelinae, Euphaniini), 13 Oct 2001,

leg. O. Vorst, slides Haelewaters 69a (holotype, 2 mature and 4 immature thalli, found on

left elytron and abdominal tergites, BR MYCO 173753-26), Haelewaters 69b (isotype, 1

mature and 2 immature thalli found on elytra, BR MYCO 173752-25), and Haelewaters

69c (isotype, 2 mature and 2 immature thalli found on elytra, BR MYCO 173751-24).

Erymo.oecy: from the Latin adjective elongatus = elongate, referring to the long,

unbranched primary appendage.

THALLUS 145-175 um long from foot to perithecial apex. Receptacle 50-60

um long, cell I obtriangular, 22-30 um long, cell I and cell IN with similar

shape, slightly pigmented, not blackened, broader than long, cell HI somewhat

longer than cell I]. PRIMARY APPENDAGE composed of a basal cell, a small

PLaTE 1. Cantharomyces elongatus from Syntomium aeneum. A. Mature thallus from elytrum

(holotype Haelewaters 69a). B-D. Immature thalli with perithecial primordium (holotype slide

Haelewaters 69a). E. Immature thallus (isotype slide Haelewaters 69b). F. Submature thallus

(isotype slide Haelewaters 69c). Scale bar = 50 um.

Cantharomyces elongatus sp. nov. (Netherlands) ... 469

“ONSEN

eee eC neey |

Pepeasrrrerrercrtcsiiiiaaes 9

G, meahecreciee:

Meese se eeee

470 ... Haelewaters & De Kesel

suprabasal cell and a series of elongate cells. Basal cell of primary appendage

in the same axis as cell II and III, apically convex, slightly more pigmented

than or concolorous with cells II and III, 23 x 30 um (length x width), basal

septum with cell III straight. Suprabasal cell relatively flattened and small [R, =

2.88 (2,77-3,17), Ry = 2.81 (2,58-2,96)], 6.5-10 um long, trapezoidal, slightly

constricted, supporting a straight, 300-415 um long unbranched series of 7-11

hyaline, thick-walled, elongate cells of 25-40 um each. ANTHERIDIUM located

around a central core in the basal cell of primary appendage, with subapical

opening on the latter’s posterior side. PERITHECIUM 72-96 x 40-57 um, ovoid,

symmetric, widest in the middle, tapering upwards, moderately pigmented;

apex pointed, hyaline, poorly differentiated. Cell VI born laterally on cell II,

41-47 x 22-30 um, hyaline. AscosporREs not seen.

ADDITIONAL SPECIMENS EXAMINED: THE NETHERLANDS, Prov. Utrecht, Utrecht

(Amelisweerd), 52°04'N 5°09’E, on Syntomium aeneum, 11 Nov 1990, leg. O. Vorst, slides

Haelewaters 145a (1 damaged thallus from right elytron, BR MYCO 173750-23) and

Haelewaters 145b (3 immature thalli found on prosternum, BR MYCO 173749-22).

Discussion

Cantharomyces elongatus is easily recognized by its very long and unbranched

primary appendage with a relatively large basal cell and small trapezoidal

suprabasal cell. Most of the time, even in a dried state, this long structure is

found undamaged, indicating its robustness. In many genera of Laboulbeniales

damaged appendages are known to regenerate in an erratic or at least atypical

way. Among all examined specimens of C. elongatus we found several thalli

with a normally regenerated, ie. unbranched, appendage. Atypical regeneration

of the appendage may be infrequent in this taxon since we found only one

specimen with a bifurcate regenerated appendage (on its fourth appendage

cell). Whenever a branched or aberrant appendage is seen in this taxon, one

should look carefully for traces of damage to the appendage.

Cantharomyces elongatus was found on the pronotum, elytra, and abdominal

tergites and does not seem to be morphologically variable on these different

regions.

Cantharomyces elongatus is related to C. orientalis Speg., a very variable

species having a primary appendage that is ramified above its suprabasal cell.

Occasionally, however, C. orientalis bears a more or less short, unbranched

appendage. In such cases, C. orientalis can still be easily distinguished from

C. elongatus by its proportionally longer and wider suprabasal cell. In C. elongatus

the basal cell is at least 2.5x longer and 2.5x wider than the suprabasal cell.

Based on data from Belgian material and illustrations in Santamaria 2003 and

Majewski 1994, this proportion is significantly smaller for C. orientalis, R, =

1.33 (0.85-1.96) and R,, = 1.24 (0.95-2.16). The combination of the appendage

Cantharomyces elongatus sp. nov. (Netherlands) ... 471

characteristics of length, branching, and relative height of the suprabasal cell

should be enough to separate C. orientalis from C. elongatus.

Moreover, although C. elongatus and C. orientalis parasitize staphylinids

from the same subfamily (Oxytelinae), they infect hosts belonging to different

tribes: C. elongatus infects hosts of tribe Euphaniini and C. orientalis hosts of

tribe Oxytelini.

Cantharomyces robustus 'T. Majewski shows many differences from C. elongatus:

the blackening on the dorsal side of cell II, the ramified, shorter appendage, the

inflated and rounded basal cell of the appendage (with antheridium), as well as

large perithecial basal cells.

The new species is easily separated from the Cantharomyces species

with an unbranched primary appendage: C. bordei F. Picard, C. denigratus

Thaxt., and C. italicus Speg., based on the extreme length and construction

of its primary appendage. Cantharomyces bordei has an unbranched, but

short primary appendage. Furthermore, it is reported from Limnichus spp.

(Coleoptera, Limnichidae) and exhibits a very short and constricted cell VI as

well as a relatively massive perithecium (124-130 um long, Santamaria 2003).

Cantharomyces denigratus and C. elongatus seem to share a similar organization

of the antheridium (see Fig. 18a and d in Santamaria 2003). Cantharomyces

denigratus, however, has a deeply pigmented receptacle. Cantharomyces italicus

has a relatively long primary appendage but its antheridium is different, i.e.

lens-shaped, small and laterally positioned within a more elongate basal cell of

the primary appendage.

Cantharomyces numidicus Maire, as well as the very similar Cantharomyces

japonicus K. Sugiy., differ in having a relatively large antheridial segment and

a dissimilar arrangement of its primary appendage’s basal cell (Thaxter 1931,

Santamaria 2003).

Cantharomyces elongatus parasitizes Syntomium aeneum, extending the

known host range of Cantharomyces to the staphylinid tribe Euphaniini

(subfam. Oxytelinae).

Acknowledgments

The authors are indebted to Dutch entomologist Oscar Vorst for putting the host

specimen and its identity at our disposal. Thanks are also due to Sergi Santamaria and

Donald H. Pfister for reviewing the manuscript. We wish to express our gratitude to

Cyrille Gerstmans for technical support and the Uyttenboogaart-Eliasen Foundation

for financing part of this research.

Literature cited

Benjamin RK. 1971. Introduction and supplement to Roland Thaxter’s Contribution towards a

Monograph of the Laboulbeniaceae. Bibliotheca Mycologica 80: 1-155.

472. ... Haelewaters & De Kesel

Bouchard P, Bousquet Y, Davies AE, Alonso-Zarazaga MA, Lawrence JF, Lyal CHC, Newton AF,

Reid CAM, Schmitt M, Slipiriski SA, Smith ABT. 2011. Family-group names in Coleoptera

(Insecta). Zookeys 88: 1-972. http://dx.doi.org/10.3897/zookeys.88.807

Frank JH. 1982. The parasites of the Staphylinidae (Coleoptera). Bulletin Florida Agricultural

Experiment Stations 824(1-7): 1-118.

