IV ... MYCOTAXON 130(3)

MY COTAXON

VOLUME ONE HUNDRED THIRTY (THREE) — TABLE OF CONTENTS

COVER SECTION

ROVICWCT SS. ch stir on PBC, Sovak ee e+: 6 PRA Mh PRG ook oe EL Gi 9 ne ck a vii

SILT ISSLOMPNOCE UNCER x Meio Se 8 rset ene ss Bt ae dh Renan. 1a ede ts ees san load, viii

EMOMRPTEE AIGOVE thy v3 gz to 9 Tae et arias torch ek bes oi, 9 RE Li, cet aT a wt ix

RESEARCH ARTICLES

Conidial fungi from the semiarid Caatinga biome of Brazil:

a new species of Selenosporella from submerged leaves

PATRICIA OLIVEIRA FIUZA, LUIS FERNANDO PASCHOLATI GUSMAO,

& RAFAEL F, CasTANEDA-Ruiz 601

New records of smut fungi. 8. Testicularia africana from Tanzania

and Mozambique CvEToMIR M. DENCHEV & TEODOR T. DENCHEV 607

Pachyphloeus depressus, a new green truffle from China

Li FAN & SONG-PIN Cal 613

A new species of Selenodriella from Brazil Marina A.G. ARAUJO,

MARCELA A. BARBOSA, MaAyRa S. OLIVEIRA,

ELAINE MALOosso, & RAFAEL E. CASTANEDA-RutIz 621

Antrodiella indica, a new species from India

GURPREET Kaur, AVNEET P. SINGH, & G.S. DHINGRA 625

Russula sichuanensis and its ectomycorrhizae from Himalayan

moist temperate forests of Pakistan M. SaBA & A.N. KHALID 629

Mycenastrum corium and gastrointestinal mycetism in México

EVANGELINA PEREZ-SILVA, TEOFILO HERRERA, & ABRAHAM J. MEDINA-ORTIZ 641

New reports of rust fungi from Malakand, Khyber Pakhtunkhwa, Pakistan

S. Hussain, A. Has1, N.S. AFSHAN, & A.N. KHALID 647

Artomyces nothofagi sp. nov. - a new clavarioid fungus from

a Chilean Nothofagus forest RICHARD KNEAL & MATTHEW E. SMITH 653

Inonotus griseus sp. nov. from eastern China

Li-We! ZHOU & XIAO-YAN WANG 661

New reports of Inocybe from pine forests in Pakistan

M. Sasa, I. AHMAD, & A.N. KHALID 671

Elmerina fragilis sp. nov. from Central China

FanG Wu, YUAN YUAN, & Hat-J1ao Li 683

A new species and new records of the lichen genus Pertusaria from China

QIANG REN 689

JULY-SEPTEMBER 2015 ... V

First record of Erysiphe howeana from China

Lu-CuHao Bal, ZHI-MIN Cao, & PENG WANG 695

Buellia, Lempholemma, and Thelidium species new for Turkey

and Asia KENAN YAZICI & ANDRE APTROOT 701

Four new records of Lecanoraceae in China

X1n ZHAO, Lu-Lu ZHANG, LI-YAN SuN, LING Hu, & ZUN-TIAN ZHAO 707

New records of smut fungi. 9. A second locality of Anthracoidea andina

CVETOMIR M. DENCHEV, HARRIE J.M. SIPMAN, & TEODOR T. DENCHEV 717

A new species and new record of Anthostomella on Alnus leaf-litter

from Argentina EsTEBAN B. Str, ANDREA I. ROMERO, & ADRIANA I. HLADKI 721

Phoma canadensis Allesch.: a synonym of Pyrenochaeta cava

LIDIANE LEAL DUARTE & ROBERT WEINGART BARRETO 731

Four new records of Rhizocarpon from China

WEI-CHENG WANG, ZUN-TIAN ZHAO, & Lu-Lu ZHANG 739

New record of Aplosporella javeedii on five hosts in China based on multi-

gene analysis and morphology

XIN-LEI FAN, QIN YANG, BIN CAo, YING-MEI LIANG, & CHENG-MING TIAN 749

Phaeostilbelloides and Velloziomyces — new dematiaceous genera from the

Brazilian Cerrado ELIANE AMARAL DE SOUZA ARMANDO,

ZULEIDE MARTINS CHAVES, & JOSE CARMINE DIANESE 757

New records of crustose Teloschistaceae and lichenicolous fungi

from Turkey MEHMET GOKHAN HALticl 769

New species of Phaeomonilia and Mirandina from dead branches

in China

YING-Ru1 Ma, JI-WEN XIA, XIU-GUO ZHANG, & RAFAEL F. CASTANEDA-RuIz 775

Species associated with cytospora canker on Populus tremuloides

JEFF B. KEPLEY, F. BRENT REEVES, WILLIAM R. JACOBI, & GERARD C. ADAMS 783

Two new species and a new record of Hansfordia from China

YuE-MING Wu, HONG-FENG WANG, JUN-JIE XU, & TIAN-YU ZHANG 807

Marthamyces chinensis sp. nov. on Fissistigma from China

Hatr-L1n Gu, YING-REN LIN, FAN PENG, QING LI, & SHI-JUAN WANG 815

Blastophragma chonggqingense sp. nov. and a new record of

Bahusutrabeeja angularis from southern China

JIAN-MEI GAO, JI- WEN XIA, YING-RuI Ma, ZHUANG LI, & XIU-GUO ZHANG 821

Sporidesmiopsis malloti sp. nov. and new records from southern China

JI-WEN X1A, YING-RuI MA, JIAN-MEI GAO, ZHUANG LI, & X1U-Guo ZHANG 827

Codinaea jianfenglingensis sp. nov. and new records from southern China

Ji- WEN X1A, YING-Rur Ma, JIAN-MEI GAo, ZHUANG LI, & X1U-GUO ZHANG 835

VI... MYCOTAXON 130(3)

Fuscoporia atlantica, a new polypore from the Brazilian Atlantic Rainforest

RICARDO MATHEUS PIRES, VIVIANA MOTATO- VASQUEZ,

& ADRIANA DE MELLO GUGLIOTTA 843

Volvariella rava sp. nov. from southern China

JIANG Xu, Tal-Hutr Li, YA-HENG SHEN, & MING ZHANG 857

First report of Leptographium pini-densiflorae causing sapstain on Pinus

densiflora in Korea Joo-Hyun Hona, YEONGSEON JANG,

SEOKYOON JANG, MIHEE MIN, & JAE-JIN Kim 867

Xylomyces acerosisporus sp. nov. from submerged leaves from Brazil

Mayra S. OLIVEIRA, ELAINE MALosso, MARCELA A. BARBOSA,

Marina A. G. ARAUJO, & RAFAEL FE. CASTANEDA-RuIz 875

Tubulicrinis indicus, a new corticioid species from India

JyvoT1 SHARMA, AVNEET P. SINGH, & G.S. DHINGRA 879

Four Rhizocarpon species new to China

WEI-CHENG WANG, ZUN-TIAN ZHAO, & Lu-LU ZHANG 883

Terriera transversa sp. nov. from Hainan, China Qtnc Lt, SHI-JUAN WANG,

Yu-XIA CHEN, YAN-PING TANG, & YING-REN LIN 893

New records of Clauzadea and Immersaria from China

Lu-Lu ZHANG, LING Hu, XIANG-XIANG ZHAO, & ZUN-TIAN ZHAO 899

Phlebiopsis punjabensis sp. nov. from India

GURPREET Kaur, AVNEET P. SINGH, & G.S. DHINGRA 907

Ocellularia lumbschii and O. saxicola spp. nov. from Vietnam

SANTOSH JOSHI, DALIP KUMAR UPRETI, THI THUY NGUYEN,

Ermy Rivas PLata, ANH DZUNG NGUYEN, SOON-OK OH, & JAE-SEOUN Hur 911

BOOK REVIEWS AND NOTICES ELSE C. VELLINGA (GUEST EDITOR) 921

REGIONAL MYCOBIOTAS NEW TO THE MYCOTAXON WEBSITE 925

Records of coprophilous fungi — a data set

MICHAEL J. RICHARDSON

A checklist of Jelly Fungi (Agaricomycotina: Basidiomycota) recorded in Brazil

RENATO LUCIO MENDES ALVARENGA & SOLANGE XAVIER-SANTOS

The lichen biota of Burdur province (Turkey)

KENAN YAZICI, ANDRE APTROOT, ALI ASLAN,

HARRIE SIPMAN, & MICHELE D. PIERCEY- NORMORE

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS PROPOSED IN

MYCOTAXON 130(3) 927

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY (2)

MYCOTAXON for APRIL-JUNE, (I-x1I + 307-600)

was issued on July 9, 2015

JULY-SEPTEMBER 2015...

REVIEWERS — VOLUME ONE HUNDRED THIRTY (3)

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 quarter.

Vladimir Antonin

A. Aptroot

Alan W. Archer

Toni Atkinson

Juliano M. Baltazar

Ann Bell

Sarah Bergemann

Uwe Braun

Ana Rosa Burgaz

Rafael F. Castafieda- Ruiz

Bao-Kai Cui

Kanad Das

Francisco Doveri

Martin Esqueda

Javier Etayo

Adam Flakus

Adriana de Mello Gugliotta

Shouyu Guo

Nils Hallenberg

Gabriela Heredia Abarca

Cheng-Lin Hou

Ze-Feng Jia

Yu-Ming Ju

Alfredo Justo

Bryce Kendrick

Paul M. Kirk

Kerry Knudsen

Shambhu Kumar

De-Wei Li

Chen Liang

Edgar Lickey

Ying-Ren Lin

Brian P. Looney

Quan Lu

Robert Liicking

Hayato Masuya

P. Brandon Matheny

Eric H.C. McKenzie

David W. Minter

Gabriel Moreno

Abdul Rehman Niazi

Lorelei L. Norvell

Shaun R. Pennycook

Omar Paino Perdomo

Ronald H. Petersen

Liliane Petrini

Victor J. Rico

Gunasekaran Senthilarasu

B.M. Sharma

Sigfrido Sierra

H.J.M. Sipman

Roger G. Shivas

Dartanha J. Soares

Einar Timdal

Ned Tisserat

Michal TomSovsky

Janice Uchida

Dalip Kumar Upreti

Kalman Vanky

Josef Vlasak

Yong Wang

Roy Watling

James J. Worrall

J.H.C. Woudenberg

Ming Ye

Meng Zhang

Li-Wei Zhou

Xudong Zhou

Mikhail Zhurbenko

VII

vill ... MYCOTAXON 130(3)

FOUR STEPS TO SUCCESSFUL MYCOTAXON PUBLICATION IN 2015

Prospective MycoTaxon authors should download instructions PDE, review and

submission forms, and other helpful templates by clicking the ‘file download page’ link

on our INSTRUCTIONS TO AUTHORS page before preparing their manuscript. Below is a

summary of our ‘4-step’ publication process.

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The Mycotaxon Webmaster <mycotaxon@gmail.com> posts general and

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MyYCOTAXON ONLINE— www.ingentaconnect.com/content/mtax/mt

Mycotaxon publishes four quarterly issues per year. Both open access and

subscription articles are offered.

JULY-SEPTEMBER 2015 ... IX

FROM THE EDITOR-IN-CHIEF

METADATA NEW TO Mycotaxon — Mike Richardson considerably expands our

mycobiota offerings by posting comprehensive specimen ‘metadata’ on the MycoTAXON

website. As data representing a lifetime of coprophilous fungal collections made

worldwide do not fit easily in PDF format, the substrates, localities, and species are

provided in Excel spreadsheet. We thank Mike for generously sharing his data, now

available to all researchers wishing to research the taxonomy or refine the nomenclature

of this difficult group of fungi.

THE OXFORD COMMA VS ‘&’ — Many authors may wonder why we regularly insert blue-

highlighted commas during our final editorial review. That is because MyCOTAXON uses

the ‘serial’ comma (also called the Oxford or Harvard comma) “before ‘and’ or ‘or’ at the

end of a series, regardless whether it is needed for clarification purposes” (Wikipedia).

Last evening, having just converted the next to last por for MycoTaxon 130(3), we

received a request from one author that we remove the ‘unnecessary’ comma from the

running header ‘Denchev, Sipman, & Denchev’ so that it CORRECTLY reads “‘Denchev,

Sipman & Denchev. Noting that Mycotaxon style dictates use of the Oxford comma,

we explained that we did not want to change headers for only one paper.

Today’s grammatical research revealed, however, that placing a serial comma

directly before the ampersand (‘&’) is controversial. While not all experts agree, the

ENGLISH USAGE AND STACK EXCHANGE advises, “In styles that use a serial comma, an

exception is made before an ampersand, which is a display element, not a word. When

an ampersand is used in a list, the serial comma is omitted because it’s visually jarring.”

We somewhat reluctantly agree that “X, Y, & Z” does seem a bit ‘weird’ and admit

that ‘serial comma + & is never found in nomenclatural authorites. So enjoy the ‘extra’

commas in 130(3) — they will not stand before ampersands in MycoTaxon 130(4)!

MYCOTAXON 130(3) presents 40 papers by 150 authors (representing 16 countries) and

revised by 69 expert reviewers.

Within its pages are 2 new genera (Phaeostilbelloides and Velloziomyces from Brazil)

and 28 species new to science representing Anthostomella from Argentina; Antrodiella,

Phlebiopsis, and Tubulicrinis from India; Artomyces from Chile; Blastophragma,

Codinaea, Elmerina, Hansfordia, Inonotus, Marthamyces, Mirandina, Pachyphloeus,

Pertusaria, Phaeomonilia, Sporidesmiopsis, Terriera, and Volvariella from China;

Fuscoporia, Phaeostilbelloides, Selenodriella, Selenosporella, Velloziomyces, and Xylomyces

from Brazil; Ocellularia from Vietnam; and Cytospora from the United States.

In addition to more detailed descriptions, range extensions, and new hosts for

previously named taxa, we also cover the toxic effects of Mycenastrum in Mexico and

offer a newly established synonymy for Pyrenochaeta cava.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

2 October 2015

MY COTAXON

http://dx.doi.org/10.5248/130.601

Volume 130, pp. 601-605 July-September 2015

Conidial fungi from the semiarid Caatinga biome of Brazil:

a new species of Selenosporella from submerged leaves

PATRICIA OLIVEIRA FruzA', Luis FERNANDO PASCHOLATI GUSMAO™*,

& RAFAEL F CASTANEDA-RUIZ ”

"Departamento de Ciéncias Biolégicas, Laboratério de Micologia, Universidade Estadual de Feira de

Santana, Avenida Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Brazil

?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

* CORRESPONDENCE TO: lgusmao@uefs.br

ABSTRACT — Selenosporella minima sp. nov., collected on submerged leaves of Calophyllum

brasiliense in the Brazilian semiarid region, is described and illustrated. It is distinguished by

verticillate branched conidiophores and navicular to acerose conidia.

KEY worRDs — asexual fungi, taxonomy, freshwater

Introduction

Selenosporella G. Arnaud ex MacGarvie comprises twelve species.

Descriptions, illustrations, and a key to Selenosporella species has been

provided by Castaneda-Ruiz et al. (2009), who divided the genus into two

groups: i) with conidiogenous cells discrete and arranged in verticils and

ii) with conidiogenous cells integrated and not arranged in verticils.

Most Selenosporella species have been described from litter or wood (Seifert

et al. 2011), except for Selenosporella aristata Kuthub. & Nawawi, which

was described from submerged leaves (Kuthubutheen & Nawawi 1988). An

interesting Selenosporella specimen recently found on decaying submerged

leaves of Calophyllum brasiliense is described here as new.

Materials & methods

During several expeditions between November 2013 and May 2014 through Serra

da Tromba in the semiarid region of northeastern Brazil, samples of submerged leaves

602 ... Fiuza, Gusmao, & Castafieda-Ruiz

of Calophyllum brasiliense were stored in plastic bags, taken to the laboratory, and

treated according to Castafeda-Ruiz (2005). Mounts were prepared in PVL (polyvinyl

alcohol, lactic acid, and phenol) and measurements made at a magnification of

1000. Microphotographs were obtained with an Olympus microscope (model BX51)

with bright field and Nomarski interference optics. The holotype is deposited in the

Herbarium of Universidade Estadual de Feira de Santana, Bahia, Brazil (HUEFS).

Taxonomy

Selenosporella minima Fiuza, Gusmao & R.F. Castafieda, sp. nov. Fics 1, 2

IF 550802

Differs from Selenosporella cymbiformis by single, verticillate branched conidiophores.

Type: Brazil, Bahia, Piata, Serra da Tromba, 13°05’S 41°50’W, on submerged leaves

of Calophyllum brasiliense Cambess. (Calophyllaceae), 1-V-2014, coll. P.O. Fiuza

(Holotype: HUEFS 210422).

ErymMo_oey: Latin, minima, meaning small, refers to conidial size.

CoLonigs on the natural substrate effuse, hairy, brown. Mycelium mostly

immersed, composed of septate, branched, brown hyphae. CONIDIOPHORES

distinct, single, erect, straight, verticillate branched, 2—-5-septate, smooth, dark

brown, 60-100 x 4-5 um. CONIDIOGENOUS CELLS holoblastic, denticulate,

sympodially elongated, discrete, brown to pale brown. Conidial secession

schizolytic. Conrp1a solitary, navicular to acerose, unicellular, smooth, hyaline,

DD el pM.

Note: Selenosporella cymbiformis B. Sutton is similar to S. minima in conidial

shape and size, but S. cymbiformis has single or loosely synnematous,

unbranched or irregularly branched, 35-55 x 3-4 um, dark to medium brown

conidiophores (Sutton 1973).

Acknowledgments

The authors express their sincere gratitude to Dr. De-Wei Li, Prof. Dr. Meng Zhang,

and Dr. Shambhu Kumar, for their critical review of the manuscript. The authors are

grateful to the “Programa de Pesquisa em Biodiversidade” - (PPBio Semi-arid/MCTI/

CNPq) for financial support. POF thanks the ‘Coordenacao de Aperfeigoamento de

Pessoal de Nivel Superior” (CAPES) and the ‘Programa de Pdés-graduacao em Botanica

PPGBot/UEFS. The authors thank the support provided by ‘Programa Ciéncia sem

Fronteiras. RFCR is grateful to and Cuban Ministry of Agriculture and ‘Programa de

Salud Animal y Vegetal; project P131LH003033 for facilities. We acknowledge the

facilities provided by Dr. PRM. Kirk and Drs. V. Robert and A. Decock through the

Fic. 1. Selenosporella minima (ex holotype, HUEFS 210422): A, B. Conidiogenous cells and conidia;

C. Conidiophore, conidiogenous cells and conidia; D, E. Conidiophores. Scale bars: A = 10 um;

B, C = 5 um; D, E= 20 um.

Selenosporella minima sp. nov. (Brazil) ... 603

604 ... Fiuza, Gusmao, & Castafieda-Ruiz

Fic 2. Selenosporella minima (ex holotype, HUEFS 210422):

A. Conidia; B. Conidiogenous cells and conidia; C. Conidiophores.

Scale bars: A, B = 5 um; C = 20 um.

Selenosporella minima sp. nov. (Brazil) ... 605

IndexFungorum and MycoBank websites. Dr. Lorelei Norvell’s editorial review and

Dr. Shaun Pennycook’s nomenclature review are greatly appreciated.

Literature cited

Castaneda Ruiz RE. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Castafieda Ruiz RE, Guerrero B, Adamo GM, Morillo O, Minter DW, Stadler M, Gené J, Guarro

J. 2009. A new species of Selenosporella and two microfungi recorded from a cloud forest in

Mérida, Venezuela. Mycotaxon 109: 63-74. http://dx.doi.org/10.5248/109.63

Kuthubutheen AJ, Nawawi A. 1988. A new species of Selenosporella (hyphomycetes) from Malaysia.

Trans. Br. Mycol. Soc. 91: 331-334. http://dx.doi.org/10.1016/S0007-1536(88)80222-5

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes.

CBS Biodiversity Series 9. 997 p.

Sutton BC. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycol. Pap. 132.

143 p.

MY COTAXON

http://dx.doi.org/10.5248/130.607

Volume 130, pp. 607-611 July-September 2015

New records of smut fungi. 8.

Testicularia africana from Tanzania and Mozambique

CVETOMIR M. DENCHEV’ & TEODOR T. DENCHEV

Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences,

2 Gagarin St., 1113 Sofia, Bulgaria

* CORRESPONDENCE TO: cmdenchev@yahoo.co.uk

AssTRACT — Testicularia africana is reported for the first time from Tanzania and

Mozambique. The new records extend its geographical range from Western Africa and

Cameroon to Eastern Africa.

Key worps — Anthracoideaceae, Cyperaceae, Rhynchospora, taxonomy, Ustilaginomycetes

Introduction

The genus Testicularia, described by Klotzsch (1832), belongs to the

Anthracoideaceae. This small genus of smut fungi includes three species:

T. cyperi Klotzsch (known from USA), T: minor (Juel) L. Ling (from the Greater

Antilles, Colombia, Brazil, and Bolivia), and T! africana (from Western Africa

and Cameroon). All three are found on plants of Rhynchospora (Cyperaceae)

(Vanky & Piatek 2006, Vanky 2010, 2011, Vanky et al. 2011). Their sori are

formed in some spikelets or around the basal part of some of the inflorescence

branches of infected plants. Initially, the sori are covered by a thick peridium

that later ruptures, becoming sac-like and exposing an agglutinated or granular

mass of spore balls. The spore balls characteristically comprise a peripheral

layer of dark, thick-walled spores surrounding central mass of thin-walled and

light-coloured sterile fungal cells.

Testicularia africana is known only from single collections from Guinea

(Macenta Pref., near Macenta; Vanky & Piatek 2006), Cameroon (Menoua

Dept., Dschang, Dschang Lake; Vanky 2009), and Sierra Leone (erroneously

recorded by Zambettakis 1970: 680 as T: cyperi; cf. Vanky & Piatek 2006, Vanky

et al. 2011).

608 ... Denchev & Denchev

In this paper we report two new records of Testicularia africana from

Tanzania and Mozambique. The collections on which these records are based

were obtained during a visit to the herbarium at the Botanic Garden and

Botanical Museum Berlin-Dahlem (B).

Material & methods

Dried specimens from the herbarium of the Botanic Garden and Botanical Museum

Berlin-Dahlem were examined under light microscope (LM) and scanning electron

microscope (SEM). For LM observations and measurements, spores were mounted in

lactoglycerol solution (w: la: gl = 1: 1 : 2) on glass slides, gently heated to boiling

point to rehydrate the spores, and then cooled. The measurements of spores are given

as min-max (mean + | standard deviation). In the description, the total number of

spores (n) from all collections (x) measured is given as ‘(n/x). 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 and photographed at 10

kV accelerating voltage using a JEOL SM-6390 scanning electron microscope. The

description given below is based entirely on the specimens examined.

Taxonomy

Testicularia africana Vanky & Piatek, Mycol. Balcanica 3: 164, 2006. Fics 1-8

Sor! in some flowers, when young concealed by the glumes that later flake

away, 1-3 mm long, or around the basal part of some of the inflorescence

branches, 3-9 mm long; globose to ovoid, covered by a thick reddish brown

peridium. The peridium of the mature sori ruptures irregularly (becoming sac-

like or forming irregular lobes at the base of the sori) to expose an agglutinated

blackish brown mass of spore balls. The surface of the peridium smooth in

young sori, densely and irregularly cracked at maturity. SPORE BALLS globose,

subglobose, broadly ellipsoidal or ovoid, (46-)50-100(-117) x 40-90 um,

composed of an outer layer of loose spores and sterile cells in the central part.

Spores often flattened, in plane view orbicular, suborbicular, broadly elliptical

or oval in outline, sometimes slightly irregular, in plane view (10.5-)11-14.5

(-15.5) x (10-)10.5-12.5(-13.5) (12.8 + 0.9 x 11.7 + 0.8) um (n, = 150), medium

reddish brown, wall evenly thickened, 0.6-0.9 um thick, with two lighter areas

in the middle of the flattened sides, 3-5 um in diameter; in LM smooth, in SEM

punctate. SPORE GERMINATION unknown.

SPECIMENS EXAMINED — On Rhynchospora corymbosa (L.) Britton: TANZANIA,

KIGOMA REGION (the locality as ‘Deutsch-Ostafrika, Uha, zwischen Tare und Bujenze’),

1640-1455 m, 11.III.1926, leg. A. Peter (the plant specimen as A. Peter, Reisen in Afrika

1925/26, no. 38 637, Exkursion V.182) (B 10 0506843); MOZAMBIQUE, SOFALA

PROVINCE, between Beira and Manga (as ‘zwischen Beira und Manga, Trollyfahrt, auf

dem Wegdamm und auf teilweise nassen Wiesen gegen Manga hin’), 27.1X.1925, leg.

A. Peter (the plant name given on the original label as Cyperus sp., A. Peter, Reisen

Testicularia africana from Tanzania and Mozambique ... 609

Fics 1-4. Sori (arrow) of Testicularia africana on Rhynchospora corymbosa. 1,3. Habit (Mozambique,

B 10 0506842); 2, 4. Habit (Tanzania, B 10 0506843). Scale bars: 1, 2 = 1 cm; 3, 4=2 mm.

610 ... Denchev & Denchev

j .

Fics 5-8. Testicularia africana on Rhynchospora corymbosa (Mozambique, B 10 0506842):

5, 6. Spore balls with an outer layer of dark, thick-walled spores, and sterile cells in the central part,

in LM; 7. A spore ball with loosely situated spores on the surface, and parts of some sterile cells

between the spores, in SEM. 8. Spores in SEM. Scale bars: 5-7 = 10 um; 8 = 5 um.

Testicularia africana from Tanzania and Mozambique ... 611

in Afrika, no. 31 094, Exkursion V.39; later identified by Schultze-Motel 1960: 511, as

R. corymbosa) (B 10 0506842).

ComMENTS — ‘The new records extend the geographical range of Testicularia

africana from Western Africa and Cameroon to Eastern Africa. This smut

fungus is known to infect only one host species, Rhynchospora corymbosa,

which is widespread and common with a pantropical distribution. We expect

that Testicularia africana is more widely distributed in Africa and elsewhere.

Acknowledgements

This research received support from the SYNTHESYS Project http://www.synthesys.

info/ which is financed by European Community Research Infrastructure Action under

the FP7 “Capacities” Program. The authors also gratefully acknowledge Dr Kalman

Vanky (Herbarium Ustilaginales Vanky, Tubingen, Germany) and Dr Roger G. Shivas

(Queensland Primary Industries and Fisheries, Australia) for critically reading the

manuscript and serving as pre-submission reviewers.

Literature cited

Klotzsch JF. 1832. Mycologische Berichtigungen. Linnaea 7: 193-204, Pls VII-X.

Schultze-Motel W. 1960. Afrikanische Cyperaceae aus den Sammlungen von A. Peter. Willdenowia

2: 495-518.

Vanky K. 2009. Ustilaginales exsiccata. Fasc. LIII-LIV (nos 1301-1350). Publications from the

Herbarium Ustilaginales Vanky (HUV) 18: 1-17.

Vanky K. 2010. The smut fungi (Ustilaginomycotina) of Rhynchospora (Cyperaceae). Mycologia

Balcanica 7: 93-104.

Vanky K. 2011 [“2012”]. Smut fungi of the world. APS Press, St. Paul, Minnesota, USA.

Vanky K, Piatek M. 2006. The genus Testicularia (Ustilaginomycetes). Mycologia Balcanica 3:

163-167.

Vanky K, Vanky C, Denchev CM. 2011. Smut fungi in Africa - a checklist. Mycologia Balcanica

8: I-27,

Zambettakis C. 1970. Recherches sur les Ustilaginales d'Afrique. Bulletin de la Société Mycologique

de France 86: 305-692.

MY COTAXON

http://dx.doi.org/10.5248/130.613

Volume 130, pp. 613-620 July-September 2015

Pachyphloeus depressus, a new green truffle from China

Li FAN* & SONG-PIN CAI

College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

* CORRESPONDENCE TO: fanli@mail.cnu.edu.cn

ABSTRACT — Pachyphloeus depressus sp. nov. is described based on specimens collected

from the valley of the Jinshajiang River in southwest China. The species can be distinguished

from all other species in Pachyphloeus by its smooth, greenish brown ascomata covered with

yellow-green hairs, and its globose ascospores with spines that have somewhat T-shaped or

capitate apices. Phylogenetic analyses of the ITS and ITS-LSU supported the erection of a

new species.

Key worps — morphology, phylogeny, Pezizaceae, taxonomy

Introduction

During recent investigations of truffles in southwestern China, some small

green truffles were collected from Qiaojia County, Yunnan Province, and Huili

County, Sichuan Province, in the soil of mixed forest with Pinus armandii

Franch. as the dominant species. Both counties are near the Jinshajiang River.

According to the local people, these green truffles are not uncommon in these

areas, although they are not found in large quantities. The species appears to

be easily confused with the Chinese black truffle Tuber pseudohimalayense G.

Moreno et al. (= T! pseudoexcavatum Y. Wang et al.) because the green truffles

also have a distinctive depression at the base of the ascomata, somewhat similar

to the excavated ascomata of T. pseudohimalayense. Macroscopically, our

collections led us to Pachyphloeus Tul & C. Tul. [nom. illeg.; = Pachyphlodes

Zobel; Pezizaceae], a green truffle genus that is well known from both Europe

and North America (Calonge et al. 2002, Colgan & Trappe 2004, Frank

et al. 2006, Fogel & States 2002, Gilkey 1954, Healy et al. 2009, Lange 1956,

Moreno-Arroyo et al. 1996, Pegler et al. 1993). After critical comparison of

the morphological and molecular characters of our collections with the known

614 ... Fan & Cai

species in both genera, we are confident that our collections represent an

undescribed species.

Materials & methods

Morphological observation

The specimens were collected from Sichuan and Yunnan Provinces of China and

deposited in the Herbarium of the Biology Department, Capital Normal University,

Beijing, China (BJTC). Macroscopic characters were described from fresh specimens.

Microscopic characters were described from razor-blade sections of fresh specimens

mounted in 3% KOH; Melzer’s reagent (Dring 1971: 98), or 0.1% (w/v) cotton blue in

lactic acid. For scanning electron microscopy (SEM), ascospores were scraped from

the dried gleba onto doubled-sided tape, which was mounted directly on an SEM stub,

coated with platinum-palladium, and examined and photographed using a HITACHI

S-4800 scanning electron microscope.

Phylogenetic studies

Herbarium samples were crushed by shaking for 3 min at 30 Hz (Mixer Mill MM

301, Retsch, Haan, Germany) in a 1.5 ml tube, together with one 3 mm diameter

tungsten carbide ball. Total genomic DNA was extracted using the PeqLab E.Z.N.A.

Fungal DNA Kit following the manufacturer's protocol. The internal transcribed spacer

ribosomal RNA gene (ITS) and the 28S large subunit ribosomal RNA gene (LSU) were

amplified using the universal fungal primers ITS1/ITS4 and LROR/LR5 (White et al.

1990, Vilgalys & Hester 1990). PCRs were performed in 50 ul reactions containing

2 ul of DNA template, 2 ul of primer (10 uM/L), and 25 ul of 2x Master Mix (Tiangen

Biotech (Beijing) Co. Ltd.). 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 purification, sequencing, and editing. The

other ITS and LSU sequences included in this study were downloaded from GenBank.

GenBank accession numbers are shown in TABLE 1.

DNA sequences were aligned using Clustal X (Thompson et al. 1997). The alignment

was manually adjusted using Se-Al v.2.03a (Rambaut 2000). Phylogenetic analysis

was conducted separately on the ITS and ITS-LSU alignments. Each aligned dataset

was analyzed with maximum parsimony (MP) using PAUP*4.0b10 (Swofford 2002).

Maximum parsimony analysis was conducted using heuristic searches with 1,000

replicates of random-addition sequence, and the 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

(BS) analysis was performed with 1000 replicates, each with 10 random taxon

addition sequences. TBR branch swapping was employed. Outgroups in these analyses

were sequences from Peziza infossa Fogel & States and Peziza cf. badioconfusa Korf,

chosen following our preliminary data on phylogenetic relationships across the genus

Pachyphloeus (Lzessoe & Hansen 2007, Healy et al. 2009)

TABLE | Specimens and sequences used in this study.

Pachyphloeus depressus sp. nov. (China) ... 615

New sequences are set in bold font.

ACCESSION NUMBER IN GENBANK

SPECIES VOUCHER ITS LsU

Pachyella clypeata (Schwein.) Le Gal - - EU543195

P we Ae a : SOC 775 AY830854 a

Meas JX414191 ws

P. carneus Harkn. RHO00 EU543199 EU543199

RH25 EU543202 EU543202

JT12818 EU543208 EU543208

P. citrinus JRWL2497 EU543196 EU543196

MIN: RH840 JN409343 JN409343

P. conglomeratus 17057 JF908512 -

MA: 29354 JN102487 =

P. depressus BEG aeNe02 KP027405 KT220750

(holotype)

BJTC FAN324 KP027406 KT220751

P. marroninus Healy et al. RH299 EU427549 EU427549

Garcia3557 EU427551 EU427551

JT32454 EU543209 EU543209

P. melanoxanthus (Berk.) Tul. & C. Tul. MM1860 EU543194 EU543194

- JF908511 =

P. thysellii JT13182 EU543197 EU543197

P. virescens RH279 EU543198 EU543198

Peziza infossa - DQ974817 DQ974817

Peziza cf. badioconfusa RH7 EU571229 EU571229

Results

Molecular phylogenetics

In our ITS analyses, the sequences of the new species, Pachyphloeus

depressus, clustered as an independent clade with bootstrap support of 100%

(Fic. 1). The MP tree of combined ITS-LSU (Fic. 2) revealed that the sequences

of P. depressus were similarly grouped in a clade with 100% bootstrap support.

Also with strong bootstrap support was their grouping in a distinctive clade

with three other Pachyphloeus species (P. citrinus Berk. & Broome known from

Europe, P. virescens Gilkey, and P. thysellii W. Colgan & Trappe both from North

America).

Taxonomy

Pachyphloeus depressus L. Fan, sp. nov. Fia. 3

MycoBank MB810700

Differs from all other Pachyphloeus spp. by its smooth, greenish-brown ascomata.

616 ... Fan & Cai

78 Pachyphloeus melanoxa JF908511

93) Pachyphloeus conglome JF908512

74 Pachyphloeus conglome JN102487

Pachyphloeus melanoxanthus EU543194

Pachyphloeus carneus EU543199

99 Pachyphloeus carneus EU543202

Pachyphloeus carneus EU543208

76 ~ Pachyphloeus marroninus EU427549

100] Pachyphloeus marroninus EU427551

Pachyphloeus marroninus EU543209

Pachyphloeus virescens EU543198

Pachyphloeus thysellii EU543197

Pachyphloeus citrinus EU543196

100

Pachyphloeus citrinus JN409343

RU Pachyphloeus depressus BJTC FAN324

100

Pachyphloeus depressus BJTC FAN302

Pachyphloeus austro-oregonensis AY 830854

100

Pachyphloeus austro-oregonensis JX414191

Peziza infossa DQ974817

+. Peziza cf.badioconfus EU571229

Fic. 1. Phylogeny derived from maximum parsimony analysis of the ITS rDNA sequences from

Pachyphloeus species using two Peziza species as outgroup. A total of 220 characters out of 676 were

parsimony-informative. Tree length (TL) = 661 steps, consistency index (CI) = 0.7655, retention

index (RI) = 0.7939, homoplasy index (HI) = 0.2345, and rescaled consistency index (RC) = 0.6077.

Bootstrap values of more than 70% are shown above the respective branches. Novel sequences are

printed in bold.

Pachyphloeus depressus sp. nov. (China) ... 617

Pachyphloeus carneus EU543202

100

toq . Pachyphioeus carneus EU543199

— Pachyphloeus carneus EU543208

Pachyphloeus melanoxanthus EU543194

100

Pachyphloeus marroninus EU427549

Pachyphloeus marroninus EU427551

400

Pachyphloeus marroninus EU543209

Pachyphloeus depressus BJTC FAN302

Pachyphloeus depressus BJTC FAN324

Pachyphloeus virescens EU543198

ay Pachyphloeus thysellii EU543197

Pachyphloeus citrinus EU543196

Pachyphloeus citrinus JN409343

Pachyella clypeata EU543195

[+ Peziza infossa DQ974817

Peziza cf.badioconfus EU571229

Fic. 2. Phylogeny derived from maximum parsimony analysis of the ITS-LSU rDNA sequences

from Pachyphloeus species and Pachyella clypeata using two Peziza species as outgroup. A total of

336 characters out of 1458 were parsimony-informative. Tree length (TL) = 993 steps, consistency

index (CI) = 0.8087, retention index (RI) = 0.7974, homoplasy index (HI) = 0.1913, and rescaled

consistency index (RC) = 0.6449. Bootstrap values of more than 70% are shown above the respective

branches. Novel sequences are printed in bold.

618 ... Fan & Cai

Type: China. Yunnan Province, Qiaojia County, in soil under forest dominated by

Pinus armandii, 16 Oct. 2013, Jin-zhong Cao 807 (Holotype, BJTC FAN302; GenBank

KP027405, KT220750).

EryMoLocy: depressus (Lat.), referring to the shape of the ascomata.

AscoMaAta oblate spheroid or cake-like, 0.9-2.1 cm diam., texture rubbery,

smooth, yellow-green, brownish green to dark brownish green, covered with

densely yellow-green hairs in places; the apical orifice more or less circular and

usually distinctively depressed, dark brown and often with flat or pyramidal

warts; basal tuft of hyphae earth brown, sometimes located in a distinctive

superficial depression. Odor pungent when ripe, somewhat like burnt potato.

Taste not tested. PERIDIUM at maturity one layer of pseudoparenchymatous

tissue, 250-500 um thick, composed of sub-angular to rectangular cells, light

brown, thin-walled, cells 10-60 um diam., more or less arranged with long axes

parallel to the outer surface; towards inside grading to a thin layer of parallel

textura intricata, hyphae hyaline, 5-10 um wide next to gleba. The outermost

cells are often irregularly elongated with ends free, creating a peridial surface

that is more or less felty or hairy. GLEBA yellow-green to brownish green when

fresh, with large and rare whitish yellow veins. Glebal hyphae 7.5-10 um wide

at septa, hyaline, thin-walled, some cells swollen to 15 um. Ascr not forming a

palisade, subglobose or broadly elliptic, 70-95 x 45-75 um, thin-walled, sessile

or with a short stalk, with 8 irregularly arranged ascospores, non-reactive

to Melzer’s solution with pretreatment in 3% KOH. Ascospores globose,

17.5-20 um excluding spines, hyaline at first, then light greenish, ornamented

with coarse, rod-like spines 2-2.5 um long, apices expanding to irregular

T-shape or capitate under SEM.

ADDITIONAL SPECIMEN EXAMINED: CHINA. SICHUAN PROVINCE, Huili County, in soil

under forest dominated by Pinus armandii, 16 Oct. 2013, Jin-zhong Cao & Li Fan 826

(BJTC FAN324; GenBank KP027406, KT220751).

ComMMENTs — The new species, Pachyphloeus depressus, is characterized

morphologically by its combination of cake-like ascomata, smooth on the

surface and covered with yellow-green hairs, subglobose asci, and ascospores

densely covered with spines that have irregularly T-shaped tips. Pachyphloeus

depressus is distinguished from the other Pachyphloeus species by its smooth

ascomata, with the exception of P conglomeratus Berk. & Broome known

from Europe. Pachyphloeus conglomeratus also lacks warts on its ascomatal

peridium, but instead has red-brown scurfy particles, and its ascomata are dark

brown (Pegler et al. 1993) rather than the yellow-green or brownish green of

P. depressus. Furthermore, the distinctively long clavate asci of P conglomeratus

also clearly separate it from P depressus. Our ITS phylogeny (Fic. 1,2) shows

Pachyphloeus depressus sp. nov. (China) ... 619

10um

Fic. 3. Pachyphloeus depressus (BJTC FAN302, holotype): a,b. ascomata; c. asci and ascospores

observed under the light microscope; d. ascospores observed under SEM.

that sequences of P. depressus group in a distinct clade with strong support (BP

= 100%), further supporting the erection of a new species.

Pachyphloeus depressus and three other species in the genus cluster in a

distinct clade (BP = 97%) in the tree constructed from the combined ITS-LSU

database (Fig. 2). The morphological trait that these species share is ascomata

that are more or less green or have yellow tints. However, P depressus has

ascomata that usually have flat or pyramid warts around the apical orifice

only. In contrast, the ascomata of all the other three species have distinctively

pyramidal warts over all their surfaces.

Pachyphloeus depressus seems not uncommon in the Jinshajiang River

valley. It has been locally called the “green female truffle” because its ascomata

superficially resemble that of Tuber pseudohimalayense, a very common truffle

species in this area.

Acknowledgments

We are grateful to Dr. Toni Atkinson, Otago Mycological Institute, New Zealand,

and Prof. Ying-Ren Lin, College of Forest and Garden, Anhui Agricultural University,

China, for serving as pre-submission reviewers. We wish to extend a special thanks

620 ... Fan & Cai

to Prof. Lei Cai, Institute of Microbiology of the Chinese Academy of Sciences, for

his critical suggestions on our manuscript. This study was supported by the National

Natural Science Foundation of China (No. 31270058) and the Beijing Natural Science

Foundation (No. 5122003).

Literature cited

Calonge FD, Garcia F, Juste P. 2002. Nuovi dati sui funghi ipogei della Spagna. IX. Pachyphloeus

macrosporus sp. nov. Bollettino Gruppo Mycologico G. Bresadola 45: 51-61.

Colgan W III, Trappe JM. 2004. NATS truffle and truffle-like fungi 10: Pachyphloeus thysellii sp.

nov. (Pezizaceae, Pezizomycotina). Mycotaxon 90: 281-284.

Dring DM. 1971 Techniques for microscopic preparation. 95-111, in: C Booth (ed.). Methods in

microbiology, Vol. 4. New York: Academic Press Inc.

Fogel R, States J. 2002. Materials for a hypogeous mycoflora of the Great Basin and adjacent

cordilleras of the western United States. VIII: Pachyphloeus lateritius sp. nov. and Cazia

quericola [sic] sp. nov. (Ascomycota, Pezizales). Mycotaxon 81: 83-89.

Frank JL, Southworth D, Trappe JM. 2006. NATS truffle and truffle-like fungi 14: Pachyphloeus

austro-oregonensis, a new species from southern Oregon. Mycotaxon 98: 253-259.

Gilkey HM. 1954. Tuberales. North American Flora, Ser. 2, 1: 1-36.

Healy RA, Bonito G, Guevara G. 2009. The truffle genus Pachyphloeus in the US.

and Mexico: phylogenetic analysis and a new species. Mycotaxon 107: 61-71.

http://dx.doi.org/10.5248/107.61

Leessoe T, Hansen K. 2007. Truffle trouble: what happened to the Tuberales? Mycological Research

111: 1075-1099. http://dx.doi.org/10.1016/j.mycres.2007.08.004

Lange M. 1956. Danish hypogeous fungi. Dansk Botanisk Arkiv 16: 1-48.

Moreno-Arroyo B, Gomez J, Calonge FD. 1996. Pachyphloeus prieguensis, sp. nov. (Ascomycotina),

encontrada en Espana. Boletin de la Sociedad Micoldgica de Madrid 21: 85-92.

Pegler DN, Spooner BM, Young TWK. 1993. British truffles: a revision of British hypogeous fungi.

Kew: Royal Botanic Gardens. 216 p., 26 pl.

Rambaut A. 2000. Estimating the rate of molecular evolution: incorporating non-contemporaneous

sequences into maximum likelihood phylogenies. Bioinformatics 16: 395-399.

http://dx.doi.org/10.1093/bioinformatics/16.4.395

Swofford DL. 2002. PAUP*, phylogenetic analysis using parsimony (*and other methods), version

4. Sunderland, MA, USA, Sinauer Associates.

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The CLUSTALX windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 24: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several species of Cryptococcus. Journal of Bacteriology 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

Methods and Applications. Academic Press, San Diego.

MY COTAXON

http://dx.doi.org/10.5248/130.621

Volume 130, pp. 621-624 July-September 2015

A new species of Selenodriella from Brazil

Marina A. G. ARAUJO', MARCELA A. BARBOSA’, Mayra S. OLIVEIRA’,

ELAINE MALOSSO”*, & RAFAEL E CASTANEDA-RuIz?3

'Programa de Pés-Graduagao em Biologia de Fungos, Universidade Federal de Pernambuco &

*Centro de Ciéncias Biologicas, Departamento de Micologia/Laboratério de Micorrizas,

Universidade Federal de Pernambuco, Avenida da Engenharia, s/n

Cidade Universitaria, Recife, PE, 50.740-600, Brazil

*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

* CORRESPONDENCE TO: elaine.malosso@ufpe.br

ABsTRACT—Selenodriella amoena sp. nov., collected on a decaying leaf of an unidentified

plant in the Brazilian Atlantic Forest, is described and illustrated. It is distinguished by

irregularly branched conidiophores and subcylindrical to slightly navicular, unicellular,

colorless conidia. A key to Selenodriella species is provided.

KEY worps — asexual fungi, leaf litter, taxonomy

Introduction

Selenodriella R.F. Castafteda & W.B. Kendr. is distinguished by distinct

single erect septate brown setae and conidiophores; polyblastic, mostly

discrete, sessile, lageniform, conidiogenous cells that become apically

swollen and inconspicuously denticulate due to sympodial elongation during

conidiation and which are mostly borne laterally (sometimes terminally) on

the conidiophore or its branches; and falcate to fusiform, unicellular, colorless,

smooth conidia (Castafieda-Ruiz & Kendrick 1990, 1991; Bhat & Kendrick

1993). After Castanteda-Ruiz et al. (2009) transferred Selenodriella perramosa

W.B. Kendr. & R.E Castafieda to Selenosporella, Selenodriella comprised four

accepted species (Index Fungorum 2014). We describe a new species described

here.

622 ... Aratijo & al.

Materials & methods

During an expedition in October 2014 through “Abreu e Lima’, Pernambuco State,

in Brazil’s Atlantic forest, samples of decaying leaves were stored in plastic bags, taken to

the laboratory, and treated according to Castafeda-Ruiz (2005). Mounts were prepared

in PVL (polyvinyl alcohol and lactic acid) and measurements made at a magnification of

1000. Photomicrographs were obtained with a Nikon microscope with bright field and

phase contrast optics. The holotype was deposited in the Herbarium of Universidade

Federal de Pernambuco, Recife, Brazil (URM).