Huldén L. 1983. Laboulbeniales (Ascomycetes) of Finland and adjacent parts of the U.S.S.R.

Karstenia 23(2): 31-136.

Lohse GA. 1964. Staphylinidae I (Micropeplinae bis Tachyporinae). Die Kafer Mitteleuropas, Band

4, 264 p.

Majewski T. 1990. Rare and new Laboulbeniales from Poland. X. Acta Mycologica 23(2): 97-108.

Majewski T. 1994. The Laboulbeniales of Poland. Polish Botanical Studies 7: 1-466.

Newton AF, Thayer MK. 2005. Catalog of austral species of Staphylinidae and other Staphylinoidea

[online]. Chicago: Field Museum of Natural History.

http://www. fieldmuseum.org/peet_staph/db_1b.html [accessed June 16 2012]

Rossi W, Santamaria S. 2000. New Laboulbeniales parasitic on Staphylinidae. Mycologia 92(4):

786-791. http://dx.doi.org/10.2307/3761436

Rossi W, Santamaria S. 2012. Rodaucea, a new genus of the Laboulbeniales. Mycologia 104(3):

785-788. http://dx.doi.org/10.3852/11-337

Santamaria S. 2003. Laboulbeniales, II. Acompsomyces-Ilyomyces. Flora Mycologica Iberica 5:

1-344.

Sugiyama K. 1973. Species and genera of the Laboulbeniales (Ascomycetes) in Japan. Tokyo,

Academia Scientific Book Inc.

Tavares II. 1985. Laboulbeniales (Fungi, Ascomycetes). Mycologia Memoir 9: 1-627.

Thaxter R. 1890. On some North American species of Laboulbeniales. Proceedings of the American

Academy of Arts and Sciences 27: 29-45. http://dx.doi.org/10.2307/20020464

Thaxter R. 1896. Contribution towards a monograph of the Laboulbeniaceae. Memoirs of the

American Academy of Arts and Sciences 12: 187-429.

Thaxter R. 1908. Contribution towards a monograph of the Laboulbeniaceae. Part II. Memoirs of

the American Academy of Arts and Sciences 13: 217-469. http://dx.doi.org/10.2307/25058090

Thaxter R. 1931. Contributions towards a monograph of the Laboulbeniaceae. Part V. Memoirs

of the American Academy of Arts and Sciences 16: 1-435. http://dx.doi.org/10.2307/25058136

MY COTAXON

http://dx.doi.org/10.5248/123.473

Volume 123, pp. 473-477 January-March 2013

Lyophyllum rhombisporum sp. nov. from China

XIAO-QING WANG’”, DE-QUN ZHOU’, YONG-CHANG ZHAO’, XIAO-LEI

ZHANG’, LIN LP? & SHU-HonG Lr?”

"Faculty of Environmental Sciences and Engineering, Kunming University of Science and

Technology, Kunming 650500, Yunnan, China

’ Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences,

Kunming 650223, Yunnan, China

*CORRESPONDENCE TO: shuhonglifungi@gmail.com

ABSTRACT — A new Species, collected from Yunnan Province, China, is here described as

Lyophyllum rhombisporum (Lyophyllaceae, Agaricales). The new species is macroscopically

and microscopically very similar to L. sykosporum and L. transforme, but its spore shape

is quite different. Molecular analysis also supports L. rhombisporum as an independent

species.

KEY worpDs — Agaricomycetes, mushroom, taxonomy, ITS, phylogeny

Introduction

Lyophyllum is a genus in the Lyophyllaceae (Agaricales) with about 40 species,

which are widespread in north temperate regions (Kirk et al. 2008). A few

Lyophyllum species are well-known edible mushrooms, such as L. shimeji

(Kawam.) Hongo, L. decastes (Fr.) Singer, and L. fumosum (Pers.) P.D. Orton.

Research in China has reported 19 Lyophyllum species (Li 2002; Li et al. 2005;

Dai & Tolgor 2007; Li & Li 2009), but more research is still needed on the

taxonomy, molecular phylogeny, and cultivation of this genus. Here we describe

a new species, L. rhombisporum from Yiliang County, Yunnan Province, in

southwestern China.

Materials & methods

Microscopic and macroscopic characters were described based on the specimen

material (L1736) following the methods of Yang & Zhang (2003). Sections were made by

hand with a razor blade, then mounted in 5% KOH solution and lactophenol cotton blue

respectively under an Olympus BH-2. The specimens were deposited in the Herbarium

of Yunnan Academy of Agricultural Sciences, Kunming, China (YAAS).

Total DNA was extracted using the modified CTAB methodology (Hofstetter et al.

2002; Li et al. 2011). The internal transcribed spacer region (ITS) was amplified using

A7A4 ... Wang &al.

Macrocybe gigantea JX193694

100 L. connatum HE819396

L. connatum EF421104

L. deliberatum JF908338

L. semitale AF357048

0 L. semitale AF357049

100 L. atratum AF357053

94 61 92 L. anthracophilum AF357054

L. anthracophilum AF357055

99 L. crassifolium JF908331

L. caerulescens AF357052

L. sykosporum AF357051

53 L. sykosporum AF357050

73 99 L. rhombisporum JX966307

L. rhombisporum JX966308

L. ambustum AF357057

L. decastes AB285107

L. decastes AB269928

98 L. decastes FJ810160

96 L. decastes AB269929

100 L. decastes AB285104

L. decastes AB285105

61 L. decastes HM572544

L. decastes AF357059

58 83 L. shimeji HM572536

50 L. shimeji AB301604

L. shimeji JX966311

L. fumosum HM572540

L. fumosum HM572538

100 L. fumosum JX966309

L. fumosum JX966310

Clade |

Subclade I

L. leucophaeatum AF357032

87 100 L. favrei AF357035

100 L. favrei AF357034

94 L. ochraceum JF908329

L. ochraceum AF357033

_10

PLATE 1. Equally weighted parsimony tree of the genus Lyophyllum based on ITS sequences from

36 ITS sequences. Numbers above branches indicate bootstrap support.

primers ITS4 and ITS5 (Hofstetter et al. 2002; Larsson & Sundberg 2011), following the

modified Li’s procedure (Li et al. 2011). Purifying and sequencing of PCR products were

conducted by Beijing Genomics Institute.

Thirty-six sequences were included in molecular analysis: six generated from this

research and another 30 retrieved from GenBank. Macrocybe gigantea (Massee) Pegler

& Lodge was selected as the outgroup. Sequences were edited and assembled using

SeqMan II (Larsson & Sundberg 2011), aligned by Muscle 3.7, and adjusted manually

using BioEdit 7.0.1. Phylogenetic analyses were carried out via PAUP* 4.0 BEAT. Equally

Lyophyllum rhombisporum sp. nov. (China) ... 475

weighted parsimony (MP) was used to search for optimal trees according to Hofstetter

et al. (2002) and Larsson & Sundberg (2011).

Results & discussion

Phylogenetic analysis

Six strict consensus trees were generated based on the MP analyses of 36

ITS sequences (L = 479, CI = 0.714, RI = 0.866, RC = 0.618) (PLATE 1). The

phylogeny supports L. rhombisporum as distinct from the sampled Lyophyllum

species, although it is closely related to L. sykosporum. In the strict consensus

tree, L. rhombisporum and L. sykosporum form Clade I with 53% bootstrap

support. This subdivides into two subclades, with Subclade I comprising L.

rhombisporum with a strong bootstrap support (bs = 99%) and separated from

Subclade II comprising L. sykosporum.