Taxonomy

Selenodriella amoena M.A.G Aratijo, M.A. Barbosa, Malosso &

R.F. Castafieda, sp. nov. Fig. 1

INDEXFUNGORUM IF550923

Differs from other Selenodriella spp. by its subcylindrical conidia that are subacute at the

base and rounded at the apex.

Type: Brazil, Pernambuco, Abreu e Lima, Sitio Sao Joao Sistema Agroflorestal, 7°53’S

34°53’W, on decaying leaf of an unidentified plant, 31-October-2014, coll. M.A.G.

Araujo (Holotype: URM87583).

EryMoLoey: Latin, amoena, meaning delightful and beautiful, in reference to the

appearance of conidiogenous cells and conidia.

CoLonizs on the natural substrate effuse, amphigenous, hairy, brown.

Mycelium mostly immersed, composed of septate, branched, pale brown to

brown hyphae, 1-2 um wide. Conrp1opHorss distinct, single, erect, straight

or flexuous, alternately or irregular branched, sometimes setiform and

sterile, 4-9-septate, smooth, dark brown below, pale brown toward the apex,

80-200 x 5-12 um. CONIDIOGENOUS CELLS polyblastic, discrete, mostly

sessile, lageniform to somewhat subulate, sympodially elongated, with small

denticles arising unilaterally or alternately from the conidiophores, from one-

celled branches or sometimes at the conidiophore apex, pale brown to almost

colorless, 12-20 x 2-4 um. Conidial secession schizolytic. Conip1A solitary,

subcylindrical or slightly navicular, subacute at the base, rounded at the apex,

unicellular, smooth, colorless, 7-11 x 1-3 um.

Key to Selenodriella species

2. Conidia subfalcate, inconspicuously truncate at the base,

rounded-at-the apex al 5a20'% 252-5 llr ee tee eats s betes bates bos S. fertilis

2% Conidiadfaleate,; endsracute 7 10k O5= 0 winks, cp ap.geu scoot ese amen eece$ S. indica

Fig.l. Selenodriella amoena (ex URM 87583). A. Conidia. B-C. Conidiogenous cells.

D. Conidiophores and conidiogenous cells.

Selenodriella amoena sp. nov. (Brazil) ... 623

624 ... Aratijo & al.

3. Conidia subcylindrical or slightly navicular, subacute at the base,

roynded abtheapex, 21k lutte lag. ve Bie ee Bh ee le ce we S. amoena

3. Conidia slightly fusiform, vermiform, or needle-shaped .....................04. 4

4. Conidia fusiform or vermiform, asymmetrical, tapered toward each end,

base curved and pointed, apex obtuse or rounded, 9-12 x 1-1.5 um

sb ts see bee suet ti or olBS sg wh se oitia aod, =: Spelt ate trode ht beast abs brags Wh S. inaequilaterospora

4. Conidia needle-shaped or fusiform, 9-15 x 0.5 um..............006- S. intermedia

Acknowledgments

The authors express their sincere gratitude to Prof Dr. Bryce Kendrick and Dr.

De-Wei Li for their critical review of the manuscript. The authors are grateful to the

“Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)” for financial

support through project 88881.062172/2014-01 and the “Programa Ciéncia sem

Fronteiras.” We are grateful to Jones Severino Pereira for allowing substrate sampling

in his property. RFCR is grateful to the Cuban Ministry of Agriculture and “Programa

de Salud Animal y Vegetal’, project P131LH003033 for facilities. We acknowledge the

facilities maintained by Dr. P.M. Kirk and Drs. V. Robert and A. Decock through the

Index Fungorum and MycoBank websites. Dr. Lorelei Norvell’s editorial review and Dr.

Shaun Pennycook’s nomenclature review are greatly appreciated.

Literature cited

Bhat DJ, Kendrick B. 1993. Twenty-five new conidial fungi from the Western Ghats and the

Andaman Islands (India). Mycotaxon 49: 19-90.

Castaneda Ruiz RE. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Castaneda Ruiz RF, Kendrick B. 1990. Conidial fungi from Cuba II. University of Waterloo Biology

Series. 33. 61 p.

Castafieda-Ruiz RE, Kendrick B. 1991. Ninety-nine conidial fungi from Cuba and three from

Canada. University of Waterloo Biology Series 35. 132 p.

Castafieda Ruiz RE, Guerrero B, Adamo GM, Morillo O, Minter DW, Stadler M, Gené J, Guarro

J. 2009. A new species of Selenosporella and two microfungi recorded from a cloud forest in

Mérida, Venezuela. Mycotaxon 109: 63-74. http://dx.doi.org/10.5248/109.63

Index Fungorum. 2014. http://www.indexfungorum.org/names/Names.asp. (Accessed: 30.X1.2014.)

MY COTAXON

http://dx.doi.org/10.5248/130.625

Volume 130, pp. 625-627 July-September 2015

Antrodiella indica, a new species from India

GURPREET KAUR, AVNEET P. SINGH’, & G.S. DHINGRA

Department of Botany, Punjabi University, Patiala 147002, India

*CORRESPONDENCE TO: avneetbot@gmail.com

ABSTRACT — A new poroid species, Antrodiella indica, is described from Chandigarh, India.

Key worps — Basidiomycota, Agaricomycetes, Phanerochaetaceae

During September 2012 Dhingra, with the help of Sagar, collected a fungus from

a dead tree of Cassia fistula from Sector 18-D Park, Chandigarh, India. After

comparison of macroscopic and microscopic characters (Thind et al. 1957, Reid

et al. 1959, Donk 1967, Thind & Dhanda 1980, Niemela 1982, David & Tortic

1986, Ryvarden & Gilbertson 1993, Nufiez & Ryvarden 2001, Bernicchia 2005,

Miettinen et al. 2006, Sharma 2012, Baltazar et al. 2014, Ryvarden & Melo 2014),

it has been identified as a species of Antrodiella (Basidiomycota, Agaricomycetes,

Phanerochaetaceae) based on basidiocarps causing white rot, dimitic hyphal

system, clamped generative hyphae, and thick-walled skeletal hyphae. It differs

from previously described Antrodiella species, A. romellii (Donk) Niemela in

particular, by its bigger, ellipsoid to subfusiform basidiospores. The new species

is here named as A. indica. A portion of the basidiocarp was sent to Dr. Leif

Ryvarden (Oslo, Norway) and Dr. Nils Hallenberg (Sweden), who confirmed

the findings.

Antrodiella indica G. Kaur, Avneet P. Singh & Dhingra, sp. nov. PiatEs 1, 2

MycoBANnk MB 811102

Differs from Antrodiella romellii by its bigger, ellipsoid to subfusiform basidiospores.

Type: India, Union Territory, Chandigarh, Sector 18-D Park, on dead tree of Cassia

fistula L. (Leguminosae), 1 September 2012, G.S. Dhingra 7072 (PUN, holotype).

EryMo.ocy: The epithet refers to the country where first collected.

Basidiocarp annual, resupinate to effused-reflexed to pileate, dimidiate, soft

leathery when fresh, tough leathery when dry; pilei up to 1.5 x 1.2 x 0.8 cm,

applanate, abhymenial surface smooth, grayish brown to light brown to reddish

brown when fresh, not changing much on drying; hymenial surface poroid,

626 ... Kaur, Singh, & Dhingra

PLATE 1. Antrodiella indica (holotype). 1. Collection of material from the dead tree of Cassia fistula;

2. Fresh basidiocarp; 3. Transverse section through a pore; 4. Basidium; 5. Basidiospores.

pale orange to grayish orange to brownish orange when fresh, reddish gray to

dull red to light brown on drying; pores angular, 4-5 per mm; dissepiments

thin, entire to lacerate; context up to 0.5 mm thick, reddish brown; pore tubes

up to 2 mm long, concolorous with hymenial surface; margins acute, irregular

on drying. Causing white rot. Hyphal system dimitic; generative hyphae up

to 3 um wide, branched, septate, clamped, thin-walled. Skeletal hyphae up to

4 um wide, rarely branched, aseptate, thick-walled. Tramal zone dominated

by skeletal hyphae. Cystidia absent, but cystidioles present in the hymenium,

9.5-14.5 x 3.5-5.5 um, fusoid, thin-walled, with basal clamp. Basidia 9-15.5

x 4.8-5.8 um, clavate, 4-sterigmate, with basal clamp; sterigmata <5 um long.

Basidiospores 5.3-6.8 x 2-2.8 um, ellipsoid to subfusiform, smooth, inamyloid,

acyanophilous.

REMARKS— Antrodiella romellii can be distinguished from the new species by

its smaller (3.5-4 x 2-2.5 um) ellipsoid spores.

Acknowledgements

The authors thank SERB, DST, Government of India for financial assistance, Head,

Department of Botany, Punjabi University, Patiala, for providing research facilities,

Dr. Leif Ryvarden (Oslo, Norway) for expert comments, Dr. Nils Hallenberg (Sweden)

for expert comments and peer review, Prof. B.M. Sharma (Department of Plant

Pathology, COA, CSKHPAU, Palampur, H.P., India) for peer review, and Mycotaxon,

Ltd. for underwriting page charges for this publication.

Antrodiella indica sp. nov. (India) ... 627

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PLATE 2. Antrodiella indica (holotype). 1. Basidiospores; 2. Basidia; 3. Generative

hyphae; 4. Skeletal hyphae; 5. Cystidioles; 6. Transverse section of basidiocarp showing

tramal hyphae, basidia, cystidioles, and basidiospores.

Literature cited

Baltazar JM, Ryvarden L, Gibertoni TB. 2014. Diplomitoporus (Polyporales, Basidiomycota) in

Brazil revisited. Mycol. Progress 13: 313-319. http://dx.doi-org/10.1007/s11557-013-0916-4

Bernicchia A. 2005. Polyporaceae s.]. Fungi Europaei 10. Edizioni Candusso, Alassio, Italia. 808 p.

David A, Tortic M. 1986. Contribution a étude de quatre polypores européens peu connus.

Cryptogamie Mycologie 7: 1-13.

Donk MA. 1967. Notes on European polypores-II. Persoonia 5: 47-130.

Miettinen O, Niemela T, Spirin W. 2006. Northern Antrodiella species: the identity of A. semisupina

and type studies of related taxa. Mycotaxon 96: 211-239.

Niemela, T. 1982. Taxonomic notes on the polypore genera Antrodiella, Daedaleopsis, Fibuloporia

and Phellinus. Karstenia 22: 11-12.

Nunez M, Ryvarden L. 2001. East Asian polypores 2. Synopsis Fungorum 14: 170-522.

Reid DA, Thind KS, Chatrath MS. 1959. The Polyporaceae of the Mussoorie hills (India) IV. Trans.

Brit. Mycol. Soc. 42: 40-44. http://dx.doi.org/10.1016/S0007-1536 (59)80064-4

Ryvarden L, Gilbertson RL. 1993. European polypores, part 1. Fungiflora, Oslo, Norway. 387 p.

Ryvarden L, Melo I. 2014. Poroid fungi of Europe. Synopsis Fungorum 31: 455 p.

Sharma JR. 2012. Aphyllophorales of Himalaya. Botanical Survey of India, Calcutta. 590 p.

Thind KS, Dhanda RS. 1980. The Polyporaceae of India-XII. Indian Phytopathology 33: 380-387.

Thind KS, Bindra PS, Chatrath MS. 1957. The Polyporaceae of the Mussoorie hills—III. Res. Bull.

Panjab University 129: 471-483.

MY COTAXON

http://dx.doi.org/10.5248/130.629

Volume 130, pp. 629-639 July-September 2015

Russula sichuanensis and its ectomycorrhizae

from Himalayan moist temperate forests 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 — Russula sichuanensis and its ectomycorrhizae were collected from monoculture

forests of Pinus wallichiana. This fungus is a new record for Pakistan and the ectomycorrhizae

are described for the first time, based on morphology and nrDNA ITS sequences. Russula

sichuanensis is a morphologically variable species especially with respect to the colour

and shape of its pileus. The taxon is characterized by small to medium-sized stipitate

basidiomata, reticulate globose to subglobose basidiospores, and pluriseptate pileocystidia.

The ectomycorrhizae are distinguished by dichotomously branched orange-brown to grayish

brown tips, its pseudoparenchymatous acystidiate mantle with irregularly shaped and often

interlocking cells, and curved and frequently septate emanating hyphae. Pinus wallichiana is

a new host for this species.

Key worps — Khyber Pakhtoonkhaw, phylogeny, taxonomy, Russulaceae, Russulales

Introduction

The genus Russula Pers. is among the most diverse genera of Agaricomycetes.

The genus includes approximately 750 species (Kirk et al. 2008), many of which

are notorious for their intergrading colour variations and obscure morphological

and anatomical discontinuities (Miller & Buyck 2002). Although thirty-three

Russula species have been recorded from Pakistan (Ahmad et al. 1997, Niazi et

al. 2006, Razaq 2013), ectomycorrhizae have been described only for Russula

brevipes Peck (Niazi et al. 2006).

The aim of the present study is to elucidate the morphology and phylogeny of

Russula sichuanensis and its ectomycorrhizae that form symbiotic associations

with forest trees and play important role in the establishment and conservation

of Himalayan forests.

During a mushroom survey in 2012 and 2013, seven different collections of

morphologically different basidiomata and their ectomycorrhizae were made

630 ... Saba & Khalid

in Khyber Pakhtoonkhaw, Pakistan, which upon molecular analysis clustered

together. Many of their morpho-anatomical features coincided with those of

the Chinese species, Russula sichuanensis, and this identification was confirmed

by ITS sequence analyses. Russula sichuanensis, a morphologically variable

species, represents a new record for Pakistan, and its association with Pinus

wallichiana has not previously been noted. Its ectomycorrhizae are described

here for the first time.

Materials & methods

Morphological examination

Basidiomata were collected in 2012 and 2013, photographed, vouchered, dried,

and characterized morphologically. All specimens are conserved in the Herbarium,

Department of Botany, University of the Punjab, Lahore, Pakistan (LAH). Sections of

specimens were mounted in 5% KOH for microscopic observations under a biological

microscope (MX4300H, Meiji Techno Co., Ltd., Japan). Tissues were mounted in

phloxine to increase contrast of the structures, and Melzer’s reagent was used to examine

amyloid reactions.

Measurements of anatomical features (basidiospores, basidia, cystidia, pileus hyphae,

and stipe hyphae) were made from at least 25 observations using an ocular micrometer

and 100x oil-immersion objective; they include x = arithmetic mean of spore length

and spore width for all spores measured, Q = spore length divided by spore width. Line

drawings were made with a camera lucida. Color designations are from Munsell (1975).

DNA extraction, PCR amplification, DNA sequencing

Genomic DNAs were extracted from small pileus pieces using a modified CTAB

method (Bruns 1995). Amplification of internal transcribed spacers (ITS1 and ITS2)

and the 5.8S region of the nuclear ribosomal RNA gene were targeted using the primer

pairs ITS1F/ITS4 (White et al. 1990; Gardes & Bruns. 1993) using the Extract-N-Amp

plant DNA extraction Kit (Sigma-Aldrich, St. Louis, MO, USA). PCR was carried out

under the following cycling parameters: initial denaturation (94°C for 1 min), 35 cycles

(94°C for 1 min, 53°C for 1 min, and 72°C for 1 min), and final extension (72°C for 8

min). Amplified PCR products were sent for purification and bidirectional sequencing

to Macrogen (Korea).

Sequence alignment & phylogenetic analysis

Sequences were manually edited and assembled using BioEdit (www.mbio.ncsu.edu/

bioedit/bioedit.html). Following Dentinger et al. (2011) for complete ITS sequences,

all sequences were trimmed with the conserved motifs 5’-( Gat) CATTA- and —GACCT

(CAAA )-3’, and the alignment portions between them were included in analysis. All

positions containing gaps and missing data were eliminated. There were a total of 406

positions in the final dataset. The analysis involved 37 nucleotide sequences. Albatrellus

ovinus was used as outgroup based on results reported by Li et al. (2013).

Seven sequences from Russula sichuanensis basidiocarps, one sequence from an

ectomycorrhizal root tip, and other related sequences retrieved from the GenBank

were included in the phylogenetic analysis. The sequences were aligned by Muscle

Russula sichuanensis ectomycorrhiza (Pakistan) ... 631

Russula sichuanensis JX391968

W@ Russula sichuanensis_KM596859

Russula sichuanensis KF002768

Russula sichuanensis KF002760

i Russula sichuanensis_KM596860

91 | Russula sichuanensis JX391969, ex holotype

@ Russula sichuanensis_KM596861

@ Russula sichuanensis_KM596862

@ Russula sichuanensis_KM596866

37] | mi Russula sichuanensis_KM596863

i Russula sichuanensis_KM596864

@ Russula sichuanensis_KM596865

Russula_cessans_AY061730

g2| | Russula_laricina_AY061685

30L Russula_nauseosa_JF908642

Russula_curtipes_AY061668

Russula_violacea_AY061725

63 Russula_caerulea_AY061661

Russula_olivacea_AY061699

Russula_decolorans_AY061670

Russula_xerampelina_AY061734

Russula_queletii_AY061711

Russula_exalbicans_AF418622

Sect. Laricinae

*L Russula_fellea_ AY061676

Russula_emetica_AY061673

95 Russula_adusta_AY061652

Russula_ingwa_JX266624

53 Russula_delica_AY061671

Russula7_foetens_ AY061677

Russula_cyanoxantha_AY061669

Russula_ilicis AY061682

96 Russula_crustosa_EU598194

Russula_mustelina_AY061693

77 Russula_aeruginea_AF418612

79 Russula_grisea_AY061679

Albatrellus_ovinus_AY198202

0.02

Fic. 1. Phylogenetic relationship of Russula sichuanensis and closely related taxa based on Maximum

Likelihood method inferred from nrITS sequences. The bootstrap consensus tree inferred from

1000 replicates is taken to represent the evolutionary history of the taxa analyzed.

632 ... Saba & Khalid

(Beta) using the default setting in Molecular Evolutionary Genetics Analysis (MEGA)

software (Tamura et al. 2011). A phylogenetic tree was constructed with the Maximum

Likelihood (ML) algorithm using a Jukes & Cantor (1969) model of nrITS sequences

and nearest-neighbor-interchange (NNI) as ML heuristic search method using MEGA5

software (Tamura et al. 2011). The topology was assessed by 1000 bootstrap replicates.

Corresponding bootstrap values >50% are cited on the tree figure. Percent Identities

(PID) and DNA divergences were calculated by DNAStar.

The Russula sichuanensis sequences generated for this study were submitted to

GenBank and the accession numbers for these and other closely related taxa used in the

phylogenetic analysis are cited in phylogenetic tree (Fic. 1).

Results

Sequencing of the PCR products from the nrITS region of seven Pakistani

specimens and one ectomycorrhizal root tip yielded fragments of 560-770 base

pairs. Initial blast analysis of nucleotide sequences revealed a 99% maximum

identity of the Pakistani sequences (GenBank KM596859-596866) with the

sequence from the Russula sichuanensis holotype (GenBank JX391969). For

phylogenetic analysis, several closely related taxa were selected from GenBank

(Fiealy.

After removing the ambiguous letters from the aligned datasheet, 1008

characters were subjected to phylogenetic analysis of which 294 characters

were conserved, 428 were variable, 270 were parsimony informative, and 143

were singletons.

Percentage similarity was calculated by MegAlign (DNA Star Inc.). The

Pakistani collections showed 98.6% identity and 0% genetic divergence with

sequence JX391969 (R. sichuanensis holotype); 98% identity and 0% genetic

divergence with sequence JX391968 (R. sichuanensis paratype), and 99%

identity and 0.3% divergence with sequences KF002760 and KF002768.

Taxonomy

Russula sichuanensis G.J. Li & H.A. Wen, Mycotaxon 124: 179 (2013). Figs 2-4

Basidiomata small to medium sized, epigeous or semi-hypogeous, russuloid

when mature. PILEUs 15-62 mm diam., when young broadly parabolic, broadly

convex with incurved margin and moderately to deeply indented centre or plane

with flattened umbo, flesh grey upon drying, when mature straight or flaring

to uplifted, tuberculate striate; surface dull, viscid, the centre very dark red

(2.5R1/4) to very deep red (2.5R1/8), gradually less intense towards margin to

dark red (2.5R3/4), the margin light grayish red (2.5R6/4) to deep red (2.5R3/10)

or light olive brown (2.5Y5/2). LAMELLAE adnate or sinuate, regular, crowded,

pale greenish yellow (7.5Y9/4) or light orange yellow (10YR9/8), margins even,

concolorous. LAMELLULAE absent. STIPE 32-75 mm long, 7-11 um wide above,

6-16 um wide below, central, clavate or subcylindrical, tapering upwards, dry,

Russula sichuanensis ectomycorrhiza (Pakistan) ... 633

Fic. 2. Russula sichuanensis basidiomes: A, MSM0019; B, MSM0020; C, MSM0021; D, MSM0022.

Scale bars-= 10 mm.

dull, longitudinally rugulose, stuffed at first, becoming hollow (cavernous)

upon maturity, white with yellowish tinge. Annulus and volva absent. Odor

and taste not recorded.

BASIDIOSPORES 8-12 x 7-10 um [x = 9 x 7.7 um, Q = 1.0-1.3], globose or

subglobose, covered with amyloid warts linked as small crests and forming a

634 ... Saba & Khalid

Fic. 3. Russula sichuanensis basidiomes: A, MSM0023; B, MSM0024. Scale bars = 10 mm.

partial network, rarely intermixed with isolated verrucae, warts often 0.5-1.5 um

high, thin-walled, yellowish in KOH, suprahilar plage well defined, 1.5-3.5 um.

Basip1a 28-40 x 11-17 um, clavate, bi- or tetra-spored, thin-walled, hyaline

in KOH; sterigmata (2.5-)4.5-6 um. TRAMA cellular, cells inflated, thin walled,

hyaline or yellowish in KOH 19-36 x 22-47 um. CHEILOCYSTIDIA absent.

PLEUROCYSTIDIA 42-78 x 8-16 um, clavate or rarely mucronate with crystal

Russula sichuanensis ectomycorrhiza (Pakistan) ... 635

Fic. 4. Russula sichuanensis (MSMO0019): A, Basidia; B, Pleurocystidia; C, Pileocystidia;

D, Caulocystidia; E, Basidiospores. Scale bars: A = 20 um; B-D = 25 um; E = 8 um.

or refringent contents, hyaline or yellowish in KOH, thin walled. PILEIPELLIs

an ixotrichoderm composed of cylindrical hyphae, 3.5-6.3 um diam., thin-

walled, hyaline or greenish in KOH. Piteocystip1a with refractive contents,

septate, clavate to cylindrical, 6.3-11.3 um. STIPITIPELLIS interwoven, hyphae

cylindrical, 3.3-5.3 um, septate, hyaline in KOH. CauLocystipia with

refractive contents, septate, clavate to cylindrical, 56-89 x 6-10 um. CLamp

CONNECTIONS absent.

MATERIAL EXAMINED (all in LAH): PAKISTAN, KuYBER PAKHTOON KHaw,

Mansehra, Chattar plain, under Pinus wallichiana A.B. Jacks., 15 September 2012,

M. Saba MSM0019 (GenBank KM596859), MSM0020 (GenBank KM596860); 22

September 2013, M. Saba, MSM0021 (GenBank KM596861), MSM0022 (GenBank

KM596862), MSM0023 (GenBank KM596863), MSM0024 (GenBank KM596864),

MSM0025 (GenBank KM596865); Shangla, ectomycorrhiza near rhizosphere of Pinus

wallichiana, 2 September 2013, M. Saba MSE003 (GenBank KM596866).

Morphology of the ectomycorrhizae Fic. 5

ECTOMYCORRHIZAL SYSTEM: Dichotomous, ramified system 2.5 mm long.

Unramified ends straight, 0.5 mm long, orange-brown to grayish brown and

0.5 mm in diameter (basal portion 0.4 mm wide; apical portion 0.5 mm wide).

Very tips orange brown. Mantle surfaces smooth, matte, rarely giving rise to

emanating hyphae and with host visible through mantle. Mycorrhizae orange-

brown when young and grayish brown when old.

636 ... Saba & Khalid

HT }

| Le

Fic. 5. Russula sichuanensis (MSE003) ectomycorrhizae: A, Morphotypes; B, Outer mantle;

C, Inner mantle; D, Emanating hyphae. Scale bars: A = 5 mm; B-D = 15 um.

OUTER MANTLE: pseudoparenchymatous (type L; Agerer 1987-2012) with

irregularly shaped, often interlocking cells. Cells olive yellow to yellowish

brown, 9.6—28.3 x 6.3-17.4 um. INNER MANTLE: pseudoparenchymatous (type

L; Agerer 1987-2012) with irregularly shaped, often interlocking cells. Cells

olive yellow to yellowish brown, 47-86.5 x 24-58 um. EMANATING HyPHAE:

curved, frequently septate, yellowish brown to olive yellow, thin walled, rarely

branched, 8-25.9 x 2.4-4.1 um. No clamp connection observed; H-type

anastomosis observed among hyphae. RHIZOMORPHS: not observed. CysTIDIA:

not observed.

Russula sichuanensis ectomycorrhiza (Pakistan) ... 637

Discussion

Russula sichuanensis was originally described by Liet al. (2013) from Sichuan

province, China. It is characterized by an uneven pileus that often covers a

white percurrent stipe and tightly crowded sinuate cream to yellow lamellae;

it was additionally described as having a cream-tinged pileus (often olivaceous

brown at the centre), a yellowish to orange sublamellate gleba, and stipitate

basidiomata (Li et al. 2013). Our collections differ morphologically from the

type description as follows: (1) curly and tightly contorted pilei were observed

in only a few young basidiomata while the majority of mature basidiomata

have broadly convex pilei with straight or uplifted margins and regular, straight

(without convolutions) lamellae; (2) the pileus color is variable, ranging from a

very dark red to very deep red centre, dark red gradually towards margin, light

grayish red to deep red margin, or light olive brown. Most of the microscopic

features, including shape, size of basidiospores, their ornamentation,

presence of ixotrichoderm pileipellis with septate pileocystidia and septate

caulocystidia, coincide with the description given by Li et al. (2013); however

our collections had basidiospores with a well defined suprahilar plage and

frequent pleurocystidia. Nonetheless, a BLAST comparison of all ITS sequences

retrieved from our seven different basidiomata and one ectomycorrhizal root

tip with the holotype of R. sichuanensis showed 99% identity and 100% query

coverage and clustered with the holotype sequence and three other Chinese

sequences in a clade with 91% support in phylogenetic reconstruction (Fic. 1).

Russula sichuanensis can now be characterized as a morphologically variable

species, especially with respect to pileus colour and shape and which produces

either asublamellate gleba or regular lamellae. In addition, it can be characterized

by partially small to medium sized stipitate basidiomata, subreticulate globose

to subglobose basidiospores, and pluriseptate pileocystidia. The morphological

variation in colour and forms result from non-genetic phenotypic plasticity

and some edaphic or substrate factors may be involved in maintaining natural

variants (Nicholl 1996; Petersen & Hughes 1999). Phenotypic plasticity in

mushrooms has been largely unexplored, no doubt due to the ephemeral nature

of most fungal basidiomata, which form only when environmental conditions

are appropriate and then last only a short time.

Our phylogeny clusters R. sichuanensis with members of Russula sect.

Laricinae, suggesting that it is closely related to the R. laricina/cessans group

as previously depicted by Li et al. (2013). Typical morphological characters

of this section include a cavernous stipe, tuberculate-striate cap margin, and

flesh that does not yellow but greys after drying, all characters observed in our

collections.

Russula mycorrhizae can be characterized by a pseudoparenchymatous

outer mantle with either angular or irregularly shaped cells and which may be

638 ... Saba & Khalid

more or less densely covered with cystidia or acystidiate (Eberhardt 2002). In

addition to the mantle features, Agerer (1999) defined a russuloid rhizomorph

type, distinguished by its appression to a substrate (often to thicker roots)

and differentiated into a peripheral layer with or without cystidia and central

layer with vessel-like hyphae. According to Eberhardt (2002), ectomycorrhizae

in R. subg. Russula, Incrustatula, Tenellula (in which R. sichuanensis falls),

Polychromidia, and Insidiosula and in R. sect. Felleinae are acystidiate with a

pseudoparenchymatous outer mantle that has irregularly shaped and often

interlocking cells.

The ectomycorrhizal characters of R. sichuanensis are same as those of Russula

subg. Tenellula, in which Li et al. (2013) described this species: dichotomously

branched and orangish brown to grayish brown tips, a pseudoparenchymatous

and acystidiate mantle with irregularly shaped and often interlocking cells and

frequently septate, curved emanating hyphae. The presence of rhizomorphs

was not noted.

Li et al. (2013) reported R. sichuanensis from conifer forests (Picea spp.);

our study describes the basidiocarp and its ectomycorrhiza from monoculture

Pinus wallichiana forests located in Himalayan moist temperate region.

Acknowledgments

We are highly indebted to Higher Education Commission (HEC), Pakistan, for

funding this project under Phase II, Batch I, Indigenous PhD fellowships program for

5000 scholars. We are cordially grateful to Dr. Kanad Das (Cryptogamic Unit, Howrah,

India), Dr. Sarah Bergemann, (Biology Department, Middle Tennessee State University,

USA), Dr. Slavomir Adam¢ik, and Dr. Shaun Pennycook for critically reviewing the

manuscript and giving their valuable comments. We are thankful to all lab fellows for

accompanying us in the field trip.

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http://doi:10.1371/journal.pone.0025081

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mycorrhizal anatomy and sporocarp features in the genus Russula. Mycol. Prog. 1(2): 201-223.

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Russula sichuanensis ectomycorrhiza (Pakistan) ... 639

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Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10" Edition. CABI

Europe-UK.

Li G, Zhao Q, Zhao D, Yue S, Li $, Wen H, Liu X. 2013. Russula atroaeruginea & R. sichuanensis

spp. nov. from southwest China. Mycotaxon 124: 173-188. http://dx.doi.org/10.5248/124.173

Miller SL, Buyck B. 2002. Molecular phylogeny of the genus Russula in Europe with a comparison

of modern infrageneric classifications. Mycological Research, 106: 259-276.

Munsell. 1975. Munsell™ soil color charts. Baltimore.

Niazi AR., Iqbal SH, Khalid AN. 2006. Biodiversity of mushrooms and ectomycorrhizas. 1. Russula

brevipes Peck. and its ectomycorrhiza - a new record from Himalayan moist temperate forests

of Pakistan. 38(4): 1271-1277.

Nicholl D. 1996. Relationships within the Pleurotus djamor species complex. Master's thesis.

University of Tennessee, Knoxville, TN.

Petersen RH, Hughes KW. 1999. Species and speciation in mushrooms. Development of a species

concept poses difficulties. Bioscience 49: 440-452.

Razaq A. 2013. Molecular characterization and identification of gilled fungi from Himalayan moist

temperate forests of Pakistan using internal transcribed spacers (ITS) of rDNA. PhD thesis,

Department of Botany, University of the Punjab, Lahore.

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. Molecular evolutionary

genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony

methods. Mol. Biol. Evol. 28: 2731-2739. http://dx.doi.org/10.1093/molbev/msr121

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, New York.

MY COTAXON

http://dx.doi.org/10.5248/130.641

Volume 130, pp. 641-645 July-September 2015

Mycenastrum corium and gastrointestinal mycetism in México

EVANGELINA PEREZ-SILVA*, TEOFILO HERRERA,

& ABRAHAM J. MEDINA-ORTIZ

Departamento de Botanica, Laboratorio de Micologia,

Instituto de Biologia, Universidad Nacional Autonoma de México,

Apartado Postal 273, Ciudad Universitaria, Coyoacan. 04510, México, Distrito Federal

* CORRESPONDENCE TO: psilva@ib.unam.mx

ABSTRACT — Several specimens of Mycenastrum corium were collected for the first time at

the Botanic Garden of the UNAM Biology Institute in the Pedregal de San Angel Ecological

Reserve (REPSA), México City. Of the five people who ingested the fungus, three showed no

signs of gastrointestinal distress while two suffered from severe flatulence and diarrhea six

hours after ingestion. The collections are described and distribution of the species in Mexico

and elsewhere is noted.

Key worps — Agaricaceae, chorology, puffball, toxic fungi

Introduction

Mycenastrum corium has been recorded from South America (Homrich &

Wright 1973), United States of America (Perreau & Heim 1971; Miller et al.

2005), and Yemen (Kreisel & Al-Fatimi 2004). The fungus is not well known

in México, where very few records can be found. In this paper we report

an abundant collection of M. corium from the core area of Botanic Garden

of National Autonomous University of Mexico (UNAM), Biology Institute,

in the Pedregal de San Angel Ecological Reserve (REPSA), Mexico City. We

describe the species based on Mexican material, and comment on its national

distribution.

Material & methods

The identified specimens were collected between May and August in the Botanic

Garden, IBUNAM, RESPA (19°17 N 99°11 W), and the species was under constant

supervision during the rainy season from June to August 2014. The material studied has

been deposited in the Fungi Collection from National Herbarium of Mexico (MEXU)

of Biology Institute. Specimen identification was based on Herrera (1964), Guzman &

642 ... Pérez-Silva, Herrera, & Medina-Ortiz

Herrera (1969), Homrich & Wright (1973), and Li et al. (2012). Tissue sections were

mounted in 10% KOH, cotton blue in lactic acid and/or Melzer’s solution. The color

codes enclosed in parenthesis are based on Kornerup & Wanscher (1978). Samples of

the gleba (MEXU 27540) were fixed in 4% glutaraldehyde. Dehydrated with graduated

alcohols, dried in an Emitech K850 Critical Point Dryer, and fixed on aluminum slides

covered with gold layer for examination with a Hitachi Electronic Scanning Microscope

(model SU1510).

Taxonomy

Mycenastrum corium (Guers.) Desv., Ann. Sci. Nat., Bot., sér. 2, 17: 147 (1842

PLATE 1

Young fructifications are white, globose, 5-27 cm in diameter. Pseudostipe

2-7 cm long, consisting of soil-covered mycelium. Peridium 1-2 cm thick,

consisting of two layers in the juvenile stage and in the adult stage, 1-2 mm.

Exoperidium is adhered to the endoperidium, also white, smooth, in its juvenile

phase. When the basidiome matures, the exoperidium ruptures forming large

deciduous scales exposing the light-brown endoperidium (4F4), which is

tasteless, smooth, and with a coriaceous consistency and a foul dirty-sock odor

that increases when dried; when mature the endoperidium opens, leaving a

star-shaped opening and exposing a dark-brown gleba (5F5) composed of the

basidiospores and capillitium; capillitial filaments thick-walled, branched with

numerous short thorn-like points of different diameter. The basidiospores are

ornamented, spherical, approximately 8 um in diameter with and exosporium

that in young specimens is thick and relatively smooth and when mature

develops a reticulate ornamentation (SEM).

SPECIMENS STUDIED— MEXICO. DistrITo FEDERAL: México City, Tlalpan, private

house, growing on grass, 6.VIII.2012, R. Martinez Flores (MEXU 27530 immature

specimens; MEXU 27531 mature specimens); UNAM- University City: Botanic Garden,

Biology Institute, 9.V.2014, R. Garibay (MEXU 27532); 13.V.2014, A. De la Cruz-

Martinez (MEXU 27534); 20.V.2014, E. Pérez-Silva, A.J. Medina-Ortiz & A. De la

Cruz-Martinez (MEXU 27533); 2.VII.2014, E. Pérez-Silva & A.J. Medina-Ortiz (MEXU

27540); 25.VII.2014, E. Pérez-Silva & A.J. Medina-Ortiz (MEXU 27561); 13.VIII.2014,

E. Pérez-Silva & A.J. Medina-Ortiz (MEXU 27551); 18.[X.2014, E. Pérez-Silva & A.J.

Medina-Ortiz (MEXU 27553); 13.X.2014, E. Pérez-Silva & A.J. Medina-Ortiz (MEXU

27554). EstaDpo DE MExiIco: Municipality of Naucalpan de Juarez, Satelite City-

Economist Circuit, on green area from Santander Bank, 24.VII.2014, E. Pérez-Silva

(MEXU 27560 immature specimens).

Discussion

The morphology and anatomy of our specimens as confirmed by SEM

examination coincide with the taxonomy description of M. corium by Homrich

& Wright (1973), and the capillitium and spores match the descriptions by

Perreau & Heim (1971) and Li et al. (2012). The species has previously been

Mycenastrum corium in Mexico ... 643

D 10.0kV 16.1mm x6.50k SE 8.oum | SU1510 10.0kV 16.2nimx6.50k SE

Pirate 1. Mycenastrum corium. 1. Young single basidiome (MEXU 27532). 2-6 (MEXU 27540):

2. Gregarious basidiomes in different stages of maturity; 3. Mature basidiome, showing cracked

endoperidium and gleba); 4. Capillitium branched with spines; 5. Subglobose immature spore;

6. Spherical mature spore.

recorded in Mexico from Chihuahua State (Moreno et al. 2010), Nuevo

Leon and San Luis Potosi States (Guzman & Herrera 1969), and Sonora State

644 ... Pérez-Silva, Herrera, & Medina-Ortiz

(Guzman 1972, Esqueda et al. 2000) growing in grassland, oak, juniper and

pine forests; its habit can be solitary or gregarious (with MEXU 27540 forming

1.5 m “fairy circles”).

The specimens examined varied in size from 5 to 27 cm in diameter and

were greatly admired by many who thought they were edible due to their

resemblance to some Calvatia species. Thus with no precaution, the fungi were

cooked with butter, garlic, salt, goat cheese, and red wine. One person who ate

two specimens (approximately 155 g each) had diarrhea 8 hours later; another

person who also ate two similar specimens had a stomachache, passed gas

throughout the night, and finally had diarrhea. Three others experienced no

symptoms.

These symptoms indicate that the two collectors suffered a kind of

gastrointestinal mycetism; such symptoms place M. corium within the

“benign species” (Heim 1963). Benjamin (1995) does not consider or mention

any symptoms. Ayala & Ochoa (1998) mention this species as having anti-

inflammatory properties, as a tonic for throat and lungs, and as hemostatic.

Children use large basidiomes to play ball because the thick exoperidium allows

them to sustain strong blows without rupturing. As fructifications enlarge,

the exoperidium thins to 2 mm and the endoperidium becomes leathery and

difficult to cut.

Acknowledgements

Thanks to Sistema Nacional de Investigadores (CONACyT), and DGAPA-UNAM

IN-207311 project for their support. The authors also thank Dr, Martin Esqueda and

Prof. Gabriel Moreno for presubmission review.

Literature cited

Ayala N, Ochoa C. 1998. Hongos conocidos de Baja California. Universidad Autonoma de Baja

California. Baja California, México. 165 p.

Benjamin DR. 1995. Mushrooms poisons and panaceas. A handbook for naturalists, mycologists,

and physicians. W.H. Freeman & Company, New York. 422 p.

Esqueda-Valle M, Pérez-Silva E, Herrera T, Coronado M, Estrada A. 2000. Composicién de

gasteromicetos en un gradiente de vegetacién en Sonora, México. Anales del Instituto de

Biologia, Universidad Nacional Autonoma de Mexico, Serie Botanica 71: 39--62.

Guzman G. 1972. Macromicetos mexicanos en el Herbario, the National Fungus Collection de

E.U.A. Boletin de la Sociedad Botanica de México 32: 31-55.

Guzman G, Herrera T. 1969. Macromicetos de las zonas aridas de México, Gasteromicetos. Anales

del Instituto de Biologia, Universidad Nacional Auténoma de México, Serie Botanica 40: 1-97.

Heim R. 1963. Les champignons toxiques et hallucinogenes. N. Boubée & Cie. Paris, France. 320 p.

Herrera T. 1964. Clasificacion, descripcion y relaciones ecoldgicas de Gasteromycetos del Valle de

México. Anales del Instituto de Biologia, Universidad Nacional Auténoma de México, Serie

Botanica 35: 9-43.

Homrich MH, Wright JE. 1973. South American Gasteromycetes. The genera Gastropila, Lanopila

and Mycenastrum. Mycologia 65: 779-794. http://dx.doi.org/10.2307/3758517

Mycenastrum corium in Mexico ... 645

Kornerup A, Wanscher JH. 1978. Methuen handbook of color. Eyre Methuen, London. 252 p.

Kreisel H, Al-Fatimi M. 2004. Basidiomycetes and larger ascomycetes from Yemen. Feddes

Repertorium 115(7-8): 547-561. http://dx.doi.org/10.1002/fedr.200411053

Li XL, Liu BF, Xie WB, Deng JC, Xu JW. 2012. The contrastive analysis of puffballs produced from

Inner Mongolia and Jilin. Chinese Journal of Applied Chemistry 29(4): 477-482.

Miller Jr OK, Brace R-L, Evenson VS. 2005. A new subspecies of Mycenastrum corium from

Colorado. Mycologia 97: 530-533. http://dx.doi.org/10.3852/mycologia.97.2.530

Moreno G, Lizarraga M, Esqueda M, Coronado M. 2010. Contribution to the study

of gasteroid and secotioid fungi of Chihuahua, Mexico. Mycotaxon 112: 291-315.

http://dx.doi.org/10.5248/112.291

Perreau, J, Heim R. 1971. A propos des Mycenastrum représentés ou décrits par N. Patouillard.

Revue de Mycologie 36: 81-95.

MY COTAXON

http://dx.doi.org/10.5248/130.647

Volume 130, pp. 647-651 July-September 2015

New reports of rust fungi from

Malakand, Khyber Pakhtunkhwa, Pakistan

S. HussaIn’, H. AHMAD’, N.S. AFSHAN?*, & A.N. KHALID?

"Department of Genetics, Hazara University, Mansehra, Pakistan

"Department of Botany &*Centre for Undergraduate Studies,

University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

*CORRESPONDENCE TO: pakrust@gmail.com

ABSTRACT — Puccinia microspora and P. nakanishikii were collected from Malakand district,

Khyber Pakhtunkhwa and are new records for Pakistan.

KEY worps — Hazar Nao, Swat, Uredinales

Introduction

The tribal region, Malakand is located at 34°35¢N 71°57¢E. It is bounded by

Dir and Swat, by Buner, Mardan, and Charsadda districts, and by Mohmand

and Bajaur Agencies (Chaghtai & Ghawas 1976). Malakand extends from

the rugged and partly glaciated mountain ranges of the Hindukush down

to northern edge of the Peshawar basin. Vegetation of the district is mostly

sub-tropical, typified as Coniferous Sub-tropical Pine Forest of Pakistan (Sheikh

1993). The dominant forest trees are Pinus roxburghii Sarg. mixed with Quercus

oblongata D. Don [= Q. incana Roxb.] and Olea ferruginea Royle, and Acacia

modesta Wall. and Dodonaea viscosa Jacq. are found at the lower elevations

(Barkatullah & Ibrar 2011).

During the exploration of Uredinales of Malakand, Pakistan, rust

infected plants were collected from different localities and examined macro-

microscopically. Among the specimens, Puccinia microspora and P. nakanishikii

were found as new records for Pakistan. Previously about 174 species of rust

fungi have been reported from Khyber Pakhtunkhwa (Afshan & Khalid 2008,

2009; Afshan et al. 2008a,b,c,d, 2010; Ishaq et al. 2013; Fiaz et al. 2015), but

Malakand district is totally unexplored uredinologically. So this is a first attempt

to study and explore this floristically rich area with respect to rust fungi.

648 ... Hussain & al.

Materials & methods

Freehand sections & scrape mounts of infected plant materials were made in lactic

acid. The plants were photographed and infected portions were observed under a

stereomicroscope. Twenty spores representing each spore state were examined under

a Nikon YS 100 microscope, and paraphyses and spore were measured using a Zeiss

eyepiece screw micrometer. Sections, paraphyses and spores were microphotographed

by Digiporo-Labomed. Spores and paraphyses were drawn with the aid of a camera

lucida (Ernst Leitz Wetzlar Germany). Specimens are conserved in the Herbarium,

Hazara University, Dhodial, Pakistan (HUP).

New records

PLATE 1: Puccinia microspora (HUP SHR-25):

A. Urediniospores. B. Teliospores. Scale bars = 10 um.

Puccinia microspora Dietel, Bot. Jahrb. Syst. 37: 101 (1905) PLATE 1

SPERMOGONIA and AECIA not found. Sori frequent on the abaxial surface.

UrREDINIA pale brown to cinnamon brown. UREDINIOSPORES 22.5—26 x 17-20

um, oval to obovoid, cinnamon brown, echinulate, 3—4 equatorial germ pores,

wall thickness 1.7—2.3 um. Tetra dark brown to blackish brown. TELIOSPORES

33-39 x 20.5-25 um, smooth, chestnut brown, wall thickness 2-3 um, pale

brown; pedicels 22—26 um, thin walled, persistent.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, District Malakand,

Qaldara, at 510 m a.s.l., on Imperata cylindrica (L.) P. Beauv. (Poaceae), stages II + III,

April, 2014, S. Hussain SHR-208. (HUP SHR-25).

New Puccinia records for Pakistan ... 649

ComMENTSs: Puccinia microspora is a new record for Pakistan. This fungus

previously been reported from Asia, North America, and South America

(Cummins 1971, Bagyanarayana & Ravinder 1988).

PLATE 2A-G: Puccinia nakanishikii (HUP SHR-26): A. Host plant. B. Uredinial and

telial sori under stereomicroscope. C, D. Urediniospores. E-G. Teliospores. Scale

bars: A, B = 10 mm; C, D = 10 um; E-G = 12 um.

Puccinia nakanishikii Dietel, Bot. Jahrb. Syst. 34: 585 (1905) PLATE 2

SPERMOGONIA and AECIA unknown. Uredinial and telial sori amphigenous.

Urepinia_ light brown to cinnamon-brown, rounded to _ globose.

UREDINIOSPORES 25-30 x 18-24 um, globose to ellipsoid, echinulate, light

brown to chestnut brown, 4—5 equatorial germ pores, wall thickness 1—2.5 um.

TELIA sub-epidermal but early exposed, dark brown to blackish, elongated,

0.2—0.5 mm. TELIOSPORES 32—38 x 20—26 um, chestnut brown to dark brown,

ellipsoidal, smooth with little granular contents, wall thickness at sides 2.3-3.5

um, and 4—6 um apically. Pedicel usually persistent, light brown, thick walled,

94-111 x 5-7 um.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, District Malakand,

Jaban, at 490 m a.s.l., on Sorghum halepense (L.) Pers. (Poaceae), stages II + III, Nov.