Taxonomy

PLATE 2. Fruit-body of Lyophyllum rhombisporum.

Lyophyllum rhombisporum Shu H. Li & Y.C. Zhao sp. nov. PLATES 2, 3

MycoBank MB 801752

Differs from Lyophyllum sykosporum and L. transforme by its rhombic basidiospores.

Type: China: Yunnan, Yiliang County, 24°42’N 103°09’E, alt. 1922 m, in forest of

Pinus yunnanensis Franch, 23.6.2011, Shuhong Li, (Holotype, YAAS L1763; GenBank

JX966307).

Erymo oey: Latin rhombisporum, referring to the rhombic or subrhombic shape of the

spores.

476 ... Wang &al.

FET He ey

20pm

PLATE 3. Lyophyllum rhombisporum (Holotype L1763). Fic. A. Basidia in lactophenol cotton blue;

Fic. B. Spores in 5% KOH; Fic. C. Spore in lactophenol cotton blue.

PiLEus 2.0-7.5 cm, convex, applanate at maturity, smooth, dry, not striate,

greyish to pale taupe. CONTEXT thin, white to whitish grey. ODOR and TASTE not

distinctive. STIPE 3.0-8.0 long, 0.6-1.5 cm thick, cylindrical, greyish or greyish

brown, slightly enlarged downwards, appressed fibrillose, pale brown at base.

LAMELLAE moderately close together, adnate or subdecurrent, broad, whitish

to greyish, blackening when bruised, edge blunt). HyYMENOPHORAL TRAMA

regular, consisting of hyaline hyphae, clamp-connections rare. SPORES rhombic

or subrhombic, hyaline, cyanophilous, with a single large oil-droplet or a few

small oil-droplets, smooth, 14.5-17 x (7.5-)10-11.5 um, Qm = 1.3. BASIDIA

36.5-46 x 10-11.3 um, 4-spored, clavate, with siderophilous granulation well

visible in lactophenol cotton blue or cotton blue, clamp connections absent.

PLEURO- and CHEILOCYSTIDIA absent.

EcoLoGy & DISTRIBUTION: Scattered or gregarious in forests of Pinus

yunnanensis. Summer to autumn. Known only from Yunnan Province, China.

Lyophyllum rhombisporum sp. nov. (China) ... 477

ADDITIONAL SPECIMEN EXAMINED: CHINA: YUNNAN, Yiliang County, 24°42'N

103°09'E, alt. 1952 m, in Pinus yunnanensis forest, 21.9.2011, Shuhong Li, (YAAS L2082;

GenBank JX966308).

ComMENTS — ‘This species is characterized by large sub-rhombic or rhombic

basidiospores, and white or greyish gills that blacken when bruised, a feature

of section Lyophyllum (Breitenbach & Kranzlin 1991; Hofstetter et al. 2002;

Larsson & Sundberg 2011). Morphologically Lyophyllum rhombisporum

is morphologically similar to L. sykosporum Hongo & Clémencon and

L. transforme (Sacc.) Singer (Breitenbach & Kranzlin 1991; Hofstetter et

al. 2002), which are distinguished by their triangular basidiospores. The

phylogenetic analysis supports L. rhombisporum as a new species.

Acknowledgements

The authors are grateful to Wang Yun, Zhao Ruilin, and Dr Ian R. Hall, who critically

reviewed the manuscript and provided invaluable suggestions. We acknowledge Ma Tao

who helped us with the microscopic technical examination. This study was financially

supported by the National Science Foundation of China (No.31160010).

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Mycotaxon 88: 447-454.

MY COTAXON

http://dx.doi.org/10.5248/123.479

Volume 123, pp. 479-489 January-March 2013

New records of corticolous lichens from Vietnam

SANTOSH JOSHI’, THI THUY NGUYEN’, NGUYEN ANH DZUNG?’,

UDENI JAYALAL’, SOON-OK OH!’ & JAE-SEOUN HurR"*

‘Korean Lichen Research Institute, Sunchon National University, Suncheon-540 950, Korea

*Department of Basic Biology, Faculty of Natural Science and Technology, Tay Nguyen University,

Buon Ma Thuot City, Dak Lak Province, Vietnam

CORRESPONDENCE TO *: jshur1@sunchon.ac.kr

ABSTRACT —Eight new records of crustose lichens are reported from Dak Lak, Dak Nong,

and Vinh Phuc provinces in Vietnam. Descriptions of Diorygma hieroglyphicum, Letrouitia

leprolytoides, L. transgressa, Ocellularia perforata, Opegrapha robusta, Platythecium

colliculosum, P. dimorphodes, and Syncesia farinacea include morphological, anatomical,

chemical characters, and comparisons with similar taxa.

Key worps—Graphidaceae, Letrouitiaceae, Roccellaceae, evergreen forest, tropical

Introduction

A recent survey suggested that lichen species richness increases towards

lower latitudes and that tropical ecosystems harbor about half of the total lichen

species known worldwide (Licking et al. 2011). Vietnam, part of the Eastern

palaeotropical region, provides a climate suitable for lichens, particularly

crustose, corticolous, and foliicolous species (Nguyen et al. 2010, 2011).

Sporadic surveys and lichen collections from different regions of this country

indicate that the majority of its land is still unexplored for its lichen diversity.

Aptroot & Sparrius (2006) provided the first checklist of Vietnamese lichens,

but no update has been published since. However, recent publications (Giao

2009; Nguyen et al. 2010, 2011) on foliose and foliicolous lichens in Vietnam

document the potential richness in lichens in this region, and suggest that

many more species have not yet been recorded.

We report here eight corticolous species representing Diorygma, Letrouitia,

Ocellularia, Opegrapha, Platythecium, and Syncesia as new for Vietnam. Species

in four of these genera have previously been recorded from Vietnam: Diorygma

tinctorium Eschw., Letrouitia domingensis (Pers.) Hafellner & Bellem.,

Ocellularia allosporoides (Nyl.) Patw. & C.R. Kulk., O. asiatica (Vain.) Hale,

A480 ... Joshi & al.

O. dolichotata (Nyl.) Zahlbr., O. papillata (Leight.) Zahlbr., O. tenuis (Hale)

Hale, O. thelotremoides (Leight.) Zahlbr, Opegrapha filicina (Mont.) Mill. Arg.,

Op. fuscovirens Mill. Arg., Op. varia var. heterocarpa Mill. Arg., and Op. vegae

R. Sant. No species of Platythecium and Syncesia were recorded in the checklist

of Vietnamese lichens (Aptroot & Sparrius 2006).

Materials and methods

Lichen specimens were collected during April 2012 from Chu Yang Sin and Yok Don

National Parks in Dak Lak province, and Dray Nur Waterfall in Dak Nong province in

the central highlands and Tam Dao National Park in Vinh Phuc province in northern

Vietnam. The specimens were conserved in the lichen herbarium of the Korean Lichen

Research Institute, Suncheon, South Korea (KoLRI).

The specimens were examined using standard light microscopy techniques. Sections

of thalli and ascocarps were mounted in water, 10% KOH, lactophenol blue, and Lugol’s

solution. All measurements were made in water. Chemical constituents were identified

by spot color reaction tests and thin layer chromatography (TLC; solvent system C)

following published protocols (Orange et al. 2010). Measurements of hymenium,

exciple, asci, and ascospores were made on 10-15 thin, hand-cut apothecial sections.