2010, S. Hussain SHR-209. (HUP SHR-26).

650 ... Hussain & al.

PLATE 2H-I: Puccinia nakanishikii (HUP SHR-26):

H. Urediniospores. I. Teliospores. Scale bars = 10 um.

ComMENTs: Puccinia nakanishikii is a new record for Pakistan. It has previously

been reported from Africa and Asia on the grass genera Andropogon,

Bothriochloa, Capillipedium, Cymbopogon, and Sorghum (Cummins 1971).

Acknowledgements

We sincerely thank Dr. Abdul Rehman Niazi (Department of Botany, University

of the Punjab, Lahore, Pakistan) and Dr. Omar Paino Perdomo (Dominican Society

of Mycology, Santo Domingo, Dominican Republic) for their valuable suggestions to

improve the manuscript and acting as presubmission reviewers.

Literature cited

Afshan NS, Khalid AN. 2008. New rust fungi on noxious weeds from Pakistan. Pakistan Journal of

Phytopathology 20(1): 82-87.

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, Niazi AR. 2008a. New records and distribution of rust fungi from Pakistan.

Mycotaxon 105: 257-267.

Afshan NS, Khalid AN, Niazi AR. 2008b. New records of graminicolous rust fungi from Pakistan.

Pakistan Journal of Botany 40(3): 1279-1283.

Afshan NS, Khalid AN, Javed H. 2008c. Further addition to the rust flora of Pakistan. Pakistan

Journal of Botany 40(3): 1285-1289.

Afshan NS, Berndt R, Khalid AN, Niazi AR. 2008d. New graminicolous rust fungi from Pakistan.

Mycotaxon 104: 123-130.

Afshan NS, Niazi AR, Khalid AN. 2010. Three new species of rust fungi from Pakistan. Mycological

Progress 9: 485-490. http://dx.doi.org/10.1007/s11557-010-0655-8

New Puccinia records for Pakistan ... 651

Bagyanarayana G, Ravinder EJ. 1988. Puccinia microspora Diet. (Uredinales)--a new record for

India from Andhra Pradesh State. Acta Botanica Indica 16(1): 101-102.

Barkatullah, Ibrar M. 2011. Plants profile of Malakand Pass Hills, District Malakand, Pakistan.

African Journal of Biotechnology 10(73): 16521-16535. http://dx.doi.org/10.5897/AJB11.1258

Chaghtai SM, Ghawas IH. 1976. The study of the effect of exposure on community setup in

Malakand Pass, NWEP., Pakistan. Sultania 2: 1-8.

Cummins GB. 1971. The rust fungi of cereals, grasses and bamboos. Springer, New York.

http://dx.doi.org/10.1007/978-3-642-88451-1

Fiaz M, Habib A, Afshan NS, Khalid AN. 2015. Some new records of Uredinales from Khyber

Pakhtunkhwa, Pakistan. Mycotaxon 130(2): 569-575. http://dx.doi.org/10.5248/130.569

Ishaq A, Afshan NS, Khalid AN. 2013. New records of Puccinia on Poaceae from Khyber

Pakhtunkhwa, Pakistan. Mycotaxon 126: 177-182. http://dx.doi.org/10.5248/126.177

Sheikh MI. 1993. Trees of Pakistan. Pictorial Printers (Pvt.) Ltd., Islamabad, Pakistan.

MY COTAXON

http://dx.doi.org/10.5248/130.653

Volume 130, pp. 653-660 July-September 2015

Artomyces nothofagi sp. nov., a clavarioid fungus

from a Chilean Nothofagus forest

RICHARD KNEAL' & MATTHEW E. SMITH"?

‘Department of Plant Pathology, University of Florida, Gainesville, FL, 32611, USA

*CORRESPONDENCE TO: trufflesmith@ufl.edu

ABSTRACT - We collected an Artomyces species on highly decayed wood of Nothofagus

dombeyi in southern Chile that did not match the description of any previously described

species. Here we describe this fungus as Artomyces nothofagi based on a combination of

unique morphological features and a distinct nrITS DNA sequence.

Key worps - Auriscalpiaceae, coral fungi, Patagonia, Russulales

Introduction

In the past, coral fungi were treated as a single taxonomic unit because

all coralloid and clavarioid fungi were thought to be closely related (e.g.,

Coker 1923, Corner 1950). However, recent molecular phylogenetic results

indicate that coralloid and clavarioid fungi are polyphyletic and are spread across

many different orders of Agaricomycotina (Pine et al. 1999, Hibbett 2007). Jiilich

(1982) erected the genus Artomyces Jiilich to accommodate clavarioid species

with simple to pyxidately branched basidiomata, hymenial gloeocystidia,

gloeoplerous hyphae, and small, hyaline, amyloid basidiospores. In a molecular

phylogenetic overview of the Russulales, Miller et al. (2006) included Artomyces

in their circumscription of Auriscalpiaceae where it comprises a clade sister

to the other genera in that family, including Auriscalpium Gray, Lentinellus

P. Karst., Gloiodon P. Karst., and Dentipratulum Domanski.

Lickey et al. (2003) provided the only comprehensive phylogenetic

and taxonomic study of Artomyces. This study supported Jilich’s (1982)

interpretation of Artomyces as a distinct genus that is separate from Clavicorona

Doty. Lickey et al. (2003) formally described seven new species, created a new

combination based on Clavicorona microspora Qiu X. Wu & R.H. Petersen,

and provisionally described two additional unnamed taxa. Overall, Lickey et

al. (2003) accepted 15 species within Artomyces, based on a combination of

654 ... Kneal & Smith

morphology, nuclear ribosomal internal transcribed spacer DNA (nrITS DNA)

sequences, and mating studies. No new Artomyces species have been described

since this comprehensive work.

During a fungal foray near the base of the Melimoyu volcano in southern

Chile, we encountered a clavarioid basidiomycete in the genus that was fruiting

on highly decayed wood of Nothofagus dombeyi (Mirb.) Oerst. This clavarioid

fungus, which did not match any described Artomyces species, had a unique

nrITS DNA sequence. We describe this fungus here as Artomyces nothofagi.

Materials & methods

Colors of fresh and dried specimens are described here in general terms. Microscopic

features were described from dried material mounted in 5% KOH, Melzer’s reagent,

Congo Red, or water for standard light microscopy. Microscopical observations were

made using a Nikon Optiphot microscope. Basidiospores were measured from mature

basidiocarps that were shaved into thin slices or crush mounted and then stained in

Melzer’s reagent. Spore statistics were calculated from measurements from 20 spores

as length x width. Length : width ratios are designated as Q; L_ = mean length; W, =

mean width; and Q, = mean Q. At least 10 structures were measured for each of the

other anatomical characteristics. Basidia, generative hyphae, gloeoplerous hyphae and

gloeocystidia were observed and measured in crush mounts with 5% KOH, Congo Red

and water. Specimens are conserved in Herbario, Museo Nacional de Historia Natural,

Santiago, Chile (SGO) and University of Florida Herbarium, Gainesville, U.S.A. (FLAS).

Fresh basidiocarp tissue was preserved in the field with CTAB buffer. DNA was

extracted by a modified CTAB method followed by PCR of the nrITS DNA region

using the primers ITSIF and ITS4 according to published protocols (Gardes & Bruns

1993). PCR products were visualized on 1.5% agarose gels stained with SYBR Green

I (Molecular Probes, Eugene, OR, USA). Successful amplicons were cleaned with

ExoSAP-IT (U.S.B. Corporation, Cleveland, OH, USA). Bidirectional sequencing was

performed with ITS1F and ITS4 by the University of Florida ICBR sequencing center

(www.biotech.ufl.edu). Sequences were edited with Sequencher v.4.1 (Gene Codes Inc.,

Ann Arbor, MI, USA).

All available Artomyces nrITS DNA sequences were downloaded from GenBank

and combined with our sequence into an alignment using Mesquite v.1.1 (Maddison

& Maddison 2006). Preliminary sequence alignments were performed with MUSCLE

(Edgar 2004) followed by manual adjustments in Mesquite. A maximum parsimony

(MP) heuristic search was conducted with PAUP* v.4.0a109 (Swofford 2002) using 100

random addition replicates and TBR branch swapping. MP bootstrapping was used to

assess support for topology using 1000 replicates with 10 random addition replicates

each. Maximum likelihood analysis was performed using GARLI (Zwickl 2006) with the

GTR+I+G model followed by ML bootstrapping with 1000 bootstrap replicates.

Results

Morphological analysis of Artomyces nothofagi indicates that this species is

distinct from all other described species based on a combination of basidiomata

Artomyces nothofagi sp. nov. (Chile) ... 655

‘ ViCvie

. eee

-—

e a_i _

FiGuRE 1. Artomyces nothofagi (holotype, SGO 164746; isotype, FLAS-F-58994): A. Basidio-

mata fruiting directly on decayed wood of Nothofagus dombeyi in southern Chile. B. Fresh

basidiomata on grey background. C. 4-spored basidia. D. 2-spored basidia. E. Gloeocystidia.

E Guttulate, amyloid basidiospores. G. Clamp connection on generative hyphae.

Images C-E and G mounted in KOH and Congo Red; image F mounted in Melzer’s reagent.

Scale bars; A = 2.5 cm; B = 1 cm; C-G=5 um.

656 ... Kneal & Smith

shape, size, and color, spore size and ornamentation, and morphology of the

gloeocystidia (Fic. 1). Additionally, molecular phylogenetic analysis shows

A. nothofagi witha unique nrITS DNA sequence, supportingit as phylogenetically

distinct from all other Artomyces species. Although A. nothofagi clearly belongs

to Artomyces, our ML and MP analyses did not provide bootstrap support

for a close relationship with any of the other species. Nonetheless, the best

ML tree (Fic. 2) and best MP trees (data not shown) consistently resolved

Artomyces nothofagi in an unsupported clade with A. colensoi (Berk.) Jiilich and

A. stephenii Lickey.

Taxonomy

Artomyces nothofagi M.E. Sm. & Kneal, sp. nov. FIG. 1

MycoBank 810369

Differs from other Artomyces species by its combination of smooth basidiospores,

mostly unbranched basidiomata, gloeocystidia that protrude well past the hymenium,

and a unique nrITS DNA sequence.

Type: Chile, XI Regién Aysén del General Carlos Ibafez del Campo, Melimoyu reserve,

near Pacific Ocean at the base of Volcan Melimoyu, Tres Lagunas trail 44°05’14.5”S

73°04’58.9’W, 14 March 2012, M.E. Smith MES559 (Holotype, SGO164746; isotype,

FLAS-F-58994; GenBank KP025706).

EryMo.ocy: The epithet nothofagi refers to the substrate species, Nothofagus dombeyi.

BASIDIOMATA lignicolous, scattered to gregarious, <50 mm tall x 15 mm

wide at the apex; generally unbranched and coronate, rarely forming cristate

branches at the apices; apices coronate-cristate to cuspidate and dull grey to

faintly brownish with occasional vinaceous tones; when dried black to caramel

brown; usually lacking obvious basal mycelium, but occasionally with whitish

basal mycelium. ODOR sweet and pleasant, reminiscent of apricots; taste not

recorded.

CONTEXT composed of: (1) generative hyphae, thin walled, usually

encrusted with yellow-brown material, 2.0—15.0 um diam.; clamp connections

present; and (2) gloeoplerous hyphae, common in the trama, aseptate, 2.5—8.0

um diam., terminating in the hymenium as gloeocystidia. GLOEOCYSTIDIA

common, mostly cylindrical to clavate but occasionally fusiform, flexuous

or subcapitate to capitate, 2.5-6.5 um diam., consistently narrowing within

the subhymenium, non-emergent to protruding 8.0-25.0(—50) um. Basip1a

(2—)4-sterigmate, narrowly clavate to cylindrical, with obvious basal clamp

connections, 20.8—25.6 x 4.7-6.5 um (L, x W = 22.5 x 5.4 um) not including

sterigmata; STERIGMATA 2.6—4.3 um long, (L, = 3.2 um). BASIDIOSPORES

4.0-5.2 x 3.4-4.5 um (L_ x W = 4.7 x 3.7 um), Q= 1.0-1.4(Q, = 1.3), globose

to subglobose, smooth, hyaline, distinctly amyloid in Melzer’s reagent; contents

with a single large guttule; hilar appendage present.

Artomyces nothofagi sp. nov. (Chile) ... 657

ECOLOGY & DISTRIBUTION — Known only from the type collection. Clustered

to gregarious on highly decayed wood of Nothofagus dombeyi in southern

Chile; March.

Discussion

Both morphology and molecular phylogenetic analysis support Artomyces

nothofagi as a distinct species. The petite, mostly unbranched basidiomata of this

species along with the smooth globose to subglobose basidiospores are unusual

features within Artomyces. The frequently encrusted generative hyphae, basidia

that are sometimes two-spored, and gloeocystidia that protrude well past the

hymenium (<50 um) are also distinctive within the genus (Fic. 1). Artomyces

nothofagi has smooth spores, which contrasts with 14 (out of 15) of the other

species characterized by spores that are clearly ornamented when viewed with

light microscopy. This new species is also unusual because its basidiomata are

mostly unbranched and coronate, only rarely forming cristate branches at the

apices. In contrast, most described Artomyces species have basidiomata that

branch regularly more than twice.

The most morphologically similar species to A. nothofagi is A. cristatus

(Kauffman) Julich, which is known from conifer wood in Canada, northern

Europe, and the USA. Both species have unbranched basidiomata and smooth

basidiospores with large guttules. However, the spores of A. cristatus are

distinctly cylindrical, not globose to subglobose as in A. nothofagi. In addition,

the gloeocystidia of A. cristatus protrude only <6 um past the hymenium

whereas those of A. nothofagi protrude <50 um. Based on the habitat and

several distinct morphological differences, it is evident that these are indeed

two different species.

The only two Artomyces species previously known from southern South

America, A. adrienneae Lickey and A. austropiperatus Lickey, are also

morphologically distinct. The basidiomata of A. adrienneae are “pure white

or white mellowing to a fleshy buff” and regularly branched whereas those of

A. nothofagi are much darker and rarely branched. Moreover, A. adrienneae

basidiospores are ornamented (asperulate) and narrower than A. nothofagi

spores. Artomyces adrienneae also lacks the encrusted generative hyphae

(Lickey et al. 2003) that are common in A. nothofagi.

Artomyces austropiperatus is the only other species described from South

America, although this species has also been documented from Tasmania and

the North Island of New Zealand. Although A. austropiperatus is similar to

A. nothofagi in the sizes of the gloeocystidia, basidia, and spores,

A. austropiperatus spores are ornamented (“obviously asperulate”) whereas

those of A. nothofagi are smooth. Also, Lickey et al. (2003) describes

658 ... Kneal & Smith

Artomyces nothofagi MES559

Artomyces colensoi AF454423

Artomyces colensoi AF454424

Artomyces stephenii AF454425

Artomyces stephenii AF454426

Artomyces sp. PDD 89068 JF714648

Artomyces austropiperatus AF454408

Artomyces austropiperatus AF454407

Artomyces dichotomus AF454406

Artomyces costaricensis AF454410

Artomyces costaricensis AF454411

Artomyces carolinensis AF454409

Artomyces piperatus AF454404

Artomyces piperatus AF454405

Artomyces tasmaniensis AF454412

Artomyces tasmaniensis AF454413

Artomyces cristatus AF454421

Artomyces cristatus AF454422

Artomyces pyxidatus AF336147

Artomyces pyxidatus AF336145

Artomyces pyxidatus AY513136

Artomyces pyxidatus AF336144

Artomyces pyxidatus AY297586

Artomyces pyxidatus AF336142

Artomyces pyxidatus AF336141

Artomyces pyxidatus AF336140

Artomyces pyxidatus AF336139

Artomyces pyxidatus AF336148

Artomyces pyxidatus AF336143

Artomyces pyxidatus AF336146

Artomyces microsporus AF336137

Artomyces microsporus AF336138

Artomyces microsporus DQ449944

Artomyces novae zelandiae AF454416

Artomyces novae zelandiae AF454417

Artomyces candelabrus HQ533045

Artomyces candelabrus GQ411509

Artomyces candelabrus AF454419

Artomyces candelabrus AF454420

Artomyces adrienneae AF454418

Artomyces turgidus AF454402

Artomyces turgidus AF454401

Artomyces turgidus AF454403

Artomyces sp. 3456 AF454415

Artomyces sp. 9905 AF454414

Gloeocystidiellum porosum KJ140772

Lentinellus subaustralis AF454427

Lentinellus vulpinus AY513230

Lentinellus sublineolatus NR 119505

— 5 changes

FicuRE 2. Maximum likelihood phylogeny (-In 3515.31912) of Artomyces based on internal

transcribed spacer ribosomal DNA. Nodes with both maximum likelihood and maximum

parsimony bootstrap support values =>75 are depicted with solid black circles whereas nodes that

are supported by only one of these two methods are depicted with grey circles. Gloeocystidiellum

porosum and three species of Lentinellus were used as outgroups.

Artomyces nothofagi sp. nov. (Chile) ... 659

A. austropiperatus as “well-branched, more or less compact and stoutish,’ and

A. austropiperatus also lacks encrusted generative hyphae.

Although A. nothofagi consistently clustered with A. stephenii and A. colensoi

in both our ML and MP analyses, there was no bootstrap support for this

relationship. Furthermore, A. nothofagi is morphologically distinct from both

species. Artomyces stephenii forms relatively large basidiomata and is known

only from hardwood substrates in Costa Rica. It is described as “reminiscent

of A. pyxidatus” and is “often profusely branched and compact” (Lickey et al.

2003). Microscopically, A. stephenii is described with “minutely asperulate,

weakly amyloid” basidiospores whereas the spores of A. nothofagi are smooth,

distinctly amyloid, and larger than those of A. stephenii (Lickey et al. 2003).

Also, the gloeocystidia of A. stephenii are described as protruding only <12 um

past the hymenium, not <50 um as occurs frequently in A. nothofagi.

Artomyces nothofagi is also morphologically distinct from A. colensoi.

Artomyces colensoi is an Australasian species known from hardwood logs

in New Zealand and southern Australia. This species was described as

“well-branched” and having a slightly peppery odor (Lickey et al. 2003) that

contrasts with the sweet, pleasant odor of A. nothofagi. Artomyces colensoi also

has smaller, more ellipsoid spores, with Q, = 1.48 (cf.Q, = 1.3 for A. nothofagi).

Finally, A. colensoi is described as having “some encrusted” generative hyphae,

“a character not observed in the type specimen” (Lickey et al. 2003), whereas in

A. nothofagi the generative hyphae are frequently encrusted.

Acknowledgements

A special thanks is due to the conservation and sustainable development organization

Patagonia Sur (www.patagoniasur.com) for facilitating access to the Melimoyu Reserve

where A. nothofagi was collected. We also thank Dr. Donald H. Pfister and Harvard

University’s David Rockefeller Center for Latin American studies for travel funding and

logistical planning that made this work possible. Funding for this work was provided

in part by National Science Foundation grant DEB-1354802 (to MES) and also by the

University of Florida's Institute for Food and Agricultural Sciences (IFAS). We would

also like to thank Dr. Ronald Petersen and Dr. Edgar Lickey for providing valuable

reviews and feedback that improved this paper.

Literature cited

Coker WC. 1923. The clavarias of the United States and Canada. The University of North Carolina

Press, North Carolina.

Corner EJH. 1950. A monograph of Clavaria and allied genera. Annals of Botany Memoirs 1.740 p.

Edgar RC. 2004. MUSCLE: a multiple sequence alignment method with reduced time and space

complexity. BMC Bioinformatics 5: 113. http://dx.doi.org/10.1186/1471-2105-5-113

Gardes M, Bruns T. 1993. ITS primers with enhanced specificity for basidiomycetes-

applications to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

660 ... Kneal & Smith

Hibbett DS. 2007. After the gold rush, or before the flood? Evolutionary morphology of mushroom-

forming fungi (Agaricomycetes) in the early 21st century. Mycological Research 111:

1001-1018. http://dx.doi.org/10.1016/j.mycres.2007.01.012

Julich W. 1982 [“1981”]. Higher taxa of Basidiomycetes. Bibliotheca Mycologica 85. 485 p.

Lickey EB, Hughes KW, Petersen RH. 2003. Phylogenetic and taxonomic studies in Artomyces and

Clavicorona (Homobasidiomycetes: Auriscalpium). Sydowia. 55(2): 181-254.

Maddison WP, Maddison DR. 2006. Mesquite: a modular system for evolutionary analysis. Version

1.11. http://mesquiteproject.org

Miller SL, Larsson E, Larsson KH, Verbeken A, Nuytinck J. 2006. Perspectives in the new Russulales.

Mycologia 98(6): 960-970. http://dx.doi.org/10.3852/mycologia.98.6.960

Pine, EM, Hibbett DS, Donoghue, MJ. 1999. Phylogenetic relationships of cantharelloid and

clavarioid homobasidiomycetes based on mitochondrial and nuclear rDNA sequences.

Mycologia 91(6): 944-963. http://dx.doi.org/10.2307/3761626

Swofford DL. 2009. Phylogenetic Analysis Using Parsimony (*and other methods) ver 4.0a109.

Sinauer Associates, Sunderland, Massachusetts.

Zwickl DJ. 2006. Genetic algorithm approaches for the phylogenetic analysis of large biological

sequence datasets under the maximum likelihood criterion. University of Texas Press, Austin,

Texas.

MY COTAXON

http://dx.doi.org/10.5248/130.661

Volume 130, pp. 661-669 July-September 2015

Inonotus griseus sp. nov. from eastern China

L1-WEI ZHOU! & XIAO-YAN WANG’?

'State Key Laboratory of Forest and Soil Ecology, Institute of Applied Ecology,

Chinese Academy of Sciences, Shenyang 110164, PB. R. China

*University of Chinese Academy of Sciences, Beijing 100049, P. R. China

* CORRESPONDENCE TO: liwei_zhou1982@163.com

ABSTRACT —Inonotus griseus is described and illustrated from Anhui Province, eastern

China. This new species is distinguished by its annual and resupinate basidiocarps with a

grey cracked pore surface, a monomitic hyphal system in both subiculum and trama, the

presence of subulate to ventricose hymenial setae, the presence of hyphoid setae in both

subiculum and trama, and ellipsoid, hyaline, slightly thick-walled cyanophilous basidiospores

(9-10.5 x 6.3-7.2 um). ITS sequence analyses support I. griseus as a distinct lineage within

the core clade of Inonotus.

Key worps — Hymenochaetaceae, Hymenochaetales, Basidiomycota, polypore, taxonomy

Introduction

Inonotus P. Karst. (Hymenochaetaceae) was typified by I. hispidus (Bull.)

P. Karst. Although more than 100 species have been accepted in this genus

in a wide sense (Ryvarden 2005), molecular phylogenies have supported

some Inonotus species in other small genera, including Inocutis Fiasson &

Niemela, Inonotopsis Parmasto, Mensularia Lazaro Ibiza, and Onnia P. Karst.

(Wagner & Fischer 2002). Dai (2010), accepting this taxonomic segregation,

morphologically emended the concept of Inonotus. Current characters of

Inonotus sensu stricto include annual to perennial, resupinate, effused-reflexed

or pileate, and yellowish to brown basidiocarps, homogeneous context, a

monomitic hyphal system (at least in context/subiculum) with simple septate

generative hyphae, presence or absence of hymenial and hyphoid setae,

and ellipsoid, hyaline to yellowish or brownish, thick-walled and smooth

basidiospores (Dai 2010).

More than 1200 wood-decaying basidiomycetes have been reported from

China (Dai 2011, 2012a). The taxonomy of Hymenochaetaceae has been

extensively studied in China (Zhou & Jia 2010, Zhou & Dai 2012, Zhou & Xue

2012, Cui & Decock 2013, Qin & Zhou 2013, Zhou 2013, 2014a,b,c,d, Zhou &

662 ... Zhou & Wang

Qin, 2013, Zhou et al. 2014a), and 38 species of Inonotus have been reported

here (Dai 2010, 2012b, Cui et al. 2011, Dai et al. 2011, Wu et al. 2012, Zhou

& Qin 2012, Tian et al. 2013, Yu et al. 2013). However, there are still many

Chinese specimens within Hymenochaetaceae that have not been identified to

species level. Of these specimens, one is described and illustrated here as a new

species of Inonotus.

Materials & methods

Morphological examination

Our specimen is deposited in the herbarium of the Institute of Applied Ecology,

Chinese Academy of Sciences, Shenyang, China (IFP). The microscopic procedure

follows Zhou et al. (2014b). Specimen sections were prepared with cotton blue (CB),

5% potassium hydroxide (KOH), and Melzer’s reagent (IKI), and then examined

using a Nikon Eclipse 80i microscope under magnifications up to 1000x and phase

contrast illumination. Measurements and drawings were made from sections stained

with CB. When describing the basidiospore size variation, the upper and lower 5%

of measurements were excluded from the range, with extreme values enclosed in

parentheses. Line drawings were made with the aid of a light tube. Special color terms

follow Petersen (1996). The following abbreviations are used: IKI- = neither amyloid

nor dextrinoid; CB+ = cyanophilous; L = mean basidiospore length (arithmetic

average of all basidiospores); W = mean basidiospore width (arithmetic average of all

basidiospores); Q = the ratio of L/W; and n = number of basidiospores measured from

given number of specimens.

Molecular sequencing

The ITS sequence of our specimen was directly amplified using Phire® Plant Direct

PCR Kit (Finnzymes Oy, Finland) in accordance with the manufacturer's instructions

with primers ITS5 and ITS4 (White et al. 1990). The PCR procedure was as follows:

initial denaturation at 98°C for 5 min, followed by 39 cycles at 98°C for 5 s, 59°C for 5s

and 72°C for 5 s, and a final extension of 72°C for 10 min. The amplicon was sequenced

in Beijing Genomics Institute, China with the same primers as in amplifications. The

newly generated sequence was deposited at GenBank (http://www.ncbi.nlm.nih.gov).

Phylogenetic analysis

To explore the phylogenetic position of our specimen, 35 additional ITS sequences of

Inonotus (Fic. 1) were retrieved from GenBank. The ITS sequence of Phellinus populicola

Niemela was selected as outgroup (Wagner & Fischer 2002). The dataset was aligned

using MAFFT 7 (Katoh & Standley 2013) with Q-INS-i option (Katoh & Toh 2008). The

resulting alignment was subjected to phylogenetic analysis using maximum likelihood

(ML) and maximum parsimony (MP) algorithms. raxmlGUI 1.2 (Stamatakis 2006,

Silvestro & Michalak 2012) with GTR + 1+ G model and auto FC option (Pattengale et

al. 2010) in bootstrap (BS) replicates was used to construct ML trees. MP analysis was

conducted using PAUP* 4.0b10 (Swofford 2002) with heuristic search. All characters

were equally weighted and gaps were treated as missing data. Other parameters were

set as follows: starting tree obtained via stepwise addition, tree-bisection-reconnection

Inonotus griseus sp. nov. (China) ... 663

branch swapping, steepest descent option not in effect, and ‘MulTrees’ option in effect.

The resulting trees were tested by 1000 BS replicates. As the topologies from ML and

MP analyses were congruent, only the topology from the ML analysis is presented along

with BS values simultaneously 250% from both analyses.

Results

An alignment of 957 characters resulted from the ITS dataset of 37 sequences.

Of these characters, 524 are constant, 116 parsimony-uninformative, and 317

parsimony-informative. A total of 250 BS replicates was performed to test the

ML tree, and two equally most parsimonious trees of 1217 steps (CI = 0.569,

RI = 0.757) were retained. The ITS-based phylogenetic tree (Fic. 1) recovered

at least three strongly supported Inonotus clades, and the sequence from our

specimen was nested as a distinct lineage within the core clade that included

the generic type, Inonotus hispidus.

65/61, Imonotus dependens KC778777

—isoaoo} Inonotus dependens KC778778

Inonotus dependens KC778779

- Inonotus cubensis JQ860324

100/100 Inonotus cubensis JQ860325

Inonotus pseudolinteus KC77878 1 g

dort0g|| ZRonotus pseudolinteus KC778780 | &

gg/g2 | onotus pseudolinteus AF534075_ | §

sas Inonotus pseudolinteus AY 558629 | =

Inonotus linteus JQ860322 S

100/100' Inonotus linteus JQ860321 ee

100/99 | Inonotus sideroxylicola KC778782 >

100/100! Inonotus sideroxylicola KC778783

;— Inonotus tropicalis KC543999

roooo| [ Lnonotus tropicalis AF534077

100/100 Leitronoend tropicalis AY 641432

100/100' Inonotus tropicalis AY 599487

Inonotus lonicericola JQ860308

-— Inonotus baumii JN642567 $

96/95 Inonotus zonatus JQ860305 =

90/82 Inonotus alpinus JQ860310 cc

59! Inonotus lonicerinus JN642575 =

'— Inonotus weigelae JQ860315 >

-————————_Inonotus weirianus AF 110989 | &

Inonotus vaninii JN642591 =

'— Inonotus sanghuang JQ860316

Inonotus tricolor JN169787

Inonotus glomeratus AF247968

80/82 Inonotus griseus KM434333 -

7] 85/59 L______ Iyonotus obliquus GU903006 S

Inonotus micantissimus JF692194 | ©

oes 89/76 —— Inonotus andersonii AM269781 =

Inonotus hispidus AY 558602 =

100/100 -—— Inonotus quercustris AY072026

goof Jnonotus patouillardii AY072024

95/78 Inonotus rickii KF 147913

Phellinus populicola GQ383706

Figure 1. Phylogenetic position of Inonotus griseus inferred from ITS sequences. Topology is from

maximum likelihood analysis, and the bootstrap values simultaneously 250% from maximum

likelihood/maximum parsimony analyses are indicated at the nodes.

664 ... Zhou & Wang

—

FIGURE 2. Inonotus griseus (holotype) in situ.

Taxonomy

Inonotus griseus L.W. Zhou, sp. nov. FIGS; 2,33

MycoBank MB 810088

Differs from Inonotus chihshanyenus by its greyish brown to dark grey pore surface,

smaller pores, shorter hymenial setae, and larger basidiospores.

Type: China. Anhui Province: Huangshan, Qimen County, Guniujiang Park, on dead

angiosperm tree, 10.VHI.2013, L.W. Zhou LWZ 20130810-20 (holotype, IFP; GenBank

KM434333).

ETYMOLOGY: griseus (Lat.): refers to the grey pore surface.

BASIDIOCARPS annual, resupinate, adnate, inseparable, without odor or taste

when fresh, woody, up to 20 cm long, 9 cm wide, and 1.5 cm thick. PORE SURFACE

greyish brown to dark grey, sometimes black at the margin, slightly glancing,

cracked; STERILE MARGIN narrow, up to 0.5 mm, cinnamon-buff; PoREs round

to angular, 4-6 per mm or even 1-2 per mm at the margin; DISSEPIMENTS thick,

entire. SuBICULUM yellowish brown, soft, very thin to almost lacking, up to

0.5 mm. TuBEs honey-yellow to cinnamon-buff, woody, up to 1.5 cm long.

HYPHAL SYSTEM monomitic; GENERATIVE HYPHAE simple septate; tissue

darkening but otherwise unchanged in KOH.

Inonotus griseus sp. nov. (China) ... 665

FiGuRE 3. Inonotus griseus (holotype).

a: Basidiospores. b: Basidia and basidioles. c: Hymenial setae.

d: Hyphae and hyphoid setae from trama. e: Hyphae and hyphoid setae from subiculum.

SUBICULUM. GENERATIVE HYPHAE yellowish, slightly thick- to thick-walled

with a wide lumen, rarely branched, straight to flexuous, interwoven, strongly

666 ... Zhou & Wang

agglutinated, CB+, 3-5 um in diam. HyPHOID SETAE present, acute, dark

brown, thick-walled, slightly flexuous, up to hundreds um long, 4-10 um wide.

TUBES. GENERATIVE HYPHAE yellowish, slightly thick-walled with a wide

lumen, occasionally branched, straight, subparallel along the tubes, CB+,

2.5-4 um in diam. HyMENIAL SETAE subulate to ventricose, acute, straight,

dark brown, thick-walled with a wide to narrow lumen, 12-32 x 5.5-11 um;

HYPHOID SETAE embedded in hymenium, never penetrating beyond hymenial

surface, acute, dark brown, thick-walled, parallel along the tubes, up to

hundreds um long, 12-22 um wide; CysTIDIA and CYSTIDIOLES absent; BASIDIA

barrel-shaped, with a simple septum and four sterigmata, 13-17 x 8-12 um;

BASIDIOLES barrel-shaped to subglobose, slightly smaller than basidia.

Basipiospores ellipsoid, hyaline, slightly thick-walled, smooth, mostly

bearinga medium tolarge guttule, IKI-,CB+,(8.5-)9-10.5(-11) x (6-)6.3-7.2 um,

L=9.7 um, W = 6.74 um, Q = 1.44 (n = 30/1).

REMARKS: Inonotus griseus has annual and resupinate basidiocarps, amonomitic

hyphal system with simple septate generative hyphae, hyphoid and hymenial

setae, and ellipsoid, hyaline, slightly thick-walled cyanophilous basidiospores.

All these characters fit well with the concept of Inonotus as emended by Dai

(2010).

Inonotus chihshanyenus 'T.T. Chang & W.N. Chou, I. magnisetus Y.C. Dai,

I. micantissimus (Rick) Rajchenb., and I. perchocolatus Corner share resupinate

basidiocarps producing a monomitic hyphal system, and both hyphoid and

hymenial setae with I. griseus. Compared with I. griseus, I. chihshanyenus has

larger pores (1-3 per mm), longer hymenial setae (25-60 um long), and smaller

basidiospores (6.5-9 x 5-6.5 um, Chang & Chou 1998) that were also considered

to be diagnostic of I. chihshanyenus by Ryvarden (2005); I. magnisetus, recently

described from Guangdong Province, southern China, has similarly sized

pores (5-6 per mm), but its basidiospores are colored and much smaller

(4-4.5 x 3.7-4 um, Dai 2010); I. micantissimus also has similarly sized pores

(5-7 per mm), but produces colored and larger basidiospores (10-13 x 8-12 um)

and is distributed in tropical America (Ryvarden 2005); I. perchocolatus differs

mainly in its smaller pores (6-9 per mm) and basidiospores (5-7 x 4.5-5.5 um,

Ryvarden 2005). Our phylogeny (Fic. 1) indicates a close relationship between

I. griseus and I. obliquus (Ach. ex Pers.) Pilat, although with low statistical

support. Inonotus obliquus differs morphologically from I. griseus by its dark

reddish-brown pore surface, smaller pores (6-8 per mm), and narrower,

hyaline to brownish basidiospores (5.5-6.5 um, Ryvarden 2005). Therefore, the

combination of phylogenetic and morphological evidence supports Inonotus

griseus as an independent Inonotus species.

Inonotus griseus sp. nov. (China) ... 667

Inonotus has been traditionally distinguished by an annual growth habit

and a monomitic hyphal system from Phellinus Quél., another large genus in

Hymenochaetaceae (Ryvarden 2005). Based on a phylogeny inferred from the

nLSU region, Wagner & Fischer (2002) transferred several Phellinus species

with perennial basidiocarps and a dimitic hyphal system (at least in the trama)

into Inonotus: I. baumii (Pilat) T. Wagner & M. Fisch., I. linteus (Berk. & M.A.

Curtis) Teixeira, I. vaninii (Ljub.) T. Wagner & M. Fisch., and I. weirianus

(Bres.) T. Wagner & M. Fisch. This classification was followed in several later

publications (Dai 2010, Wu et al. 2012, Zhou & Qin 2012, Tian et al. 2013,

Vlasak et al. 2013), placing newly described and combining similar species

in Inonotus. However, based on their ITS and mitochondrial SSU sequence

analyses, Jeong et al. (2005) did not accept the above mentioned species with

perennial basidiocarps and dimitism in Inonotus, considering them to have a

sister relationship with Inonotus. The generic circumscription of Inonotus is still

under debate. In our phylogeny (Fic. 1), species (including I. griseus) with a

monomitic hyphal system in both context/subiculum and trama form the core

Inonotus clade, while the other sampled Inonotus species that are dimitic (at

least in trama) form the residual clades A and B. More sampling and additional

loci are needed to determine a more exact circumscription of Inonotus.

Acknowledgements

We express our gratitude to Drs. Bao-Kai Cui (Beijing Forestry University, China)

and Josef Vlasak (Biology Centre of the Academy of Sciences of the Czech Republic,

Czech Republic) who reviewed the manuscript before final submission. The research was

financed by the National Natural Science Foundation of China (Project No. 31200015)

and the Youth Fund for Creative Research Groups, Institute of Applied Ecology, Chinese

Academy of Sciences.

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Cui BK, Decock C. 2013. Phellinus castanopsidis sp. nov. (Hymenochaetaceae) from southern

China, with preliminary phylogeny based on rDNA sequences. Mycol. Prog. 12: 341-351.

http://dx.doi.org/10.1007/s11557-012-0839-5

Cui BK, Du P, Dai YC. 2011. Three new species of Inonotus (Basidiomycota, Hymenochaetaceae)

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MY COTAXON

http://dx.doi.org/10.5248/130.671

Volume 130, pp. 671-681 July-September 2015

New reports of Inocybe from pine forests in Pakistan

M. SABA!*, I. AHMAD’, & A.N. KHALID?

"Department of Botany, University of the Punjab

Quaid-e-Azam Campus, Lahore, 54590, Pakistan

*Department of Botany, University of Peshawar, Peshawar, Pakistan

* CORRESPONDENCE TO: rustflora@gmail.com

ABSTRACT — During investigations of ectomycorrhizal fungi associated with pine trees in

Pakistan, Inocybe amicta and I. mimica were collected underneath Pinus wallichiana. Both

morphological and nrDNA ITS molecular evidence support the identity of these species.

They represent new records from pristine forested areas of Pakistan, and from Asia. Pinus

wallichiana is recorded as a new host for both species. Twenty-six Inocybe taxa have now been

reported from Pakistan.

Key worps — Agaricales, Basidiomycota, Inocybaceae, Shangla

Introduction

The family Inocybaceae Julich, containing the genera Inocybe (Fr.) Fr.,

Auritella Matheny & Bougher, and Tubariomyces Esteve-Rav. & Matheny, is a

highly diverse monophyletic group of ectomycorrhizal fungi that comprises

between 500 and 700 species worldwide (Matheny et al. 2009, Alvarado et al.

2010). Between 70% and 80% of species in the family have been described from

the north temperate zone in association primarily with the ectomycorrhizal

(ECM) plant families Pinaceae, Fagaceae, and Salicaceae. However, most

studies on fungal ecology and biodiversity have focused on taxa outside the

tropics, a region where ECM fungal taxa deserve more attention (Alexander &

Selosse 2009, Smith et al. 2011).

Inocybe is a large genus of gilled fungi that includes many fairly small

and inconspicuous brown species (Kokkonen & Vauras 2012). Matheny

(2009) identified seven clades within Inocybaceae: /inocybe, /nothocybe,

/pseudosperma, /mallocybe, /inosperma, /mallocybella, and /auritella.

Previously, I. adaequata (Britzelm.) Sacc., I. argillacea (Pers.) Singer,

I. asterospora Quél., I. bongardii (Weinm.) Quél., I. dulcamara (Pers.) P. Kumm.,

672 ... Saba, Ahmad, & Khalid

I. fastigiata (Schaeff.) Queél., I. fibrosa (Sowerby) Gillet, I. flocculosa Sacc.,

I. fuscidula Velen., I. geophylla (Bull.) P. Kumm., I. glabripes Ricken, I. hirtella

Bres., I. microspora J.E. Lange, I. napipes J.E. Lange, I. nitidiuscula (Britzelm.)

Lapl., I. oblectabilis (Britzelm.) Sacc., I. patouillardii Bres., I. posterula

(Britzelm.) Sacc., I. praetervisa Quél., I. pyriodora (Pers.) P. Kumm., I. splendens

R. Heim, I. splendentoides Bon, I.‘ squamata J.E. Lange, and I. vaccina Kithner

have been reported from Pakistan (Ahmad et al. 1997, Sultana et al. 2011, Ilyas

et al. 2013). During our present exploration of ectomycorrhizal fungi from pine

forests of the western Himalayas, I. amicta and I. mimica were found associated

with Pinus wallichiana A.B. Jacks. Both morphological and molecular evidence

were used to confirm the identity of these species. These records raised the

number of Inocybe spp. reported from Pakistan to twenty-six.

Materials & methods

Morphological evaluation

Basidiomata were collected, photographed, vouchered in the Herbarium,

Department of Botany, University of the Punjab, Lahore, Pakistan (LAH), dried, and

characterized morphologically. Sections of specimens were mounted in 5% KOH for

microscopic observations under a biological microscope (MX4300H, Meiji Techno Co.,

Ltd., Japan). Phloxine was used to increase contrast of the structures. Measurements

of anatomical features (basidiospores, basidia, cystidia, pileus, and stipe hyphae) are

presented from at least 25 measurements made with an ocular micrometer and 100 oil-

immersion objective, and include x = arithmetic mean of spore length and spore width

for all spores measured, Q = spore length divided by spore width. Line drawings were

made with a camera lucida. Color designations are from Munsell (1975).

DNA extraction, PCR amplification, DNA sequencing

Genomic DNA was extracted from a small piece of pileus by a modified CTAB

method (Bruns 1995). Amplification of internal transcribed spacers (ITS1 & ITS2) and

the 5.8S region of the nuclear ribosomal RNA gene were targeted using the primer pairs

ITS1F and ITS4 (White et al. 1990, Gardes & Bruns 1993) using the Extract-N-Amp

plant DNA extraction Kit (Sigma-Aldrich, St. Louis, MO, USA). PCR was carried out

under the following cycling parameters: initial denaturation (94°C for 1 min), 35 cycles

(94°C for 1 min, 53°C for 1 min, and 72°C for 1 min), and final extension (72° C for

8 min). Amplified PCR products were sent for purification and bidirectional sequencing

to Macrogen (Republic of Korea).

Sequence alignment and phylogenetic analysis

Sequencing of the nrITS region of our four Pakistani Inocybe collections yielded

fragments of 640-670 base pairs. GenBank sequences for alignment and phylogenetic

analysis were selected based on the top 100 BLAST search results for our four new

sequences.

Inocybe spp. new to Pakistan ... 673

aN SS ¢ >

A “\ A Ke

WA WW ‘ \\y

Fic. 1. Basidiomata: A. Inocybe amicta (MSM0010); B. Inocybe mimica (MSMO0011).

Scale bars: A = 7 mm; B = 15 mm.

674 ... Saba, Ahmad, & Khalid

Sequences were manually edited and assembled using BioEdit (www.mbio.ncsu.edu/

bioedit/bioedit.html). Following Dentinger et al. (2011) for complete ITS sequences,

all sequences were trimmed with the conserved motifs 5’-(...GAT) CATTA- and -

GACCT (CAAA...)-3’ and the alignment portion between them was included in analysis.

Simocybe serrulata (Murrill) Singer was used as outgroup based on results reported by

Larsson et al. (2009).

One sequence of Inocybe amicta, three sequences of I. mimica, and other related

sequences retrieved from the GenBank were aligned by Muscle using the default setting

in Molecular Evolutionary Genetics Analysis (MEGA) software (Tamura et al. 2011).

A phylogenetic tree was constructed with the Maximum Likelihood (ML) algorithm

using a Jukes & Cantor (1969) model of nrITS sequences and nearest-neighbor-

interchange (NNI) as ML heuristic search method using MEGAS software (Tamura

et al. 2011). The topology was assessed by 1000 bootstrap replicates. Corresponding

bootstrap values >50 % are cited on the trees (Fics 5, 6).

Sequences of I. amicta and I. mimica generated for this study were submitted to

GenBank. Accession numbers for the taxa retrieved from GenBank used in the

phylogenetic analysis are cited in the phylogenetic trees. Percent identities and DNA

divergences were calculated by DNAStar.

Taxonomy

Inocybe amicta Kokkonen & Vauras, Mycol. Progr. 11: 323 (2012). Fis 1A, 2, 4A,B

PILEUS 25-35 mm diam., conical or conic-convex, with an obtuse umbo;

margin reflexed, rimulose, surface dull, smooth at the disc, torn or cracked

towards the margin; brown, dark brown or reddish brown (2.5YR1/4) at the

center, fading to grayish brown or strong brown (5YR4/6) towards the margin.

LAMELLAE adnexed, moderately crowded, with one tier of lamellulae, pale

brown (5YR4/6), edge even, concolorous. STIPE 50-84 mm long, 6 mm diam. at

the apex, 10 mm diam. at the base, which is rounded subbulbous; central, solid;

surface not pruinose, white felted or floccose, striate; dark brown (5YR4/6).

BASIDIOSPORES 8-9.6 x 5.6-7 um [x = 8.9 x 64 um, Q = 1.20-1.56],

weakly to strongly nodulose, nodules vary in size, pale yellowish brown in

KOH, inamyloid. Basrp1a 22.8-32 x 6.1-12.5 um, clavate, four-spored, thin-

walled, hyaline in KOH; sterigmata 3.3-6.3 um. PLEUROCYSTIDIA 49-68 x

15-21 um, metuloid, lageniform with short neck or utriform rarely fusiform,

apex encrusted with crystals, tapered at the base. CHEILOCYSTIDIA 47-66 x

19-28 um, similar to pleurocystidia. PILEIPELLIS a cutis, hyphae cylindrical,

thin-walled, hyaline in KOH. Cautocystipia 24-59 x 7-18 um, present at

the stipe apex, cylindrical or subfusiform, hyaline in KOH, hyphoid cystidia

common, often with septa, wall thick at base or with protuberances in center.

CLAMP CONNECTIONS present.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTOONKHAW, Shangla, under Pinus

wallichiana, 2 September 2013, coll. M. Saba & A.N. Khalid MSM0010 (LAH; GenBank

KJ686344).

Inocybe spp. new to Pakistan ... 675

Fic. 2. Inocybe amicta (MSM0010):

A. Basidiomata; B. Basidia; C. Pleurocystidia; D. Cheilocystidia; E. Basidiospores.

Scale bars: A = 7 mm; B = 12 um; C = 15 um; D = 20 um; E= 8 um.

Inocybe mimica Massee, Ann. Bot. 18: 492 (1904). FIGS 1B, 3, 4C,D

PILEUS 31-35 mm diam., conical, obtusely umbonate, margin reflexed,

bent downward; surface radially fibrillose, striate, shiny, with a few scattered

scales, cracked towards the center; pale greenish/brownish yellow (7.5Y9/4).

LAMELLAE sinuate or adnexed, close, pale greenish yellow (7.5Y9/4), edge

wavy, white fimbriate. STIPE 55-72 mm long, 6-8 mm diam. at the apex, 9-14

mm diam. at the base, central, equal, with a rounded subbulbous base, solid;

676 ... Saba, Ahmad, & Khalid

surface longitudinally striate, fibrillose, off-white in the upper part and with

pale greenish yellow in the lower part.

BASIDIOSPORES (9.2—)10-14.1 x 6.6-8 um [x = 12.5 x 9.8 um, Q = 1.4-1.96],

mostly ellipsoid, apiculus present, smooth, yellowish brown or pale brown in

KOH, inamyloid. Bastp1a 9.9-55.4 x 9.6-17.2 um, clavate, four-spored, thin-

walled, hyaline in KOH; sterigmata up to 5.1 um long. PLEUROCYSTIDIA absent.

CHEILOCYSTIDIA 33.2-74.5 x 10.8-23.6 um, narrowly clavate or cylindrical,

smooth, hyaline in KOH. PILerPeLtis a cutis, hyphae cylindrical, 6.6-21 um,

thin-walled, hyaline in KOH, some with encrusting pigment. CAULOCYSTIDIA

33.7-65 x 9.9-15.5 um, cylindrical or clavate, hyaline in KOH, thin-walled.

CLAMP CONNECTIONS present.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTOONKHAW, Shangla, under Pinus

wallichiana, 2 September 2013, coll. M. Saba & A.N. Khalid MSM0011 (LAH; GenBank

KJ546158); 2 September 2013, I. Ahmad & A.N. Khalid P-48 (GenBank 726737), P-69

(KJ700456).

Fic. 3. Inocybe mimica (MSM0011):

A. Basidiomata; B. Basidia; C. Cheilocystidia; D. Basidiospores.

Scale bars: A = 15 mm; B = 10 um; C = 18 um; D = 6 um.

Inocybe spp. new to Pakistan ... 677

all

| fe

DP suemnssaat

Fic. 4. Inocybe amicta (MSM0010): A. Caulocystidia; B. Pileipellis.

Inocybe mimica (MSM0011): C. Caulocystidia; D. Pileipellis.

Scale bars: A = 20 um; B-D = 15 um.

Results

After removing and editing ambiguous positions in the alignment, a total

of 740 characters was subjected to phylogenetic analysis of which 377 were

conserved, 335 were variable, 230 were parsimony informative, and 100 were

singletons.

678 ... Saba, Ahmad, & Khalid

The ITS sequence of our Pakistani collection MSM0010 grouped in a clade

with four GenBank sequences of Inocybe amicta with 100% support (Fic. 5);

and the ITS sequences of our Pakistani collections MSM0011, P-48, and P-69

grouped in a clade with two GenBank sequences of Inocybe mimica with 98%

support (FIG. 6).

Discussion

In general, Inocybe spp. show a high sequence divergence in the ITS region.

Closely related species often deviate in several substitutions and insertion/

deletion events and are therefore easy to identify using simple sequence

comparison (Altschul et al. 1997).

Initial BLAST analysis of ITS sequences of our Pakistani collection

MSMO0011 showed 98% identity with Inocybe amicta sequences JF908263 from

Italy and JN580861 from Finland. Our phylogenetic reconstruction based on

ITS sequences shows authentic I. nitidiuscula well separated from I. amicta.

Morphologically, MSM0011 has the nodulose spores and white felted or floccose

stipe described for I. amicta (Kokkonen & Vaurus 2011), in contrast to the

smooth basidiospores and whitish pruinose upper third of stipe characterizing

I. nitidiuscula (Obase et al. 2006). We therefore conclude that HQ604600 is a

sequence from a misdetermined specimen of I. amicta.

Inocybe amicta has been described from three localities in Finland from

southern and northern boreal zones on richer but mainly sandy soils with

Betula, Picea abies, and Pinus sylvestris (Kokkonen & Vauras 2012). The species

is grouped in clade /inocybe, which is characterized by a distinct apiculus on the

spores and presence of pleurocystidia. Species with nodulose spores probably

evolved independently on numerous occasions (Matheny 2005). The /inocybe

clade includes species that are distributed primarily in temperate areas. ‘This is

undoubtedly the most evolutionarily diverse clade and contains approximately

85% of the species in Inocybaceae (Matheny 2009).

To our knowledge, this is the first report of I. amicta from Asia and the

first record of its association with Pinus wallichiana. We also believe this

is the first report of this species since it was first described from Finland in

2012 (Kokkonen & Vaurus 2012). This study demonstrates I. amicta has a

wide distribution outside European countries. It is closely related to I. silvae-

herbaceae (Fic. 5), which is distinguished by the finely fibrillose to delicately

floccose stipe turning pale brown (often with a reddish tinge), and the presence

of multiform caulocystidia. The ITS sequences of these two species differ only

by 14 bases and 4 insertions/deletions (each 1 base in length) (Kokkonen &

Vauras 2012).

Initial BLAST analysis of ITS sequences of our three Pakistani collections

MSMO0010, P48, and P69 revealed 99% maximum identity with Inocybe mimica

0.02

Inocybe spp. new to Pakistan ... 679

Wi Inocybe_amicta_KJ686344

Inocybe_amicta_JN580859

Inocybe_amicta_JN580861

Inocybe_amicta_JN580860

Inocybe_amicta_JF908263

Inocybe_silvae-herbaceae_JN580854

Inocybe_silvae-herbaceae_JN580855 Clade

100] Inocybe_silvae-herbaceae_NR_119991 /inocybe

Inocybe_silvae-herbaceae_JN580858

Inocybe_purpureobadia_JN580876

Inocybe_purpureobadia_JN580875

Inocybe_boreocarelica_JN580835

100

Inocybe_boreocarelica_NR_119989

Simocybe_serrulata_DQ494696 | Outgroup

Fic. 5. Phylogenetic relationship of Inocybe amicta with other related Inocybe spp. based on

Maximum Likelihood inferred from nrITS sequences.

0.02

Inocybe_mimica_KF056319

97 | Mi Inocybe_mimica_KJ700456

Hl Inocybe_mimica_KJ726737

Ml Inocybe_mimica_KJ546158

Inocybe_mimica_FJ904124

Inocybe_arenicola_FJ904134

Inocybe_arenicola_FJ904133

100; Inocybe_dulcamaroides_FJ904127

Inocybe_dulcamaroides_FJ904126

100, Inocybe_spuria_FJ904139

Inocybe_spuria_JF908260

Inocybe_flavella_ JQ724026

100! Inocybe_flavella JQ724025

Simocybe_serrulata_DQ494696 | Outgroup

100

Clade

/inosperma

Fic. 6. Phylogenetic relationship of Inocybe mimica with other related Inocybe spp. based on

Maximum Likelihood method inferred from nrITS sequences.

sequences KF056319 from India and FJ904124 from Finland, both from

coniferous forests.

Inocybe mimica was described by Massee (1904) from Yorkshire, UK.

Larsson et al. (2009) reported this species under Betula, Picea, and Pinus trees

680 ... Saba, Ahmad, & Khalid

on calcareous soils from Northwest Europe (Sweden). Our collections represent

the first report of I. mimica from Asia and the first record of its association with

Pinus wallichiana. According to Larsson et al. (2009), I. mimica is a very rare

and poorly known species that is closely related to I. arenicola (Fic. 6). Both

are characterized by rather large spores and a preference for calcareous soils,

but other morphological and ecological traits are highly variable. Larsson et al.

(2009) placed I. mimica in clade /inosperma. Molecular analysis confirms that

most species in this clade bear phaseoliform spores for the most part and/or

have reddening context. A rimose pileus appears to be symplesiomorphic for

the clade as some species of I. sect. Cervicolores, which are derived within the

group, bear a squamulose pileus (Matheny 2009).

This study raises the total number of known taxa of Inocybe to twenty-six

in Pakistan.

Acknowledgments

We are cordially grateful for Higher Education Commission for funding this

project under Phase II, Batch I, Indigenous fellowship program. We are thankful to

Dr. P. Brandon Matheny (Department of Ecology and Evolutionary Biology, University

of Tennessee, Knoxville) and Dr. Vladimir Antonin (Moravian Museum, Czech

Republic) for critically reviewing the manuscript. We are thankful to all lab fellows for

accompanying us on field excursions. We are cordially thankful to Ms. Tayyaba Qasim

for illustrating basidiomata.

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MY COTAXON

http://dx.doi.org/10.5248/130.683

Volume 130, pp. 683-688 July-September 2015

Elmerina fragilis sp. nov. from Central China

FANG Wu’, YUAN YUAN’, & Hal-J1ao Li'’?*

"Institute of Microbiology, RO. Box 61, Beijing Forestry University, Beijing 100083, China

*National Institute of Occupational Health and Poison Control,

Chinese Center for Disease Control and Prevention, Beijing 100050, China

*CORRESPONDENCE TO: lihaijiao715@126.com

Asstract — Elmerina fragilis (Auriculariales) is described and illustrated from Central

China, based on morphological characters. It is characterized by an annual growth habit,

pileate basidiocarps (gelatinous and flesh when fresh, becoming fragile when dry), a

monomitic hyphal system with clamp connections on generative hyphae, vertically septate

basidia, broadly ellipsoid basidiospores measured as 3-4 x 2-2.7 um, and growth on a

decorticated trunk of Quercus.

Key worps — Basidiomycota, polyporoid fungi, taxonomy, wood-inhabiting fungi

Introduction

Elmerina Bres. is a polyporoid genus with longitudinally septate basidia,

thus belonging to Auriculariales. The genus, typified by Elmerina cladophora

(Berk.) Bres., is characterized by annual and corky basidiocarps, poroid to

lamellate hymenophores, a dimitic hyphal system with clamp connections on

generative hyphae, vertically septate basidia, and hyaline, thin-walled, allantoid

to ellipsoid basidiospores (Reid 1992, Wei & Dai 2008, Zhou & Dai 2013).

Several new species of wood-decaying fungi have been found previously in

Central China (Li et al. 2007, Dai et al. 2009, Zhou & Dai 2012, Zhao & Cui

2013, Shen & Cui 2014). During a continuation of polyporoid fungal research in

Central China, a specimen was collected that exhibited an annual growth habit,

pileate basidiocarps, a monomitic hyphal system with clamp connections on

generative hyphae, vertically septate clamps, and broadly ellipsoid hyaline thin-

walled basidiospores. This specimen also had gelatinous and flesh basidiocarps

when fresh but became fragile when dry. Several attempts to extract its DNA

have failed. Soft fruiting bodies and a monomitic hyphae system are not typical

684 ... Wu, Yuan, & Li

for Elmerina, but the polyporoid hymenophores and vertically septate basidia

make this fungus a new species closest to Elmerina. We describe and illustrate

the new species here.

Materials & methods

Morphological studies

Sections were studied microscopically according to Dai (2010) at magnifications

<1000x using a Nikon Eclipse 80i microscope with a phase contrast illumination.

Drawings were made with the aid of a drawing tube. Microscopic features,

measurements, and drawings were made from sections stained with cotton blue and

Melzer’s reagent. Basidiospores were measured from sections cut from the tubes.

To present spore size variation, the 5% of measurements excluded from each end of

the range are given in parentheses. Basidiospore spine lengths were not included in

the measurements. Abbreviations include IKI = Melzer’s reagent, IKI+ = amyloid,

IKI- = negative in Melzer’s reagent, KOH = 5% potassium hydroxide, CB = cotton

blue, CB- = acyanophilous, L = mean spore length (arithmetic average of all spores),

W = mean spore width (arithmetic average of all spores), Q = the L/W ratio, and n

= number of spores measured from given number of specimens. Special color terms

follow Petersen (1996). The type specimen is deposited in the herbarium of Beijing

Forestry University, Beijing, China (BJFC).

Taxonomy

Elmerina fragilis F. Wu & Hai J. Li, sp. nov. FIGURES 1, 2

MycoBAnk MB 811274

Differs from other Elmerina species by its gelatinous and flesh basidiocarp when fresh, a

monomitic hyphal system, and amyloid basidiospores.

Type: China. Henan Province: Neixiang County, Baotianman Nature Reserve, on fallen

decorticated trunk of Quercus, 22.1X.2009, Dai 11267 (holotype, BJFC007241).

EryMo_oey: fragilis (Lat.) referring to the fragile basidiocarps when dry.

FruiTBopy. Basidiocarps annual, pileate, flesh and gelatinous when fresh,

strongly crumpled and fragile when dry. Pileus circular when fresh, irregularly

shaped when dry, projecting up to 1.8 cm, 2 cm wide, and 1 mm thick at

center. Pileal surface orange-yellow to yellowish brown when fresh, becoming

orange-brown to reddish brown and full of wrinkles when dry, glabrous. Pore

surface yellowish brown when fresh, becoming brown to dark brown when dry;

pores round, 5-7 per mm; dissepiments thin, entire. Context orange-brown to

reddish brown, fragile when dry, <0.9 mm thick. Tubes brown to dark brown,

fragile when dry, <0.1 mm long.

HyYPHAL STRUCTURE. Hyphal system monomitic; generative hyphae bearing

clamp connections, IKI-, CB-; tissue unchanged in KOH.

CONTEXT. Generative hyphae hyaline to pale yellowish, thin-walled,

moderately branched, interwoven, 2.8-8.5 um in diam; gloeoplerous hyphae

Elmerina fragilis sp. nov. (China) ... 685

Ficure 1. Elmerina fragilis (holotype): Fruiting bodies.

dominant, hyaline to pale yellowish, thin-walled, frequently branched, 1-4 um

in diam.

TuBES. Generative hyphae hyaline to pale yellowish, thin-walled, parallel

along the tubes, mostly collapsed, rarely branched, 3.5-7.2 um in diam.

Gloeoplerous hyphae common, hyaline to pale yellowish, thin-walled, parallel

along the tubes, unbranched, 3-7 um in diam. Cystidia and cystidioles absent.

Basidia clavate, thin to slightly thick-walled, with a vertical septate, which

divides basidia into two parts, each part with two sterigmata, a basal clamp

connection present, 11.5-14 x 3.7-5 um; basidioles similar to basidia in shape,

but without vertical septate and slightly smaller.

Spores. Basidiospores broadly ellipsoid, hyaline, thin-walled, smooth,

IKI+, CB-, (2.8-)3-4 x 2-2.7(-2.8) um, L = 3.51 um, W = 2.35 um, Q = 1.49

(n= 30/1).

Discussion

Elmerina was previously treated in Tremellales because of its tremelloid

basidia (Bandoni et al. 1984), but phylogenetic analyses indicate that the genus

is a member of the Auriculariales (WeifS & Oberwinkler 2001, Larsson et al.

2004, Zhou & Dai 2013).

Elmerina fragilis is distinct in the genus because of its monomitic hyphal

system with both normal generative hyphae and gloeoplerous hyphae and

686 ... Wu, Yuan, & Li

FiGuRE 2. Elmerina fragilis (holotype):

a. Basidiospores. b. Basidia and basidioles.

c. Hyphae from tube trama. d. Hyphae from context.

Elmerina fragilis sp. nov. (China) ... 687

its amyloid basidiospores. As we noted above, these characters are not found

in accepted Elmerina species. DNA extraction from our type material was

attempted several times, but failed; therefore, the phylogenetic relationships of

E. fragilis with other taxa in Auriculariales remain unknown. Elmerina fragilis

definitely belongs to Auriculariales based on its morphological characters,

poroid hymenophores, and vertically septate basidia. For the time being, we

place this species in Elmerina for the reason that the species is mostly close

to that genus. Protomerulius Méller is morphologically similar to Elmerina

(Ryvarden 1991, Reid 1992), but the two genera are phylogenetically divergent

(Zhou & Dai 2013). Protomerulius has gloeocystidia and pear-shaped or

subglobose basidia, while Elmerina lacks gloeocystidia and its basidia are

clavate (Zhou & Dai 2013). In addition, Protomerulius species occur in tropical

Africa and South America, whereas almost all Elmerina species (including the

type) have been described from tropical to temperate localities in eastern and

southeastern Asia. Thus, Elmerina fragilis has a morphology and a geographical

distribution closer to Elmerina than to Protomerulius.

Acknowledgements

We express our gratitude to Drs. Li- Wei Zhou (Institute of Applied Ecology, Chinese

Academy of Sciences, China) and De-Wei Li (The Connecticut Agricultural Experiment

Station Valley Laboratory, USA), who reviewed the manuscript. The research was

financed by the National Natural Science Foundation of China (Project No. 31372115).

Literature cited

Bandoni RJ. 1984. The Tremellales and Auriculariales: an alternative classification. Trans. Mycol.

Soc. Japan 25: 489-530.

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 131-343.

http://dx.doi.org/10.1007/s13225-010-0066-9

Dai YC, Yuan HS, Wang HC, Yang F, Wei YL. 2009. Polypores (Basidiomycota) from Qin Mts. in

Shaanxi Province, central China. Ann. Bot. Fennici 46: 54-61.

Larsson KH, Larsson E, Ko6ljalg U. 2004. High phylogenetic diversity among corticioid

homobasidiomycetes. Mycol. Res. 108: 983-1002.

http://dx.doi.org/10.1017/S0953756204000851

Li J, Xiong HX, Zhou XS, Dai YC. 2007. Polypores (Basidiomycetes) from Henan Province in

central China. Sydowia 59: 125-137.

Petersen JH. 1996. The Danish Mycological Society's colour-chart. Foreningen til

Svampekundskabens Fremme, Greve. 6 p.

Reid DA. 1992. The genus Elmerina (Tremellales) with accounts of two species from Queensland,

Australia. Persoonia 14: 465-474.

Ryvarden L. 1991. Genera of polypores, nomenclature and taxonomy. Synop. Fung. 5. 363 p.

Shen LL, Cui BK. 2014. Morphological and molecular evidence for a new species of Postia

(Basidiomycota) from China. Crypt. Mycol. 35: 199-207.

http://dx.doi.org/10.7872/crym.v35.iss2.2014.199

688 ... Wu, Yuan, & Li

Wei YL, Dai YC. 2008. Notes on Elmerina and Protomerulius (Basidiomycota). Mycotaxon 105:

349-354.

WeifS M, Oberwinkler F. 2001. Phylogenetic relationships in Auriculariales and related groups—

hypotheses derived from nuclear ribosomal DNA sequences. Mycol. Res. 105: 103-415.

http://dx.doi.org/10.1017/S095375620100363X

Zhao CL, Cui BK. 2013. Three new Perenniporia (Polyporales, Basidiomycota) species

from China based on morphological and molecular data. Mycoscience 54: 231-240.

http://dx.doi.org/10.1016/j.myc.2012.09.013

Zhou LW, Dai YC. 2012. Phylogeny and taxonomy of Phylloporia (Hymenochaetales): new species

and a worldwide key to the genus. Mycologia 104: 211-222. http://dx.doi.org/10.3852/11-093

Zhou LW, Dai YC. 2013. Phylogeny and taxonomy of poroid and lamellate genera in the

Auriculariales (Basidiomycota). Mycologia 105: 1219-1230. http://dx.doi.org/10.3852/12-212

MY COTAXON

http://dx.doi.org/10.5248/130.689

Volume 130, pp. 689-693 July-September 2015

A new species and new records of the lichen genus Pertusaria

from China

QIANG REN*

College of Life Science, Shandong Normal University, Jinan, 250014, China

*CORRESPONDENCE TO: rendagiang@hotmail.com

ABSTRACT — Pertusaria alticola sp. nov. is described and illustrated from the Qingzang

Plateau in Southwestern China; it is characterized by an isidiate thallus with fertile verrucae,

8-spored asci with uniseriate, ellipsoid ascospores, and the presence of psoromic acid. Three

other species, Pertusaria borealis, P. leucosora, and P. leucosorodes, are reported from China

for the first time.

Key worps — Pertusariaceae, lichenized fungi, taxonomy

Introduction

Pertusaria is a genus of crustose lichens with a high biodiversity; diagnostic

characters for identifying species are the apothecial structure, number of

ascospores per ascus, spore structure, and chemistry (Dibben 1980, Archer

1997, Schmitt & Lumbsch 2004). Since the publication of an annotated key to

the lichen genus Pertusaria in China (Zhao et al. 2004), additional specimens

have been collected and studied. During the survey of Chinese Pertusaria

species, the author has encountered a new species and, in addition, three

Pertusaria species are reported here for the first time from China.

Materials & methods

The present study is based mainly on collections preserved in the herbarium,

Shandong Normal University, Jinan, China (SDNU) and some specimens in the Lichen

Section of Herbarium, Institute of Microbiology, CAS, Beijing, China (HMAS-L). The

morphology and anatomy of all specimens were studied using an Olympus SZX16

stereomicroscope and an Olympus BX61 compound microscope and photographed

using an attached Olympus DP72 digital camera. Color reactions (spot tests) and the

chemical constituents were identified using standard methods (Orange et al. 2001).

690 ... Ren

Taxonomy

The new species

Pertusaria alticola Q. Ren, sp. nov. PLATE 1

MycoBank MB811504

Differs from Pertusaria wulingensis by its isidiate thallus and smaller ascospores.

Type: China, Yunnan, Yulong County, Lijiang Alpine Botanical Garden, alt. 3590 m, on

Pinus densata Mast., 16 Aug. 2011, Ren 2011180 (holotype, SDNU).

Erymo ocy: from the Latin altus, high and cola, dweller, a reference to the high altitudes

at which the specimens were collected.

THALLUsS gray orash-gray, thin, the margin entire and unzoned; sURFACE smooth,

generally matt, continuous; sOREDIA absent, but IsIp1A present and generally

prolific, papillate or granulate, rarely coralloid-branched, and fragile, 0.1-0.3

mm wide and 0.1-0.5 mm tall, concolorous with thallus. FERTILE VERRUCAE

uncommon, lecanorate, well dispersed, rarely fused, concolorous with the

thallus, borne between isidia, 0.5-1.0(-1.5) mm in diam. Discs yellowish, level

and lightly pruinose when mature. APOTHECIA | or 2-3 per verruca, the fruit

center hyaline. HypoTHEcIUM hyaline or brownish, EPITHECIUM brownish,

K-. Ascr 8-spored, cylindric; ascosporgs strictly uniseriate, hyaline, ellipsoid

or round, 12-20 x 10-12 um; sPORE WALL smooth, thin, c. 1-2 um thick and

the end wall not trimmed; Pycnip1a not seen.

CHEMISTRY — Cortex K-, C-, KC-, Pd-; medulla K+ yellow, C-, KC-,

Pd+ yellow. TLC: psoromic acid.

ECOLOGY & DISTRIBUTION — Pertusaria alticola is an isidiate corticolous

species, found at several localities on the Qingzang Plateau. This new species

grows mainly on bark of conifers, especially Pinus and Picea, at altitudes

between 2450 m and 4100 m.

ADDITIONAL SPECIMENS EXAMINED — CHINA. YUNNAN: Yulong County, Lijiang

Alpine Botanical Garden, on Pinus densata, alt. 3460 m, 2011, Ren 2011448 (SDNU); alt.

3590 m, 16 Aug. 2011, Ren 2011179 (HMAS-L). X1zANG: Linzhi County, Mt. Sejilashan,

alt. 3300 m, on bark, 2007, Han 20073032 (SDNU). Gansu: Wenxian County, Qiujiaba,

on bark, alt. 2450 m, 2007, Yang 20070021, 20070048 (SDNU)); alt. 2940 m, 2007, Yang

20070056 (SDNU); Zhouqu County, Huacaopo, alt. 3400 m, on Picea, 2006, Shi 061058

(SDNU).

ComMENTs — Pertusaria alticola is chemically identical to P wulingensis

Z.T. Zhao & Z.S. Sun, which differs by its larger ascospores (27-40 um long),

pink discs, and lack of isidia (Ren et al. 2009). Pertusaria alticola also has the

same chemistry as P wauensis Elix & A.W. Archer, a sterile species that differs

by its generally wider, more irregularly shaped isidia, and which is known only

from Papua New Guinea, Sarawak, and Australia (Elix et al. 1997).

Pertusaria alticola sp. nov. (China) ... 691

PLATE 1. Pertusaria alticola (Ren 2011180, SDNU).

A. thallus; B. Apothecium cross section. Scale bars: A = 1 mm; B = 20 um.

692 ... Ren

New records for China

Pertusaria borealis Erichsen, Ann. Mycol. 36: 354 (1938).

This species is characterized by a corticolous thallus with white soralia and

the presence of fumarprotocetraric and protocetraric acids. It grows mainly on

old bark of pine trees (Pinus), and sometimes on deciduous tree bark (Quercus

and Betula) in China, and has a temperate distribution in Europe (Smith et al.

2009) and North America (Dibben 1980). This is a new record for China.

SPECIMENS EXAMINED — CHINA. YUNNAN: Chuxiong City, Mt. Zixishan, alt. 2460 m,

on Pinus armandii Franch., 2011, Ren 2011337 (HMAS-L), 2011346, 2011360 (SDNU);

Yulong County, Mt. Laojunshan, alt. 3075 m, on bark, 2002, Ren 14 (HMAS-L 091314);

Yulong Snow Mountain, near Yufeng Temple, alt. 2800 m, on Pinus sp., 2002, Ren 124

(SDNU); Lijiang Alpine Botanical Garden, alt. 3590 m, on Quercus pannosa Hand.-

Mazz., 2011, Ren 2011397 (SDNU); Jianchuan County, Mt. Shibaoshan, alt. 2640 m,

on Quercus acutissima Carruth., 2011, Ren 2011264 (SDNU). X1zane: Linzhi County,

Lulang Town, Sejilashan Observatory Station, alt. 3300 m, on bole of Betula, 2001, Han

20073067 (SDNU); the east slope of Mt. Sejilashan, alt. 4100 m, on bark, 2007, Han

20073250 (SDNU).

Note — Morphologically, Pertusaria borealis resembles P. amara (Ach.) NyL.,

which differs by having KC+ violet soralia and containing picrolichenic acid.

Pertusaria leucosora Nyl., Flora 60: 223 (1877).

This rare, saxicolous species is characterized mainly by a thick, cracked,

areolate thallus with a zoned margin; hemispherical white soralia (0.8-1.5

mm in diam.); and the production of protocetraric acid. It is reported from

Europe (Smith et al. 2009), Japan (Oshio 1968), India (Awasthi 1991), and Iran

(Valadbeigi & Sipman 2010). This is a new record for China.

SPECIMENS EXAMINED — CHINA. SHAANXI: Mt. Taibai, on the roadside between

Yingtou Police Station and Shangbaiyun, alt. 1150 m, on rock, 2005, Wang & Yang

TBW377 (SDNU). YUNNAN: Luquan County, Jiaozi Snow Mountain, alt. 3800 m, on

rock, 2008, Wang 20082256, 20081172, 20082298 (SDNU), Sun 20082162, 20082377

(SDNU).

Note — In ecology and morphology, P leucosora can be confused with the

European Pertusaria aspergilla (Ach.) J.R. Laundon, but P. aspergilla contains

fumarprotocetraric acid and has isidia and pale yellow-whitish soralia (Smith

et al. 2009).

Pertusaria leucosorodes Nyl., Acta Soc. Sci. Fenn. 26(10): 16 (1900).

= Pertusaria scaberula A.W. Archer, Mycotaxon 41: 240 (1991).

This sterile species is characterized by a thallus with abundant disciform

soralia and the presence of thamnolic acid and lichexanthone (cortex K+

yellow, UV+ orange, medulla K+ vivid yellow, Pd+ yellow). This corticolous

species occurs in Sri Lanka, Australia, Papua New Guinea, and Norfolk Is.

Pertusaria alticola sp. nov. (China) ... 693

(Archer 1997, as Pertusaria scaberula) and India (Shyam et al. 2011). This is a

new record for China.

SPECIMENS EXAMINED — CHINA. Fujian: Wuyishan City, Dawangfeng, alt. 540 m, on

bark, 2013, Ren 2012576 (SDNU); Jianou City, Wanmulin, alt. 370 m, on bark, 2007,

Jia 20073478 (SDNU); Longyan City, Huanglianyu, alt. 1400 m, on bark, 2010, Jiang

20105510 (SDNU). YuNNaN: Shilin County, near Shilin Scenic Area, alt. 1909 m, on

bark, 2002, Ren 316 (HMAS-L); Menghai County, Mangao Nature Reserve Area, alt.

1350 m, on Quercus, 2011, Ren 2011811 (SDNU). GUANGDONG: Xinyi City, Datianding,

alt. 1700 m, on bark, 2010, Wang 20107049 (SDNU); Fengkai County, Heishiding Shan,

alt. 220 m, on dead branch, 1998, Guo 2193 (HMAS-L). JIANGx1: Mt. Lushan, Sanbaoshu,

alt. 890 m, 2008, Zhao 20090129 (SDNU). Harnan: Ledong County, Jianfengling, alt.

900 m, on bark, 2007, Meng M314 (HMAS-L).

Acknowledgements

The author thanks Drs. Alan W. Archer (National Herbarium of New South

Wales, Australia) and André Aptroot (ABL Herbarium, Soest, The Netherlands) for

presubmission review. This project is supported by the National Natural Science

Foundation of China (31100011), and the Excellent Young Scholars Research Fund of

Shandong Normal University. The author is grateful to Ms. Hong Deng (HMAS-L) for

loan of specimens during the study.

Literature cited

Archer AW. 1997. The lichen genus Pertusaria in Australia. Bibliotheca Lichenologica 69. 249 p.

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

Lichenologica, 40. 332 p.

Dibben MJ. 1980. The chemosystematics of the lichen genus Pertusaria in North America north of

Mexico. Publications in Biology and Geology, Milwaukee Public Museum 5. 162 p.

Elix JA, Aptroot A, Archer AW. 1997. The lichen genus Pertusaria (lichenized Ascomycotina) in

Papua New Guinea & Australasia: twelve new species and thirteen new reports. Mycotaxon

64: 17-35.

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

London: British Lichen Society.

Oshio M. 1968. Taxonomical studies on the family Pertusariaceae of Japan. Journal of Science of

the Hiroshima University, Series B, Div. 2 (Botany) 12: 81-151.

Ren Q, Sun ZS, Zhao ZT. 2009. Pertusaria wulingensis (Pertusariaceae), a new lichen from China.

Bryologist 112(2): 394-396. http://dx.doi.org/10.1639/0007-2745-112.2.394

Schmitt I, Lumbsch HT. 2004. Molecular phylogeny of the Pertusariaceae supports secondary

chemistry as an important systematic character set in lichen-forming ascomycetes. Molecular

Phylogenetics and Evolution 33: 43-55. http://dx.doi.org/10.1016/j.ympev.2004.04.014

Shyam KR, Thajuddin N, Upreti DK. 2011. Diversity of lichens in Kollihills of Tamil Nadu, India.

International Journal of Biodiversity and Conservation 3(2): 36-39.

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA (eds). 2009. The

lichens of Great Britain and Ireland. Second edition. London: British Lichen Society.

Valadbeigi T, Sipman HJM. 2010. New records of lichens and lichenicolous fungi from Iran and

their biogeographical significance. Mycotaxon 113: 191-194. http://dx.doi.org/10.5248/113.191

Zhao ZT, Ren Q, Aptroot A. 2004. An annotated key to the lichen genus Pertusaria in China.

Bryologist 107(4): 531-541.

http://dx.doi.org/10.1639/0007-2745(2004)107[531:AAKTTL]2.0.CO;2

MY COTAXON

http://dx.doi.org/10.5248/130.695

Volume 130, pp. 695-700 July-September 2015

First record of Erysiphe howeana from China

Lu-CHAO Bat’, ZHI- MIN CAO", & PENG WANG?

‘College of Forestry, Northwest A&F University,

Taicheng Road, Yangling, Shaanxi 712100 China

°Xining Forestry Station, Xining, Qinghai 810000 China

* CORRESPONDENCE TO: zmcao@nwsuaf.edu.cn

AsBstTRACT — Erysiphe howeana (Erysiphe sect. Erysiphe) was identified on Oenothera

biennis. This is the first record of this powdery mildew from China. The Chinese specimen

is described, illustrated, and discussed, and its identity is confirmed by means of molecular

methods.

KEY woRpDs — common evening primrose, morphology, molecular phylogeny, distribution

data, Asia

Introduction

In the autumn of 2014, Oenothera biennis (Onagraceae) heavily infected by

a powdery mildew was found in Shaanxi, China. Based on the morphological

characteristics of the asexual morph, it was classified as Erysiphe sp. Molecular

analyses were applied to resolve the species identity. The internal transcribed

spacer (ITS) and 28S rDNA regions were sequenced and compared with

sequences in corresponding depositories. Based on sequence analysis of the

rDNA ITS region, this species has been identified as Erysiphe howeana.

This species had not previously been reported from China (Zheng &

Yu 1987, Wu & Wu 1991, Chen 1993, Wang et al. 2002, Liu 2010), and our

collection constitutes the first Chinese record.

Materials & methods

Living leaves of Oenothera biennis bearing the anamorph of a powdery mildew

were collected in October 2014 in the Qinling Mountains within Shangbahe Forest

Park in China. A herbarium specimen was deposited in the Mycological Herbarium

of Forestry College, Northwest A & F University, Yangling, Shaanxi Province, China

(HMNWAFU - CR).

696 ... Bai, Cao, & Peng

A sample was mounted in distilled water and examined using light microscopy

(Olympus, CX31RTSF, Japan). A scanning electron microscope (JEOL, JSM-6360LV)

was used to observe the anamorph ultrastructure, particularly the surface features of

conidia (according to Cook et al. 1997) and hyphal appressoria of this fungus.

Genomic DNA was extracted from mycelium and the asexual morph using

Chelex-100 (Walsh et al. 1991, Hirata & Takamatsu 1996). The ITS region of the nuclear

rDNA (including 5.88) and 28S rDNA sequences with domains D1 and D2 were

amplified via polymerase chain reaction (PCR); PM5 and PM6 (Takamatsu & Kano

2001) were used to amplify the ITS region, while LSU1 and LSU2 (Scholin et al. 1999)

were used to amplify the 28S rDNA sequence.

The PCR assays were conducted in a 50 uL final volume (Hirata & Takamatsu 1996)

containing 27 uL of 2x BoisTaq PCR MasterMix, 1 uL of each of primer, 1 uL of the

extracted DNA, and 20 uL of ddH20 (Hirata & Takamatsu 1996). Thermal cycling in

a PTC-200 thermal cycler (BioRad) comprised an initial denaturation step at 95°C for

5 min, 35 cycles of 94°C for 1 min + 60°C for 1 min + 72°C for 1 min, and a final

elongation step at 72°C for 8 min. A negative control for each set of reactions replaced

template DNA with ddH,O. The PCR products were separated by electrophoresis on a

2% agarose gel in TAE buffer and purified using the Zymoclean™ Gel DNA Recovery

Kit, according to the manufacturer's instructions. The purified DNA products were

ligated into the pMD18-T vector (Takara) and transformed into E. coli DH5a cells. The

cloned fragments were sequenced by Sangon Biotech (Shanghai) Co., Ltd.

All DNA sequences were aligned using Clustal X 1.81 (Thompson et al. 1997), and

the alignments were adjusted following Nei & Kumar (2000). All positions containing

gaps or missing data were eliminated from the dataset. Cladistic trees were constructed

using the neighbor-joining method with the Kimura 2-parameter substitution model in

MEGA 4.0 (Tamura et al. 2007). Branch robustness was assessed by bootstrap analysis

with 1000 replicates.

Taxonomy

Erysiphe howeana U. Braun, Mycotaxon 14: 373, 1982. PLATE 1

SPECIMEN EXAMINED: CHINA, SHAANXI, Qinling Mountains, Shangbahe Forest Park,

33°21’42”N 108°27'07’E, alt. 1472 m, on living leaves of Oenothera biennis L., Oct. 2014,

L.C. Bai (HMNWAFU-CF 2014001; GenBank KP247216, KP247417).

Mycelium mainly foliicolous, amphigenous, also on stems and fruits, effuse

or in irregular patches, often covering the entire leaf surface, white, persistent.

Hyphal appressoria lobed, solitary or in opposite pairs; hyphae smooth or

almost smooth, 3-7 um wide, colorless. Conidiophores arising from the

upper surface of mother cells, erect, straight or somewhat curved, foot-cells

cylindrical, about 23-42 x 6-11 um, followed by 1-2 shorter cells, forming

conidia singly. Conidia ellipsoid-ovoid, doliiform or subcylindrical, 22-33 x

10-17 um, with a length/width ratio of 1.3-2.2.

Erysiphe howeana new for China... 697

“eg a

~ cet

y ¢ t

’ oA z y

. : y

[ y ) = ees

iM . r Piya ] i 4

> : / | ~

Cte. Oka if ss *

— 7 MT 8 . * S-4800 10.0kV 8.6mm x1.80k

* Le Z ==

_ f

~ *

: /D

PLATE 1. Erysiphe howeana (HMNWAFU-CF 2014001): A & B. Oenothera biennis infected by a

powdery mildew; C & D. Conidiophore, conidia, and hyphae.

Phylogeny

28S ANALYSIS PLATE 2

The 28S rDNA sequence comprising 597 total characters was deposited

in GenBank under accession number KP247216. The sequence was aligned

with 15 sequences representing the five tribes of Erysiphaceae. The 28S rDNA

phylogenetic tree places the Erysiphe howeana sequence in one clade (bootstrap

value = 98%) where it clusters with E. paeoniae R.Y. Zheng & G.Q. Chen, clearly

supporting its placement in the tribe Erysipheae. Byssoascus striatosporus

(G.L. Barron & C. Booth) Arx was used as outgroup.

ITS ANALYSIS PLATE 3

The ITS rDNA sequence comprising 398 total characters was deposited

in GenBank under accession number KP247217. The sequence was aligned

with 16 sequences representing Erysiphe R. Hedw. ex DC. (emend. Braun

& Takamatsu 2000). Erysiphe adunca (Wallr.) Fr. var. adunca was used as

outgroup. The ITS phylogenetic tree placed the Chinese specimen and Erysiphe

howeana (GenBank AF011301, based on North American material) in a

strongly supported clade with 100% bootstrap support, indicating that these

two sequences represent the same species.

Discussion

The teleomorph of Erysiphe howeana was not found in the Chinese collection.

Powdery mildew on Oenothera spp., including worldwide collections of

698 ... Bai, Cao, & Peng

Erysiphe adunca var. adunca AB022374

irysiphe carpinicola AB252472

Erysiphe paeoniae AB257438

Erysiphe howeana KP247416

Erysipheae

Arthrocladiella mougeotii AB022379 Golovinomycetea

Neoerysiphe galeopsidis ABO22369

Pleochaeta turbinata AB218773

Phyllactinia moricola ABO22401

96 Leveillula taurica ABO22387

Phyllactinieae

Golovinomyces cichoracearum AB022360 |

Cystotheca wrightii AB022355

100 ——s Sawadaea tulasnei ABO022366

as Podosphaera fusca AB525914 ala ai

|| 99 Podosphaera tridactvla AB022393

Blumeria graminis AB022399 Blumerieae

Byssoascus striatosporus 017912

0.01

PLATE 2. Neighbor-joining tree derived from 28S rDNA sequences from 15 taxa of Erysiphaceae

with Byssoascus striatosporus as outgroup. The bar indicates a distance of 0.01.

Erysiphe weigelae AB015932

Erysiphe ligustri AB015917

58 75 Erysiphe pulchra KF601334

Erysiphe paeoniae AB257436

Erysiphe sedi JX173288

99— Erysiphe macleayae AB016048

Erysiphe lespedezae AB015923

100 | Erxysiphe howeana AF011301

Erysiphe howeana KP247417

62 Erysiphe heraclei AB104512

95 Erysiphe betae AF011290

74 Erysiphe polygoni AF011307

Erysiphe convolvuli AB104518

Erysiphe alphitoides AB237799

85 Erysiphe quercicola AB193590

Erysiphe liriodendri AF011302

Erysiphe adunca var. adunca D84383

0.02

PLATE 3. Neighbor-joining tree derived from ITS1 + 5.8S rDNA + ITS2 sequences from 16 Erysiphe

taxa with Erysiphe adunca var. adunca as outgroup. ‘The bar indicates a distance of 0.02.

Erysiphe howeana new for China ... 699

Pseudoidium-like asexual morphs, has been generally referred to E. howeana,

originally described from North America (Braun 1987, Braun & Cook 2012).

However, Bereczky et al. (2015), who sequenced powdery mildew on Oenothera

spp. from different parts of the world, found surprising results. Two different

species with morphologically minimally distinguishable anamorphs have

been identified on Oenothera spp., namely the common E. howeana and the

oak powdery mildew E. alphitoides (Griffon & Maubl.) U. Braun & S. Takam.,

which may occur on evening-primrose. Therefore, identification of Oenothera

powdery mildew based on the oidium alone is barely possible. Thus molecular

methods (ITS rDNA sequences) were applied to confirm the identity of the

Shaanxi specimen as E. howeana. This species has been reported from South

Africa, North America, and Europe, but not previously from Asia. This is the

first record of E. howeana for China.

Acknowledgments

This paper was supported by the Natural Science Foundation of Shaanxi Province

(No. 2010JZ003). We wish to thank U. Braun (Institute of Biology, Herbarium, Martin

Luther University, Germany) for valuable suggestions and critical comments to the

manuscript. We also thank Chen Liang (College of Agronomy and Plant Protection,

Qingdao Agricultural University, China) and Uwe Braun for their expert reviews.

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Occurrence of morphologically barely distinguishable powdery mildew anamorphs on

Oenothera spp. in different parts of the world. Mycoscience 56: 267-272.

http://dx.doi.org/10.1016/j.myc.2014.08.001.

Braun U. 1987. A monograph of the Erysiphales (powdery mildews). Beihefte zur Nova Hedwigia

89: 1-700.

Braun U, Cook RTA. 2012. Taxonomic manual of the Erysiphales (powdery mildews). CBS

Biodiversity Series 11.707 p.

Braun U, Takamatsu S. 2000. Phylogeny of Erysiphe, Microsphaera, Uncinula (Erysipheae) and

Cystotheca, Podosphaera, Sphaerotheca (Cystotheceae) inferred from rDNA ITS sequences -

some taxonomic consequences. Schlechtendalia 4: 1-33.

Chen ZX. 1993. Fujian Erysiphaceae. Fuzhou, Fujian Science and Technology Press. (in Chinese).

Cook RTA, Inman AJ, Billings C. 1997. Identification and classification of powdery mildew

anamorphs using light and scanning electron microscopy and host range data. Mycol .Res. 101:

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Hirata T, Takamatsu S. 1996. Nucleotide sequence diversity of rDNA internal transcribed spacers

extracted from conidia and cleistothecia of several powdery mildew fungi. Mycoscience 37:

283-288. http://dx.doi.org/10.1007/BF02461299

Liu TZ. 2010. Powdery mildew in Inner Mongolia. Hohhot, Inner Mongolia Science and

Technology Press. (in Chinese).

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http://dx.doi.org/ 10.1046/j.1529-8817.1999.3561356.x

Takamatsu S, Kano Y. 2001. PCR primers useful for nucleotide sequencing of rDNA of the powdery

mildew fungi. Mycoscience 42: 135-139. http://dx.doi.org/10.1007/BF02463987

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(MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596-1599.

http://dx.doi.org/10.1093/molbev/msm092

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The CLUSTAL X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Res. 24: 4876-4882. http://dx.doi.org/10.1093/nar /25.24.4876

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PCR-based typing from forensic material. Biotechniques 10: 506-513.

Wang RS, Dou YX, Zhu KG. 2002. The notes on Erysiphales in the Guiging Mountain of

Gansu Province. Journal of Gansu Agricultural University 37(4): 416-420. (in Chinese).

http://dx.doi.org/10.3969/j.issn.1003-4315.2002.04.004

Wu MX, Wu MQ. 1991. Guizhou plant powdery mildew. Guizhou, Guizhou Science and

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Press. (in Chinese).

MY COTAXON

http://dx.doi.org/10.5248/130.701

Volume 130, pp. 701-706 July-September 2015

Buellia, Lempholemma, and Thelidium species

new for Turkey and Asia

KENAN YAZICI **& ANDRE APTROOT?”

"Biology Department, Faculty of Science, Karadeniz Technical University,

61080, Trabzon, Turkey

?ABL Herbarium G.v.d.Veenstraat 107 NL-3762 XK Soest, The Netherlands

* CORRESPONDENCE TO: kcagri_1997@yahoo.com

ABSTRACT — Three lichen species - Buellia triseptata, Lempholemma isidioides, and Thelidium

fontigenum — were determined as new to Turkey and Asia following a recent lichenological

survey in the Burdur region (Turkey). Descriptions of the collections accompany notes on

geographic distribution, substrate, chemistry, and comparisons with morphologically similar

taxa.

Key worps — Ascomycota, biodiversity, Lichinaceae, Physciaceae, Verrucariaceae

Introduction

Studies on the lichen biota of Turkey during the last four years have

contributed many new records for the country (Aptroot & Yazici 2012; Arslan

et al. 2011; Karagéz & Aslan 2012; Karago6z et al. 2011; Kinalioglu & Aptroot

2011; Osyczka et al. 2011; Vondrak et al. 2012, Yazici et al. 2012, 2011a,b,

2010a,b,c). Within our project “Lichen and Bryophyte flora of Burdur Province”

we have found some lichenized fungi as new records for Turkey. Nonetheless,

more research is needed in the remaining lichenologically unexplored areas of

the country before we complete the lichen flora of Turkey.

Although approximately 1650 lichen taxa (including 35 taxa of Buellia,

7 of Lempholemma, and 3 of Thelidium) have been recorded in Turkey, only 37

lichenized fungi have thus far been reported from Burdur (Aslan & Yazici 2013,

Cobanoglu 2005, Oztiirk et al. 2005, Pisut & Guttova 2008, Senkardesler 2009,

Yazici et al. 2013). The present report on fieldwork in southwestern Turkey’s

Burdur region presents three new records for Turkey and Asia.

Burdur has a continental Mediterranean climate with cold snowy winters

and long hot dry summers. The mean annual temperature is 15°C, and the

temperature ranges from -16 °C to 39 °C. The mean annual rainfall is 468 mm,

and the average humidity is 51.2 % (Akman 1999).