The following literature was used for species identification: Aptroot & Sparrius 2006;

Kalb et al. 2004; Tehler 1997; Staiger 2002; Frisch et al. 2006; Archer 2009; Elix 2009;

Ertz 2009; Mangold et al. 2009; Rivas Plata et al. 2010; Joshi et al. 2011; Nguyen et al.

2010, 2011.

Taxonomy

Diorygma hieroglyphicum (Pers.) Staiger & Kalb, Symb. Bot. Upsal.

34(1): 1151, 2004. PL. 1A

Thallus corticolous, epiphloeodal, ecorticate, <200 um thick, + verrucose

due to emerging ascocarps, marginally smooth, dull, whitish to pale green;

prothallus distinct, whitish; algal layer well developed, continuous; medulla

distinct, white. Vegetative propagules absent. Ascocarps numerous, lirellate,

immersed, straight to bend, branched, stellate, <2 x 0.25 mm; disc exposed,

flat to concave, pale brown to brown, densely white pruinose. Thalline margin

prominent, 90-100 um wide. Proper exciple hyaline to pale brown or brown,

divergent, poorly developed, 15-20 um thick. Epihymenium pale to brownish,

granular, crystalline, 10-20 um high. Hymenium hyaline, not inspersed (but

disintegrated ascospore locules sometimes have the appearance of large oil

droplets), 100-135 um high. Subhymenium hyaline to pale at the base, 30-35(-

50) um high. Paraphyses simple with branched anastomosing apices, 1-2 um

thick. Asci clavate, 1-spored, 90-130 um, I-. Ascospores hyaline, muriform,

ellipsoidal, with round ends, 65-85 x 17-29 um, I+ blue.

CHEMISTRY— Thallus K+ yellow orange, Pd+ reddish, C-, KC-. Stictic acid

detected with TLC.

Corticolous lichens new to Vietnam ... 481

DISTRIBUTION & ECOLOGY— his corticolous species is widely distributed

in the Eastern palaeotropical region. It is known from Australia, Cameroon,

Tanzania, Indonesia, Singapore, Papua New Guinea, Philippines, New

Caledonia, Vanuatu, Solomon Islands, and Norfolk Island (Archer 2009; Kalb

et al. 2004). In Vietnam, the species was collected on trunks of trees in an

evergreen forest at an altitude of about 1000 m.

SPECIMEN EXAMINED — VIETNAM. VINH PHUC PROVINCE: Tam Dao National Park,

21°27'10.8"N 105°38'58.2"E, alt. 987 m, on tree trunk, 25 April 2012, Hur, Oh & Nguyen

VN120351 (KoLRI).

REMARKS—Diorygma hieroglyphicum resembles D. pruinosum (Eschw.) Kalb

et al. in having open, pruinose apothecial discs, 1-spored asci, and muriform

ascospores, but D. pruinosum contains protocetraric acid in the thallus whereas

D. hieroglyphicum contains stictic acid.

Letrouitia leprolytoides S.Y. Kondr. & Elix, Australas. Lichenol. 62: 17,2008. PL. 1B

Thallus corticolous, epiphloeodal, corticate, thin (ca. 150 um thick)

inconspicuous to well developed, pale greenish, greenish yellow to pale

yellow or orange; algal layer well developed, continuous; medulla indistinct

to endophloeodal, white, crystalline. Isidia coralloid, spread throughout

the thallus. Ascocarps numerous, apothecioid, biatorine (but appearing

lecanorine), + rounded to more often irregular, solitary to marginally fused,

emergent, sessile, orange, <2.5 mm diam.; disc exposed, + flat, epruinose,

reddish to dark brown, entire, ca. 2 mm diam.; margin prominent, smooth,

shiny, brighter than the disc, orange, entire, <0.3 mm. Thalline margin reduced

to absent. Proper exciple hyaline to pigmented, made up of radiating hyphae,

<130 um thick. Epihymenium orange brown to brownish, pigmented, 10-15

um high. Hymenium hyaline, not inspersed, <70-100 um high. Paraphyses

slightly branched, + interwoven or anastomosing, 1-1.5 um thick. Hypothecium

hyaline to slightly reddish in lower side, <70 um high. Asci clavate, 8-spored,

70-95(-100) x13-16 um, outer wall I+ blue. Ascospores hyaline, transversely

septate, fusiform to oblong or ellipsoidal, with narrowly rounded to subacute

ends, non-halonate, 20-27 x 6-12 um, with 5-9 locules; locules + rounded to

slightly angular, + lentiform, septa moderately thick, I- .

CHEMISTRY— Thallus K+ purple, Pd-, C-, KC-; apothecia K+ purple violet.

Anthraquinones detected with TLC (greenish gray spot at Rf 5 and yellow at

7).

DISTRIBUTION & ECOLOGY—Letrouitia leprolytoides is common in monsoon

forests in Asia and has a pantropical distribution (Kondratyuk & Elix 2008; Elix

2009). In Vietnam, it was reported from the tree trunks in evergreen forests at

300-800 m altitude.

SPECIMENS EXAMINED — VIETNAM. Dak NONG PROVINCE: Dray Nur Waterfall,

12°33'06.3"N 107°53'19.6"E, alt. 371 m, on tree trunk, 19 April 2012, Hur, Oh & Nguyen

482 ... Joshi & al.

VN120006 (KoLRI). Dak LAK PROVINCE: Yok Don National Park, 12°51'20.0"N

107°45'58.1"E, alt. 760 m, on tree trunk, 22 April 2012, Hur, Oh & Nguyen VN120205,

VN120212, VN120218 (KoLRI).

REMARKS— This species can be distinguished from most other Letrouitia species

by its isidiate thallus. It resembles L. leprolyta (Nyl.) Hafellner and L. corallina

(Mull. Arg.) Hafellner, which also have an isidiate thallus, but L. leprolyta has

erumpent to flat, wart-like isidia and L. corallina has flattened and unbranched

isidia and 1-2-spored asci.

Letrouitia transgressa (Malme) Hafellner & Bellem.,

Nova Hedwigia 35: 710, 1983. PL ILE

Thallus corticolous, epiphloeodal, corticate, thin, 55-80 um, smooth to

verrucose, pale green to olive green or olivaceous; prothallus distinct, whitish;

algal layer well developed, continuous; medulla indistinct to white, crystalline.

Vegetative propagules not seen. Ascocarps numerous, apothecioid, biatorine

(but appearing lecanorine), round to irregular, solitary to marginally fused,

emergent, sessile, <1.9 mm in diam.; disc exposed, flat to convex, epruinose,

reddish brown to brown, <1.7 mm in diam; margin prominent, smooth, shiny,

paler than the disc, pigmented, ca. 0.2 mm thick. Thalline margin absent. Proper

exciple hyaline to pigmented, made up of radiating hyphae, 90-150 um thick.

Epihymenium orange brown, pigmented, 10-25 um high. Hymenium hyaline,

not inspersed, 70-125 um high. Hypothecium reddish brown, <105 um high.

Paraphyses thin, septate, rarely branched and anastomosed, conglutinate, 1-1.5

um thick. Asci clavate, 2-4(-8)-spored, 100-135 x 20-27 um, I-. Ascospores

hyaline to pale when mature, primarily 6-9 transversely septate, becoming

progressively submuriform (1-3 vertical septa), ellipsoidal, with round to

subacute ends, non halonate, 35-40(-50) x 13-17(-20) um, I-.

CuEmistry—Thallus K-, Pd-, C-, KC-; apothecia K+ purple.

Anthraquinones detected with TLC (greenish gray spot at Rf 5 and yellow at 7).

DISTRIBUTION & ECOLOGY— This corticolous species is common in Africa,

Asia, Australia, New Caledonia, Papua New Guinea, and South America

(Hafellner 1983; Elix 2009). In Vietnam, it is widespread on trees in evergreen

forests at an altitude 600-1100 m.

SPECIMENS EXAMINED - VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National

Park, 12°27'57.0"N 108°20'34.9"E, alt. 780 m, on tree trunk, 21 April 2012, Hur, Oh &

Nguyen VN120272 (KoLRI); 12°28'04.4"N 108°20'39.0"E, alt. 622 m, on tree trunk, 20

April 2012, Hur, Oh & Nguyen VN120286 (KoLRI); 12°28'12.3"N 108°20'59.9"E, alt.

763 m, on tree branch, 20 April 2012, Hur, Oh & Nguyen VN120242 (KoLRI). Vinu

PHUC PROVINCE: Tam Dao National Park, 21°27'21.7"N 105°39'00.8’E, alt. 1092 m, on

tree trunk, 25 April 2012, Hur, Oh & Nguyen VN120333 (KoLRI).

REMARKS—Letrouitia transgressa is similar to L. parabola (Nyl.) R. Sant. &

Hafellner in having submuriform ascospores at maturity but differs in having

Corticolous lichens new to Vietnam ... 483

transgressa. D. Ocellularia perforata. E. Opegrapha robusta. F. Platythecium colliculosum.

G. Platythecium dimorphodes. H. Syncesia farinacea. Scale bars: A, B, G = 3 mm; C = 1.5 mm;

D, E=1mm;F=10mm;H=5mm.

ascospores with transverse septation and lens-shaped lumina in the primary

stage. In contrast, the ascospores of L. parabola havea spiral septation. Moreover,

L. parabola has slightly smaller ascospores with fewer septa (6-8 locules).

484 ... Joshi & al.

Letrouitia Hafellner & Bellem. comprises ca. 18 species world-wide. Most

Letrouitia species have similar morphological and chemical features and have

been segregated according to ascospore size and septation (Hafellner 1983;

Elix 2009). Generally three septation types are found in this genus: transverse,

spiral, and muriform. More frequently, ascospores with either transverse or

spiral septation become (sub-)muriform at maturity. As the taxonomic value

of ascospore septation in lichenized fungi has been questioned (Salisbury

1978; Galloway & Guzman 1988), molecular data is needed for the revision of

Letrouitia.

Ocellularia perforata (Leight.) Mill. Arg., Hedwigia 31: 284, 1892. PL. 1D

Thallus corticolous, epiphloeodal, corticate, <400 um thick, continuous to +

verrucose, smooth to rough, dull to + glossy, greenish gray or pale to olivaceous

green; algal layer well developed, continuous; medulla thin to indistinct, white,

with small to moderately large, clustered or scattered calcium oxalate crystals.

Vegetative propagules absent. Ascocarps numerous and conspicuous, porinoid,

becoming apothecioid, hemispherical, + rounded to irregular at maturity,

solitary to fused, immersed to + raised, rarely strongly emergent, <1 mm diam.;

disc with the columella visible at maturity, epruinose to slightly pruinose, flesh-

colored to pale or brownish; pore formed by the rim of the thalline margin,

round to irregular at maturity, entire, to 0.5 mm diam. Thalline margin has

the same color as the thallus or slightly paler, thin to thick, + incurved. Proper

exciple fused, brownish to dark brown, ca. 120 um thick. Epihymenium

hyaline, with grayish or brownish granules, <15 um high. Hymenium hyaline,

not inspersed, <150 um high. Paraphyses straight to bent, unbranched, with

slightly thickened tips, + interwoven, moderately to distinctly conglutinated,

1-1.5 um thick. Columellar structure well developed at maturity, entire,

brownish to carbonized, + covered by grayish or brownish granules, to 300 um

thick. Subhymenium indistinct to hyaline. Asci clavate, 8-spored, (75-)100-

120 um, I-. Ascospores hyaline, transversely septate, oblong to ellipsoidal, with

+ rounded to narrowly rounded or subacute ends, thick walled, non halonate,

20-32 x 6-9 um, with 8-10 locules, I+ blue violet; locules + rounded to slightly

angular, oblong to lentiform, with hemispherical to conical end cells.

CHEMISTRY— thallus K+ yellowish brown, Pd+ orange red, C-, KC-.

Protocetraric acid detected with TLC.

DISTRIBUTION & ECOLOGY— Ocellularia perforata has a pantropical and

southern-temperate distribution (Mangold et al. 2009) and has been reported

from rainforests. In Vietnam, it was collected from tree trunks in evergreen

forests at an altitude of ca.1000 m.

SPECIMEN EXAMINED — VIETNAM. VINH PHUC PROVINCE: Tam Dao National Park,

21°27'21.7"N 105°39'00.8"E, alt. 1092 m, on tree, 25 April 2012, Hur, Oh & Nguyen

VN120323 (KoLRI).

Corticolous lichens new to Vietnam ... 485

REMARKS—Ocellularia perforata, which is highly morphologically variable, is a

widespread species. The morphologically similar O. papillata and O. terebrata

(Ach.) Mill. Arg., can be distinguished by their thallus chemistry: the thallus

of O. terebrata produces psoromic acid, whereas O. papillata lacks secondary

metabolites.

Opegrapha robusta Vain., Bot. Tidsskr. 29: 137, 1909. PL. 1E

Thallus corticolous, endo- to epiphloeodal, ecorticate (cortex not clearly

distinguishable in section), thin (<150-200 um thick), + inconspicuous,

spread in large patches, dull to slightly glossy, pale-green to green, olive green

or dark brownish; prothallus distinctly dark brown to black; algal layer well

developed, continuous; medulla indistinct, white. Vegetative propagules

absent. Ascocarps lirellate, numerous and evenly distributed on the thallus;

lirellae simple to slightly furcate, sessile, black, epruinose, straight to curved,

+ flexuose, reaching <7 x 0.25 mm. Thalline exciple basal to absent. Proper

exciple completely carbonized, convergent, K+ olivaceous, entire, 130-160

um thick. Epihymenium indistinct to pale. Hymenium hyaline, not inspersed,

gelatinous, ca. 150 um high. Hypothecium hyaline to yellowish, <25 um high.

Paraphyses richly anastomosing, 1-1.5 um thick. Asci ellipsoid to clavate,

8-spored, 80-115 x 20-24 um, K/I+ blue apical ring distinct. Ascospores

hyaline, fusiform, transversely 9-15-septate, 40-70 x5-9 um (including a

perispore <2 um thick), not constricted at septa, I-.

CHEMISTRY— thallus K-, Pd-, C-, KC-. Confluentic acid detected with

TEC

DISTRIBUTION & ECOLOGY—Opegrapha robusta is known from Africa

(Gabon, Zaire), Asia (Indonesia, Malaysia, Papua New Guinea, The Philippines,

Thailand), Australia, and Solomon Islands (Ertz 2009). In Vietnam, it was

recorded on trees in dense, tropical evergreen forests at an altitude of 600-800

SPECIMENS EXAMINED —- VIETNAM. DAKLAK PROVINCE: Chu Yang Sin National

Park, 12°28'04.4"N 108°20'39.0"E, alt. 622 m, on tree trunk, 20 April 2012, Hur, Oh &

Nughen VN120290 (KoLRI); 12°27'57.0"N 108°20'34.9"E, alt. 780 m, on tree trunk, 21

April 2012, Hur, Oh & Nughen VN120275 (KoLRI).