702 ... Yazici & Aptroot

The Bucak and Altinyayla districts are mountainous with much forest

dominated by Abies, Cedrus, Ficus, Fraxinus, Juniperus, Olea, Pinus, Pistacia,

Prunus, Quercus (especially in the Altinyayla district), and Rhus alternating

with streams, lakes, and dams, such as the Yaprakli Dam (Altinyayla district)

and Karacaéren Dam (Bucak district) (Baytop & Denizci 1963). Additionally,

the understory flora is very abundant and the mountains are covered with

much marble rock.

Materials & methods

Lichen samples were collected by K. Yazici during 28-29 June 2012. Air-dried

samples were examined with a Nikon SMZ1500 stereomicroscope and a Nikon Eclipse

80i compound light microscope. The specimens were identified by consulting relevant

keys (Ahti et al. 2007, Giralt et al. 2010, Giralt & Nordin 2002, Gowan & Ahti 1993,

Nordin 2000, Pykala 2010, Smith et al. 2009, Thues & Nascimbene 2008). Thin layer

chromatography (TLC) analyses were performed according to Orange et al. (2001).

Vouchers are stored in the Herbarium of the Biology Department, Karadeniz Technical

University, Trabzon, Turkey (KTUB).

The descriptions are based on Turkish specimens; additional details are provided

from appropriate literature (Ahti et al. 2007, Nash et al. 2001, Nordin 2000, Smith et al.

2009).

The species

Buellia triseptata A. Nordin Fic 1

For a full description of the species, see Bungartz et al. (2008)

Thallus crustose, white to grey-white, <3 cm diam, thin, + rounded, +

verrucose and rimose-areolate, prothallus absent; apothecia numerous, crowded,

aggregated or spreading, up to 0.45 mm diam., at first immersed and flat later

convex, epruinose, margin distinct, later excluded; excipulum 15-55 um thick,

pale to dark brown, dark brown in outermost part; hymenium 60-70 um

tall, without oil droplets; asci 45-60 x 13-20 um. Paraphyses tips <5.5 um

wide, apical caps distinct; ascospores 3-septate, ellipsoid, (14-)16 -19(-21) x

(5.5-) 6.5 -7.5(-9) um, rarely with longisepta dividing the middle cells,

sometimes curved; without uneven wall thickenings , perispore c. 0.2 um thick,

rugulate. Pycnidia rare. Conidia c. 4-6 x 1 um, bacilliform.

SPECIMEN EXAMINED — TURKEY. Burpur: ALTINYAYLA, exit of Corten village,

36°59'06.50’N 29°29'06.33”E, 1114 m, on Quercus sp., 11.06.2013, leg. K.Yazici.

(KTUB-2430).

REMARKS—Buellia triseptata is similar to B. triphragmioides with yellowish

thallus but develops a grayish white thallus, smaller apothecia and spores,

and longer conidia. Associated species include Candelariella aurella, Collema

nigrescens, Lecanora dispersa, Melanelia subaurifera, Melanelixia glabra,

Melanohalea exasperata. Physcia adscendens, and Physconia distorta.

Lichens new for Turkey ... 703

Fic. 1. Buellia triseptata, habitus. Scale = 1 mm

HABIT & DISTRIBUTION—Buellia triseptata occurs in continental

submontane to montane areas within a wide range of altitudes. The lichen

grows on bark and wood (primarily on branches and twigs) of broad-leaved

and coniferous trees in open situations (Burgaz et al. 2002). It prefers solitary

trees in open situations or along streams or forest edges. Previously known

from southwestern Europe (Spain), Morocco, and western North America.

Lempholemma isidioides (Nyl. ex Arnold) H. Magn. Fic 2

For a full description of the species, see Ahti et al. (2007)

Thallus squamulose, forming irregular rosettes, <2.5mm in diam., pale

brown to black brown, with cylindrical isidia-like lobes, <0.25 mm long;

squamules 0.009-0.01 mm in diam.; apothecia laminal, subimmersed, pale

orange to orange, in groups of 1-4, <0.01 mm in diam; hymenium I + red

brown; ascospores simple, colourless, ellipsoid, 10-13 x 7-8 um. Pycnidia

immersed, rare; conidia simple, bacilliform, colourless, 1-2 x 1 um. Photobiont

Nostoc.

SPECIMEN EXAMINED — TURKEY. Burpur: Bucak, between Cobanpinar and Telciler

villages, 37°21'04.53”N 30°46'11.09”E, 357 m, on calcareous rock, 26.06.2013. leg.

K.Yazici. (KTUB-2432).

REMARKS— Associated species include Collema tenax and Leptogium diffractum.

HABIT & DISTRIBUTION—Lempholemma isidioides grows on irrigated sun-

exposed calcareous rocks. Previously known from Europe, North America, and

Scotland (Ahti et al. 2007). New to Turkey and Asia.

704 ... Yazici & Aptroot

Fic. 2. Lempholemma isidioides, habitus. Scale = 1 mm

Thelidium fontigenum A. Massal. Fig 3

For a full description of the species, see Smith et al. (2009)

Thallus crustose, thin, yellowish, pale white to grey-white or grey,

continuous, <4 cm in diam., often with patches of a ae K + violet-red

Fic. 3. Thelidium fontigenum, habitus. Scale = 1 mm

Lichens new for Turkey ... 705

pigment; perithecia <0.20 mm diam., + % immersed in pits or in projections,

0.10-0.35 mm wide; involucrellum thin, diverging from the apex of exciple,

15-35 um wide; ascospores 3-septate, 25-38 x 12-16 um. Thallus (yellow

areas) K + purple.

SPECIMEN EXAMINED — TURKEY. Burpwr: Bucak, Kuyubasi village, 37°22’12.25”N

30°39’22.76’E, 990 m, on calcareous rock, 24.08.2012, leg. K. Yazici. (KTUB-2429).

REMARKS— Thelidium fontigenum resembles T. papulare but develops smaller

perithecia, involucrellum, and spores and has K+ red pigment patches (Orange

2013). Associated species include Caloplaca aurantia and Verrucaria nigrescens.

HABIT & DISTRIBUTION—Thelidium fontigenum occurs on limestone,

lime-containing sandstone, mortar, tufa, dolomite, calcareous sandstone, and

silt-impregnated wood as well as mica schist and siliceous stones at varying

altitudes from low mountain ranges to (sub-)alpine areas. Previously known

from Europe and North America. New to Turkey and Asia.

Acknowledgements

We are grateful to Dr. Ana Rosa Burgaz and Dr. Victor J. Rico (Complutense

University, Madrid, Spain) for revisions and helpful comments on an earlier draft of this

manuscript. This study was supported by TUBITAK (Project 111T857).

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http://www.mycotaxon.com//resources/checklists/Yazici-v116-checklist.pdf

Yazici K, Aptroot A, Aslan A. 2011b. Lecanora wrightiana and Rhizocarpon inimicum, rare lichens

new to Turkey and Middle East. Mycotaxon 117: 145-148. http://dx.doi.org/10.5248/117.145.

Yazici K, Aptroot A, Aslan A. 2012. Candelariella, Ochrolechia, Physcia, and Xanthoria species new

to Turkey. Mycotaxon 119: 149-156. http://dx.doi.org/10.5248/119.149

Yazici K, Aslan A, Aptroot A. 2013. New lichen records from Turkey. Bangladesh Journal of Plant

Taxonomy 20(2): 207-211. http://dx.doi.org/10.3329/bjpt.v20i2.17395

MY COTAXON

http://dx.doi.org/10.5248/130.707

Volume 130, pp. 707-715 July-September 2015

Four new records of Lecanoraceae in China

XIN ZHAO’, Lu-LU ZHANG?”, LI-YAN SUN?, LING Hu’, & ZUN-TIAN ZHAO'*

‘College of Life Sciences, Suandong Normal University, Jinan, 250014, P. R. China

School of Mathematical Sciences, Shandong Normal University, Jinan 250014, P. R. China

College of Pharmacology, Taishan Medical University, Taian 271016, P. R. China

* CORRESPONDENCE TO: ztzhao@sohu.com

ABSTRACT — Four lichen species of Lecanoraceae, Clauzadeana macula, Lecidella oceanica,

Lecidella tumidula, and Miriquidica obnubila, are reported for the first time from China.

Detailed descriptions of morphology with high resolution photographs, chemistry,

comments, and distribution are provided.

Key worps — Lecanorales, Asia, taxonomy

Introduction

Lecanoraceae Korb. (Lecanorales, Lecanoromycetes, Ascomycota) contains

about 27 genera (Lumbsch & Huhndorf 2010) with more than 800 species

worldwide; the largest genus of the family is Lecanora Ach., including about

552 species (Kirk et al. 2008). Only 27 lecanoraceous species have been

reported from China: two of Calvitimela Hafellner, three of Carbonea (Hertel)

Hertel, eleven of Lecidella Korb., two of Miriquidica Hertel & Rambold, three

of Pyrrhospora K6rb., and six of Rhizoplaca Zopf (Wei 1991, Abbas & Wu

1998, Aptroot & Seaward 1999, Abbas et al. 2001, Aptroot & Sparrius 2003,

Obermayer 2004, Zhang et al. 2012a, Zhao et al. 2013). During our research

on the lecideoid taxa of Lecanoraceae in China, we identified four species

new to the country: Clauzadeana macula, Lecidella oceanica, L. tumidula, and

Miriquidica obnubila.

All three genera (Clauzadeana Cl. Roux, Lecidella, Miriquidica) have

Lecanora-type asci. Clauzadeana was created by Roux (1983) to accommodate

a species that differed from Lecidea Ach. in several microscopic characters,

such as the presence of algal cells in the exciple tissue and below the hymenium,

the richly branched and anastomosing paraphyses, and the Lecanora-type

708 ... Zhao & al.

ascus structure. Lecidella is characterized by dark brown to black lecideine

apothecia with a green or olive to brown epithecium, lax and simple paraphyses,

simple ascospores, and abundant xanthones as major secondary metabolites.

Miriquidica can be distinguished from Lecidella by a thallus that often has an

epinecral layer and different secondary metabolites, usually miriquidic acid,

lobaric acid and/or norstictic acid.

Materials & methods

The specimens studied were collected from Liaoning, Sichuan, Xinjiang, and

Yunnan provinces in China and are preserved in the Lichen Section of Botanical

Herbarium, Shandong Normal University, Jinan, China (SDNU) and the Herbarium,

Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China (KUN).

Morphological and anatomical characters were examined using a Nikon SMZ 745T

stereomicroscope and Olympus CX21 compound microscope. Lichen substances were

identified using the standardized thin layer chromatography techniques (TLC) with

solvent system C (Orange et al. 2001). Morphological and anatomical photos of the

specimens were taken under an Olympus SZX16 stereomicroscope and an Olympus

BX61 compound microscope with an Olympus DP72 camera.

Taxonomic descriptions

Clauzadeana macula (Taylor) Coppins & Rambold, Biblioth. Lichenol. 34: 85

(1989) PLATE 1

THALLUS crustose, areolate, + 0.25 mm thick, without isidia or soredia;

AREOLES angular to irregular, 0.2-0.5 mm in diam., continuous, brown to

dark-brown, plane, smooth, with epinecral layer; PROTHALLUS usually present

at the thallus margin and between the areoles, black. APOTHECIA rounded,

scattered, immersed, 0.15-0.3 mm in diam.; pisc black, plane to slightly

convex, epruinose, margins not obvious; EPITHECIUM brown to bluish green,

with crystals, N+ purple red; HYMENIUM hyaline, I+ blue, 30-50 um high;

PARAPHYSES Often branched and anastomosing, 2 um wide below, 3-4 um wide

apically; HYPOTHECIUM colorless, 50-60 um thick. Asci clavate, 30-40 x 10-11

um, Lecanora-type, 8-spored; ascospores hyaline, simple, ellipsoid, 7-10 x

4.5-5.5 um. Pycnip1a: not observed.

CHEMISTRY — Containing an unknown substance.

SPECIMENS EXAMINED — CHINA. SICHUAN: Xiangcheng county, Rewu commune,

Rechong dairy farm, on siliceous rock, alt. 4400 m, 9.VIII.1981, J.K. Yang 2359 (KUN).

XINJIANG: Urumdi city, Tianshan mountains, Glacier No.1, on siliceous rock, alt.

3100 m, 26.VHI.2011, L.L. Zhang 20126151 (SDNU); alt. 3800 m, 27.VIII.2012, L. Li

20126471 (SDNU).

COMMENTS — Clauzadeana macula is characterized by its saxicolous habit,

small and immersed aspicilioid apothecia, colorless hypothecium, and

Clauzadeana, Lecidella, & Miriquidica spp. new for China... 709

eS i

SY 9g

E oom om} LG co

PLaTE 1. Clauzadeana macula (Li 20126471, SDNU). A. Thallus; B. Prothallus; C. Apothecium

section; D. Crystals of apothecium section visualized with polarized light; E. Paraphyses; F. Ascus

apex reaction; G. Ascospores. Scale bars: A, B = 1 mm; C, D = 50 um; E-G = 10 um.

low hymenium. Aspicilioid apothecia also occur in Amygdalaria Norman,

Aspicilia A. Massal., Immersaria Rambold & Pietschm., and Schaereria Korb.,

but Clauzadeana differs from all of these genera by its Lecanora-type ascus

structure.

DISTRIBUTION — Clauzadeana macula has been reported from Europe,

North America, Asia, and Australasia (Rambold 1989, Nash et al. 2004,

Galloway 2007, Smith et al. 2009). New to China.

710... Zhao & al.

Lecidella oceanica Lu L. Zhang & Xin Y. Wang, Bryologist 115: 330 (2012) PLATE 2

THALLUS crustose, rimose to areolate, thin, <0.15 mm thick, yellow-white,

grey-white to greenish grey, smooth to lightly rough, without soredia or isidia;

medulla I-; PROTHALLUs absent or present (if present black). APOTHECIA

brownish black to black, semi-immersed to sessile, 0.2-0.5 mm in diam.; Disc

flat, epruinose; margin distinct to reduced, sometimes lighter than disc; EXCIPLE

brown, without algal cells; PARATHECIUM light brown to hyaline, with a small

PLATE 2. Lecidella oceanica (Cheng 20120798, SDNU). A. Thallus; B. Apothecia; C. Apothecium

section; D. Crystals of apothecium section visualized with polarized light; E. Ascus; E Ascus

apex reaction; G. Paraphyses; H. Ascospores. Scale bars: A = 1 mm; B = 250 um; C, D = 50 um;

E-H = 10 um.

Clauzadeana, Lecidella, & Miriquidica spp. new for China... 711

amount of crystals; EPIHYMENIUM Olive, olive-brown to brown; HYMENIUM

50-70 um tall, with dispersed crystals; PARAPHYSES simple, rarely branched or

anastomosing, c. 2 um wide, slightly thickened apically; HyPOTHECIUM hyaline.

Asci clavate, Lecanora-type, 8-spored; ascospores ellipsoid to broadly

ellipsoid, 7.5-13 x 5-7um. Pycnip1iA not observed.

CHEMISTRY — Containing atranorin (+), 2,5,7-trichloro-3-O-

methylnorlichexanthone (capistratone), thiophanic acid, and isoarthothelin.

SPECIMENS EXAMINED — CHINA. LIAONING: Beipiao city, Mt. Dahei, alt. 1300 m, on

siliceous rock, 14.VIII.2011, Y.L. Cheng 20120942, 20120798, 20120933, 20120912B

(SDNU); Lvshunkou District, Dongjiacun, alt. 200 m, on siliceous rock, 27.VI.2012,

G.L. Zhou 20128775, 20128770 (SDNU).

Comments — Lecidella oceanica is morphologically close to L. enteroleucella

(Nyl.) Hertel in having small apothecia, colorless hypothecium, and similar

habitats. Lecidella enteroleucella is distinguished by its hymenium without

crystals and its secondary metabolites: atranorin, thuringione, and arthothelin.

Our specimens, which represent only the second report of L. oceanica, were

all collected in Liaoning province near the southwest coast of South Korea (the

original collection area). The South Korean specimens have a smoother and

lighter-colored thallus (usually yellowish white) and apothecia that are slightly

smaller (<0.3-0.4 mm) than our Chinese specimens. However, the microscopic

characters of the Korean and Chinese specimens are nearly the same, and the

secondary metabolites are the same.

DISTRIBUTION — Lecidella oceanica has previously been reported only from

South Korea (Zhang et al. 2012b). New to China.

Lecidella tumidula (A. Massal.) Knoph & Leuckert, Biblioth. Lichenol. 68: 131

(1997) PLATE 3

THALLUS crustose, continuous, rimose to granulose, thin, 0.6-1 mm thick;

PROTHALLUS absent; UPPER SURFACE yellowish white, greenish white to greenish

grey, dull, without soredia or isidia. APOTHECrIA sessile, slightly constricted at

base, (0.3—)0.5-0.9 mm in diam.; pisc black, flat to convex, epruinose; margin

distinct when young, then excluded; EXcCIPLE c. 65 um thick, green to black

green exteriorly, violet brown interiorly, without crystals; EPIHYMENIUM green

to blackish green; HYMENIUM hyaline, 50-90 um tall, with dispersed crystals;

PARAPHYSES simple, rarely branched or anastomosing, slightly thickened

apically; HYPOTHECIUM reddish brown. Asci clavate, Lecanora-type, 8-spored;

ASCOSPORES hyaline, simple, broadly ellipsoid to ovoid, 10-13 x 5.5-7 um.

PyYcNIDIA not observed.

CHEMISTRY — Containing lichexanthone and diploicin (+).

SPECIMENS EXAMINED — CHINA. XINJIANG: Mulei county, Dongdonggou, on bark,

alt. 1800 m, 2.VIII.2013, C. Li XL0047 (SDNU); Tianshan mountains, Tianchi, on bark,

alt. 2100 m, 23. VIII.2011, L.L. Zhang 20126562 (SDNU); alt. 1900 m, 23.VII.2011, L.L.

Zhang 20126663 (SDNU); alt. 1950 m, 23.VIII.2011, L.L. Zhang 20126512, 20126096

712... Zhao & al.

—,*

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E hel 10 um F 10 um 10 um H | 10 ym

PiatE 3. Lecidella tumidula (Li XL0047, SDNU). A. Thallus; B. Apothecia; C. Apothecium section;

D. Crystals of apothecium section visualized with polarized light; E. Ascus; F Ascus apex reaction;

G. Paraphyses; H. Ascospores. Scale bars: A = 500 um; B = 200 um; C, D = 50 um; E-H = 10 um.

(SDNU); Urumqi, Nanshan-Xiaoquzi, on bark, alt. 2700 m, 28.VIII.2011, L. Li

20125798 (SDNU); alt. 1900 m, 28.VIII.2011, L.L. Zhang 20126673 (SDNU); alt. 2200

m, 28.VIII.2011, L. Li 20126464, 20125877 (SDNU).

ComMENTs — It is difficult to distinguish Lecidella tumidula from

L. elaeochroma (Ach.) M. Choisy by morphology. However, L. elaeochroma has

larger (c. 1 mm) apothecia, a thicker hypothecium, larger spores (10-17 x 6-9

um), and a different chemistry, usually containing several xanthones but never

lichexanthone (Knoph & Leuckert 1997, Nash et al. 2004).

Clauzadeana, Lecidella, & Miriquidica spp. new for China... 713

DISTRIBUTION — Lecidella tumidula has been reported only from the

northern hemisphere in temperate areas of Asia, Europe, and North America

(Knoph & Leuckert 1997, Nash et al. 2004). New to China.

Miriquidica obnubila (Th. Fr. & Hellb.) Hertel & Rambold, Mitt. Bot. Staatssamml.

Munchen 23: 389 (1987) PLATE 4

THALLUs crustose, dispersed areolate, c. 0.45 mm thick; AREOLEs irregularly

shaped, more or less sublobate, grey, dark grey to rusty colored, plane to

E 4 nea F ' ’ . oun G ~ 40pm

Piate 4. Miriquidica obnubila (Zhang 20100347, SDNU). A. Thallus; B. Pycnidium section;

C. Apothecium section; D. Crystals of apothecium section visualized with polarized light; E. Ascus

apex reaction; F. Paraphyses; G. Ascospores. Scale bars: A = 1 mm; B = 20 um; C, D = 50 um;

E-G = 10 um.

714 ... Zhao & al.

irregularly convex, with rough surface, 0.6-1.25 mm diam.; epinecral layer

absent; medulla white, I-; ayPoTHALLUs black, obvious at the thallus margin and

between the areoles. APOTHECIA roundish, dispersed to crowed, subimmersed

to immersed, 0.4-2 mm diam; pisc plane to convex, black-brown to black,

epruinose; EXCIPLE distinct, ectal zone blackish brown, inner zone brown

to reddish brown; EPIHYMENIUM brown to olive-brown; HYMENIUM hyaline,

with crystals; PARAPHYSES rarely branched and anastomosing, 1.5-2 um wide,

c. 3 um wide apically; HypoTHECIUM dark reddish brown. Asc clavate,

Lecanora-type, 8-spored; ascospores hyaline, 8-11 x 4.5-7 um. PycNnIpDIA

immersed, + globular; conrp1a filiform, curved.

CHEMISTRY — Containing miriquidic acid.

SPECIMENS EXAMINED — CHINA. YUNNAN: Lijiang city, Mt. Laojun, on siliceous rock,

alt. 4000 m, 7.X1.2009, L.L. Zhang 20100347 (SDNU); Luquan county, Mt. Jiaozixue,

on siliceous rock, alt. 3800 m, 27.X.2008, H.Y. Wang 20082271 (SDNU); alt. 3700 m,

10.1V2010, L.S. Wang 10-31285 (KUN).

ComMENTs — Miriquidica obnubila and M. molybdochroa (Hertel) Hertel &

Rambold are not easily separated. Both species have grey thallus, dark brown

hypothecium, and miriquidic acid, but M. molybdochroa differs by its non-

sublobate areoles and wider spores (usually 6-8 um wide; Hertel & Rambold

1987)

DISTRIBUTION — Miriquidica obnubila has been reported from extra-

tropical Asia, Europe, and North America (Rambold et al. 2015, Andreev

2004). New to China.

Acknowledgments

The project was financially 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). We are grateful to Prof. Dr. Gerhard

Rambold (University of Bayreuth, Germany) and Dr. Alan Fryday (Herbarium, Dept

Plant Biology, Michigan State University, USA) for assistance in specimen identification.

We also thank Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Institute of

Microbiology, Chinese Academy of Sciences, Beijing, China) and Dr. Einar Timdal

(Natural History Museum, University of Oslo, Norway) for presubmission reviews.

Literature cited

Abbas A, Wu JN. 1998. Lichens of Xinjiang. Urumgi, Sci-Tech & Hygiene Publishing House of

Xinjiang (K).

Abbas A, Mijit H, Tumur A, Wu JN. 2001. A checklist of the lichens of Xinjiang, China. Harvard

Papers in Botany 5(2): 359-370.

Andreev MP. 2004. Notes on the lichen genus Miriquidica (Lecanorales, Lecanoraceae) in Russia.

Bibliotheca Lichenologica 88: 15-42.

Aptroot A, Seaward MRD. 1999. Annotated checklist of Hong Kong lichens. Tropical Bryology 17:

57-101.

Clauzadeana, Lecidella, & Miriquidica spp. new for China... 715

Aptroot A, Sparrius LB. 2003. New microlichens from Taiwan. Fungal Diversity 14: 1-50.

Galloway DJ. 2007. Flora of New Zealand Lichens. 2nd Edition. Lincoln, Manaaki Whenua Press.

Hertel H, Rambold G. 1987. Miriquidica genus novum Lecanoracearum (Ascomycetes

lichenisati). Mitteilungen der Botanischen Staatssammlung Miinchen 23: 377-392.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th edition.

Wallingford, CAB International.

Knoph JG, Leuckert C. 1997. Chemotypes and distribution patterns of the non-saxicolous species

of Lecidella (Lecanoraceae, Lecanorales). Bibliotheca Lichenologica 68: 129-134.

Lumbsch HT, Huhndorf SM. 2010. Myconet volume 14. Part one. Outline of Ascomycota — 2009.

Fieldiana, Life and Earth Sciences 1: 1-42. http://dx.doi.org/10.3158/1557.1

Nash TH, Ryan BD, Diederich P, Gries C, Bungartz F. 2004. Lichen flora of the Greater Sonoran

Desert Region, vol. 2. Tempe, Lichens Unlimited.

Obermayer W. 2004. Additions to the lichen flora of the Tibetan region. Bibliotheca Lichenologica

88: 479-526.

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens. 2nd

edition. London, British Lichen Society.

Rambold G. 1989. A monograph of the saxicolous lecideoid lichens of Australia (excl. Tasmania).

Bibliotheca Lichenologica 34: 1-345.

Rambold G, Davydov E, Elix JA, Nash III TH, Scheidegger C, Zedda L (eds). 2015. LIAS light - a

database for rapid identification of lichens. http://liaslight.lias.net/ (viewed online on 28 July

2015).

Roux C. 1983. Clauzadeana Roux, nova likengenro (Clauzadeana Roux, nouveau genre de lichen).

Bulletin de la Société Linnéenne de Provence 35: 99-102.

Smith, CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA. 2009. The

lichens of Great Britain and Ireland. London, British Lichen Society.

Wei JC. 1991. An enumeration of lichens in China. Beijing, International Academic Publishers.

Zhang LL, Wang LS, Wang HY, Zhao ZT. 2012a. Four new records of lecideoid lichens from China.

Mycotaxon 119: 445-451. http://dx.doi.org/10.5248/119.445

Zhang LL, Wang XY, Wang HY, Wang LS, Hur JS. 2012b. A new species and new records of

saxicolous species of the genus Lecidella (Lecanoraceae) from South Korea. Bryologist, 115:

329-332. http://dx.doi.org/10.1639/0007-2745-115.2.329

Zhao X, Zhang LL, Zhao ZT. 2013. A new species of Miriquidica from China. Mycotaxon 123:

363-367. http://dx.doi.org/10.5248/123.363

MY COTAXON

http://dx.doi.org/10.5248/130.717

Volume 130, pp. 717-720 July-September 2015

New records of smut fungi. 9.

A second locality of Anthracoidea andina

CVETOMIR M. DENCHEV’, HARRIE J.M. SIPMAN? & TEODOR T. DENCHEV'

‘Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences,

2 Gagarin St., 1113 Sofia, Bulgaria

*Botanic Garden & Botanical Museum Berlin-Dahlem, Free University of Berlin,

6-8 K6nigin-Luise-St., D-14195 Berlin, Germany

* CORRESPONDENCE TO: cmdenchev@yahoo.co.uk

AsstRrAct — Anthracoidea andina, known only from the type collection in the Argentine

part of Tierra del Fuego, is reported from a new locality in Chilean Tierra del Fuego.

Key worps — Anthracoideaceae, Chile, Cyperaceae, Schoenus andinus, taxonomy,

Ustilaginomycetes

Introduction

There are two species of Anthracoidea on Schoenus: A. schoenus (G. Cunn.)

Vanky on Schoenus antarcticus (Hook. f.) Dusén, S. maschalinus Roem. & Schult.,

and S. pauciflorus (Hook. f.) Hook. f., reported from South America (Chile)

and New Zealand (Vanky & Websdane 1995, Vanky & McKenzie 2002,

Vanky 2011), and Anthracoidea andina on Schoenus andinus (Phil.) H. Pfeiff.

(Roivainen 1977, Vanky 2011). Anthracoidea andina is known only from the

type collection (Roivainen 1977, Vanky 2011).

We report here a new record of Anthracoidea andina from Chilean Tierra

del Fuego. The collection on which this record is based was obtained during

a visit to the herbarium at the Botanic Garden and Botanical Museum Berlin-

Dahlem (B).

Material & methods

A dried specimen from the herbarium of the Botanic Garden and Botanical Museum

Berlin-Dahlem was examined under light microscope (LM) and scanning electron

microscope (SEM). For LM observations and measurements, spores were mounted in

lactoglycerol solution (w: la: gl = 1: 1:2) on glass slides, gently heated to boiling point

718 ... Denchevy, Sipman, & Denchev

to rehydrate the spores, and then cooled. The 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 sputter coated with gold. The surface

structure of spores was observed and photographed at 10 kV accelerating voltage using

a JEOL SM-6390 scanning electron microscope. The height of warts was measured in

SEM. The type of spore ornamentation is in accordance with Denchev et al. (2013). The

description given below is based entirely on the specimen examined.

Taxonomy

Anthracoidea andina (Kukkonen) Vanky, Mycotaxon 56: 219, 1995. FIGs 1-4

SoRI in ovaries, scattered in the inflorescence, as broadly ellipsoidal to

ovoid, black, hard bodies, up to 2.5 mm long, when young covered by a thin

membrane, later becoming exposed; spore mass of the mature sori powdery

on the surface. Sporss flattened, in plane view slightly irregular, suborbicular,

broadly elliptical or oval in outline, occasionally small protuberances present,

in plane view (15.5-)16.5-23(-24.5) x (14.5-)15.5-20.5(-21.5) (19.6 + 1.5 x

17.7 + 1.2) um (n = 100), in side view 9-13 um thick, dark reddish brown,

wall slightly uneven, 1.0-2.2(-2.8) um thick, light refractive areas and internal

swellings absent, in LM minutely verruculose, spore profile not affected or

nearly so. In SEM minutely verruculose, warts 0.1-0.3 um high, often fused in

small groups or short rows. SPORE GERMINATION unknown.

SPECIMEN EXAMINED — On Schoenus andinus (det. G. Kiikenthal): CHILE, PROVINCIA

DE TIERRA DEL FUEGO, Rio Fontaine, 1 March 1908, leg. C. Scottsberg, no. 195 (B 10

0506615).

CoMMENTS — Schoenus andinus is distributed in Chile and Argentina - in

central Chile and Patagonia, including Tierra del Fuego and Cape Horn

Archipelago. The distribution of Anthracoidea andina is restricted to Tierra

del Fuego. This species has been previously known only from the type locality

(Argentine Tierra del Fuego, south of Fagnano Lake, Lago Escondido, Cerro

Garibaldi, alt. 390 m, 3 Dec. 1969, H. Roivainen 721b). The present record

is based on an infected plant specimen from the Chilean Tierra del Fuego,

collected near Fontaine River (which flows into Almirantazgo Fjord), during

Scottsberg’s Expedition in Patagonia (cf. Scottsberg 1911: 44).

In both sites where Anthracoidea andina occurs, its host, Schoenus andinus,

grows in wetlands, mostly occupied by peat-forming plant communities,

which are threatened by human impact (peat mining, urban development,

road construction, recreational activities, and overgrazing) and the invasion

of the Canadian beaver. Anthracoidea andina is preliminarily assessed as a

Near Threatened species and submitted for inclusion in the IUCN Red List

(Denchev & Denchev 2015).

Anthracoidea andina in Chile ... 719

Fics 1-4. Anthracoidea andina on Schoenus andinus. 1. Habit. 2. Spores in LM. 3, 4. Spores in SEM.

Scale bars; 1 = 1 mm; 2 = 10 pm; 3, 4= 5 pm.

720 ... Denchev, Sipman, & Denchev

The other Anthracoidea species on Schoenus, A. schoenus, differs from

A. andina by a thicker wall (1-4 um) with light refractive areas and internal

swellings.

Acknowledgements

This research received support from the SYNTHESYS Project http://www.synthesys.

info/ and financed by European Community Research Infrastructure Action under

the FP7 “Capacities” Program. The authors also gratefully acknowledge Dr Kalman

Vanky (Herbarium Ustilaginales Vanky, Tiibingen, Germany) and Dr Roger G. Shivas

(Department of Agriculture and Fisheries, Queensland, Australia) for critically reading

the manuscript and serving as pre-submission reviewers.

Literature cited

Denchev CM, Denchev TT. 2015. Anthracoidea andina (Kukkonen) Vanky. In: The Global Fungal

Red List Initiative. http://iucn.ekoo.se/iucn/species_view/412475/

Denchev TT, Denchev CM, Michikawa M, Kakishima M. 2013. The genus Anthracoidea

(Anthracoideaceae) in Japan and some adjacent regions. Mycobiota 2: 1-125.

http://dx.doi.org/10.12664/mycobiota.2013.02.01

Roivainen H. 1977. Resultados micoldgicos de la expedicion a Argentina y Chile en 1969-1970.

Karstenia 17: 1-18.

Scottsberg C. 1911. The wilds of Patagonia. A narrative of the Swedish Expedition to Patagonia,

Tierra del Fuego, and the Falkland Islands in 1907-1909. The Macmillan Company, New York

& Edward Arnold, London.

Vanky K. 2011 [“2012”]. Smut fungi of the world. APS Press, St. Paul, Minnesota, USA.

Vanky K, McKenzie EHC. 2002. Smut fungi of New Zealand. Fungi of New Zealand. Vol. 2.

Fungal Diversity Press, Hong Kong.

Vanky K, Websdane K. 1995. Ustilaginales of Schoenus (Cyperaceae). Mycotaxon 56: 217-229.

MY COTAXON

http://dx.doi.org/10.5248/130.721

Volume 130, pp. 721-729 July-September 2015

A new species and a new record of Anthostomella

on Alnus leaf-litter from Argentina

ESTEBAN B. Str*?’, ANDREA I. ROMERO?, & ADRIANA I. HLADKI’

' Laboratorio Criptogdmico, Fundacion Miguel Lillo,

Miguel Lillo 251, San Miguel de Tucumdan (CP 4000), Tucumdn, Argentina

3 Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET)

* PRHIDEB-CONICET, Dep. de Biodiversidad y Biologia Experimental,

Facultad de Ciencias Exactas y Naturales (UBA),

Ciudad Universitaria, Pabellon II, 4to. Piso, CP1428EHA Buenos Aires, Argentina

* CORRESPONDENCE TO: sirestebanbenjamin@gmail.com

AsstrRact — A study on saprobic fungi occurring on litter of Alnus acuminata from

northwestern Argentina yielded an unknown Anthostomella species (described here as

Anthostomella saltensis) and the first record of A. scotina for Argentina. A table is provided

comparing A. saltensis with similar species.

Key worps — Ascomycota, Betulaceae, Xylariaceae, Yungas

Introduction

There have been few investigations on the genus Anthostomella in Argentina.

The main publications are those by Spegazzini (1880, 1881, 1887, 1899, 1910,

1915) recording eight species, of which only four are currently accepted:

A. achira (Speg.) Speg., A fuegiana Speg., A. sphaeroidea Speg., and A. sepelibilis

(Berk. & M.A. Curtis) Sacc. Lu & Hyde (2000) synonymized or excluded the

remaining species, treating A. nummularioides Speg. and A. phoenicicola Speg.

as synonyms of A. tenacis (Cooke) Sacc. and A. platensis Speg. as a synonym

of A. tumulosa (Roberge ex Desm.) Sacc. and excluding A. lonchosperma

Speg. from the genus. Additionally, they synonymised Phaeophomatospora

argentinensis Speg. with Anthostomella limitata Sacc., Anthostoma yatay

Speg. with Anthostomella dilatata (Berk. & Broome) Petch, and Entosordaria

fuegiana Speg. (nom. illeg.) with Anthostomella phaeosticta (Berk.) Sacc.

Lu & Hyde (2000) also cited a collection from Argentina on a Bromelia sp.

722 ... Sir, Romero, & Hladki

when they proposed the new species Anthostomella rhaphidophylli B.S. Lu &

K.D. Hyde. An erroneous record of A. tomicoides Sacc. from Argentina (Lu &

Hyde 2000: 188, as A. italica Sacc. & Speg.) is based on a Spegazzini collection

from Bellano, Italy.

Recently, Capdet (2012) has reported Anthostomella nitidissima (Durieu &

Mont.) Sacc., A. palmaria B.S. Lu & K.D. Hyde, A. puiggarii Speg., and A. spiralis

K.D. Hyde & B.S. Lu on fallen woody parts of native palms from Argentina.

Overall, fourteen species growing on herbaceous plants (Cannaceae,

Juncaceae, Agavaceae, Apiaceae, Smilacaceae, Asparagaceae) and woody parts

of palms (Arecaceae) (Lu & Hyde 2000, Capdet 2012) have been reported so far

from Argentina.

Alnus acuminata Kunth (“aliso del cerro”; Betulaceae) is a tree species present

from northern Mexico to northwest Argentina (www.theplantlist.org). It is the

dominant plant of the Argentina Yungas at the highest altitudes between 700

and 1500 m.s.l. (Zuloaga & Morrone 1999). Little is known about Xylariaceae

on A. acuminata. Sir et al. (2012) recently reported Hypoxylon rubiginosum

var. microsporum, Xylaria luxurians, and X. mellissii from Tucuman province

(Argentina), but no Anthostomella species have been recorded on this host.

The present work deals with Anthostomella species collected in leaf-litter

of Alnus acuminata during a survey of the Xylariaceae within the Argentine

Yungas. We propose a new species and record A. scotina for the first time for

the country.

Materials & methods

The sampling area (23-29°S 64-68°W) is part of the “Yunga’ phytogeographic region

(Cabrera 1971) in northwest Argentina, involving the Salta, Jujuy, Catamarca, and

Tucuman provinces (Sir et al. 2012: fig. 1). Microscopic preparations and observations,

terminology, and the criteria used for descriptions follow Lu & Hyde (2000) and Francis

C1875):

Taxonomy

Anthostomella saltensis Sir, Hladki & A.I. Romero, sp. nov. PLATE 1

INDEXFUNGORUM IF550513

Differs from Anthostomella clypeata by its slightly smaller, inequilaterally ellipsoidal

ascospores with a germ slit extending the full length of the spore.

Type: Argentina, Salta, Department Santa Victoria, National Reserve El Nogalar de los

Toldos, 27-XH-2011, Sir & Hladki 118 (Holotype, LIL).

ETyMoLoGey: “saltensis” refers to the name of the Salta province.

AscoMarta peritheciate, immersed, clustered or solitary, visible as black and

conical areas, dome-shaped in vertical sections (183—)186-250(-274) x (188-)

Anthostomella saltensis sp. nov. (Argentina) ... 723

Plate 1 (Fics 1-8). Anthostomella saltensis (Holotype). 1: Clypeus on surface of Alnus acuminata

leaf; 2: Longitudinal section of immersed perithecium; 3: Peridium (arrow); 4: Asci; 5: Ascal apical

apparatus; 6, 8: Ascospores; 7: Ascospores showing germ slit. Bars: 1 = 200 um; 2, 3 = 100 um;

4-8 = 10 um.

193-250(-273) um (x= 225 x 222.8 um,n=11).CLypeEus black, (160-)162.5-250

(-314) um diam. (x = 208.6 um, n = 11); ostiolar papilla (30-)37-60(-61) um

diam. (x = 47.7 um, n = 11), periphysate ostiolar canal 27 um broad. PERIDIUM

7 2A ... Sir, Romero, & Hladki

10-16.5 um thick (x = 14 um). Asci cylindrical, unitunicate, 8-spored, short-

pedicellate (64—)68-97(-98) x (5-)5.5-7.5(-8) um (x = 83.2 x 6.1 um, n = 50),

apically rounded with IK+ apical ring inverted hat-shaped, 2-3 x 1-2 um (x = 2.1 x

1.7 um, n = 50). PARAPHYSES septate, hyaline, 2-5(-5.5) um diam. (x = 3.5 um,

n=50). ASCOSPORES (9-)9.5-13(-14) x 4-5.5(-6) um (x= 11.2 x 4.9 um,n=100),

two-celled with a hyaline globose basal cell, 1-2 x 1.5-2(-2.5) um (x = 1.4 x

2 um), and a larger brown cell, 8-11(-12) x 4-5.5(-6) um (x = 9.5 x 4.8 um),

inequilaterally ellipsoidal, usually with narrowly rounded ends, with one end

narrowly rounded and the other end truncate; germ slit straight extending over

the full length, ventral. Mucilaginous sheath lacking.

Host — leaf-litter of Alnus acuminata.

DISTRIBUTION — Argentina (Salta province).

ADDITIONAL MATERIAL EXAMINED — ARGENTINA, Satta, Department Santa

Victoria National Reserve El Nogalar de los Toldos: near the river, 26-VI-2013, Sir &

Hladki 499 (LIL); road to the National Park Baritt, 28-XII-2011, Sir & Hladki 119 (LIL);

27-VI-2013 Sir & Hladki 500 (LIL). CHILE, Los Rios, Valdivia, on leaves and stems

of Uncinia erinacea, Jan. 1909, C. Spegazzini (LPS 6537, holotype of Paranthostomella

unciniicola).

Notes — TABLE 1 summarizes the morphological differences between

Anthostomella saltensis and the three most similar Anthostomella species.

Anthostomella saltensis is close to Anthostomella clypeata (De Not.) Sacc.

as described by Lu & Hyde (2000), but our examination of the holotype of

Paranthostomella unciniicola Speg. (considered a synonym of A. clypeata by Lu

& Hyde 2000) revealed differences between the Argentine collection and the

accepted concept of A. clypeata. Anthostomella clypeata differs from A. saltensis

by its ascospores being equilaterally ellipsoidal, without a germ slit, and slightly

larger (x = 13.3 x 5.3 um; Lu & Hyde 2000). Another allied species, A. clypeoides

Rehm, has ascospores of a similar size and shape, but they lack a germ slit, have

brown cells with one conic end, and are surrounded by a thin mucilaginous

sheath (Lu & Hyde 2000). Anthostomella sabinianae S.M. Francis is also similar

but has larger ascospores with one end widely rounded.

As Anthostomella spores do not easily germinate, cultures are not available

and few sequences are registered at GenBank (http://www.ncbi.nlm.nih.gov/),

making molecular comparisons impractical. There is no phylogenetic

study on the genus available at present. Most new taxa have been based on

morphological characters (Lu & Hyde 2000; Lee & Crous 2003). There are a few

general studies on some Anthostomella sequences such as Zhang et al (2006)

and Pelaez et al. (2008), and sequences have been generated for five species by

Lu & Hyde (2000) and for two other species by Crous et al. (2006) and Crous

& Groenewald (2010).

Anthostomella saltensis sp. nov. (Argentina) ... 725

TABLE 1. Comparison of ascospores and the ascal apical ring in Anthostomella saltensis

and similar species.

ASCOSPORES

(all two-celled)

SPECIES

APICAL

HYALINE RING SOURCE

TOTAL CELL

LENGTH

(um) SIZE

(um)

+ ellipsoid,

one end Francis

broadly 1975

A. rounded

sabiniana iad

Feaieat: Lu & Hyde

2000

base

(wm)

Lu & Hyde

age 2000;

A. clypeata 12-15 ellipsoid Fiandis

1975

oval-

ellipsoid, L Francis,

one end 1975

A. conic

clypeoides <s

+ ellipsoid, Lu & Hyde

one side 2000

flattened

9-14 + ellipsoid,

A. saltensis one end

narrowly

rounded

*G = germ slit; M = mucilaginous sheath; +/- = present/absent.

Anthostomella scotina (Durieu & Mont.) Sacc., Syll. Fung. 1: 288. 1882. PLATE 2

ASCOMATA immersed, clustered or solitary, visible as black and conical

areas, dome-shaped, in adaxial and abaxial leaf, globose or subglobose in

vertical sections, (220-)235-300(-310) x (180-)200-295(-300) um (x = 258.1 x

248.3 um, n= 13). Ctypeus black (100-)200-255(-280) um diam. (x = 225.1 um,

n = 13); conical ostiolar papilla 50-98 um diam. (x = 63.7, n = 13), periphysate

ostiolar canal 14-37.5 um diam. (x = 25.7 um). PERrDIUM (8-)9-16(-17) um

diam. (x = 13.1 um). ASCI cylindrical, unitunicate, short-pedicellate, 8-spored,

(85-)87.5-117.5 x 4.5-7 um (x = 101.7 x 5.4 um, n = 50), apically rounded, with

a J+ cylindrical-shaped apical ring (1.5-)2-3-x 1-2 um (x = 2.4 x 1.3 um,

n = 50). PARAPHYSES septate, hyaline (3-)3.5-6(-7) um diam. (x = 4.7 um, n = 50).

ASCOSPORES (20—)21-29(-30) um total length (x = 25.7 um, n = 100), fusiform,

three cells, with inequilaterally ellipsoidal or cylindrical brown central cell,

726 ... Sir, Romero, & Hladki

Plate 2 (Fics 9-14). Anthostomella scotina (Sir & Hladki 120). 9: Clypei on surface of Alnus

acuminata leaf; 10: Longitudinal section of immersed perithecium; 11: Peridium (arrow); 12: Ascal

apical apparatus; 13: Ascospores; 14: Ascospores showing germ slit (arrows). Bars: 9 = 450 um;

10, 11 = 100 um; 12-14: 10 um.

with one broadly rounded basal end and one narrowly rounded distal end

(9-)9.5-14(-15) x (3-)3.5-4.5(-5) um (x = 11.3 x 4.1 um), straight and faint

germ slit. Hyaline globose basal cell on the widest part of the brown cell

(6-)6.5-8(-9) x 2-3 um (x = 7.4 x 2.5 um) and hyaline curved distal cell

(5-)6-9(-10) x 1-2.5 (x = 7.1 x 1.9 um), both becoming narrower towards the

end, lacking mucilaginous sheath.

MATERIAL EXAMINED — ARGENTINA: CatTamMarca: Department Aconquija,

near road, 13-V-2013, leg. Sir & Hladki 405-406 (LIL). Juyuy: Department Ledesma,

entering the National Park Calilegua, 12-V-2012, leg. Sir & Hladki 120 (LIL). TUCUMAN:

Department Yerba Buena, Cerro San Javier, Memorial Park, Aliso forests, 27-X-2011,

leg. Sir & Hladki 366 (LIL); Villa Nougues, near road, 25-XI-2011, leg. Sir & Hladki 362

(LIL); Department Monteros, Las Azucenas, 27-X-2011, leg. Sir & Hladki 364 (LIL); La

Heladera, 27-X-2011, leg Sir & Hladki 365 (LIL); Department Chicligasta, Provincial

Park El Cochuna, near road, 26-XI-2011, leg. Sir & Hladki 363 (LIL); La Banderita, Aliso

Anthostomella saltensis sp. nov. (Argentina) ... 727

forests, 13-V-2013, leg. Sir & Hladki 404 (LIL); Las Lenguas, Aliso forests, 13-V-2013,

leg. Sir & Hladki 403 (LIL); Department Tafi del Valle, near the los Sosa river, 7-VI-2012,

leg. Sir & Hladki 361 (LIL). CHILE: Bio Bio, Concepcion, Caracol, on Persea lingue,

Jan. 1909, C. Spegazzini (LPS 7098, holotype of Entosordaria perseicola).