REMARKS—Opegrapha robusta is well separated from the two other similar

species Op. prosodea Ach. and Op. viridis (Ach.) Behlen & Desberger in having

robust lirellate apothecia and confluentic acid. Opegrapha prosodea and Op.

viridis have smaller apothecia (1-3 mm long) and lack secondary compounds.

Platythecium colliculosum (Mont.) Staiger, Biblioth. Lichenol. 85: 380, 2002. PL. 1F

Thallus corticolous, epiphloeodal, corticate, <120 um thick, continuous,

smooth or irregularly to scarcely verrucose (due to the substrate texture),

486 ... Joshi & al.

+ glossy, olive to grayish green; prothallus distinct, white; algal layer well

developed, continuous; medulla distinct, white. Vegetative propagules absent.

Ascocarps lirellate, numerous and scattered on the thallus; lirellae emergent,

straight to bend, radially to irregularly branched, with acute ends, 10-20

x 0.25 mm (but the frequent branching increases the length of lirellae <150

mm); labia closed to open, white, crenate in mature apothecia, <133 um thick;

disc reddish brown to brown (not well exposed in the examined specimen).

Thalline margin complete, 30-35 um thick. Proper exciple indistinct to

brownish, + striate. Epihymenium brownish, crystalline to granular, 9-10 um

high, I+ blue. Hymenium hyaline, not inspersed, 80-90 um high, I+ slightly

blue. Subhymenium well developed, hyaline to pale yellow, <45 um high, I-.

Paraphyses simple at the base, but apically irregularly branched, 1-2 um thick.

Asci clavate, 8-spored, 60-70 x 12-15 um, I-. Ascospores hyaline, submuriform,

ellipsoidal to ovoid, transversely 4-locular with one vertical septum in middle

locules, 9-13 x 6-8 um, I+ blue violet.

CHEMISTRY— thallus K+ red, Pd+ yellow orange, C-, KC-. Salazinic acid

detected with TLC.

DISTRIBUTION & ECOLOGY—Platythecium colliculosum occurs most

commonly on bark. It is distributed mostly in tropical and subtropical regions

of the world. In Vietnam, the species was reported from trees in an evergreen

forest at an altitude between700-800 m.

SPECIMEN EXAMINED — VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National Park,

12°27'57.0"N 108°20'34.9E, alt. 780 m, on tree trunk, 21 April 2012, Hur, Oh & Nguyen

VN120135 (KoLRI).

REMARKS— The Vietnamese specimen of P. colliculosum shows a slight chemical

variation. Staiger (2002) described the species as producing both salazinic and

protocetraric acids, but we detected only salazinic acid in our material. The

morphologically similar species P. dimorphodes differs in containing norstictic

acid in the thallus.

Platythecium dimorphodes (Nyl.) Staiger, Biblioth. Lichenol. 85: 383, 2002. PL. 1G

Thallus corticolous, epiphloeodal, corticate, 200-500 um thick, smooth

to uneven, irregularly to scarcely verrucose, shiny to + matt, grayish-green

to greenish; algal layer well developed; medulla distinct, white. Vegetative

propagules absent. Ascocarps lirellate, numerous, emergent, straight to

bend, irregularly branched, 10 x 0.25-0.7 mm; labia open, white, crenate

in mature apothecia; disc reddish brown to brown, exposed, flat to slightly

concave, occasionally with a thin whitish pruina. Thalline margin complete,

+ undulating, 50-75 um thick. Proper exciple pale brown to brown, simple to

+ divided laterally in mature apothecia. Epihymenium brownish, crystalline

to granular, 12-14 um high, I-. Hymenium hyaline, not inspersed, 80-90 um

Corticolous lichens new to Vietnam ... 487

high, I-. Subhymenium well developed, hyaline to yellowish, 40-45 um high,

I-. Paraphyses+ anastomosed, apically irregularly branched, 1-1.5 um thick.

Asci clavate, 8-spored, 70-80 x 13-15 um, I-. Ascospores hyaline, ellipsoidal

to ovoid, submuriform, with three transverse septa and one vertical septum in

the middle locule, 15-17 x 7-8 um, I+ blue violet.

CHEMISTRY— Thallus K+ yellowish, Pd+ yellow orange, C-, KC-. Norstictic

acid detected with TLC.

DISTRIBUTION & ECOLOGY—The species is common in the Eastern

palaeotropical countries and is distributed in tropical and subtropical regions

of Australia, Malaysia, Java, Papua New Guinea, Philippines, and Sri Lanka

(Staiger 2002; Archer 2009). In Vietnam, it was reported from trees in an

evergreen forest at an altitude of 600-800 m.

SPECIMENS EXAMINED — VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National Park,

12°27'57.0"N 108°20'32.9’E, alt. 780 m, on tree trunk, 20 April 2012, Hur, Oh & Nguyen

VN120123 (KoLRD; 12°27'57.0"N 108°20'34.9”E, alt. 780 m, on tree trunk, 21 April

2012, Hur, Oh & Nguyen VN120176 (KoLRI); 12°28'04.4"N 108°20'32.0"E, alt. 622 m,

on tree trunk, 20 April 2012, Hur, Oh & Nguyen VN120284 (KoLRI).

ReMARKS—Platythecium colliculosum, which superficially resembles P

dimorphodes, differs in containing salazinic and protocetraric acids as thallus

compounds. Platythecium dimorphodes, which has immature entire labia

and apothecial disc color similar to those in P grammitis (Fée) Staiger and

P. leiogramma (Nyl.) Staiger, differs in its lack of secondary metabolites. The

Vietnamese collections differ from P dimorphodes as originally described

(Staiger 2002) in lacking isidia-like structures.

Syncesia farinacea (Fée) Tehler, Flora Neotropica, Monogr. 74: 25, 1997. PL. 1H

Thallus corticolous, epiphloeodal, ecorticate, 100-200 um thick, tomentose

to dense, coherent, homoiomerous, water-absorbent, + rimose, smooth to

slightly verrucose, slightly pruinose, creamy-white to + greenish, UV+ cream

to yellowish; prothallus dark brown; calcium oxalate crystals absent; surface

gel absent; medulla usually indistinct; hypomedullary plectenchyma absent.

Ascomata numerous, pseudo-monocarpocentral, sessile with constricted base,

<3 mm diam.; discs flat to convex, <0.5 mm diam., tomentum sometimes

present, but usually absent. Thalline margin at the same level than the disc,

ecorticate, but with algae. Proper exciple + inconspicuous to ca. 50 um thick.

Epihymenium dark brown, hyphae richly branched, not separable, verrucose,

brownish, 13-20 um high, I+ intense blue, KI+ blue. Hymenium hyaline, clear,

70-100 um high, I+ blue. Paraphyses rather lax, 2-3 um thick, branched, I-,

KI+ blue. Hypothecium dark brown, + carbonaceous, extending down to the

substrate, 200-400 um high, K+ olive green, I-, KI-. Asci, clavate, 8-spored, 60-

70 x 10-15 um, I+ blue. Ascospores hyaline, transversely 3-septate, fusiform, +

curved, 25-30 x 3-5 um, I-.