Hosts — Alnus (Betulaceae), Betula (Betulaceae), Cladium (Cyperaceae),

Lithocarpus (Fagaceae); Persea (Lauraceae).

DISTRIBUTION — Algeria, Argentina (Catamarca, Jujuy, and Tucuman

provinces), Belgium, Chile, UK, USA (California).

Notes — There are some discrepancies between our ascospores and those

described by Francis (1975) and Lu & Hyde (2000). We follow Lu & Hyde (2000)

in interpreting the ascospores as 3-celled, while Francis (1975) interpreted the

distal hyaline cell as a gelatinous appendage, consequently citing a much smaller

“total” spore length; the total ascospore length measured by Lu & Hyde (2000)

is similar to ours. The length of the brown cell measured by Francis (1975) is

similar to ours, but Lu & Hyde (2000) cite a shorter length. Lu & Hyde (2000)

described a very thin gelatinous sheath and absence of a germ slit; whereas both

Francis (1975) and we observed a faint germ slit but no mucilaginous sheath.

Provisional key to species of Anthostomella from northwestern Argentina

1. Ascospores with a single hyaline basal dwarf cell .................... A. saltensis

Pe ASCOSPOLES WILY TWO. hiyaliiic-Celis. Satur. ta tur, cag rke eter Melylee Mea teeters ge, «Seeder Ree 2

2. Brown cell 9-15 x 3-5 um, inequilaterally ellipsoidal,

hyaline cells 5-10 x 1-3 um, apical ring cylindrical shaped........... A. scotina

2. Brown cell 12-16.5 x 5-7 um, inequilaterally ellipsoidal,

dwarf cells 1.5-2.5 x 2-3 um apical ring wedge shaped ............ A. sepelibilis

Discussion

Eleven xylariaceous genera have been reported in the protected areas of

the Argentine Yungas (Sir et al. 2012) but the only report of Anthostomella

species from this region is A. sepelibilis, cited from Tucuman (Lu & Hyde 2000).

Our survey in the Argentine Yungas yielded two additional Anthostomella

species. The characters of one did not match those of any previously described

Anthostomella. Its morphology differs enough that the specimens can be

described as a new species, A. saltensis. Future molecular research will test

our hypothesis and contribute more information about the affinities among

Anthostomella species. The other species found, A. scotina, is incorporated into

the mycobiota of Argentina.

Anthostomella species are found on the stems and leaves of 82 plant families

(Lu & Hyde 2000), but only A. scotina was cited on Betulaceae (Betula alba L.,

B. pendula Roth). Our study reports for the first time an Anthostomella species

on another genus of this family, Alnus.

728 ... Sir, Romero, & Hladki

Alnus acuminata is a tree of great ecological importance for Argentina's

Yungas. It is a pioneer species of forest succession, appearing mainly after

landslides or large fires (Arturi et al. 1998) and is associated with an unusual

and diverse mycobiota. Walker et al. (2012) reported a new member of

Gnomoniaceae on Alnus acuminata leaves from Tucuman (Argentina) and our

research added two species of a xylariaceous genus, one new to science and one

new to the country.

Acknowledgements

The authors express their appreciation to the authorities of Fundation Miguel Lillo,

Tucuman, for financial support. Dr. Andrea I. Romero thanks the Consejo Nacional de

Investigaciones Cientificas y Técnicas (CONICET-Argentina), PROPLAME-PRHIDEB

(publication N° 201). The authors also thank the pre-submission reviewers Dr. Liliane

Petrini and Dr. Yu-Ming Ju for valuable comments and suggestions.

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the family Xylariaceae based on ribosomal DNA sequences. Fungal Diversity 31: 111-134.

Sir EB, Parrado MF, Romero AI, Hladki AI. 2012. Biodiversity of Xylariaceae (Ascomycota) and

their hosts in protected areas from Tucuman (Argentina). Kurtziana 37(2): 35-48.

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Argentina. 12: 97-117, 174-189.

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11: 135-308.

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Ophiognomonia (Gnomoniaceea, Diaphortales); including twenty-five new species in this highly

diverse genus. Fungal Diversity 57: 85-147. http://dx.doi.org/10.1007/s13225-012-0200-y

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MY COTAXON

http://dx.doi.org/10.5248/130.731

Volume 130, pp. 731-738 July-September 2015

Phoma canadensis Allesch.:

a synonym of Pyrenochaeta cava

LIDIANE LEAL DUARTE & ROBERT WEINGART BARRETO*

Universidade Federal de Vicosa, Departamento de Fitopatologia,

Vicosa, Minas Gerais, 36570-000, Brazil.

*CORRESPONDENCE TO: rbarreto@ufv.br

ABSTRACT — The isotype of Phoma canadensis on Conyza canadensis was investigated to

clarify its dubious taxonomic placement and taking the broad rearrangement of Phoma taxa

into consideration. Phylogenetic and morphological analyses show that this species is, in fact,

misplaced in Phoma and actually belongs in Pyrenochaeta as a synonym of Pyr. cava.

KEY worps — asexual morph, coelomycete, Cucurbitariaceae, molecular phylogeny,

reappraisal

Introduction

Phoma isa large genus of filamentous fungi, widely distributed and occupying

diverse ecological niches (Boerema et al. 2004, Aveskamp et al. 2008). Due to its

problematic and confusing taxonomy it has been the subject of intense studies

aimed at generating a more natural delimitation for the taxa within this genus

and to uncover its phylogenetic relationship with other allied genera (Boerema

et al. 2004; Aveskamp et al. 2010; Gruyter et al. 2009, 2010, 2013). Even after

this recent and significant mycological effort, an estimated more than half

of the herbarium Phoma specimens worldwide still need reexamination and

proper classification (Aveskamp et al. 2008).

One example is Phoma canadensis Allesch., an obscure fungus known only

from its original and brief description on dead stems of Conyza canadensis (L.)

Cronquist [= Erigeron canadensis L.; Asteraceae) in Germany and subsequently

recorded only once from Los Angeles (USA) (Allescher 1896, Farr et al. 2015).

Considering that the limited information in the original description makes its

position in Phoma questionable, this paper attempts, through morphological

and phylogenetic analyses of the isotype specimen, to clarify the placement of

Ph. canadensis.

732 ... Duarte & Barreto

TABLE 1. Species and isolates included in the phylogenetic analyses of

Phoma canadensis and Pyrenochaeta cava

* new sequence from Phoma canadensis isotype

SPECIES STRAIN Host ORIGIN GENBANK #

Chaetodiplodia sp. CBS 568.88 rock Israel EU754142

Chaetopyrena penicillata CBS 199.89 Medicago sativa South Africa = GQ387573

CBS 498.72 soil Turkey GQ387574

Chaetosphaeronema coonsii CBS 141.84 Malus sylvestris Japan GQ387575

Chaetosphaeronema hispidulum CBS 216.75 Anthyllis vulneraria Germany EU754144

Coniothyrium carteri CBS 101633 Quercus sp. Netherlands GQ387593

CBS 105.91 Quercus robur Germany GQ387594

Coniothyrium dolichi CBS124140 Dolichos biflorus India GQ387611

CBS124143 Dolichos biflorus India GQ387610

Coniothyrium glycines CBS 124141 Glycine max Zimbabwe GQ387598

CBS 124455 Glycine max Zambia GQ387597

Coniothyrium telephii CBS 101636 — Glycine max Zimbabwe GQ387601

CBS 188.71 air Finland GQ387599

CBS 856.97 mineral wool Finland GQ387600

Herpotrichia parasitica CBS 218.77 Abies alba Germany GQ387618

CBS 451.73 Abies alba Germany GQ387617

Neosetophoma samararum CBS 138.96, Phlox paniculata Netherlands =GQ387578

CBS 139.96 Poaceae Netherlands GQ387579

CBS 568.94 Urtica dioica Netherlands GQ387580

Paraconiothyrium minitans CBS 122788 —— U. Kingdom = EU754173

Paraphoma chrysanthemicola CBS 172.70 Chrysanthemum morifolium Germany GQ387583

CBS 522.66 Chrysanthemum morifolium U.Kingdom GQ387582

Paraphoma radicina CBS 102875 — Solanum esculentum Germany EU754190

CBS 111.79 Malus sylvestris Netherlands = EU754191

Phoma betae CBS 109410 _—_— Beta vulgaris Netherlands = EU754178

Phoma gardeniae CBS 302.79 airborne Netherlands GQ387596

CBS 626.68 Gardenia jasminoides India GQ387595

Phoma herbarum CBS 615.75 Rosa multiflora Netherlands = _EU754186

Pleurophoma pleurospora CBS116668 Sarothamnus scoparius Netherlands = JF740326

CBS130329 Lonicera sp. Netherlands JF740327

Pyrenochaeta cava BPI 356246* Conyza canadensis Germany KC841911

CBS257.68 wheat-field soil Germany EU754199

CBS 115953 Quercus cerris Italy GQ387607

Pyrenochaeta corni CBS 102828 Olea europaea Italy GQ387609

CBS 234.92 Olea europaea Italy EU754176

CBS 248.79 Fraxinus excelsior Netherlands GQ387608

Pyrenochaeta nobilis CBS 292.74 Ilex aquifolium Netherlands = GQ387615

CBS 407.76 Laurus nobilis Italy EU754206

CBS 566.75 Buxus sempervirens Netherlands GQ387616

Pyrenochaeta quercina CBS 115095 Quercus robur Italy GQ387619

CBS 297.74 sea water Montenegro GQ387620

Pyrenochaeta unguis-hominis CBS 378.92 fingernail U. Kingdom GQ387621

CBS 112.79 airborne U. Kingdom GQ387622

CBS 111112 = Agapornis sp. Netherlands GQ387623

Pyrenochaetopsis decipiens CBS 165.89 Hordeum vulgare Netherlands GQ387625

Pyrenochaetopsis indica CBS 244.54 Saccharum officinarum India GQ387626

Pyrenochaetopsis leptospora CBS 536.66 wheat field soil Germany GQ387628

Pyrenochaetopsis microspora CBS 102876 water Montenegro GQ387631

Setophoma sacchari CBS 333.39 Saccharum officinarum Brazil GQ387586

Setophoma terrestris CBS 335.29 Allium sativum USA GQ387587

CBS 335.87 Allium cepa Senegal GQ387528

CBS 377.52 Allium cepa —— GQ387588

Phoma canadensis: a synonym of Pyrenochaeta cava ... 733

Material & methods

The type material (isotype) from the packet labeled BPI 356246, originally collected

in Germany, as well as two other samples (BPI 356244, BPI 356245; PLATE 1) loaned

by the U.S National Fungus Collections were carefully examined under a dissecting

microscope; slides containing fungal structures were prepared with the help of a

freezing microtome (Microm HM 520) and mounted in lactophenol or lactofuchsin.

Fungal structures were observed, measured, and illustrated with an Olympus BX 51

light microscope fitted with an Olympus E330 camera and a drawing tube.

After acquiring permissing to extract DNA directly from the stem samples,

total genomic DNA was isolated using the Wizard® Genomic DNA Purification Kit

(Promega Corporation, WI, USA) according to the manufacturer's instructions. The

primers LROR and LR5 (Vilgalys & Hester 1990) were used to amplify a part of the

large ribosomal subunit (LSU). The 25 uL PCR reaction and PCR conditions were

the same as described by Fernandes et al. (2013). The nucleotide sequence data were

obtained by DNA sequencing (Macrogen Inc., Korea) employing the same primers used

for PCR amplification. The sequence fragments were assembled from the forward and

reverse sequences with the help of Staden program 2003.0.1 (Staden et al. 2000). The

generated sequence was aligned with closely related and outgroup published sequences

retrieved from GenBank with the MEGA 5.0 software package and manually adjusted

for improvement by eye where necessary (TABLE 1). Bayesian inference analyses were

conducted and the best-fit evolutionary model was determined by comparing different

evolutionary models via the Akaike information criterion using PAUP (version 4.0b10,

Sinauer Associates, Sunderland, MA) and MrModeltest 2.2 (Nylander 2004). Five

analyses of two Markov Chain Monte Carlo (MCMC) chains were run from random

trees for 1,000,000 generations and sampled every 1000 generations using MrBayes v3.1

(Huelsenbeck & Ronquist 2001), which also calculated the consensus trees after the

program Tracer v1.5 (Rambaut & Drummond 2003) was used to ensure the convergence

of the chains. The first 25% of sampled trees were discarded as “burnin”. Phylogenetic

trees were visualized with the program FigTree v1.3.1 (Rambaut 2009).

Results

Phylogenetic analysis

An amplicon of 1294 bases of the LSU region was generated (KC841911);

a standard nucleotide BLAST search against the nucleotide collection on

GenBank showed 100% identity between KC841911 and the LSU sequence

of Pyrenochaeta cava CBS 257.68 (EU754199). For the phylogenetic analysis,

closely related LSU sequences published by Gruyter et al. (2010) were retrieved

from GenBank (TABLE 1). The resulting alignment data matrix included 52 taxa

(including the outgroup sequences) and 1301 characters, of which 158 sites

were variable and 125 sites were parsimony informative. The Ph. canadensis

isolate fell within the Cucurbitariaceae, where it grouped with the Pyr. cava

isolates with maximum support (PLATE 2).

734 ... Duarte & Barreto

Taxonomy

Pyrenochaeta cava (Schulzer) Gruyter, Aveskamp & Verkley,

Mycologia 102: 1076, 2010. PLATE 1

= Phoma cava Schulzer, Verh. Zool.-Bot. Ges. Wien 21: 1248, 1871.

= Aposphaeria cava (Schulzer) Sacc. & Schulzer, Syll. Fung. 3: 174, 1884.

= Coniothyrium cavum (Schulzer) Kuntze, Revis. Gen. Pl. 3(2): 459, 1898.

= Pleurophoma cava (Schulzer) Boerema, Loer. & Hamers, Persoonia 16: 172, 1996.

= Phoma canadensis Allesch., Ber. Bayer. Bot. Ges. 4: 32, 1896.

ORIGINAL DESCRIPTION OF PHOMA CANADENSIS (Allescher 1896): “Peritheciis

minutis, tectis, dense gregariis, in maculas griseas, longas, saepe longissimas

dispositis, poro pertusis, atro-fuscis, ca. 40-60u diam.; sporulis minutissimis,

oblongis, utrinque obtusis, biguttulatis, continuis, hyalinis, ca. 3-5 x 0,5-1;

basidiis non visis.

Hab. in caulibus emortuis Erigerontis canadensis. Miinchen: in silva prope

«Ludwigshohe». IV. 95 ipse legi.”

External mycelium absent. Internal mycelium indistinct. Pycnidia

subcuticular, crowded, forming vertical linear strands over the grayish

loosening cuticle of the host tissue, subglobose, glabrous, 40-84 x 69.5-158.5

um, walls composed of pale to chestnut brown textura angularis, 1-2 layers,

9-10 um thick. One central ostiole per ascoma, circular, 9.5-12 um diam.

Conidiogenesis indistinct. Conidia ellipsoid to subcylindrical, straight to

slightly curved, 2-5 x 1 um, aseptate, ends rounded, with 2 small polar guttules,

hyaline to subhyaline, smooth.

SPECIMENS EXAMINED: GERMANY, Bavaria, Munich, near Ludwigshoehe, on dead

stems of Erigeron canadensis [= Conyza canadensis], April 1895, Andreas Allescher

(BPI 356246, isotype; GenBank KC841911). U.S.A., CALIFORNIA, Los Angeles Co.,

Glendale, on stems of Erigeron canadensis, November 20 1931, O.A. Plunkett (BPI

356244, BPI 356245).

Discussion

In addition to Ph. canadensis Allesch. 1896, there is an illegitimate later

homonym, Ph. canadensis Voglino 1910, described from Populus canadensis

in Italy (Saccardo & Trotter 1913). Regrettably, we were not able to obtain

herbarium specimens of Ph. canadensis and have therefore been unable to

assess its relationship (if any) with Ph. canadensis Allesch. Consequently, all

data presented and discussed in this paper pertains only to the fungus described

by Allescher.

The fungal morphometric data in BPI 356244 and BPI 356245 were

consistent with that of the isotype BPI 356246, indicating that the specimens

represent the same species. The isotype specimen, as well as the extra material,

consists of stem fragments fully colonized by a pycnidial-producing fungus,

Phoma canadensis: a synonym of Pyrenochaeta cava ... 735

us0356245

SOUTHERN CALIFORNIA FUNGI

PLaTE 1. Phoma canadensis Allesch. on Conyza canadensis. A-C. Herbarium packets containing:

(A) BPI 356244; (B) BPI 356245; (C) BPI 356246 (isotype); D-F Densely crowded bands

of pycnidia of Ph. canadensis as seen on: (D) BPI 356244; (E) BPI 356245; (F) BPI 356246;

G-L. Pycnidia. M. Hyaline conidia. Scale bars: G-L = 20 um; M = 2 um.

showing numerous dark colored pycnidia distributed along a horizontal

band and with aseptate and hyaline spores (PLATE 1). In 1896, Andreas

Allescher placed his newly found fungus in the genus Phoma as a new species,

Ph. canadensis. This was in agreement with the accepted 19th century

736 ... Duarte & Barreto

1.00 > Chaetopyrena penicillata CBS 199.89

Chaetopyrena penicillata CBS 498.72

Phoma herbarum CBS 615.75 Didymel laceae

Phoma gardeniae CBS 302.79

Phoma gardeniae CBS 626.68

1.00

Chaetosphaeronema coonsii CBS 141.84

Chaetosphaeronema hispidulum CBS 216.75

Setophoma sacchari CBS 333.39

Setophoma terrestris CBS 335.29

100 Setophoma terrestris CBS 335.87

Setophoma terrestris CBS 377.52

Neosetophoma samararum CBS 138.96

Neosetophoma samararum CBS 568.94

Phaeosphaeriaceae

0.72 Neosetophoma samararum CBS 139.96

0.507 Paraphoma chrysanthemicola CBS 172.70

Paraphoma chrysanthemicola CBS 522.66

Paraphoma radicina CBS 102875

0.814 Paraphoma radicina CBS 111.79

1.00 -Herpotrichia parasitica CBS 218.77 * .

sy Lierporihia parasitica CBS 451 oe | Cucur b ltarlaceae

Phoma betae CBS 109410 ff Pleosporaceae

Coniothyrium telephii CBS 101636

ot} Coniothyrium telephii CBS 188.71

Coniothyrium telephii CBS 856.97

0.72 0.88 Chaetodiplodia sp. CBS 568.88

Coniothyrium carteri CBS 101633 * .

Phat; ‘oniothyrium carteri CBS 105.91 Coniothyriaceae

1.00 - Coniothyrium glycines CBS 124141

Coniothyrium glycines CBS 124455

Coniothyrium dolichi CBS 124140

4 0.80 Coniothyrium dolichi CBS 124143

1.00 - Pyrenochaetopsis decipiens CBS 165.89

Pyrenochaetopsis indica CBS 24454

Pyrenochaetopsis leptospora CBS 536.66

0.90'— Pyrenochaetopsis microspora CBS 102876

Pyrenochaeta nobilis CBS 292.74

Pyrenochaeta nobilis CBS 407.76

Pyrenochaeta nobilis CBS 566.75

Pyrenochaeta corni CBS 102828

1.00 Pyrenochaeta corni CBS 234.92

Pyrenochaeta corni CBS 248.79

Pyrenochaeta unguis-hominis CBS 1111112

Cucurbitariaceae

Pyrenochaeta unguis-hominis CBS 112.79

0.94 Pyrenochaeta unguis-hominis CBS 378.92

Pyrenochaeta quercina CBS 115095

Pyrenochaeta cava BPI 356246

Pyrenochaeta cava CBS 115953

Pyrenochaeta cava CBS 257.68

1.00 Paraconiothyrium minitans CBS 122788

0.99 Pleurophoma pleurospora CBS 116668 Outgroup

Pleurophoma pleurospora CBS 130329

PLaTE 2. Phylogenetic relationship of species of Pyrenochaeta and related genera inferred by

Bayesian analysis of a part of the large subunit ribosomal region. Bayesian posterior probabilities

are indicated at the nodes. In bold, the sequence generated in this study from the Phoma canadensis

isotype [= Pyrenochaeta cava].

concept for Phoma. Sutton (1964) introduced conidiogenesis as an important

taxonomic feature for Phoma and related genera. Unfortunately, Allescher did

not provide any detail about conidiogenesis of Ph. canadensis, nor were we

able to observe the necessary details in such old herbarium specimens. Since

established by Saccardo (1880), Phoma has grown to accommodate more than

3000 species that form pycnidia with aseptate hyaline conidia (Aveskamp et al.

Phoma canadensis: a synonym of Pyrenochaeta cava ... 737

2008). More recently several publications have reappraised this fungal group

(Boerema et al. 2004; Aveskamp et al 2010; Gruyter et al. 2009, 2010, 2013) and

split Phoma into several new genera. Regrettably, Ph. canadensis as well as other

Phoma spp. that are obscure and unavailable in culture, were not included

in these recent studies. Here, in order to elucidate the generic position of

Ph. canadensis, the LSU sequence of the isotype (BPI 356246) was analyzed within

a dataset covering a number of Phoma species and other closely related genera

(TABLE 1). Since our attempts to obtain sequences from BPI 356244 and

BPI 356245 failed, the phylogenetic analysis covered only the sequence from

specimen BPI 356246.

The sequence from isolate BPI 356246 grouped within the clade of Pyr. cava

with maximum posterior probability (PLATE 2). Although the genus

Pyrenochaeta presents, among other morphological features, setae around

the ostiole, Schneider (1979) pointed out that this attribute is not a stable

generic character, and Gruyter et al. (2010) confirmed that the presence of

setose pycnidia is rare in Pyr. cava. This both agrees with what we observed

and explains why setose ostioles were not reported in the original description

(Allescher 1896). The fungus found on Conyza canadensis also shares with

Pyr. cava the characteristic minute conidia and the undifferentiated

conidiogenous cells. Based on their phenotypic and genotypic similarities, we

propose that Ph. canadensis Allesch. and Pyr. cava are conspecific.

Acknowledgments

The authors wish to acknowledge Fundacao de Amparo a Pesquisa do Estado de Minas

Gerais (FAPEMIG), Conselho Nacional de Desenvolvimento Cientifico e Tecnoldgico

(CNPq) and Coordenagao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)

for financial support, BPI for the loan of specimens and Dr. JHC Woudenberg and Dr.

DJ Soares for having reviewed the manuscript and for their valuable suggestions.

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Allescher A. 1896. Diagnosen einiger neuer, im Jahre 1895 gesammelter Arten bayerischer Pilze,

aus der Abteilung der Fungi imperfecti. Berichte der Bayerischen Botanischen Gesellschaft

4: 31-40.

Aveskamp MM, Gruyter J, Crous PW. 2008. Biology and recent developments in the systematics of

Phoma, a complex genus of major quarantine significance. Fungal Divers. 31: 1-18

Aveskamp MM, Gruyter J de, Woudenberg JHC, Verkley GJM, Crous PW. 2010. Highlights of the

Didymellaceae: a polyphasic approach to characterize Phoma and related pleosporalean genera.

Stud Mycol 65: 1-60. http://dx.doi.org/10.3114/sim.2010.65.01

Boerema GH, Gruyter J de, Noordeloos ME, Hamers MEC. 2004. Phoma identification manual.

Wallingford, UK: CABI publishing.

Farr DE, Rossman AY, Palm ME, McCray EB. 2015. Fungal databases, systematic botany and

mycology laboratory, ARS, USDA. [http://nt.ars-grin.gov/fungaldatabases/ (viewed online on

5 August 2015)].

738 ... Duarte & Barreto

Fernandes AF, Miranda BEC, Duarte LL, Barreto RW. 2013. Passalora stromatica sp. nov. associated

with leaf spots of Tithonia diversifolia in Brazil. IMA Fungus 4: 201-204.

http://dx.doi.org/10.5598/imafungus.2013.04.02.05

Gruyter J de, Aveskamp MM, Woudenberg JHC, Verkley GJM, Groenewald JZ, Crous PW. 2009.

Molecular phylogeny of Phoma and allied anamorph genera: towards a reclassification of the

Phoma complex. Mycol. Res. 113: 508-519. http://dx.doi.org/10.1016/j.mycres.2009.01.002

Gruyter J de, Woudenberg JHC, Aveskamp MM, Verkley GJM, Groenewald JZ, Crous PW. 2010.

Systematic reappraisal of species in Phoma section Paraphoma, Pyrenochaeta and Pleurophoma.

Mycologia 102: 1066-1081. http://dx.doi.org/10.3852/09-240

Gruyter J de, Aveskamp MM, Verkley GJM, Groenewald JZ, Crous PW. 2013. Redisposition of

Phoma-like anamorphs in Pleosporales. Stud. Mycol. 75: 1-36.

http://dx.doi.org/10.3114/sim0004

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

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Uppsala University:

Evolutionary Biology Centre.

Rambaut A. 2009. Figtree V1.3.1. [http://tree.bio.ed.ac.uk/ (downloaded on 5 May 2009)].

Rambaut A, Drummond AJ. 2003. Tracer v1.5. [http://evolve.zoo.ox.ac.uk/ (downloaded on 5 May

2009)].

Saccardo PA. 1880. Conspectus generum fungorum Italiae inferiorum nempe ad Sphaeropsideas,

Melanconieas et Hyphomyceteas pertinentium systemate sporologico dispositorum. Michelia

2: 1-38.

Saccardo PA, Trotter A. 1913. Supplementum universale, pars IX, sect. 1. Ascomycetae. Sylloge

Fungorum 22(1): 1-822.

Schneider R. 1979. Die Gattung Pyrenochaeta De Notaris. Mitt. BiolBundesanst. Land.-Forstw. 189:

1-73.

Staden R, Beal KF, Bonfield JK. 2000. The Staden package, 1998. Methods Mol. Biol. 132: 115-30.

Sutton BC. 1964. Phoma and related genera. Trans. Brit. Mycol. Soc. 47: 497-509.

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several Cryptococcus species. J. Bacteriol. 172: 4238-4246.

MY COTAXON

http://dx.doi.org/10.5248/130.739

Volume 130, pp. 739-747 July-September 2015

Four new records of Rhizocarpon from China

WEI-CHENG WANG, ZUN-TIAN ZHAO, & LU-LU ZHANG

College of Life Sciences, Shandong Normal University, Jinan, 250014, P. R. China

* CORRESPONDENCE TO: lichenzll@aliyun.com

ABSTRACT — Four new lichen records are reported from China: Rhizocarpon hochstetteri,

R. parvum, R. polycarpum, and R. pusillum. Detailed taxonomic descriptions with photos and

comments are provided.

Key worps — Rhizocarpaceae, Asia, taxonomy

Introduction

Rhizocarpon Ramond ex DC. is a large genus of c. 200 species included

in the family Rhizocarpaceae. It is widely distributed in all parts of the world

(Matwiejuk 2008). Rhizocarpon species frequently occur on siliceous rocks

(some on basic substrata) in alpine and polar regions, while other species are

parasitic on other lichens (Timdal & Holtan-Hartwig 1988).

The main characters of Rhizocarpon are an areolate or rimose thallus,

black lecideine apothecia between areoles, Rhizocarpon-type asci, 1-septate

to muriform ascospores with haloes, brown hypothecium, and anastomosing

and branched paraphyses (Feuerer & Timdal 2004, Fletcher et al. 2009). The

genus is usually divided into two subgenera: R. subg. Rhizocarpon, containing

species characterized by a yellow thallus containing rhizocarpic acid and

R. subg. Phaeothallus for species with a white, ashy, smoky, or brown thallus

lacking rhizocarpic acid (Thomson 1967).

Thirty-one Rhizocarpon species have been reported from China (Aptroot &

Sparrius 2003, Guo 2005, Li et al. 2013, Mahire et al. 2015, Wei 1991, Zhao et

al. 2013). The four new records reported here are from different parts of China;

R. parvum and R. pusillum are yellow lichenicolous species with rhizocarpic

acid in the thalli, whereas R. hochstetteri and R. polycarpum are non-yellow

non-lichenicolous species without rhizocarpic acid.

740 ... Wang, Zhao, & Zhang

Materials & methods

The specimens are preserved in the Lichen Section of Botanical Herbarium,

Shandong Normal University, Jinan, China (SDNU). Their morphological and

anatomical characters were examined under a stereomicroscope (Olympus SZ) and a

polarizing microscope (Olympus CX21). Both thallus and medulla were tested with

K (a 10% aqueous solution of potassium hydroxide), C (a saturated solution of aqueous

sodium hypochlorite), I (a 10% aqueous solution of aqueous potassium iodide), and

P (a saturated solution of p-phenylenediamine in 95% ethyl alcohol) for identification.

The lichen substances were identified using standardized thin layer chromatography

techniques (TLC) with system C (Orange et al. 2010). Photos of these lichens were taken

under Olympus SZX16 and BX61 with DP72.

Taxonomic descriptions

Rhizocarpon hochstetteri (K6rb.) Vain.,

Acta Soc. Fauna Fl. Fenn. 53(1): 280 (1922) Fic. 1

MorpHotocy — Saxicolous, thallus grey-brown, areolate or rimose,

0.1-0.3 mm thick; medulla I-; prothallus visible along the margin; areoles flat

to convex, sometimes verrucose, 0.35-0.75 mm diam. APOTHEcIA black, flat

to slightly convex, 0.5-1.0 mm diam., irregular, round to angular, sometimes a

few small ones clustered in groups, sessile or innate among the areoles; margin

prominent, flexuous; exciple dark brown, poorly formed; hyphae radiating;

epihymenium Macrocarpa-green (K-, N+ red); hymenium colorless, I+

blue, 100-150 um high; hypothecium dark brown, paraphyses branched and

anastomosing, apices swollen with brown cap; no crystals in the apothecia.

Asci clavate, Rhizocarpon-type; ascospores colorless, but sometimes pale

brown when old, 1-septate, halonate, (19—)22-27 x 9-12 um, 8 per ascus.

CHEMISTRY — Medulla K-, C-, P-. Stictic acid detected by TLC.

SPECIMENS EXAMINED: CHINA. JILIn, Baishan, Mt. Changbaishan, alt. 2300 m, on rock,

22 Jul. 2014, W.C. Wang 20141058, 20141092, 20141115 (SDNU).

DIsTRIBUTION — Rhizocarpon hochstetteri has been reported from Europe, North

America, New Zealand, Ukraine, and Japan (Fletcher et al. 2009, Fryday 2002, Galloway

2007, Matwiejuk 2008, Thomson 1997, Timdal & Holtan-Hartwig 1988). New to China.

ComMENts — Rhizocarpon richardii and R. polycarpum are two other species

with large, colorless, 1-septate ascospores like R. hochstetteri but differ in

having an I+ blue medulla. Rhizocarpon cinereovirens resembles R. hochstetteri

but is distinguished by possession of norstictic acid and a more areolate thallus.

Rhizocarpon lavatum and R. reductum are usually difficult to distinguish from

R. hochstetteri because they share the same habitat and their thallus are all grey

to grey-brown, but R. hochstetteri has 1-septate ascospores while R. lavatum

and R. reductum both have muriform ascospores.

Rhizocarpon spp. new for China... 741

FiGuRE 1. Rhizocarpon hochstetteri (Wang 20141058, SDNU). A: Thallus; B: Apothecium;

C: Apothecium section; D: K reaction; E: Paraphyses; F: Ascus; G: Ascospores; Scale bars: A = 1 mm;

B = 500 um; C, D = 50 um; E, G = 10 um.

Rhizocarpon polycarpum (Hepp) Th. Fr., Lich. Scand. 1(2): 617 (1874) FIG. 2

MorpHotocy — Saxicolous, thallus crustose, areolate, thin, 0.3-0.4 mm

thick; medulla I+ blue; prothallus black, evident between the areoles and

apothecia and along the thallus margin; areoles dark brown, 0.4-0.6 mm

diam., contiguous or scattered. APOTHECIA black, epruinose, round to angular,

0.4-0.75 mm diam., flat, occasionally flexuose, with a thin, prominent proper

742 ... Wang, Zhao, & Zhang

C: Apothecium section; D: K reaction; E: Paraphyses; F: Ascus; G: Ascospores; Scale bars:

A, B = 500 um; C, D = 20 um; E, G = 10 um.

margin especially when old; exciple brown-black, 45-60 um; epihymenium

red-brown, K+ red-violet; hymenium colorless, 100-110 um high; hypothecium

dark brown, K-, paraphyses branched and anastomosing, no crystals in the

apothecia. Ascr clavate, Rhizocarpon-type; ascospores colorless or sometimes

becoming pale brown with age, 1-septate, halonate, 22-26 x 10-11 um, 8 per

ascus.

CHEMISTRY — Medulla K-, C-, P-. No lichen substances detected.

Rhizocarpon spp. new for China... 743

SPECIMENS EXAMINED: CHINA. JrLin, Baishan, Mt. Changbaishan, alt. 2300 m, on

rock, 22 Jul. 2014, W.C. Wang 20141068 (SDNU). X1zANG, Linzhi, Mt. Sejila, alt. 4600

m, on rock, 20 Jul. 2011, Y.L. Cheng 20118355 (SDNU). NEIMENGGU, Aershan, Mt.

Jiguanshan, alt. 1500 m, on rock, 29 Aug. 2011, D.F. Jiang 20124660 (SDNU).

DISTRIBUTION — R. polycarpum has been reported from Europe, North America,

Australia, New Zealand, Argentina, Asia, and Antarctica. (Anderson 1965, Fletcher et

al. 2009, Fryday 2002, Fryday & Kantvilas 2012, Golubkov & Matwiejuk 2009, Ovstedal

et al. 2009, Timdal & Holtan-Hartwig 1988). New to China.

ComMENts — ‘The I+ blue medulla and the red-brown, K+ red-violet

epihymenium separate R. polycarpum from other species with 1-septate and

colorless ascospores. Rhizocarpon richardii also has an I+ blue medulla but

differs from R. polycarpum in having apothecial crystals and a dark olive-

green, K- epihymenium. Rhizocarpon umense, another species similar to

R. polycarpum, is distinguished by its brown, distinctly smaller ascospores

(12-15 x 5-6 um) and more reduced thallus.

Rhizocarpon pusillum Runemark, Opera Bot. 2(1): 63 (1956) Fic. 3

MorpuHotoey — Lichenicolous, thallus small, 0.3-1.0 cm diam., areolate,

forming patches on the host's thallus; medulla I-; prothallus absent or indistinct;

areoles bright yellow, round to angular, flat to convex, 0.25-0.75 mm diam..

ApoTHEcia black, round to angular, 0.5-1.0 mm diam., flat to convex, margin

present or sometimes indistinct; exciple and epihymenium red-brown, K+

red, no crystals; hymenium colorless, 80-100 um high; hypothecium brown;

paraphyses branched and anastomosing, capitate. Asci clavate, Rhizocarpon-

type; ascospores blue-brown to dark olive-brown, 1-septate, halonate, 9-14(-15)

x (4-)6-7 um, 8 per ascus.

CHEMISTRY — Medulla K-, C-, P+ yellow. Rhizocarpic acid and psoromic

acid detected by TLC.

SPECIMENS EXAMINED: CHINA. XINJIANG, Urumgi, Mt. Tianshan, Glacier No. 1, alt.

3700 m, on Sporastatia testudinea (Ach.) A. Massal. on rock, 17 Aug. 2012, D.F Jiang

20129131 (SDNU); alt. 3560m, on rock, 8 Sep. 2013, Q. Ren 2013897 (SDNU)..

DISTRIBUTION — Rhizocarpon pusillum has been reported from Asia, Europe, North

America, and New Zealand (Feuerer & Timdal 2004, Galloway 2007, Matwiejuk 2008,

Poelt 1988, Thomson 1997, Timdal & Holtan-Hartwig 1988). New to China.

Comments — Rhizocarpon pusillum and R. parvum both belong to the yellow

Rhizocarpon group with rhizocarpic acid in thallus. They are similar in having

a small thallus, 1-septate brown small ascospores, and a lichenicolous habit.

However, R. pusillum is parasitic on Sporastatia testudinea, has I- medulla, an

indistinct prothallus, and two secondary products (rhizocarpic acid, psoromic

acid) detected by TLC while R. parvum is parasitic on Tremolecia atrata, has

744 ... Wang, Zhao, & Zhang

FIGURE 3. Rhizocarpon pusillum (Jiang 20129131, SDNU). A: Thallus; B: Apothecium;

C: Apothecium section; D: K reaction; E: Paraphyses; F: Ascus; G: Ascospores; Scale bars:

A, B = 500 um; C, D = 20 um; E, G = 10 um.

an I+ violet medulla, a distinctly black prothallus, and rhizocarpic acid without

psoromic acid.

Rhizocarpon parvum Runemark, Opera Bot. 2(1): 64 (1956) Fic. 4

MorpuHo.oey — Lichenicolous, thallus crustose, areolate, small, 0.25-0.75

cm diam.; medulla I+ violet; prothallus distinct; areoles pale yellow, round to

angular, 0.1-0.4 mm diam., flat to slightly convex, dispersed on dark prothallus.

Rhizocarpon spp. new for China... 745

Y . 2,

, : F . — G

10 yer (ys 10um ie 10 pm

rf) 4

FiGuRE 4. Rhizocarpon parvum (Jiang 20129131, SDNU). A: Thallus; B: Apothecium;

C: Apothecium section; D: K reaction; E: Paraphyses; F: Ascus; G: Ascospores; Scale bars:

A = 500 um; B = 200 um; C, D = 20 um; E, G = 10 um.

ApoTHEcia black, flat or slightly convex, 0.25-0.6mm diam.; margin distinct;

exciple and epihymenium red-brown, K+ red, no crystals; hymenium colorless,

70-90 um high; hypothecium brown; paraphyses branched and anastomosing,

capitate. Asci clavate, Rhizocarpon-type; ascospores dark green, 1-septate,

halonate, 9-15 x 6-7 um, 8 per ascus.

CHEMISTRY — Medulla K-, C-, P-. Rhizocarpic acid detected by TLC.

746 ... Wang, Zhao, & Zhang

SPECIMEN EXAMINED: CHINA. XINJIANG, Urumgi, Mt. Tianshan, Glacier No. 1, alt.

3700 m, on Tremolecia atrata (Ach.) Hertel on rock, 17 Aug. 2012, D.F. Jiang 20129131

(SDNU).

DISTRIBUTION — R. parvum has been reported from Scandinavia (Runemark 1956,

Poelt 1988, Timdal & Holtan-Hartwig 1988). New to China.

CoMMENTS — Rhizocarpon parvum differs from other lichenicolous species

with rhizocarpic acid and 1-septate ascospores in having an I+ blue medulla

and distinct prothallus.

Acknowledgements

The authors thank Dr. A. Aptroot (ABL Herbarium, Soest, The Netherlands) and

Prof. Shou-Yu Guo (Institute of Microbiology, Chinese Academy of Sciences) for

presubmission reviews. This study was supported by the National Natural Science

Foundation of China (31170187, 31400015), and the Scientific Research Innovation

Fundation of Shandong Normal University (SCX1525).

Literature cited

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MY COTAXON

http://dx.doi.org/10.5248/130.749

Volume 130, pp. 749-756 July-September 2015

New record of Aplosporella javeedii on five hosts in China

based on multi-gene analysis and morphology

XIN-LEI FAN’, QIN YANG’, BIN CAO’,

YING-MEI LIANG’, & CHENG-MING TIAN”

' The Key Laboratory for Silviculture and Conservation of Ministry of Education,

Beijing Forestry University, Beijing 100083, China

? Museum of Beijing Forestry University, Beijing 100083, China

* CORRESPONDENCE TO: chengmt@bjfu.edu.cn

ABSTRACT —Aplosporella javeedii (Aplosporellaceae) was collected from branch cankers

in China, identified by multigene phylogenetic analyses and morphological characters,

described, and illustrated. This species is newly recorded for China, and the five tree species

(Albizia julibrissin, Broussonetia papyrifera, Gleditsia sinensis, Juniperus chinensis, and

Styphnolobium japonicum) represent new host records for A. javeedii.

Key worps — Ascomycota, Botryosphaeriales, phylogeny, taxonomy

Introduction

Aplosporella Speg. is characterized by solitary, globose, unilocular

conidiomata in malt extract agar (MEA) and multi-locular branched

conidiomata with a single ostiole and brown aseptate conidia.

Jami et al. (2014) described Aplosporella javeedii as an endophyte from

healthy wood sections of Celtis africana Burm. f. (Cannabaceae) and Searsia

lancea (L. f.) FA. Barkley (Anacardiaceae) in South Africa. During our recent

taxonomic study of botryosphaeriaceous fungi associated with dieback

diseases in China, isolates from five different Chinese hosts were identified

as Aplosporella javeedii based on morphological and multi-gene phylogenetic

analyses. ‘These represent the first records of A. javeedii from China.

Materials & methods

COLLECTION OF SAMPLES AND ISOLATES. — Six Aplosporella isolates were collected

from five hosts: Albizia julibrissin Durazz. (Fabaceae), Broussonetia papyrifera (L.) Vent.

750 ... Fan & al.

(Moraceae), Gleditsia sinensis Lam. (Fabaceae), Juniperus chinensis L. (Cupressaceae),

and Styphnolobium japonicum (L.) Schott [= Sophora japonica L.] (Fabaceae). Isolations

were made directly from conidiomata on the host and single-spore purification

according to a protocol slightly modified from Fan et al. (2014). Specimens are deposited

in the Museum of Beijing Forestry University, Beijing, China (BJFC), and single-spore

cultures are maintained in the China Forestry Culture Collection Center, Beijing, China

(CFCC).

MoRPHOLOGICAL STUDIES. — Specimens observed on branch tissues consisted of

size of fruiting bodies and spores. More than 20 fruiting bodies were sectioned, and

50 randomly selected spores were measured using a Leica light microscope (DM 750).

Cultural characteristics of isolates incubated on MEA in the dark at 25°C were recorded.

These included colony characters and pigment production, at 3, 7, and 30 days.

DNA EXTRACTION, PCR AMPLIFICATION, AND SEQUENCING. — Genomic DNA was

extracted from pure cultures grown in MEA with cellophane at 25°C for 2 weeks.

Genomic DNA was extracted using a modified CTAB method (Doyle & Doyle

1990). DNA concentrations were estimated by electrophoresis in 1% agarose gels and

NanoDrop™ 2000 (Thermo, USA) according to Desjardins et al. (2009). The Beijing

Genomics Institute sequenced the DNA. Amplifications were primed by ITS1 & ITS4

(nITS region; White et al. 1990), NL1 & NL4 (LSU region; O’Donnell 1993), and

EF1-688F & EF1-1251R (EF-1a region; Alves et al. 2008). PCR amplification products

were estimated visually by electrophoresis in 2% agarose gels. DNA sequencing was

performed using an ABI PRISM® 3730XL DNA Analyzer with BigDye® Terminator Kit

v.3.1 (Invitrogen) at the Shanghai Invitrogen Biological Technology Company Limited

(Beijing, China).

DNA SEQUENCE ANALYSIS. —DNA sequences generated by forward and reverse primers

were used to obtain consensus sequences using SeqMan v.7.1.0 inthe DNAStar Lasergene

Core Suite software (DNAStar Inc., Madison, WI, USA). Sequences were aligned using

MAFFT v.6 (Katoh & Toh 2010) and edited manually using MEGA 6 (Tamura et al.

2013). A partition homogeneity test with heuristic search and 1000 homogeneity

was performed using PAUP v.4.0b10 to test the discrepancy among the three loci in

reconstructing phylogenetic trees. Phylogenetic analysis was run using PAUP v.4.0b10

for maximum parsimony (MP) analysis (Swofford 2003), MrBayes v.3.1.2 for Bayesian

analysis (Ronquist & Huelsenbeck 2003), and RAXxML v.7.2.8 for maximum likelihood

(ML) analysis (Stamatakis 2006). A phylogeny using Aplosporella javeedii sequences

from the current study and other reference sequences in GenBank was generated based

on the multilocus alignment (ITS, LSU, EF-1la). Fusicladium effusum STE-U 4525 and

Fusicladium oleagineum CBS 113427 were selected as outgroup (Slippers et al. 2013).

Trees were visualized using FigTree v.1.3.1 (Rambaut & Drummond 2010).

MP analysis was run using a tree-bisection and reconnection branch-swapping

algorithm with heuristic search option of 1000 random-addition sequences. Zero

length branches were collapsed and all equally parsimonious trees were saved. Other

calculated parsimony scores calculated were tree length (TL), consistency index (CI),

retention index (RI) and rescaled consistency (RC). ML analysis was also performed

Aplosporella javeedii new to China... 751

using a GTR model of site substitution, including estimation of gamma-distributed rate

heterogeneity and a proportion of invariant sites (Stamatakis 2006). The MP and ML

support values were evaluated with a bootstrapping method of 1000 replicates (Hillis

& Bull 1993). Bayesian inference was calculated using a Markov Chain Monte Carlo

(MCMC) algorithm with Bayesian posterior probabilities (Rannala & Yang 1996).

A nucleotide substitution model was estimated by MrModeltest v.2.3 (Posada &

Crandall 1998), and a weighted Bayesian analysis was considered. Two MCMC chains

were run from random trees for 1,000,000 generations, and trees were sampled every

100th generation, resulting in 10,000 total trees. The first 25% of trees were discarded as

the burn-in phase of each analysis and the posterior probabilities (PP) were calculated

using the remaining 7500 trees.

Sequence data is deposited in GenBank, and the accession numbers are listed

in TABLE 1. The multilocus sequences alignment file is deposited in TreeBASE

(www.treebase.org) as accession $16645.

TABLE 1 — Newly sequenced strains of Aplosporella javeedii used in the phylogenetic

analyses.

GENBANK ACCESSION NUMBERS

SPECIMEN Host STRAIN

ITS LSU EF-la

BJFC-S867 Albizia CFCC 89657 KM030579 KM030586 KM030593

BJFC-S884 Broussonetia CFCC 89658 KM030580 KM030587 KM030594

BJFC-S920 Albizia CFCC 89659 KM030581 KM030588 KM030595

BJFC-S1022 Gleditsia CFCC 50052 KP208838 KP208841 KP208844

BJFC-S1042 Styphnolobium CFCC 50053 KP208839 KP208842 KP208845

BJFC-S1044 = Juniperus CFCC 50054 KP208840 KP208843 KP208846

Phylogeny

The combined multi-gene region dataset included sequences from 71 ingroup

strains representing 48 species in Botryosphaeriales. A partition homogeneity

test was not significant (P = 0.145) indicating that the individual datasets

were congruent and produced trees with the same topologies. The dataset had

an aligned length of 1468 characters of which 772 characters are constant,

74 are variable parsimony-uninformative, and 622 are parsimony-informative.