A488 ... Joshi & al.

CuEmIsTRY— thallus K+ reddish, Pd-, C-, KC-. Traces of protocetraric

and roccellic acids detected with TLC.

DISTRIBUTION & ECOLOGY— This variable species occurs mostly on bark

of trees and shrubs but also sometimes on rocks. Reported from both the

northern and southern hemispheres, S. farinacea is known from Mexico, the

West Indies, Colombia, Venezuela, and Brazil (Tehler 1997). In Vietnam, the

taxon was found growing luxuriantly over tree trunks at an altitude between

600-800 m in an evergreen forest.

SPECIMENS EXAMINED — VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National

Park, 12°27'57.0"N 108°20'34.9"E, alt. 780 m, on tree trunk, 21 April 2012, Hur, Oh &

Nguyen VN120175, VN120177 (KoLRI); 12°28'04.4"N 108°20'39.0"E, alt. 622 m, on

tree trunk,21 April 2012, Hur, Oh & Nguyen VN120285 (KoLRI).

REMARKS—We identify our specimens as Syncesia farinacea, despite thallus

color and texture differences from the type description that may be due to

different environmental conditions. Syncesia graphica (Fr.) Tehler resembles

S. farinacea but differs in having lecanoric acid (C+ red thallus) and erythrin

as secondary compounds; moreover, in S. farinacea the synascomata are more

rounded and constricted at the bases and the ascospores are somewhat smaller.

Another similar species, S. effusa (Fée) Tehler, can be distinguished by its UV+

bright light yellow thallus and the production of atranorin as an additional

compound. Syncesia farinacea can sometimes be confused with S. rhizomorpha

Tehler, which also has rhizomorph-like structures but which is distinguished

by smaller ascospores, calcium oxalate crystals in the thalline margin, and

schizopeltic acid in the thallus.

Acknowledgments

This work was supported by a grant from the National Research Foundation of

Korea (#2011-0031494). Authors are grateful to the reviewers Dr. Cécile Gueidan and

Dr. Khwanruan Papong for their valuable comments on manuscript. Santosh Joshi

also thanks Dr. Andreas Aptroot for providing the lichen checklist of Vietnam, and

Dr. Laszl6 Lék6s for helpful suggestions.

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MYCOTAXON

http://dx.doi.org/10.5248/123.491

Volume 123, pp. 491-492 January-March 2013

Regional annotated mycobiotas new to the Mycotaxon website

ABSTRACT — MycoTAxon is pleased to announce five new species distribution lists to

our ‘web-list’ page covering gasteroid fungi (by Hernandez-Caffot & al.) and Paxillaceae,

Gomphidiaceae, Boletaceae and Russulaceae (by Niveiro & Alberté) in Argentina; lichens in

Armenia (by Gasparyan & Sipman); corticioids in the Canary Islands, Spain (by Beltran-

Tejera & al.); and the lichens of Igdir province, Turkey (by Yazici & al.). This brings to 105

the free access mycobiotas now available on the MycoTaxon website:

<http://www.mycotaxon.com/resources/weblists.htm|>.

SOUTH AMERICA

Argentina

Maria L. HERNANDEZ-CAFFOT, GERARDO ROBLEDO & LAURA S. DOMINGUEZ.

Gasteroid mycobiota (Basidiomycota) from Polylepis australis woodlands of

central Argentina. (12 p.)

ABSTRACT — Gasteroid fungi associated with Polylepis australis forests from Cordoba

Mountain range, central Argentina, were studied. Samples were collected during

April-May 2007. Twenty-nine species were identified among which Bovista pila,

Geastrum morganii and Tulostoma xerophilum are new records for Argentina and

the distribution range of Bovista nigrescens, Cyathus olla, Geastrum fornicatum and

Lycoperdon pyriforme are expanded to high altitude woodlands from Cordoba.

N. NIVEIRO & E. ALBERTO. Checklist of the Argentinean Agaricales 6. Paxillaceae,

Gomphidiaceae, Boletaceae and Russulaceae. (12 p.)

Asstract— A checklist of species belonging to families Paxillaceae, Gomphidiaceae,

Boletaceae, and Russulaceae was made for Argentina. The list includes all species

published up to 2011. Sixteen genera and 54 species are recorded: 15 species from

Paxillaceae, 3 from Gomphidiaceae, 11 from Boletaceae, and 25 from Russulaceae.

EUROPE

Armenia

ARSEN GASPARYAN & HARRIE J. M. SIPMAN. New lichen records from Armenia.

(5 p.)

ABSTRACT — Nineteen species are added to the known lichen mycota of Armenia.

Three of these, Lecanora wetmorei, Lecanora percrenata and Lecanora flowersiana,

are of particular interest because they are currently predominantly known from

North America, and one, Gyalecta truncigena, because it represents a genus new for

Armenia.

492 ... New regional mycobiotas online

Spain

ESPERANZA BELTRAN-TEJERA, J. LAURA RODRIGUEZ-ARMAS, M. TERESA TELLERIA,

MARGARITA DUENAS, IRENEIA MELO, M. JONATHAN Diaz-ARMAS, ISABEL

SALCEDO & JOSE CARDOSO. Corticioid fungi from arid and semiarid zones of

the Canary Islands (Spain). Additional data. 2. (27 p.)

ABSTRACT — A study of the corticioid fungi collected in the arid, semiarid, and dry

zones of the Canary Islands is presented. A total of eighty species, most of them growing

on woody plants, was found. Nineteen species are reported for the first time from

the archipelago (Asterostroma gaillardii, Athelia arachnoidea, Botryobasidium laeve,

Byssomerulius hirtellus, Candelabrochaete septocystidia, Corticium meridioroseum,

Crustoderma longicystidiatum, Hjortstamia amethystea, Hyphoderma malenconii,

Leptosporomyces mutabilis, Lyomyces erastii, Peniophora tamaricicola, Phanerochaete

omnivora, Phlebia albida, Radulomyces rickii, Steccherinum robustius, Trechispora

praefocata, Tubulicrinis incrassatus, and T: medius). The importance of endemic

plants, such as Rumex lunaria, Euphorbia lamarckii, E. canariensis, Kleinia neriifolia,

Echium aculeatum, and Juniperus turbinata ssp. canariensis, inter alia, as substrates

for corticioid fungi was analyzed. The distribution of these fungi in different types

of xerophytic plant communities and bioclimatic belts are discussed, as well as their

ecological implications.

MID-EAST

Turkey

KENAN YAZICI, ANDRE APTROOT & ALI ASLAN. The lichen biota of Igdir province

(Turkey). (26 p.)

ABSTRACT — As a result of lichenological exploration in the Igdir province (Turkey),

a total of 292 lichenized fungi belonging to 96 genera in the Ascomycota, including 2

subspecies and 6 varieties, were determined from 174 different localities. Caloplaca

glomerata, Caloplaca kansuensis, Dermatocarpon miniatum var. cirsodes, Lecanora

bicincta var. sorediata, Lecanora prophetae-eliae, Phaeophyscia hispidula, Physcia

scopulorum, Placopyrenium iranicum and Pyrenopsis subareolata are new to Turkey,

and also Asia except Pyrenopsis subareolata and Lecanora prophetae-eliae.