Maximum parsimony analysis yielded three equally parsimonious trees

(TL = 2540, CI = 0.490, RI = 0.809, RC = 0.396). The ML analysis produced

the same topology as the MP tree, with Bayesian analysis producing the same

topology, with an average standard deviation of split frequencies = 0.007233.

The phylogeny inferred from ITS, LSU, and EF-la sequences (Fic. 1)

indicated six major clades: Aplosporellaceae, Botryosphaeriaceae, Melanopsaceae,

752... Fan & al.

54/-

hyllostictaceae

60.0

accharataceae

A. javeedii CFCC 89657

A. javeedii CFCC 89658

A. javeedii CFCC 89659

A. javeedii CFCC 50052

A. javeedii CFCC 50053

93/90" A. javeedii CFCC 50054

A. javeedii CMW 38167

A. javeedi CMW 38165

A. prunicola STE-U 6327

A. prunicola CBS 121167

A. yalgorensis MUCC 512

A. yalgorensis MUCC 511

A. africana CBS 121777

A. longipes CFCC 89660

A. longipes CFCC 89661

A. hesperidica CBS 208.37

A. papillata CBS 121780

sgsa- B. fusca CBS 174.26

B. iraniana IRAN 1448

100/109 P. mamane CPC 12440

Aplosporellaceae

eee7 P. cupressi IRAN 1458

P. citrigena ICMP 16818

P. eucalypti MFLUCC 11-0579

P. porosa CBS 110496

L. crassispora WAC 12533

L. crassispora CBS 110492

L. gonubiensis CBS 115812

sig D. stevenii CBS 230.30

pe D. africana CBS 120835

100/199 N. phoenicum CBS 169.34

N. phoenicum CBS 122528

N. palmicolaMFLUCC 100822

N. palmicola MFLUCC100823

B. fusicoccum MFLUCC 110657

E. tritici CBS 118719

N. mangiferae CMW 7024

N. luteum CMW 10309

N. grevilleae CPC 16999

N. australe CMW 6837

N. eucalypticola CBS 115679

S. citricola |CMP 16827

S. viticola CBS 117009

S. viticola UCP 105

s7/99, B. corticis CBS 119047

B. fabicerciana CMW 27106

-- C. eucalypti MFLUCC 11-0655

M. phaseolina CBS 22733

N. dimidiatum CBS 251.49

N. novaehollandiae CBS 122071

N. dimidiatum CBS 499.66

gsiag P. adansoniae WAC 12689

P. adansoniae CMW 26147

P. ardesiacum CBS 122062

P. kimberleyense CBS 122058

E. endophytica CBS 120397

E. microspora CBS 353.97

too/10g M. tulasnei CBS 116805

. M. tulasnei CBS 116806

K. confusa CBS 131723

K. yuccigena CPC 20623

K. yuccigena CBS 131727

K. dasylirionicola CBS 131720

53/-

Melanopsaceae

= Planistromellaceae

S. capensis CBS 122693

eo S. kirstenboschensis CBS 123537

S. proteae CBS 115206

; P. braziliana LGMF 334

P. braziliana LGMF 330

P. braziliana LGMF 333

(— F. effusum STE-U 4525

— F, oleagineum CBS 113427

Aplosporella

Barriopsis

Phaeobotryon

Phaeobotryosphaeria

Lasiodiplodia

Diplodia

Neodeightonia

Botryobambusa

Eutiarosporella

Neofusicoccum

Spencermartinsia

Botryosphaeria

Cophinforma

Macrophomina

Neoscytalidium

Pseudofusicoccum

Endomelanconiopsis

Melanops

Kellermania

Saccharata

Phyllosticta

Fusicladium

Aplosporella javeedii new to China... 753

Phyllostictaceae, Planistromellaceae, and Saccharataceae, representing six

families in Botryosphaeriales. The sequences from our six Chinese collections

clustered within Aplosporellaceae, where they formed a highly supported clade

(MP = 93; ML = 90; BBP = 0.99) with the South African holotype and paratype

collections of Aplosporella javeedii.

Taxonomy

Aplosporella javeedii Jami, Gryzenh., Slippers & M.J. Wingf., Fungal Biology 118:

174, 2014. FIG. 2

SEXUAL STATE: unknown.

ASEXUAL STATE: Conidiomata immersed in bark, erumpent from bark

surface, separate, discoid, dark-brown to black, multi-locular, <1.5 mm diam,

wall consisting of 4-10 layers, thick outer layers composed of dark-brown

textura angularis, becoming hyaline and thin-walled towards the inner region.

Disc brown to black, circular to ovoid, (0.43-)0.48-0.66(-0.75) mm (average =

0.58 mm, n = 20), with 1-5 ostioles per disc. Ostioles multiple, conspicuous,

level with the disc surface, sometimes covered below disc by lighter entostroma,

(52.8—)61.0-85.2(-89.9) um (average = 77.6 um, n = 20) in diam. Locules

multiple, irregularly arranged, subdivided frequently by invaginations with

common walls, (0.84—)0.93-1.40(-1.52) mm (average = 1.26 mm, n = 20)

in diam. Paraphyses hyaline, smooth-walled, 31.4-80.1 um long, 3.1-5.5 um

wide. Conidiogenous cells formed from the cells lining the inner walls of

the locules, hyaline, smooth, inconspicuous. Conidia aseptate, ellipsoid to

oblong, smooth, ends rounded, initial hyaline, becoming brown when mature,

(18.8—)19.5-24.6(-25.3) x (8.3-)9.6-11.7(-12.9) um (average = 22.8 x 10.6 um,

n = 50). Cultures in PDA white, uniform, felty with abundant aerial mycelium.

Cultures in MEA white at first, then grey-olivaceous, similar to reverse side

after 7-10 days. Colony uniform with appressed aerial mycelium. After

3 weeks, conidiomata solitary, globose, unilocular, immersed to semi-immersed

on MEA.

MATERIAL EXAMINED: CHINA, GANSU PROVINCE, Qingyang City, municipal

government, 35°42’31.02”N 107°38’24.94’E, elev. 1400 m asl, on infected branches

of Albizia julibrissin, 14 July 2013, X.L. Fan (BJFC-S867; culture CFCC 89657);

Ficure 1. Phylogram of the combined genes of ITS, LSU, and EF-1a based on MP, ML, and Bayesian

analyses of botryosphaerialean taxa. Numbers above the branches indicate bootstrap values.

Thickened branches indicate PP > 0.95 from the Bayesian inferences. Scale bar = 60 nucleotide

substitutions. The newly sequenced Chinese collections of Aplosporella javeedii (designated by

CFCC #s) are the top six sequences of the tree. Ex-holotype sequences are in bold.

754 ... Fan & al.

FicuRE 2. Aplosporella javeedii (BJFC-S867). A: Habit of conidiomata on a twig. B: Multiple ostioles

on the discs of the conidiomata. C, D: Transverse sections through the conidiomata. E: Longitudinal

sections through the conidiomata. F: Paraphyses and conidiogenous cells. G: Conidia. H: Colonies

on PDA at 3 days (left) and 30 days (right). I: Conidiomata on MEA. J: Transverse sections through

the conidiomata on MEA. K: Colonies on MEA at 3 days (left) and 30 days (right). Scale bars:

A =2 mm; B-E, J = 0.5 mm; K, G = 20 um; I= 1 mm.

Aplosporella javeedii new to China... 755

Qingyang City, Heshui County, 35°46’51.46”N 107°59’43.42’E, elev. 1291 m asl, on

infected branches of A. julibrissin, 13 July 2013, X.L. Fan (BJFC-S920; culture CFCC

89659); Tianshui City, Baozi Valley, 34°34’00.21”N 105°42’21.94”E, elev. 1007 m

asl, on infected stems of Broussonetia papyrifera, 17 July 2013, X.L. Fan (BJFC-S884;

culture CFCC 89658). SHANXI PROVINCE, Datong City, Children’s Park, 40°04’44.05”N

113°16’42.89’E, elev. 1051 m asl, on infected branches of Gleditsia sinensis, 14 April 2014,

X.L. Fan & B. Cao (BJFC-S1022; culture CFCC 50052); Jinzhong City, Jinshang Park,

37°43'07.26"N 112°43’52.00’E, elev. 818 m asl, on infected branches of Styphnolobium

japonicum [= Sophora japonica], 17 April 2014, X.L. Fan & B. Cao (BJFC-S1042; culture

CFCC 50053); Jinzhong City, Jinshang Park, 37°43’08.11”N 112°43’52.20”E, elev. 809

m asl, on infected branches of Juniperus chinensis, 17 April 2014, X.L. Fan & B. Cao

(BJFC-S1044; culture CFCC 50054).

HaBITAT/DISTRIBUTION: Known from Albizia julibrissin, Broussonetia

papyrifera, Gleditsia sinensis, Juniperus chinensis, and Styphnolobium japonicum

in China and from Celtis africana and Searsia lancea in South Africa.

Discussion

Our six Chinese botryosphaeriaceous strains are identified as Aplosporella

javeedii based on morphological observations and multi-gene phylogenetic

analyses. Morphologically, they are characterized by solitary globose unilocular

conidiomata on MEA and erumpent discoid conidiomata, multiple ostioles in

branches with ellipsoid to oblong aseptate conidia averaging 22.8 x 10.6 um.

Aplosporella javeedii was previously reported as an endophyte from healthy

trees of two dicotyledonous host species in South Africa (Jami et al. 2014).

Our Chinese records extend the host range to two additional dicotyledonous

families, and also to a coniferous host; moreover, our specimens were

collected from branch cankers, raising the question whether this species may

have some pathogenic potential. Slippers & Wingfield (2007) suggested that

botryosphaeriaceous fungi might shift slightly from endophytic to pathogenic

habit when hosts are subjected to stress.

Acknowledgments

This study was financed by the National Natural Science Foundation of China (Project

No. 31170603) and the Fundamental Research Funds for the Central Universities

(Project No. BLYJ201404). The authors thank Quan Lu (Research Institute of Forest

Ecology, Chinese Academy of Forestry, Beijing) and Janice Uchuda (Department of

Plant and Environmental Protection Sciences, University of Hawaii, Honolulu) for

helpful comments and presubmission review.

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MY COTAXON

http://dx.doi.org/10.5248/130.757

Volume 130, pp. 757-767 July-September 2015

Phaeostilbelloides and Velloziomyces —

new dematiaceous genera from the Brazilian Cerrado

ELIANE AMARAL DE SOUZA ARMANDO,

ZULEIDE MARTINS CHAVES, & JOSE CARMINE DIANESE”*

Departamento de Fitopatologia, Universidade de Brasilia,

Campus Darcy Ribeiro, Asa Norte, 70910-900 Brasilia, DF, Brazil

* CORRESPONDENCE TO: jcarmine@gmail.com

Asstract —Phaeostilbelloides velloziae gen. & sp. nov. and Velloziomyces ramosiconidialis

gen. & sp. nov. are described and illustrated. Both new fungi infect Vellozia squamata,

endemic to the Brazilian Cerrado.

KEY worDs — neotropical fungi, asexual ascomycete morphs, mycodiversity, Velloziaceae

Introduction

Starting in 1993, an extensive inventory of fungi associated with plants

native to the Brazilian savanna, designated as the Cerrado, (Dianese 2000,

Dianese et al. 1993b, 1997) yielded important records of unusual hyphomycetes

such as those inhabiting the trichomes of Cerrado plants (Cantrell et al. 2011,

Pereira-Carvalho et al. 2009b). To date, 13 new ascomycete genera have been

described, ten as asexual morphs (Dornelo-Silva & Dianese 2004, Pereira-

Carvalho et al. 2009b) and three as sexual morphs (Dianese & al. 2001, Pereira-

Carvalho 2009a, Pereira-Carvalho et al. 2010). In addition, two new genera

of rust fungi, Batistopsora and Kimuromyces (Cummins & Hiratsuka 2003,

Dianese et al. 1993a, 1995), have been described and a third, Mimema, has

been reinstated (Cummins & Hiratsuka 2003, Dianese et al. 1994). Several

cercosporoid fungi (Dianese et al. 2008, Dornelo-Silva et al. 2007, Furlanetto

& Dianese 1999, Hernandez-Gutierrez & Dianese 2008, 2009, 2014, Inacio

& Dianese 2006, Medeiros & Dianese 1994) have been described and other

new ascomycetes have been included in other publications (Dornelo-Silva &

Dianese 2003, Inacio et al. 2011, 2012, Soares & Dianese 2014, Souza et al.

2008). This paper expands the number of genera among the asexual ascomycete

758 ... Armando, Chaves, & Dianese

morphs from the Cerrado, with the description and illustration of two new

genera, Phaeostilbelloides and Velloziomyces.

Materials & methods

Fungi associated with plants native in the Cerrado Biome were extensively collected.

Observations under a stereomicroscope revealed the presence of dematiaceous

hyphomycetes on leaves of Vellozia squamata Pohl (Velloziaceae), which is a Brazilian

Cerrado endemic. Squash preparations were mounted on semi-permanent slides stained

with lacto-glycerol/cotton blue or glycerol-KOH/basic phloxine. Mounting media

without stains were also used to determine the color of different structures. Sections

about 15-25 um thick were produced using a Leica CM 1520 freezing microtome.

The structures observed in the squash preparations and sections were measured,

described, and documented using a Leica DM 2500 microscope coupled with a Leica

DFC 490 digital camera connected to a microcomputer. Image capture, editing, and

measurements were made with Leica QWin V3 software. Whenever possible 20-50

measurements were made and expressed as range of length by width, with the mean

measures indicated in parentheses.

Taxonomy

Phaeostilbelloides Armando, Z.M. Chaves & Dianese, gen. nov.

MycoBank MB808833

Differs from Phaeostilbella by its lack of rigid setae on the conidiomata.

TYPE SPECIES: Phaeostilbelloides velloziae Armando et al.

ErymMo oey: Latin Phaeostilbelloides, meaning similar to the genus Phaeostilbella.

CONIDIOMATA synnematous, scattered, erect, determinate, dark brown to

brown. CONIDIOPHORES macronematous, cylindrical, branched just below

the apex, septate, straight or flexuous, brown. CONIDIOGENOUS CELLS

monophialidic, integrated, terminal, determinate. CoNrDIA acrogenous,

cylindrical to ellipsoid, unicellular, brown or pale brown.

Phaeostilbelloides velloziae Armando, Z.M. Chaves & Dianese, sp.nov. = PLATE 1

MycoBank MB808834

Differs from Phaeostilbella species by its lack of rigid curved setae on the conidiomatal

top and differs from Myrothecium species by its dark brown or brown conidiophores and

conidiogenous cells and its lack of a sporodochial phase.

Type: Brazil. Distrito Federal: Brasilia, West Sector of the IBGE-Roncador Reserve, on

living leaves of Vellozia squamata, 5 Apr. 2001, J.C. de Castro 82 (Holotype, UBMC

18404).

ErymMo_oey: Latin velloziae, referring to the host genus.

LESIONS necrotic, linear, elongate, 5-10 x 0.5 mm, following parallel leaf

veins. MycELIUM immersed; HYPHAE light brown, septate 3-5 um wide.

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 759

FicurE 1 Phaeostilbelloides velloziae (holotype, UBMC 18404) on leaf of Vellozia squamata.

A. Synnema showing stipe with textura intricata; B. Synnema top showing subapical branching

(white arrows), and abundant hyaline proliferations of the conidiophores (black arrow);

C. Conidiogenous cells and conidia; D. Conidia; E. Conidia with clearer area (arrow) at the conidial

base. Scale bars: A = 50 um; C = 10 um; B, D, E= 5 um.

CONIDIOMATA 194—(212)—294 x 54-(56)—-60 um, synnematous cylindrical,

stipe textura intricata, smooth, dark brown, expanded to a 150—(166)—175 um

wide brush-like capitulum, containing large number of immature

conidiophores that show up in squash preparations. CONIDIOPHORES 3—5 tm

760 ... Armando, Chaves, & Dianese

in diam., cylindrical, smooth, originating as sub-terminal branches at

the capitulum, reduced to a single cell bearing two conidiogenous cells.

CONIDIOGENOUS CELLS monophialidic, integrated, terminal, cylindrical,

10—(15)—20 um long, 1-3 um in diam., showing clear periclinal thickenings but

not a collarette. Conrp1a 8-(7.5)—11 x 2—(2.5)—-3 um, noncatenate, aseptate,

sub-cylindrical, with an obtuse apex and truncate base, brown to dark brown,

with a slightly clear band just above the hilum, accumulating in mucilaginous,

dark brown masses.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. DISTRITO FEDERAL: PLANALTINA,

Reserva Ecolégica Aguas Emendadas, on living leaves of Vellozia squamata, 25 Mar.

1995, Mariza Sanchez 530 (UBMC 7668).

ComMEntTs - Following the concepts by Seifert (1985) and Seifert et al. (2011),

it is not possible to place the specimen in any of the known genera producing

synnematous conidiomata, 1-celled brown to dark-brown conidia, and

phialidic conidial ontogeny. The closest genus, Myrothecium Tode [= Solheimia

E.E. Morris] as described and illustrated by Seifert et al. (2011), shares some

characteristics in common with the new taxon. However, the differences between

Phaeostilbelloides velloziae and Myrothecium species are sufficient to place

them in two separate genera. All Myrothecium species produce dark smooth

conidia, except for one species that shows conidia with strongly striate surface

and fusiform with acute ends. In addition, the synnematous conidiomata are

unbranched, and sporodochial conidiomata may also be present. In contrast,

P. velloziae lacks sporodochia, produces brown to dark brown smooth-walled

cylindrical conidia with an obtuse apex and a pale area just above the truncate

base, and synnemata that are sometimes branched. Another synnematous

species described from Brazil, Chaetantromycopsis bambusae H.P. Upadhyay et

al., produces black slimy spore heads but clearly differs from Phaeostilbelloides

by its hyaline to creamy white conidiophores and the presence of erect acute

hyaline to creamy white setae encircling the base of the synnemata (Upadhyay

et al. 1986). Finally, the Phaeostilbelloides type species is easily separated from

Phaeostilbella species because the hyaline cylindrical conidiophore branches

seen at the top in squash preparations differ completely from the rigid slightly

curved appendices of Phaeostilbella. Furthermore, Phaeostilbella synnemata

typically show textura prismatica, not textura intricata as in Phaeostilbelloides.

Also, the only species currently accepted in Phaeostilbella was reported on

grasses in Europe (Seifert et al. 2011) while Phaeostilbelloides velloziae is present

in a dicotyledonous host endemic to the Cerrado.

The dichotomous key below, modified from Seifert et al. (2011), segregates

Phaeostilbelloides from all other genera with synnematous conidiomata forming

phialidic conidiogenous cells bearing 1-celled conidia.

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 761

Key to the dematiaceous genera of phialidic synnematous anamorphs

forming 1-celled conidia

1. Conidia hyaline, globose to ellipsoidal, ovoid, fusoid,

in clear or bright colored mucilaginous false heads....................000008 2

1. Conidia brown to dark brown, cylindrical, fusoid,

imadariebrown-terblack-taleedheddsts Mele sect pratt tarsi terse taal SERS gol BES yw oleh 7

2. Synnemata originating from broad well developed stromata, found on wood

ede ccagt Nac Spt Cth tte cng ea eegh tation ah b agate eng Stromatographium Hohn.

. Synnemata lacking a well-developed basal stroma. ............... 0. cece eee eee 3

. Synnemata with ornamenting cells on stipe

EMM ethin: cial ett bas weet geSte anamorphic Stilbocrea Pat. (= Gracilistilbella Seifert)

3. Synnemata without ornamenting cells on the stipe............ eee ee eee ee 4

4. Synnematous heads with sterile hyphae among the conidiogenous cells

SR oe ee eee ee anamorphic Holwaya Sacc. (= Crinula Fr.)

4, Synnematous heads lacking sterile hyphae .............. cece eee eee eee eee 5

5. Conidiophores penicillate occurring in pustules on the upper portion of the stipe

Wai eited's apecd d Saale as Mel'g wbkal dcaipsulacabaul'g gab -sils ub cing at Cornutostilbe Seifert

5. Stipe withoutsconidiGphores in PuUStules:.. yes + ct yn on ot onmgtestern oBehs.0n epsoy on gh oh ane 6

6. Conidiogenous cells lageniform with characteristically narrow elongated necks,

CON 1a CO OV ONG is soda ti bratl abe ala. best abe beagle by etet ah soles Dennisographium Rifai

6. Conidiogenous cells holoblastic or phialidic terminal at the tip of

each conidiophore, lacking narrow neck........... Synnemellisia N.K. Rao et al.

7. Synnemata with setose capitulum, showing a crown of dark setae,

CONICS TUSK RAS eat. hs ie Pinar ak 2h AS ees ee 8, Phaeostilbella Hohn.

7. Synnemata lacking setae at capitulum or with subhyaline hyphae ................ 8

8. Capitulum without sterile hyphae, conidia fusiform, with smooth or striate surface,

sporodochial phase often present.................. Myrothecium (= Solheimia)

8. Capitulum non-setose but with subhyaline hyphae, conidia cylindrical,

with obtuse apex and pale band just above the truncate base,

sporoddchialsphasé- absent ten. fap ln heel waiter lcs sty em a MS oaks Phaeostilbelloides

Velloziomyces Armando, Z.M. Chaves & Dianese, gen. nov.

MycoBank MB808835

Differs from Minimidochium by its holoblastic conidial ontogeny and from

Dictyodochium by its schizolytic conidial secession.

TYPE SPECIES: Velloziomyces ramosiconidialis Armando et al.

EryMo.ocy: From the Latin Velloziomyces for its occurrence on Vellozia.

CONIDIOMATA sporodochial scattered or confluent, setose, dark brown, SETAE

cylindrical, erect, straight or flexuous, continuous or septate, brown to dark

brown. CONIDIOPHORES undifferentiated CONIDIOGENOUS CELLS monoblastic,

cylindrical to lageniform, indeterminate, with several enteroblastic percurrent

762 ... Armando, Chaves, & Dianese

extensions, integrated, terminal. Conip1a solitary, obclavate or Y-shaped,

0—2-septate, simple or branched, brown to pale brown.

Velloziomyces ramosiconidialis Armando, Z.M. Chaves & Dianese, sp. nov.

MycoBank MB808836 PLATES 2, 3

Differs from Minimidochium species by its holoblastic conidial ontogeny, from

Dictyodochium species by its schizolytic conidial secession of phragmospores, and from

Iyengarina species by its setose sporodochial conidiophores and association with living

leaves.

Type: Brazil. Distrito Federal: Planaltina, Estagao Ecolégica Aguas Emendadas, on

living leaves of Vellozia squamata, 13 Jun. 1995, Arenildo Soares Alves 215 (Holotype,

UBMC 8849).

EryMo_oey: From the Latin ramosiconidialis, referring to conidial branching.

CoLonigs amphigenous, light brown, erumpent, setose. Mycelium mostly

immersed, composed of septate, light brown, smooth hyphae. CONIDIOMATA

sporodochial suprastomatal, scattered or confluent, setose, dark brown,

10—(42)—55 um diam. SETAE cylindrical, erect, straight or flexuous, sometimes

slightly curved, obtuse or acerose at the apex, continue or septate, brown to dark

brown, 60—(112)—138 x 3-(4)—6 um. CONIDIOGENOUS CELLS monoblastic,

indeterminate, with several enteroblastic percurrent extensions, terminal,

short cylindrical, lageniform 3—9 x 3-5 um. Conrp1a solitary, 1—2-septate,

more or less Y-shaped, with a lateral branch, obclavate at main axis (7—(18)—25

x 3-(8)—16 um) and one shorter lateral branch, light brown, with paler conical

apical cell, truncate base.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. DisTRITO FEDERAL: BRaSf{LiA, Jardim

Botanico de Brasilia, on living leaves of Vellozia squamata, 24 May 2011, Eliane

Armando 129 (UBMC 21928).

ComMENTS - Sporodochial conidiomata are distributed in several genera

of dematiaceous anamorphs (Ellis 1971, Ellis 1976, Kiffer and Morelet 2000,

Seifert 1985, Seifert et al. 2011). However, there is no published genus described

as producing setose sporodochia on top of an obconical stromatic structure

that grows through the stomata. As illustrated here, the stroma originates

deep within the leaf parenchyma, grows through the stoma to expand at the

surface into a multicellular plate formed by a layer of tightly packed annellidic

conidiogenous cells surrounded by dark brown elongated setae. In addition, the

light brown conidia are unusual because at maturity they have one unicellular

conical branch growing from the basal cell, which is shorter than the main

2-3-celled axis of the spore. Sometimes the branch is not present.

Two genera, Minimidochium B. Sutton and Dictyodochium Sivan., as shown

in Seifert et al. (2011), have setose sporodochial conidiomata that originate

from internal mycelium emerging from plant tissue, as in Velloziomyces.

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 763

FiGurRE 2 Velloziomyces ramosiconidialis (holotype, UBMC 8849) on leaf of Vellozia squamata.

A. Setose stromatic sporodochium; B. Sporodochium detail of the superficial distribution of the

conidiogenous cells; C. Setose sporodochium bearing conidia; D. Setae detail and a group of free

conidia; E. Setose sporodochium; F. Conidiogenous cell; G-H. Conidia focused in different depths.

Scale bars: A = 20 um; B-E = 10 um; F-H = 5 um.

However, all eight species of Minimidochium: M. crepuscolare C. Ciccar.,

M. eucalypti Vittal & Dorai, M. indicum Varghese & V.G. Rao, M. microsporum

Matsush., M. monoseptatum G.C. Zhao, M. nipponicum Matsush., M. parvum

764. ... Armando, Chaves, & Dianese

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 765

Cabello et al., and M. setosum B. Sutton (Cabello et al. 1998, Ciccarone 1988,

Matsushima 1995, Sutton 1969, Varghese & Rao 1980, Vittal & Dorai 1991, Zhao

& Zhao 2012) produce phialidic conidiogenous cells bearing typically setulose

hyaline conidia. On the other hand, in the monotypic genus Dictyodochium,

the muriform brown conidia have rhexolytic secession. Also, the three species

belonging in Iyengarina Subram. (Kuthubutheen & Nawawi 1992, Subramanian

1958) are easily segregated from V. ramosiconidialis due to clear differences in

conidial morphology, inability to form setose sporodochial conidiomata, and

association with plant debris instead of living leaves. Thus, species belonging in

all three genera clearly differ from V. ramosiconidialis, which is characterized by

its holoblastic annellidic conidiogenous cells, non-setulose branched conidia

showing schizolytic secession, and setose sporodochial conidiomata.

Acknowledgements

The authors thank Drs. R.E Castafieda-Ruiz (INIFAT, Cuba) and Paul Kirk

(Kew Gardens, UK) for critical commentaries that greatly improved our manuscript

and CNPq-Brazil for the graduate fellowship provided to the senior author and for

financing the execution of the entire research project through funds provided by the

PPBIO-Cerrado/CNPgq. Finally, gratitude is expressed to Prof. Mariza Sanchez for

assistance and friendship.

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MY COTAXON

http://dx.doi.org/10.5248/130.769

Volume 130, pp. 769-773 July-September 2015

New records of crustose Teloschistaceae

and lichenicolous fungi from Turkey

MEHMET GOKHAN HALICI

University of Erciyes, Faculty of Science, Department of Biology 38039 Kayseri, Turkey

CORRESPONDENCE TO: mghalici@gmail.com

ABsTRACT — Two lichenized fungi (Flavoplaca austrocitrina, E dichroa) and two

lichenicolous fungi (Tremella caloplacae, Xanthoriicola physciae) are reported from Turkey

for the first time. Comments on their habitats, substrata, and key anatomical features are

provided for each species.

Key worps — biodiversity, lichens, hyphomycetes, Ascomycota, Basidiomycota

Introduction

There is still no checklist of Turkish lichens and lichenicolous fungi although

intensive lichenological studies have been carried especially in the last two

decades in the country and numerous papers have been published (John 2007,

Senkardesler 2011). Approximately 1200 lichenized fungal species are known

from Turkey, but at least 2000 lichenized fungal species might reasonably be

expected to be present in the country, considering its size and diversity of

phytogeographical regions and habitats (Halic: et al. 2007).

A key to lichenicolous Ascomycota and mitosporic fungi for Turkey

prepared by Halici (2008) listed 118 taxa from the country. Currently, 182 taxa

of lichenicolous fungi are known from Turkey (Halici et al. 2014), but even very

common lichenicolous basidiomycetes have not been reported.

During 2012-14 many crustose teloschistacean specimens and lichenicolous

fungi were collected from Turkey. In this paper I discuss two lichenized and

two lichenicolous species that are new to Turkey.

Material & methods

Sections were prepared by hand and examined in Lugol’s MERCK 9261, 10% KOH,

cotton blue, and water. The measurements of ascospores taken in water are given as:

770 ... Halici

(min-)[X-sd]-X-[X+sd](-max.), where ‘min’ and ‘max’ represent the extreme values,

‘X’ the arithmetic mean, and ‘sd’ the corresponding standard deviation. Measurements

are presented in the classical way where ten or fewer measurements were made. The

specimens are deposited in Erciyes University Herbarium, Kayseri, Turkey (EUH).

Taxonomy

Flavoplaca austrocitrina (Vondrak, Riha, Arup & Sochting) Arup, Sochting &

Frédén

A detailed description is provided by Vondrak et al. (2009).

Thallus yellow to somewhat brownish orange, areolate; most areoles convex,

almost subsquamulose; 275—400 um thick and 0.35—1.3 mm in diam. Areoles

with marginal soralia; but in the old thalli areoles are usually totally covered

by soredia. Apothecia sometimes present; (0.2—)0.2-0.6-1.0(-1.5) mm in

diam., zeorine, with thalline exciple prominent especially in the old apothecia.

Hymenium (50-)58-—73-87 (-90) um tall. Paraphyses tips (4-)5-5.5-6.5(-7)

um wide. Ascospores colourless and polarilocular, (9-)11-12.5-14(-16) x

(4.5-)5.5-6.5-7.5(-8.5) um; septa wide, (2-)3-4-5(-6.5) um, length/width:

(1.44—)1.71-2.04-2.36(-2.89); septum/length: (0.14-)0.23-0.31-0.4(-0.44).

Pycnidium not observed.

SPECIMENS EXAMINED: TU’RKEY, ADANA, Ceyhan, Kizildere Village, 36°56’N 35°41’E,

alt. 200 m, on mortar, 28 August 2013, M.G. Halici (ERC CL 1.689); Ice, Camlryayla,

near Giizeloluk, Pinus brutia forest, on limestone, 37°10’N 34°48’E, alt. 620 m, on

limestone, 25 February 2012, M.G. Halici (ERC CL 0.262); Izmir, Karamiirsel, Tepekoy,

40°41’N 29°39’E, alt. 230 m, on mortar, 21 May 2012, M.G. Halici (ERC CL 1.832);

KIRKLARELI, Center, west of Inece, near Ulukonak Village, on limestone, 41°32’N

27°06’E, alt. 100 m, on limestone, 29 May 2012, M.G. Halici (ERC CL 0.383).

According to Vondrak et al. (2009), Flavoplaca austrocitrina is common

both inland and in coastal areas of Europe on lime-rich artificial substrata

such as concrete and mortar but also on limestone. In Turkey we reported it

on both natural limestone and mortar in suburban areas near the coast and

inland mostly below 250 m. A related species, E flavocitrina (Nyl.) Arup et al.,

has similar soralia and ecology but differs in its thinner and smaller areoles

(Vondrak et al. 2009). New to Turkey and to Asia.

Flavoplaca dichroa (Arup) Arup, Frédén & Sochting

Detailed descriptions are provided by Arup (2006), Vondrak et al. (2009),

and Wilk (2011).

Thallus continuous, areolate, of two color forms, yellow and orange.

Areoles 0.3—0.45 mm in diam., and more or less totally covered by blastidia/

soredia. Cortex indistinct, with 1-2 layers of cells. Apothecia common in most

specimens, 0.2—0.8 mm in diam. In mature apothecia disc slightly concave or

sometimes convex, zeorine, thalline exciple mostly covered by blastidia/soredia

Flavoplaca and lichenicolous fungi new for Turkey ... 771

and clearly visible. Hymenium (45-)51-64-77(-85) um tall. Tips of paraphyses

(4-)4.5-5.55-6.5(-8.5) um wide. Ascospores colorless, polarilocular, thin-

walled, (9-)10-12-14(-16) x (4.5-)5.5-6.5-7.5(-8.5) um, septa (1.5-)2.5-

3.5-4.5(-5.5) um, length/width: (1.25-)1.6-1.93-2.26(-2.8), septum/length:

(0.16—)0.21-0.29-0.36(-0.5).

SPECIMENS EXAMINED: TURKEY, Amasya, Borabay, north of Borabay Village, Pinus

sylvestris forest, 40°09’N 36°08’E, alt. 1480 m, on calcareous rocks, 28 August 2012, M.G.

Halici (ERC CL 0.224); Ayp1n, Nazilli, Toygar Village, 37°49’N 28°21’E, alt. 90 m, on

weakly calcareous rocks, 2 July 2012, M.G. Halici (ERC CL 0.776); Samsun, Ladik, east

of Akdag Ski Center, 40°54’N 35°53’E, alt. 960 m, on calcareous rocks, 28 August 2012,

M.G. Halici (ERC CL 1.571); ToxKat, Camlibel, southwest of Tekreli Village, 40°10’N

36°30°E, alt. 1450 m, on calcareous rocks, 29 August 2012, M.G. Halici (ERC CL 0.695).

Flavoplaca dichroa can easily be recognized in the field by its yellow and

orange thallus forms growing side by side (Arup 2006; Vondrak et al. 2009).

It differs from E austrocitrina (see above) by a thinner thallus. According to

Arup (2006) the thick-walled ascospores (sand-clock type) are diagnostic for

the Nordic specimens but in the Turkish specimens the ascospores are regularly

thin-walled ascospores as it was mentioned by Vondrak et al. (2009) for the

specimens from the Black Sea Region. Wilk (2011) also stated that Polish

specimens of this species have thick-walled ascospores. This species is common

in Europe (Arup 2006; Vondrak et al. 2007) and seems to be widely distributed

in Turkey on sun exposed calcareous rocks mostly over 1000 m altitude. New

to Turkey and to Asia.

Tremella caloplacae (Zahlbr.) Diederich

A detailed description is provided by Diederich (1996).

No distinct basidiomata visible; induces formation of hardly visible galls

inside the hymenium of Calogaya biatorina (A. Massal.) Arup et al. Concolorous

with the host thallus. Basidia separated by one septum, 16—25 x 7-10 um. No

visible damage was seen but in the infected apothecia ascospore production

was suppressed.

SPECIMEN EXAMINED: TURKEY, ExaziG, Harput, Buzluk Cave, 38°44’N 39°16’E, alt.

1570 m, in the hymenium of Calogaya biatorina on calcareous rocks, 08 July 2013, M.G.

Halici (ERC CLL 0.051).

Actually we were not initially aware of this fungus as no distinct galls were

formed in the host lichen. However, ITS analyses of several apothecia from

the host revealed the existence of Tremella caloplacae. Thereafter, we made

additional apothecial sections and observed the T: caloplacae basidia. ‘This

species is known from Algeria and several European countries including the

island of Crete in Greece in the hymenia of Calogaya arnoldii (Wedd.) Arup et

al., Caloplaca saxicola (Hoftm.) Nordin, Rufoplaca arenaria (Pers.) Arup et al.,

772 ... Halici

Rusavskia elegans (Link) S.Y. Kondr. & Karnefelt, Xanthoria sorediata (Vain.)

Poelt, Variospora aurantia (Pers.) Arup et al., and Usnochroma carphineum (Ft.)

Sochting et al. (e.g., Sérusiaux et al. 2003; Diederich 2007; Alstrup et al. 2009;

Etayo 2010; Millanes et al. 2014). All the specimens having two-celled basidia

on teloschistalean hosts are currently classified under this species (Millanes et

al. 2014). New to Turkey and to Asia.

Xanthoriicola physciae (Kalchbr.) D. Hawksw.

A detailed description is provided by Hawksworth & Punithalingam (1973).

The hyphomycete grows in the hymenium of the host apothecia, which

becomes blackish and thus easily seen in the field. Conidiophores brown,

immersed, branched, smooth-walled. Conidiogenous cells broadly cupulate.

Conidiogenesis enteroblastic. Conidia dark brown to almost black, globose,

3—6 um in size, aseptate with warted wall, produced singly, not in chains,.

SPECIMEN EXAMINED: TURKEY, Bursa, Iznik, on the highway to [hsaniye, 40°27’N

29°48’E, alt. 710 m, on the apothecia of Xanthoria parietina, 23 May 2012, M.G. Halici

(ERC CLL 0.071).

According to LSU sequence analyses (Ruibal et al. 2011), Xanthoriicola belongs

to the order Capnodiales. This conspicuous species, which grows almost

exclusively on Xanthoria parietina (L.) Beltr., is widely distributed in Europe

and known also in Asia and Africa but has not previously been found in Turkey.

Acknowledgements

The manuscript was reviewed prior to submission by Kerry Knudsen (Czech

Republic & USA) and Mikhail Zhurbenko (Russia). This study was financially supported

by TUBITAK (111T927 coded project) and FDA-2013-4422 coded Erciyes University

project. The help of Jan Vondrak in the identification of the lichenized fungal species is

gratefully appreciated.

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lichenicolous fungi of South Greenland. Folia Cryptog. Estonica 46: 1-24.

Arup U. 2006. A new taxonomy of the Caloplaca citrina group in the Nordic countries, except

Iceland. Lichenologist 38: 1-20. http://dx.doi.org/10.1017/S0024282905005402

Diederich P. 1996. The lichenicolous heterobasidiomycetes. Bibliotheca Lichenologica 61: 1-98.

Diederich P. 2007. New or interesting lichenicolous heterobasidiomycetes. Opuscula Philolichenum

4: 11-22.

Etayo J. 2010. Liquenes y hongos liquenicolas de Aragon. Universidad del Pais Vasco: 1-501.

Halici MG. 2008. A key to the lichenicolous Ascomycota (including mitosporic fungi) of Turkey.

Mycotaxon 104: 253-286.

Halic1 MG, Hawksworth DL, Aksoy A. 2007. Contributions to the lichenized and lichenicolous

fungal biota of Turkey. Mycotaxon 102: 403-414.

Halici MG, Candan M, Giillii M, Ozcan A. 2014. Phoma recepii sp. nov. from the Caloplaca cerina

group in Turkey. Mycotaxon 129: 163-168. http://dx.doi.org/10.5248/129.163

Flavoplaca and lichenicolous fungi new for Turkey ... 773

Hawksworth DL, Punithalingam E. 1973. New and interesting microfungi from Slapton, South

Devonshire: Deuteromycotina. Transactions of the British Mycological Society 61: 57-69.

http://dx.doi.org/10.1016/S0007-1536(73)80088-9

John V. 2007. Lichenological studies in Turkey and their relevance to environmental interpretation.

Bocconea 21: 85-93.

Millanes A, Diederich P, Westberg M, Knutsson T, Wedin M. 2014. Tremella rhizocarpicola sp.

nov. and other interesting Tremellales and Filobasidiales in the Nordic countries. MycoKeys 8:

31-41. http://dx.doi.org/10.3897/mycokeys.8.8176

Ruibal C, Millanes A, Hawksworth DL. 2011. Molecular phylogenetic studies on the

lichenicolous Xanthoriicola physciae reveal Antarctic rock-inhabiting fungi and Piedraia

species among closest relatives in the Teratosphaeriaceae. IMA Fungus 2: 97-103.

http://dx.doi.org/10.5598/imafungus.2011.02.01.13

Sérusiaux E, Diederich P, Ertz D, van den Boom P. 2003. New or interesting lichens and lichenicolous

fungi from Belgium, Luxembourg and northern France. IX. Lejeunia 173: 1-48.

Senkardesler A. 2011. Das die Turkei betreffende lichenologische Schrittum icin ek kayitlar. Tiirk

Liken Toplulugu Biilteni 9: 9-10.

Vondrak J, Kocourkova J, Palice Z, Liska J. 2007. New and noteworthy lichens in the Czech Republic

- genus Caloplaca. Preslia 79: 163-184.

Vondrak J, Riha,P. Arup U, Sechting, U. 2009. The taxonomy of the Caloplaca citrina group

(Teloschistaceae) in the Black Sea region; with contributions to the cryptic species concept in

lichenology. Lichenologist 41: 571-604. http://dx.doi.org/10.1017/S0024282909008317

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115: 83-98. http://dx.doi-org/10.5248/115.83

MY COTAXON

http://dx.doi.org/10.5248/130.775

Volume 130, pp. 775-781 July-September 2015

New species of Phaeomonilia and Mirandina

from dead branches in China

YING-Rur Ma’, JI-WEN X1IA', XIU-GUO ZHANG’,

& RAFAEL EF. CASTANEDA-RUIZ?

"Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

"Instituto de Investigaciones Fundamentales en Agricultura Tropical “Alejandro de Humboldt”

(INIFAT), Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Two new hyphomycetes, Phaeomonilia guangxiensis and Mirandina inaequalis,

are described and illustrated from specimens collected on dead branches in Guangxi Province,

China. Phaeomonilia guangxiensis is distinguished by its globose unicellular colorless conidia

formed by thallic disarticulation of fertile branches on macronematous conidiophores.

Mirandina inaequalis differs from other described Mirandina species by its mostly 3-septate

cylindrical conidia and conspicuous denticles on the conidiogenous cell.

KEY worps — anamorphic fungi, taxonomy

Introduction

There is enormous diversity of anamorphic fungi growing on rotten wood

and dead branches in tropical forests, and several novel taxa have been recently

discovered from forest ecosystems of southern China (Dai et al. 2009; Zhang et

al. 2009, 2012; Dai & Li 2010; Ma et al. 2011, 2012a,b; Ren et al. 2012). During

ongoing mycological surveys in a nature reserve of Guangxi Province, China,

two interesting hyphomycetes were collected on dead branches. Morphological

and developmental characteristics suggest their placement in Phaeomonilia

and Mirandina. Both differ from previously described taxa and therefore are

proposed as new to science.

Materials & methods

Samples of decaying wood were collected from subtropical forests of Guangxi

Province, China, placed in separate zip-lock plastic bags, and taken to the laboratory.

They were then incubated at 27°C for more than 2 weeks in an artificial climate box in 9 cm

diameter plastic Petri dishes containing moistened filter paper. Samples were examined

through an Olympus SZ61 dissection microscope. All microscopic characteristics

776 ... Ma &al.

were determined at 60x and 100x magnification (Olympus BX51 microscope) and are

based on 50 mature conidia and 30 conidiophores mounted in lactophenol. Specimens

were deposited in the Herbarium of Shandong Agricultural University, Taian, China

(HSAUP) and the Mycological Herbarium, Institute of Microbiology, Chinese Academy

of Sciences, Beijing, China (HMAS).

Taxonomy

Phaeomonilia guangxiensis Y.R. Ma & X.G. Zhang, sp. nov. FIG. 1

MycoBAnk MB 814054

Differs from Phaeomonilia pleiomorpha by its larger conidia; and from P. corticola by its

wider, globose conidia.

Type: China, Guangxi Province: Dayaoshan Nature Reserve, 23°40’N 109°50’E, alt. 550

m, on dead branches of an unidentified broadleaf tree, 9 Oct. 2012, Y.R. Ma (Holotype,

HSAUP H4040; isotype, HMAS 243455).

ETyMOLoGy: refers to the province where the type was found.

Cotonigs on the natural substratum effuse, brown, consisting of individual

brown conidiophores over the substratum surface. Mycelium mostly

superficial but some immersed; superficial hyphae pale brown, cylindrical,

smooth, septate, branched. CONIDIOPHORES macronematous, mononematous,

erect, straight or flexuous, cylindrical, septate, smooth, <405 um tall, 7.5-11.5

um wide at the base, brown to dark brown, mostly dichotomously branched

towards the apex. CONIDIOGENOUS CELLS enterogenous, determinate, inwardly

curved, colorless, smooth, lageniform, to subulate. RAMOCONIDIA AND

conipiA thallic-arthric, catenate, ellipsoid to globose, broadly Y-shaped, 7-11

x 11-14.5 um, unicellular, colorless, dry, smooth, forming by disarticulation of

the conidiogenous branches.

Comments - The genus Phaeomonilia was established by Castaneda-Ruiz et

al. (2007) with two species: P. pleiomorpha R.F. Castaneda et al. (type species)

and P. corticola (R.F. Castafeda) R.F. Castaneda et al. [= Monilia corticola

R.F. Castafieda]. Phaeomonilia was mainly characterized by macronematous

conidiophores with globose, unicellular, colorless conidia formed by thallic

disarticulation of fertile branches and by the Stylaspergillus-like synanamorph.

The genus is similar to Botryomonilia Goos & Piroz. (Goos & Pirozynski 1975)

in its branching system, narrowing progressively towards the apex before

the maturation and fragmentation or disarticulation process that produces

the thallic-arthric conidia, as described by Kendrick (2003). Botryomonilia,

however, has conidia connected by an isthmus (Castafeda-Ruiz et al.

2007). Phaeomonilia pleiomorpha differs from P guangxiensis by its smaller

conidia (4.0-5.8 x 3.9-4.4 um) and its Stylaspergillus-like synanamorph, and

P. corticola differs by its narrower ellipsoid to doliiform conidia (5.5-16.0 x

4-5 um; Castafieda-Ruiz et al. 2007).

Phaeomonilia & Mirandina spp. nov. (China) ... 777

Fic. 1. Phaeomonilia guangxiensis (holotype, HSAUP H4040). a. Conidiophore and conidia.

b, c. Conidiogenous cells and conidia. d. Conidia.

Key to species of Phaeomonilia

1. Stylaspergillus-like synanamorph present ............... 000 e eee eee P. pleiomorpha

LANG -SvOanarnOrpPrESentee Aas eras eae ce Roe ce eae cee eee se so ee 2

2, Omid ON LOria OF S1IPSOIS , ssa. Fabs scn Fa bere ta tare 8 asee ah ect. ath cheb P. corticola

PELLONIGIA CLODOSE LW obi tak ph tw vb Gl b Gh Seal gal sh tae ay gt 4 Gee P. guangxiensis

Mirandina inaequalis Y.R. Ma & X.G. Zhang, sp. nov. FIG. 2

MycoBank MB 814055

Differs from other Mirandina species by its cylindrical and mostly 3-septate conidia.