MYCOTAXON

http://dx.doi.org/10.5248/123.493

Volume 123, pp. 493-495 January-March 2013

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 123

Acaulospora endographis B.T. Goto, p. 405

Alternaria xiaochaidanensis F. Wang, A.N. Li, D.M. Dai, X.X. Xu & D.C. Li, p. 251

Aureoboletus zangii X.F. Shi & P.G. Liu, p. 452

Cantharomyces elongatus Haelew. & De Kesel, p. 468

Chloridium xigazense Y.M. Wu & T.Y. Zhang, p. 277

Circinoconiopsis A. Hern.-Gut., p. 108

Circinoconiopsis amazonica A. Hern.-Gut., p. 108

Coccomyces pinicola R.H. Lei & C.L. Hou, p. 125

Cordochaete Sanyal, Samita, Dhingra & Avneet P. Singh, p. 103

Cordochaete cystidiata Sanyal, Samita, Dhingra & Avneet P. Singh, p. 105

Cortinarius athabascus Bojantchev, p. 382

Cortinarius cisqhale Bojantchev, p. 378

Cortinarius eldoradoensis Bojantchev, p. 384

Cortinarius gualalaensis Bojantchev, p. 394

Cortinarius miwok Bojantchev, p. 399

Cortinarius ohlone Bojantchev, p. 396

Cortinarius truckeensis Bojantchev, p. 387

Cortinarius tuolumnensis Bojantchev, p. 391

Ellisembia mercadoi G. Delgado, p. 446

Endocarpon maritimum Y. Joshi & Hur, p. 164

Endophragmiella bicolorata Hern.-Rest., J. Mena, Guarro & Gené, p. 222

Endophragmiella cantabrica J. Mena, Hern.-Rest., Guarro & Gené, p. 225

Entocybe melleogrisea T.J. Baroni & Y. Lamoureux, p. 355

Fusticeps lampadiformis J.S. Monteiro & Gusmao, p. 432

Fusticeps papillatus J.S. Monteiro & Gusmao, p. 434

Gautieria xinjiangensis T. Bau, p. 289

Gymnopus aporpohyphes (Singer) Tkaléec & MeSi¢, p. 427

494 ... MYCOTAXON 123

Gymnopus bactrosporus (Singer) Mesi¢ & Tkaléec, p. 427

Gymnopus brevistipitatus (Antonin) Tkalcéec & MeSi¢, p. 428

Gymnopus campinaranae (Singer) Me8i¢ & Tkaléec, p. 428

Gymnopus nigroimplicatus (Corner) MeSi¢ & Tkal€ec, p. 420

Gymnopus pacificus (Singer) Tkaléec & MeSi¢, p. 428

Gymnopus rhizomorphicola (Antonin) Mesi¢ & Tkaléec, p. 428

Gymnopus rigidichorda (Petch) Tkaléec & Me8&i¢, p. 428

Gymnopus subrigidichorda (Corner) Tkaléec & MeSic¢, p. 424

Gymnopus thiersii (Desjardin) Mesi¢ & Tkaléec, p. 428

Gymnopus tomentellus (Berk. & M.A. Curtis) Tkaléec & MeSi¢, p. 428

Heliocephala vietnamensis Melnik & R.F. Castafieda, p. 281

Hygrocybe americana Bessette, A.R. Bessette, Roody & W.E. Sturgeon, p. 92

= Hygrophorus acutus A.H. Sm. & Hesler 1942

non Hygrocybe acuta F.H. Moller 1945

Hygrocybe atro-olivacea (A.H. Sm. & Hesler) Bessette, A.R. Bessette, Roody &

W.E. Sturgeon, p. 92

Hygrocybe auratocephala (Ellis) Bessette, A.R. Bessette, Roody & WE. Sturgeon, p. 92

Hygrocybe basidiosa (Peck) Bessette, A.R. Bessette, Roody & W.E. Sturgeon, p. 92

Hygrocybe marginata var. concolor (A.H. Sm.) Bessette, A.R. Bessette, Roody &

W.E. Sturgeon, p. 92

Hygrocybe marginata var. olivacea (A.H. Sm. & Hesler) Bessette, A.R. Bessette,

Roody & W.E. Sturgeon, p. 92

Hygrocybe recurvata (Peck) Bessette, A.R. Bessette, Roody & WE. Sturgeon, p. 92

Hygrocybe subaustralis (A.H. Sm. & Hesler) Bessette, A.R. Bessette, Roody &

W.E. Sturgeon, p. 92

Hygrocybe tahquamenonensis (A.H. Sm. & Hesler) Bessette, A.R. Bessette, Roody

& W.E. Sturgeon, p. 92

Hymenoscyphus magnicellulosus H.D. Zheng & W.Y. Zhuang, p. 20

Hymenoscyphus uniseptatus H.D. Zheng & W.Y. Zhuang, p. 23

Hymenoscyphus yunnanicus H.D. Zheng & W.Y. Zhuang, p. 24

Inocephalus hypipamee Largent, p. 309

Inocephalus parvisporus Largent, p. 314

Inocephalus plicatus Largent, p. 304

Inocybe caroticolor T. Bau & Y.G. Fan, p. 170

Inonotus tramisetifer Har. Kaur, Avneet P. Singh & Dhingra, p. 15

Lecanora gansuensis L. Lu & H.Y. Wang, p. 286

Lichenochora tertia Etayo, Flakus & Rodr. Flakus, p. 10

Lignosus cameronensis Chon S. Tan, p. 198

Lignosus tigris Chon S. Tan, p. 196

NOMENCLATURAL NOVELTIES & TYPIFICATIONS ... 495

Lobothallia crassimarginata X.R. Kou & Q. Ren, p. 243

Lobothallia helanensis X.R. Kou & Q. Ren, p. 245

Lobothallia pruinosa X.R. Kou & Q. Ren, p. 246

Lyophyllum rhombisporum Shu H. Li & Y.C. Zhao, p. 475

Marasmius subviridiphyllus Chun Y. Deng, Yi H. Yang & T.H. Li, p. 272

Miriquidica yunnanensis Lu L. Zhang & X. Zhao, p. 364

Neolinocarpon attaleae Vitoria & J.L. Bezerra, p. 142

Ophiodothella angustissima (Peck) Hanlin & M.C. Gonzalez, p. 333

Passalora rolliniae Firmino & O.L. Pereira, p. 348

Pestalotiopsis furcata Maharachch. & K.D. Hyde, p. 54

Pestalotiopsis theae (Sawada) Steyaert 1949

(lectotypified, epitypified), p. 51

Phellodon mississippiensis R.E. Baird, L.E. Wallace & G. Baker, p. 185

Pseudocercospora aurelianae Firmino & O.L. Pereira, p. 344

Pseudocercospora vicosae Firmino & O.L. Pereira, p. 346

Scheffersomyces amazonensis (Cadete, M.A. Melo, M.R. Lopes, Zilli, M.J.S. Vital,

EC.O. Gomes, Lachance & C.A. Rosa) Urbina & M. Blackw., p. 233

Scheffersomyces ergatensis (Santa Maria) Urbina & M. Blackw., p. 233

Scleroderma suthepense Kumla, Suwannarach & Lumyong, p. 2

Sporormiella octomegaspora Doveri & Sarrocco, p. 130

Stropharia rubrobrunnea Senthil. & S.K. Singh, p. 214

Tuber formosanum H.T. Hu & Y. Wang, p. 296

Tuber subglobosum L. Fan & C.L. Hou, p. 98

Tuber wenchuanense L. Fan & J.Z. Cao, p. 99

Yunchangia L. Guo & B. Xu, p. 262

Yunchangia puccinelliae L. Guo & B. Xu, p. 262