778 ... Ma &al.

Type: China, Guangxi Province: Guposhan Nature Reserve, 24°34’N 111°30’E, alt. 600

m, on dead branches of an unidentified broadleaf tree, 9 Oct. 2012, Y.R. Ma (Holotype,

HSAUP H4039; isotype, HMAS 243456).

EryMo_oey: refers to the unequal length of the conidia.

Cotoniss effuse on the natural substratum, brown, hairy. Mycelium partly

superficial, partly immersed in the substratum, composed of branched, septate,

pale brown to brown, smooth-walled hyphae. CONIDIOPHORES macronematous,

mononematous, single or in small groups, unbranched, erect, straight or

flexuous, 5—7-septate, smooth, brown at the base and becoming almost colorless

towards the apex, thin-walled, 75-220 x 3.5-6.5 um. CONIDIOGENOUS CELLS

polyblastic, integrated, terminal, becoming intercalary, almost colorless, with

short-cylindrical denticles in the upper region, conical at the apex. Conidial

secession schizolytic. Conrp14 holoblastic, solitary, acropleurogenous, dry,

narrow, obclavate, fusiform or cylindrical, subhyaline to hyaline, 1-3-septate,

mostly 3-septate, 16-24.5 x 2.5-5 um wide in the broadest part, apex rounded,

base truncate with a protuberance.

ComMENTs - Arnaud (1952) introduced the genus Mirandina but did not

provide a Latin diagnosis. Matsushima (1971) provided a Latin diagnosis

but failed to name a type species. The generic name was finally validated by

Matsushima (1975), who designated M. corticola as the type species. Thirteen

taxa are included in the genus, but only nine have validly published names

(Index Fungorum 2014; MycoBank 2014; Matsushima 1971, 1975, 1980, 1983,

1987; Kirk 1986; Castafieda-Ruiz & Kendrick 1991; Castafeda-Ruiz et al. 1997).

Mirandina inaequalis fits the genus Mirandina, which has brown, erect

conidiophores, conidiogenous cells with short-cylindrical denticles towards

the apex, and conidia that are colorless or nearly so, cylindrical and long but

narrow (Paulus et al. 2003). The material studied is similar to M. speciosa,

“M. cylindrospora, M. dactylelloides, and M. fusiformis in conidial shape, but

M. speciosa has aseptate conidia, “M. cylindrospora” and M. dactylelloides

have very short conidiophores, and the denticles of M. fusiformis are much

denser. Mirandina inaequalis is also morphologically similar to Dactylaria

plovercovensis (Goh & Hyde 1999), D. triseptata (Matsushima 1975; Castafieda-

Ruiz et al. 1991), and D. lakebarrinensis (Hyde & Goh 1998). However,

the conidiogenous cells of D. plovercovensis have more conspicuous and

cylindrical denticles and conidia lack a protuberance at the base. D. triseptata

conidia are broadly rounded at the apex, without a protuberance at the base.

Dactylaria lakebarrinensis is most similar to our species; however, it has shorter

inconspicuous denticles, the conidiophore apex is broad, and its conidia are

olivaceous. In M. inaequalis the conidiophore apex is conical and conidia are

subhyaline to hyaline. We place the fungus in Mirandina rather than Dactylaria

based on its brownish tapered conidiophores and obclavate to cylindrical

Phaeomonilia & Mirandina spp. nov. (China) ... 779

Fic. 2. Mirandina inaequalis (holotype, HSAUP H4039). a. Conidiophores and conidia.

b. Conidiogenous cells with conidia. c. Conidiophores with denticles. d. Conidia.

conidia (Hoog 1985). Because of the confused relationship between Mirandina,

Dactylaria, and related genera, further work, including the use of molecular

tools to classify this complex of fungi, is necessary.

780 ... Ma &al.

Key to species of Mirandina

TACO mic lie SWAG Se phere «cee: Pex 3 eee Peps cee Bap act ae pa range ep Gece er peaeree epee ps ae oc Ne cn 2

Ie@onidiasyith Seperate Fa en PE soe MPL ae, EE a's gE oat igh Br ea ark sa ba eae +

2. Conidiogenous cells denticulate, denticles 1-1.5 um high............. M. arnaudii

2. Conidiogenous cells bearing inconspicuous denticles................. 000. e eee ee 3

3. Gonidia-curved orhooked at base: : 5.2 s04snuk yemee yee neem eh M. fragilis

Sac onictafaleate; teuncatedh Waste yy. fe oaks encehe fe eet. eee hee eee fle tees M. speciosa

A AOMCIODIGECS: YVAN choy athe ol ae oS AR aR oA LA! oN ER BR og GN os 5

A ACORICIOP MOTESIDU OWT. +. cape he taal goths eden Ral eee Rad ee SOR rea eo eed 6

5. Conidia narrowly obclavate, rostrate........... cc eee ee eee eee M. flagelliformis

5. Conidia narrowly fusiform, without rostrum.................... M. dactylelloides

6. Conidia pale brown centrally, subhyaline end cells ................. M. taiwanensis

abs OXes sVPeE Mine CNET e Rupr Um RR ere ane gre ae Rais ANS, AIRC AI A On, ACTOR, EE Ayr APR Acer. 3

7. Conidiogénous loci being scarred... c i ikaw ste on stew eaten t “M. cylindrospora”

7Gonidiosenous lect bemedenticul ate: 4... teta-g~ wron s een «Aan ee Sie eae 8

8. Conidiophores without basal septa, conidia long, slender, flexuous.... “M. breviphora”

8. Conidiophores with basal septa, conidia fusiform.......... 0.0... cece eee eee eee 9

9. ComidEAeMOLe aD OeGe Pp ta «soles A, soos es per ese gel deep oag! Sem peed dem ee die eee BLAH bse 10

PECOniceatesea an O=SE pla Te ss ES 5 ngs e aroce aA +A pam ween #-Agem eters Rete ete 11

10. Conidiophores bear a cluster of short-cylindrical denticles.................... 12

10. Distribution of denticles on conidiophores sparse.................. M. inaequalis

Il Conidia: bixgets 40-6052 Syn Ms tes le te scl bee's F tecatg beside otha “M. typica”

Pl, Conidia smaller, 23:5 =S8 2x15 2500s *, sold oes eb ts no Swa wbSs abs M. fusiformis

12 -Conidiasomietinesscutved 00 fa. lt te ee M. yushanensis

[2 pEOnidiaetrad OT chs wt se pees 4s pete ea atte oe At os Ee Rte cod Bile os Ets we M. corticola

Acknowledgments

The authors express gratitude to Dr Eric H.C. McKenzie (Auckland, New Zealand)

and Dra Gabriela Heredia (Xalapa, Mexico) for serving as pre-submission reviewers and

for their valuable comments and suggestions. This project was supported by the National

Natural Science Foundation of China (Nos. 31093440, 31230001) and the Ministry of

Science and Technology of the People’s Republic of China (Nos. 2006FY120100).

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MY COTAXON

http://dx.doi.org/10.5248/130.783

Volume 130, pp. 783-805 July-September 2015

Species associated with cytospora canker

on Populus tremuloides

JEFF B. KEPLEyY’, F. BRENT REEVES’,

WILLIAM R. JACOBI**, & GERARD C. ADAMS?

' Department of Biology, Colorado State University, Ft. Collins, CO 80523 USA

? Dept. of Bioagricultural Sciences & Pest Management, Colorado State University,

Ft. Collins, CO 80523 USA

° Department of Plant Pathology, University of Nebraska, Lincoln, NE 68583-0722 USA

* CORRESPONDENCE TO: William.jacobi@colostate.edu

ABSTRACT — A new species that is often associated with Cytospora chrysosperma was

found commonly on stems of Populus tremuloides in Colorado. This fungus is illustrated

and described as Cytospora notastroma sp. nov., and morphological and molecular data

demonstrate that the new species is distinct from C. chrysosperma, C. nivea, C. translucens,

and other species found on Populus. Diagnostic features are superficially visible, darkly

pigmented conceptacles circumscribing the ascostromata and conidiomata and surrounding

the white to grayish-white discs; some isolates produce a Phialocephala-like synanamorph in

vitro in addition to the Cytospora anamorph.

KEY worDs — aspen, coelomycete, pathogen, systematics, Valsa

Introduction

Cytospora canker attacks branches, stems, and roots of many woody species

worldwide. Estimates of the number of woody host species found with the

disease vary, but 85 or more have been cited by some authors (Sinclair et al.

1987, Farr et al. 1989, Adams et al. 2006). Cytospora Ehrenb. is recommended

as a holomorphic genus that includes the former genera Leucocytospora

(Hohn.) Hohn., Leucostoma (Nitschke) Hohn., Valsa Fr., Valsella Fuckel, and

Valseutypella Hohn. (Rossman et al. 2015).

In Colorado, Hinds (1964) found cytospora canker in 97% of the native

aspen (Populus tremuloides) stands that he sampled. The mortality and

dieback of aspen have been reported across western North America and

Rocky Mountains from Canada to Arizona (Frey et al. 2004, Hogg et al. 2005,

2008; Fairweather et al. 2008, Worrall et al. 2008, 2010; Zegler et al. 2012).

784 ... Kepley & al.

In Colorado, results of aerial surveys in 2006 indicated that as many as 140,000

acres were impacted, with as much as 10% of the aspen affected in some areas

(Bartos & Shepperd 2006). Mortality in some areas may also affect lateral

roots, which would hinder vegetative regeneration via root suckering. Death

of mature trees can be rapid (a year or two) and is believed often to begin at

epicenters followed by radial spread throughout an aspen stand. Studies by

Worrall et al. (2008) in southwestern Colorado concluded that agents such as

Encoelia pruinosa (Ellis & Everh.) Tork. & Eckblad and Ganoderma applanatum

(Pers.) Pat., which typically kill mature trees in aspen stands, are unimportant

in the present mortality. Rather, a group of secondary agents is involved,

among which are Cytospora species causing cytospora canker. This canker on

aspen is usually caused by Cytospora chrysosperma (Pers.) Fr. [= Valsa sordida

Nitschke], which in the current epidemic is often present and believed to play

a major role in mortality (Marchetti et al. 2011). Sinclair et al. (2005) stated

that C. chrysosperma has been inconsequential in natural forests but can

cause devastating losses in nursery seedbeds, storage, newly established forest

plantations, and landscape or shelterbelt plantings of Populus spp. A study by

Jacobi et al. (1998) concluded that aspen sprout mortality following harvesting

in Colorado results from drought and/or drought coupled with root flooding-

induced stress followed by infection by the canker fungi C. chrysosperma and

Dothiora polyspora Shear & R.W. Davidson. It is apparent that despite the

predisposing and inciting factors associated with aspen mortality, Cytospora

species are often involved in aspen disease in natural or commercial forests in

the western United States, and in many instances C. chrysosperma is believed

to be the specific agent (Hinds 1964, Krebill 1972, Hinds & Krebill 1975, Ross

1976, Juzwik et al. 1978, Walters et al. 1982, Jacobi et al. 1998). Additionally,

aspen growing in urban forests in the Rocky Mountain region are often found

with the disease (Kepley & Jacobi 2000).

Problems associated with species identification of Cytospora

A thorough understanding is lacking of the Cytospora species occurring

on aspen and other Populus spp. in North America in Colorado, the southern

Rocky Mountains, and the Great Plains regions as well as elsewhere in the

United States. Spielman (1983, 1985) lists several species and the national host

index (http://nt.ars-grin.gov/fungaldatabases/fungushost/FungusHost.cfm)

records five on aspen (Cytospora chrysosperma, C. leucosperma (Pers.) Fr.,

C. leucostoma (Pers.) Sacc., C. nivea Fuckel, C. translucens Sacc.). The causal

organism responsible for cytospora canker on aspen in Colorado is typically

reported to be C. chrysosperma, while several investigative studies have

employed C. chrysosperma isolated from aspen in Colorado (Guyon et al. 1996,

McIntyre et al. 1996, Kepley & Jacobi 2000). Fungal identification has mostly

been based on morphological characteristics and host association.

Cytospora notastroma sp. nov. (U.S.A.) ... 785

Use of morphological characters (e.g., stromatic tissues, locular size, shape,

and arrangement, conidiogenesis, and spore characteristics of the conidioma)

for accurate identification of Cytospora spp. is quite problematic (Adams et al.

2005), given the plasticity known to occur with diagnostic features (Adams et al.

2002). Spielman (1983) believed that for taxa within Valsa, most morphological

characters could be modified to some degree. Such morphological variation

combined with a poor understanding of host ranges has hindered species

delimitation (Spielman 1985). Furthermore, because the current identification

system relies on key morphological characteristics forming on bark tissues of

woody plants in nature, no workable system exists for identifying Cytospora

species in vitro or on inoculated host tissues (Adams et al. 2005).

Species causing cytospora canker on Populus tremuloides

During examinations of cankers on aspen stems, we found that another

morphologically distinct Cytospora species is frequently present with

C. chrysosperma. This species forms a conidioma of rosette-like to labyrinthine

locules and leucocytosporoid form (e.g., having entostromatic tissue separated

from host tissues by a dark line of delimiting conceptacle; Adams et al. 2005). It

typically co-occurs with and superficially resembles C. chrysosperma, a species

that forms large labyrinthine locules and cytosporoid form (e.g., without

delimiting conceptacle). In addition to morphology, isoenzyme analyses

and vegetative compatibility studies readily separate isolates of the unknown

Cytospora species from those of C. chrysosperma (Kepley 2009). Almost certainly

the two taxa have been confused in past reports, and cytospora canker on

aspen in Colorado is most likely caused by more than one species of Cytospora,

contradicting what is typically reported in the literature. Three Cytospora species

have been reported on aspen in the southern Rocky Mountains and adjacent

regions: C. nivea (Ellis & Everhart 1892, Gilbertson et al. 1979, Shaw 1973),

C. translucens (Eslyn 1960), and C. chrysosperma. Cytospora nivea from North

America was segregated as “C. pseudonivea” |= Leucostoma pseudoniveum Lat.

N. Vassiljeva] based on the smaller ascospore size compared with European

specimens including the type of C. nivea (Vasilyeva et al. 2008). In the eastern

states, C. leucosperma and C. leucostoma have also been reported on aspen. The

leucocytosporoid form of Cytospora is shared by the unknown species from

Colorado and C. nivea, C. translucens, and C. leucostoma. Additionally, the

unknown species shares with C. translucens a superficially visible ring of the

conceptacle surrounding the disc at the bark surface. This characteristic is well

illustrated and described by Hubbes (1960) for his specimens of C. translucens.

As the presence of C. nivea and C. translucens is so rarely reported on aspen in

the southern Rocky Mountains that the records may be misidentifications, we

have endeavored to verify the reports during this study.

786 ... Kepley & al.

Molecular methods for inferring fungal phylogeny and fungal identification

Molecular phylogenetic analysis has been beneficial in identifying

Cytospora species because many species have overlapping morphology and

the morphology can vary with bark thickness, environment, and host factors

(Adams et al. 2005). Approximately 110 Cytospora species have been described

(Kirk et al. 2008). Many were described based on host range, and host range is

no longer considered a reliable characteristic for species delineation (Adams

et al. 2005). Anamorph and teleomorph morphological data and other types

of phenotypic character data (e.g., culture morphology, growth rates, substrate

utilization, pigment production, isozyme analysis, pathogenicity) when

correlated with molecular phylogenetic studies should provide insight with

respect to taxonomically informative characters. Sorting out species complexes

and properly identifying pathogens are critical in designing control strategies,

plant disease quarantine regulation, breeding disease resistance, understanding

disease biology and epidemiology, and facilitating communication among

plant pathologists, mycologists, and quarantine specialists (O'Donnell et al.

2004). Cytospora canker on aspen in the Rocky Mountains is currently thought

to be caused by a complex of fungi; in the absence of genetic or morphological

analyses of these fungi, our primary goal was to use sequence, cultural, and

morphological analyses to clarify species relationships among Cytospora spp.

on aspen in Colorado.

Materials & methods

Isolates and specimens studied

Information on isolates is listed in TABLE 1 and previous publications (Adams et

al. 2005, 2006). Isolates are usually linked to a herbarium specimen except those from

international culture collections (Adams et al. 2005, 2006). Whenever possible, isolates

believed to represent a common species were chosen from different continents, differing

ecoregions, and different hosts to support international species concepts. The examined

herbarium specimens represented species concepts of C. nivea [=Valsa nivea] and

C. translucens [ Valsa translucens], particularly those from Colorado, the southern

Rocky Mountains, and adjacent regions: NYBG Herbarium J.B. Ellis Collection: #68 &

#1543 V. nivea on P. tremuloides, Short Creek, Custer Co., Colorado, coll. D.E. Cockerell;

#204 V. nivea on P. tremuloides, Ten Mile Creek, Clarke [sic, see Lewis & Clark] Co.,

Montana, coll. EW. Anderson; ISC Herbarium L.H. Tiffany Collection: #320903

V. translucens on P. xcanadensis, Holst St. For., lowa; #326295 V. nivea on P. tremuloides,

Iowa City, lowa; MICH Herbarium: #71305-71315 V. translucens on Salix spp. from:

Little Laramie, Wyoming, USA; South Dakota, USA; London, Ontario, Canada; Vienna,

Austria; Munich and Westphalia, Germany; and Riga, Latvia.

DNA extractions

Mycelia for genomic DNA extractions were obtained from cultures grown for

one week in potato-dextrose broth under ambient light and temperature conditions.

Cytospora notastroma sp. nov. (U.S.A.) ... 787

Approximately 1 cm? of mycelium was ground, extracted, and purified using the

MasterPure™ Yeast DNA Purification Kit (EPICENTRE Biotechnologies, Madison, WI)

following the manufacturer’s instructions. DNA yields were calculated on the basis of

UV absorbance and dilution, and purity estimated by the ratio of UV absorbance at

Peat bgy:

PCR amplification, gel electrophoresis, and sequencing

For PCR studies, template DNA was diluted with sterile, double-distilled water

as needed to provide a final concentration of ca. 200 ng/ul. The primer pairs for

amplification of the ITS1+5.8S+ITS2 region were ITS1f F (forward primer) and ITS4

R (reverse primer) (Gardes & Bruns 1993, White et al. 1990). EF1 F and EF2 R (Geiser

et al. 2004), EF1-526 F and EF1-1567 R (Rehner 2001), and EF1-728 F and EF1-986 R

(Carbone & Kohn 1999) were used for amplification of the EF-la gene. Protocols of

the FailSafe™ PCR System with PreMix choice (Epicentre) were used for amplification

reactions. PCR cycle conditions were those of Touchdown PCR (Korbie & Mattick

2008) that started with an annealing temperature of 60°C and was reduced by 1°C for

each cycle until 50°C was reached.

Amplified samples (10 ul ea.) were fractionated by electrophoresis on 1% agarose

gels buffered in sodium boric acid (Brody & Kern 2004). Electrophoresis was conducted

until major bands in staggered sets of samples were well separated as suggested by Rehner

(2001) for isolating desired PCR products. Upon completion of electrophoresis, gels

were placed in the refrigerator at 4°C to firm up the gel texture followed by rapid band

excision with a sterile scalpel. PCR products in gel slices were purified prior to DNA

sequencing using the QIAquick Gel Extraction Kit (Qiagen Sciences, Inc., Germantown,

MD) according to the manufacturer's instructions when a microcentrifuge was used

for gel extraction. Both purified (exposed to UV light) and unpurified PCR products

were submitted to Macrogen USA, Inc., (Rockville, MD) for sequencing. PCR products

were sequenced in both directions using the same primer pairs that were used in the

amplification reactions.

Phylogenetic analyses

Automatic alignment of the individual ITS1-5.8S-ITS2 rDNA (ITS) and partial

EF-la sequential data sets of sequences was performed in MUSCLE 3.8.31 (Edgar

2004) followed by refinement via direct examination and editing in MEGA version 5.0

(Tamura et al. 2011). Insertions/deletions (indels) and gaps introduced for alignment

purposes were handled as pairwise-deletions, a process that removed the gaps from the

analysis if the gaps had a higher percentage of ambiguous sites than the site coverage

cutoff parameter, which we specified at 95% in MEGA (100% cutoff is no ambiguous

sites) (Tamura et al. 2011).

To infer phylogenetic relationships, 87 taxa datasets were constructed separately

from the ITS and EF-1a sequences. The partition-homogeneity test (ILD test, Farris et al.

1994) was performed in PAUP 4b10 (Swofford 2003) to assess the validity of combining

the two molecular datasets. Concordance of the ITS and EF-1a datasets was evaluated

using 1000 bootstrap replications (Felsenstein 1985) and 1000 random additions of

taxa replicates per partition replicate with tree bisection-reconnection (TBR) branch

swapping and MulTrees active.

788 ... Kepley & al.

TABLE 1. Cytospora and Diaporthe taxa used for phylogenetic studies.

SPECIES

C. abyssinica

C. acaciae

C. annulata

C. berkeleyi

C. ceratosperma

C. chrysosperma [allele 1]

[allele 2]

C. cincta

C. coenobitica

C. curreyi {a}

C. diatrypelloidea

C. diatrypoides

C. eriobotryae

C. eucalypticola

C. eugeniae

C. friesii

C. germanica {a}

C. kunzei/pini

[allele 1] {a}

[allele 2] {a}

C. leucosperma

C. leucostoma

C. magnoliae

C. mali

C. melanodiscus

ISOLATE

CBS116819

CBS117004

CBS468.69

CBS118089

CBS116824

CBS116825

AR98007

CBS116.21

CO_C14

NE_TFR3w

NE_A48

CBS283.74

CBS148.42

CBS116826

CBS120062

NE_Healy1-2

NE_JacLeuco

NE_ESPAInus2

NE_StanMoist

CBS116846

CBS116853

CBS116851

IMI062499

CBS116837

IMI057979

CBS113.81

CBS196.42

ATCC20502

NE_Waterloo

NE_BogueScots

CBS118094

CBS118093

CBS197.42

CBS191.42

CBS116809

NE_RCommon

NE_HigLake4

NE_Lp8

ATCC74091

IMI259790

ATCC56632

NE_Worrall4

NE_JimsLand2

NE_Worrall2b

ORIGIN

Ethiopia

Spain

NY, USA

CA, USA

MA, USA

Netherlands

CO, USA

MT, USA

MI, USA

Netherlands

Switzerland

Australia

AK, USA

CO, USA

AK, USA

India

South Africa

Malaysia

Indonesia

Tanzania

Germany

Switzerland

Canada

MI, USA

Switzerland

NJ, USA

FL, USA

MI, USA

WV, USA

LA, USA

Japan

CO, USA

AK, USA

CO, USA

Host

Eucalyptus globulus

Ceratonia siliqua

Acer rubrum

Eucalyptus globulus

Vaccinium sp.

Fagus sylvatica

Populus tremuloides

Malus xdomestica

Betula verrucosa

Larix sp.

Eucalyptus globulus

Alnus tenuifolia

Eriobotrya japonica

Eucalyptus saligna

Eucalyptus dunnii

Eugenia aquea

Eugenia sp.

Anacardium occidentale

Abies alba

Unknown

Pinus contorta

Pinus strobus

Pinus sylvestris

Picea pungens

Picea glauca

Pinus sylvestris

Taxus baccata

Acer rubrum

Unknown twig

Alnus rugosa

Prunus serotina

Betula alleghaniensis

Magnolia sp.

Malus xdomestica

Alnus tenuifolia

NCBI ACCESSION NO.

[ITS/EF-1la sequence]

AY347352/JX439558

AY347354/JX438559

DQ243804/JX438560

AY347345/JX438576

AY34734/JX438561

AY347351/JX438562

AY188992/AY188991

AY347335/JX438577

JX438635/JX438548

JX438641/JX438549

JX438641/JX438550

AF191170/JX438579

JX438610/JX438578

AF191172/JX438580

AY347368/JX438563

JX438612/JX438584

JX438611/JX438583

JX438613/JX438584

JX438614/JX475137

AY347327/JX438564

AF260265/JX438590

AY347360/JX438591

AY347348/JX438587

AY347344/JX438586

AY347347/JX438589

AY347318/JX438592

AY347325/JX438593

JX438615/JX438594

JX438616/JX438598

JX438617/JX438597

AY347320/JX438595

AY347320/JX438596

AY347332/JX438546

AY347332/JX438547

AY347330/JX438576

AY347339/JX438576

JX463524/JX438599

JX475137/JX438600

AF191177/JX438601

JX438618/JX438602

JX438623/JX438565

AF192326/JX438571

JX438619/JX438608

JX438605/JX438621

JX438606/JX438620

C. mougeotii

C. nitschkei

C. nivea

C. nivea/translucens

C. notastroma

“C. parapersoonii”

C. pinastri {a}

C. pinastri-western NA

C. pruinosa

C. pruinosa {a}

C. punicae

C. rhizophorae

C. sacchari

C. schulzeri

C. sp. undetermined

C. translucens {a}

C. valsoidea

Cytospora sp. {b}

D. ampelina

D. vaccinii

CBS198.50

CBS116854

CBS118562

CBS259.34

CBS109489

NE_OSUAInus

Norway

Ethiopia

South Africa

Switzerland

Russia

OR, USA

NE_Cottonwd16 MI, USA

NE_HigginLake5 MI, USA

NE_NiveaPR

CO_K3

CO_LI

CO_K16

CO_K20

NE_TFR8

NE_TFR5

NE_LCN

NE_T28.1

CBS185.42

CBS118567

CBS118092

CBS196.50

CBS118555

PPRI6334

CBS201.42

CBS199.50

ATCC38475

ATCC66924

CBS160.33

CBS118559

CBS118570

NE_W7b

NE_FlorWP

NE_Pisqua

CBS118566

CBS152.42

NE_JacTissue5

CBS117003

CBS116814

CBS116816

CBS160.32

MI, USA

CO, USA

MT, USA

MI, USA

CA, USA

Switzerland

BC, Canada

Italy

South Africa

Switzerland

Turkey

LA, USA

HI, USA

India

South Africa

MI, USA

wy, USA

WI, USA

NC, USA

South Africa

Switzerland

CO, USA

Indonesia

CA, USA

South Africa

MA, USA

Picea abies

Eucalyptus globulus

Malus xdomestica

Populus nigra

Populus sp.

Alnus tenuifolia

Populus deltoides

Alnus rugosa

Populus xcanadensis

‘Robusta’

Populus tremuloides

Prunus persica

Prunus simonii

Abies alba

Pseudotsuga menziesii

Chamaecyparis sp.

Thuja sp.

Olea europaea subsp.

africana

Olea europaea

Syringa vulgaris

Punica granatum

Rhizophora mangle

Haliclona caerulea

Saccharum officinarum

Malus xdomestica

Alnus tenuifolia

Pinus strobus

Tsuga canadensis

Acacia nilotica

Salix sp.

Eucalyptus urophylla

Sequoia sempervirens

Eucalyptus paniculata

Vitis vinifera

Oxycoccus macrocarpos

Cytospora notastroma sp. nov. (U.S.A.) ... 789

AY347329/JX438566

AY347356/JX438567

DQ243796/JX438607

AF191174/JX438532

JX438624/DQ862035

JX438625/JX438534

JX438626/JX438535

JX438627/JX438536

JX438537/JX438628

JX438631/JX438539

JX438634/JX438538

JX438630/JX438540

JX438629/JX438541

JX438542/JX438633

JX438543/JX438632

AF191181/JX438544

AF191176/JX438545

AY347336'JX438572

AF192551/JX438573

AF192550/JX438574

AF192311/JX438575

DQ243790/JX463522

DQ243789/JX438581

DQ243801/JX438582

JX438622/JX438568

DQ996040/JX438609

DQ092502/JX438609

DQ243811/JX438569

DQ243792/JX438603

DQ243802/JX438604

JX438554/JX438638

JX438639/JX438555

JX438640/JX438556

DQ243800/JX438557

AF191182/JX438552

JX438637/JX438553

AF192312/JX438570

AY347340/JX438585

AY347365/JX438585

JQ038888/AY 745084

GQ250326

{a} = Reference cultures deposited by Défago as standards for her 1935 European species concepts.

{b} = Valsa eucalypti Cooke & Harkn.

Collection acronyms: ATCC = American Type Culture Collection, Manassas VA, USA; AR = Amy Rossman

collections, USDA-ARS, Beltsville MD, USA; CBS = Centraalbureau voor Schimmelcultures, Utrecht,

The Netherlands; CO = William R. Jacobi collections, Colorado State University, Ft. Collins CO, USA;

IMI = International Mycological Institute, CABI Bioscience, Egham, Surrey, UK; NE = Gerard Adams

collections, University of Nebraska, Lincoln NE, USA; PPRI = South African NCE, Plant Protection

Research Institute, Pretoria, South Africa.

The arbitrarily numbered alleles indicated under C. chrysosperma and C. kunzei/pini refer to two EF-1la

sequences present in the hyphae.

790 ... Kepley & al.

Evolutionary analyses were conducted using MEGA. Stochastic models for

estimating evolutionary distance between sequences with maximum likelihood (ML)

(Felsenstein 1981) were calculated for the combined data sets of DNA nucleotides of the

ITS and EF-1la regions. The optimal models for the pattern of nucleotide substitution,

the evolutionary rate differences among sites, and indel evolution were selected for

phylogenetic reconstructions (Nei & Kumar 2000). The EF-la dataset was treated as

non-coding rather than protein coding nucleotides because only a small percentage

of the sequence included an exon region. A phylogeny tree was inferred using the

ML heuristic method and the Close-Neighbor-Interchange search method with

initial random addition of 100 trees in MEGA. Strains of Diaporthe species including

D. vaccinii GQ250326 strain CBS160.32 and D. ampelina JQ038888 strain STEU 7005,

and AY745084 strain OH-9 served as outgroup for ITS and EF-1a data sets. Nucleotide

sequences of species of Valsa and Cytospora deposited in NCBI by other researchers

were not included in this study because of problems in verifying the identification. Final

results were summarized as the ML tree with the best negative log likelihood values,

branch support values of clade credibility, and bootstrap confidence limits calculated

with 1000 bootstrap replications.

Morphological studies

For examination of fruiting bodies on bark (natural state), conidiomata and

ascostromata were excised from aspen tissues by cutting deep enough into the bark

tissues to remove entire fruiting bodies. For examination of fruiting bodies from

cultural specimens, mature pycnidia-like conidiomata (those oozing spore masses)

were cut out with a portion of the modified Leonian’s agar medium (20 ml per 9 cm

Petri dish; Leonian 1923). These cultures were seven weeks old and grown at 25°C in

the dark. Protocols for fixing, embedding, and sectioning were modified from Adams

et al. (2005). Measurements of characteristic structures from fruiting bodies on bark

were derived from 20 observations when possible. Distilled water and several stains:

phloxine-KOH, lactophenol-cotton blue, and Melzer’s solution, were used as mounting

media for observing general morphological characteristics, e.g., stromatal tissues,

perithecial wall tissues, conidiogenous cells, asci, and spores. Sections were stained with

0.001% aqueous toluidine blue.

Cultural characteristics were determined in triplicate from isolates (C1, C14, K3,

K16, K20, and L1) grown in 9 cm Petri plates (wrapped with wax-film) containing 20

ml modified Leonian’s agar under 12 hours continuous light and 12 hours continuous

darkness at 25°C for 28 days. Colors were determined according to Rayner (1970).

Measurements of conidiomata, conidia, and hyphae were based on 20 observations

each per isolate. Distilled water, and the aqueous stains listed prior were again used as

mounting media for microscopic examinations.

Results

Molecular phylogenetic characterization

PCR products of the ITS were approximately 540 base pairs (bp) and those

of the EF-1la were approximately 300 bp, but the length of the intron varied

among isolates. As the partition homogeneity test indicated phylogenetic

Cytospora notastroma sp. nov. (U.S.A.) ... 791

congruence between the two data sets (p = 0.003), the data sets were combined.

Alignment of the combined ITS and EF-1a sequence data set included 87 taxa

and 633 sites (excluding gaps introduced for alignment) with 490 parsimony-

informative characters out of 1121 total sites (ITS = 626 sites with gaps, EF-

la = 495 sites with gaps). Maximum parsimony analysis yielded 12 equally

parsimonious trees with tree length of 1370 steps and scores of CI (consistency

index) = 0.391971, RI (retention index) = 0.719150, and RCI (rescaled

consistency index) = 0.281886 (for all sites) (Farris 1989). The final tree was

drawn in MEGA and Microsoft PowerPoint version 2010 (Microsoft, USA).

Evaluation of models of nucleotide substitution in MEGA yielded the

General-Time-Reversible (GTR) model (Tavaré 1986) as optimal. Substitution

pattern and rates of evolution for DNA nucleotide sequences were estimated

under the GTR model with a discrete Gamma distribution (GTR+G) used to

model evolutionary rate differences among sites (5 categories, 4 nucleotides,

and indel gaps as partial-deletions at 95%). The 50% majority rule bootstrap

values are displayed on the strict consensus tree (PLATE 1) with tree-length (as

sum of branch lengths, SBL) of 1.97608505, Ln likelihood of -7741.06, and a

transition/transversion ratio of 1.5418. Terminal isolates on the phylogenetic

tree were designated with names in regular fonts that include location and in

italic fonts for host of origin followed by a number. The number represents

a unique DNA sequence that corresponds to one or more isolates given in

TABLE 1 or other publications. Names to the right of the vertical bars represent

described species and species complexes.

Phylogenetic analysis (PLATE 1) strongly supports (100% bootstrap

confidence) a cluster ofisolates from aspen within a holophyletic (monophyletic)

clade. We describe the unique morphology of the fungi in this clade as a new

species, C. notastroma. It forms a sister group to the clade containing C. nivea,

C. translucens, C. leucostoma, and “C. parapersoonii’ [= Leucostoma parapersoonii

G.C. Adams et al.]. Two subgroups are inferred and supported by 94-98%

bootstrap confidence within /notastroma clade. Colorado isolates from aspen

exhibiting dieback or mortality cluster in both C. notastroma subgroups and

also in the clade of C. chrysosperma with 100% bootstrap support. Cytospora

nivea and C. translucens are not readily distinguished from one another by the

combined ITS and EF-la sequence and intermediate isolates appear to exist

(e.g., Oregon_Alnus1). “Cytospora pseudonivea” represented by isolate Russia_

Populus| (AR 3413 = CBS 109489; Vasilyeva et al. 2008), is conspecific with

Switzerland_Populus1, an isolate that G. Défago (1935) designated as typical

for the species concept of C. nivea in Europe.

A well-supported clade of mixed nomenclatural species is treated as

representing a single species complex that is named after the oldest or most

792 ... Kepley & al.

Switzerland Populus nigra ([Défago]

South Africa Populus sp.

Russian Poptits sp.

OR USA Alnus tenuffolia

Switzerland Sa/ix sp. [Défago]

g5mCO USA Sajix sp.

CA USA Prunus simonii

MIUSA Prunus persica

MIUSA Prunus serotina

WY USA Setula alleghaniensis

FL USA unknown branch

MI USA A/nus rugosa

87 Ml USA Populus daltoides

SOREN USA Populus x canadensis ‘Robusta’

fl USA Affus rugosa

CO USA Popiius tremixoidas

99mC0 USA Populus tremuloides

CO USA Popitue tremuofdes

CO USA FPopitus trenisoides

7 CO USA Panuius tremiuoides

gm CO USA Fopiulue fremuloldas

WI1USA Pinus strobus

NC USAT suga carolinjana

85ySouth Africa O/ea europaea subsp. africana

South Africa O/ea europaea

Switzerland Syringa vidgaris [Défago]

WY USA A/nus tenuifolia

CO USA Alnus tenuifolia

08 CO USA Alnus tenuifolia

88 AK USA A/nus tentifolia

59mMI USA Picea pungens

MI USA Picea glauca

MIUSA Pinus sylvestris

MI USA Pinus strobus

BO Canada Pinus contorta

87 Switzerland Pinus sy/vestris-1 [Défago]

4004 Switzerland Pinus sy/vestris-2 [Défago]

Switzerland Larixsp. [Défago]

MI USA Malus xdomestica

AK USA Afnus tenuifolia

CO USA Al/nus tenuifolia

CO USA A/nus tenuifolia

AK USA A/nus tenusfolia

India Erfobotrya japonica

Japan Malus xdomestioa

Turkey Punica granatum

CO USA Populus tremuloides

MT USA Paopults fremuflojdes-|

NIT USA Popttus trenniojdes-2

100m South Africa Ma/us domestica

MI USA Malus domestica

100gmm NY USA Acer rubrum

109 NJ USA Acer rubrum

Switzerland Taxus baccata

Norway Picea abies

Switzerland Sa/ixsp. [Défago]

Germany Aéjes alba

Netherlands Setula verrucosa

Switzerland Aéjes a/ba [Défago]

Indonesia Eucalyptus urophylia

63guMalaysia Eugenia aguea

Indonesia Eugenia sp.

96)

86)

100

100)

50)

100

100}

66)

South Africa Eucalyptus satigna

South Africa Eucalyetus dunnij

91gCA USA Eucalyptus globulus

CA USA Eucalyptus globulus

4008 § Australia Eucalyptus globulus

gg@ Australia Etrcalyptus globulus

4100g= Netherlands Fagus sy/vatioa

MA USA Vacoinium sp.

BO Canada Chamaecyparis sp.

BC Canada Pseudotsuga menziesii

ltaly TAujasp.

100) CAUSA Eucalyptus globosa

CAUSA Sequoia sempervirens

LA USA Magnolia sp.

Spain Ceratonia siligua

South Africa Acacia nilotioa

100m Ethiopia Etca/yptus sp.

Ethiopia Eucalyotus sp.

Ethiopia Eucalyptus sp.

LA USA Rhizophora mangle

400M2HI USA Hafliclona caertiea

India Saceharum officinarum

MA USA Oxycocetus macrocarpos

98)

South Africa Vitis vinifera

Tanzania Anacardium occidentale

Cytospora nivea

complex

| Cytospora translucens

| “Cytospora parapersoonii

”

Cytospora leucostoma

Cytospora notastroma

| Cytospora sp. undetermined

Cytospora pruinosa

Cytospora sp. undetermined

Cytospora melanodiscus

Cytospora kunzei & C. pini complex

Cytospora cincta & C. curreyi complex

Cytospora diatrypoides

| Cytospora mali & C. eriobotryae

Cytospora punicae

Cytospora chrysosperma

| Cytospora schulzeri

Cytospora leucosperma

Cytospora mougeotii

ie ied a ermanica

ips pore riesii

ytospord coenobitica

Cytospora pinastri

Cytospora valsoidea

Cytospora eugeniae

| Cytospora eucalypticola

| Cytospora berkeleyi

| Cytospora diatrypelloidea

| Cytospora ceratosperma

Cytospora pinastri -

western North America

Cytospora sp. [=Valsa eucalypti]

Cytospora magnoliae

Cytospora acaciae

Cytospora sp. undetermined

Cytospora abyssinica

Cytospora nitschkei

Cytospora rhizophorae

Cytospora sacchari

Diaporthe vaccinii

Diaporthe ampelina

Cytospora notastroma sp. nov. (U.S.A.) ... 793

common species until further study reveals otherwise (Adams et al. 2005,

2006). For example, the clade referred to as the C. chrysosperma species

complex in earlier studies (Adams et al. 2006) was shown to include specimens

identified as C. eutypelloides Sacc., C. hariotii Briard, C. minuta Thiim., and

C. tritici Punith.

Polytomy has been lessened in the combined ITS and EF-1la tree (PLATE

1) compared to single gene trees (see Fig. 1 in Adams et al. 2005); however

evolutionary relationships still cannot be fully resolved among several clades.

Taxonomy

Cytospora notastroma Kepley & EB. Reeves, sp. nov. PLATES. 2, 4, 6C,D, 7

MycoBAnk MB 801154

Differs from Cytospora chrysosperma by its prominent dark conceptacles visible from

the bark surface delimiting the stroma and disc of both ascostromata and conidiomata.

Type: USA, Colorado: Upper Poudre Canyon east of Cameron Pass on dead bole of

Populus tremuloides in grove, 20 Aug 2004, collectors J. Kepley & EB. Reeves (Holotype,

NEB318541).

ETYMOLOGY — notastroma, a shortened form of the Latin word notabilistromatica,

referring to the notable encircling zone of conceptacle tissue superficially visible around

the disc of the ascostroma and the disc of the conidioma.

ASCOSTROMATA immersed in bark, erumpent, ovoid to circular 2.0-3.0

x 1.2-1.8 mm, leucostomoid circinateous, 6-15 perithecia arranged

circinately in well developed orangish, cinnamon, olive-gray to creamish-

white entostroma composed of cells forming textura angularis and intricata,

conceptacles prominent, olive-black to black, apparent on the surface of the

bark. Discs prominent, snowy-white to grayish-white, nearly flat, circular to

ovoid 0.4-0.55(-0.65) mm diam, furfuraceous, composed of cells forming a

textura angularis and intricata, 2-10 laterally to vertically inserted ostioles,

surrounded by an ovoid to circular zone of olive-black to black conceptacle

tissue apparent on the surface of the bark. OsTIOLEs olive-black to black

(45-)60-100(-120) um diam, nearly level to slightly above disc surfaces.

PERITHECIA Olive-black to black, globose (0.25-)0.3-0.40(-0.50) mm diam,

inclined, walls of textura epidermoidea. Asci free, clavate to obclavate (33-)

37-43 x 8-11 um, apical apparatus non-amyloid, 8-spored. AscosPOREs

biseriate, allantoid, thin-walled, hyaline (salmon-colored in mass), and

PiateE. 1. Reconstructed phylogeny of Cytospora species based on maximum likelihood analysis

of non-coding ITS and EF-la DNA sequences. ‘The tree is a bootstrap consensus tree of 50%

majority rule with —Ln likelihood score = 7741.06, SBL tree length = 1.97608505 steps). Branch

lengths correspond to inferred genetic distances with the scale bar representing a 2% nucleotide

divergence. The numbers at the nodes represent bootstrap support values based on 1000 resamplings

(values >50% are shown).

794 ... Kepley & al.

PLATE 2. Cytospora notastroma ascostromata. A. Erumpent ascostroma with prominent ovoid

snowy-white disc and emerging black ostioles (white arrow), obscure black conceptacle tissue

(yellow arrow), and black conceptacle (black arrow). B. Excised ascostroma showing teleomorph

and anamorph in same stroma; perithecium (red arrow) and locular chambers of anamorph (green

arrow). C. Horizontal cross section showing circinate perithecia surrounded by well developed

cinnamon to creamish-white entostroma, globoid perithecium (red arrow), and conceptacle (black

arrow). D. Vertical section with ostiole emerging through disc (white arrow), black conceptacle

tissue surrounding the disc (yellow arrow), laterally inclined perithecium surrounded by

entostroma (red arrow), and conceptacle (black arrow). E. Clavate asci (with ascospores) floating

freely in perithecial centrum. Scale bars: A~D = 1.0 mm, E= 15 um.

aseptate (7.5-)8.0-9.5 x 1.5-2.0 um. ANAMORPH conidiomata usually

interspersed amongst teleomorphs but sometimes present in the same stromata

as the teleomorphs. CONIDIOMATAL STROMATA immersed in bark, erumpent,

labyrinthine to rosette-like leucocytosporoid, ovoid to circular 1.5-2.5 x

1.0-1.5 mm, conceptacles prominent, olive-black to black. Discs prominent,

white to grayish-white, nearly flat, circular to ovoid 0.25-0.40 mm diam,

furfuraceous, composed of amorphous material and cells forming a textura

angularis, 1-3 ostioles, surrounded by an ovoid to circular zone of olive-

black to black conceptacle tissue. OsTIOLEs olive-gray, olive-black to black,

75-150(-170) um diam, nearly level to slightly above disc surfaces. LocULES

multi-chambered, subdivided by invaginations into regular to irregular radially

arranged chambers sharing common walls, 100 x 300 um diam, surrounded

Cytospora notastroma sp. nov. (U.S.A.) ... 795

PLATE 3. Cytospora chrysosperma ascostromata. A. Vertical section with ostiole at disc margin

(white arrow); stromatic tissue below disc (black arrow); globose perithecium (red arrow)

lacking well developed entostroma (note lack of conceptacle delimiting stroma). B. Erumpent

ascostroma with emerging ostioles (white arrow) at margin of prominent tan to gray circular disc.

Scale bars: A-B = 0.5mm

with well developed cinnamon, olive-gray to creamish-white stromata of

textura angularis and textura intricata. CONIDIOMA CONIDIOPHORES hyaline

and branched 6.0-10.0 x 1.0-1.5 um, inclusive of phialides, arise from basal

cells (2.0-)3.0-4.5 x 1.5-3.0 um, embedded in a continuous gelatinous matrix.

CONIDIOMA CONIDIOGENOUS CELLS enteroblastic phialidic, cylindrical,

tapering to the apices, minute collarettes. CONIDIOMA CONIDIA hyaline

(salmon-colored in mass), eguttulate, allantoid, aseptate 3.0-5.0 x 1.0 um.

Phialocephala-like hyphomycetous anamorph formed in culture on modified

Leonian’s agar (20 ml/9 cm Petri dish) at 25°C with 12 hours continuous light

and 12 hours continuous darkness; synanamorphs located in older regions of

young cultures (7-10 d old) where pigmentation is forming and hyphae are

aggregated into ball-like clusters. HyPHOMYCETOUS CONIDIOPHORES arising

from main hyphae, mononematous, darkly pigmented, variable in length (short

to quite long) and numbers of septa (three or more), often subtended by basal

cells. Branching variable (dichotomous to three or more), occurring in series,

initiated at or near septa. HYPHOMYCETOUS CONIDIOGENOUS CELLS phialidic,

cylindrical (5.0-12.0 x 2.0-3.0 um), taper to apices. HYPHOMYCETOUS CONIDIA

hyaline, allantoid, aseptate.

CULTURE CHARACTERISTICS — Colony growth olivaceous-black (top

and reverse) on modified Leonian’s agar (20 ml/9 cm Petri dish) after 28 d

at 25°C with 12 hours continuous light and 12 hours continuous darkness.

Hyphae generally appressed and growing down into the agar. Pycnidium-

like conidiomata greenish-black, often covered with white, smoke-grey to

olivaceous-grey hyphae, and typically forming vertically oriented beaks/necks.

Exuded cirrhi milky-white.

796 ... Kepley & al.

PLATE 4. Cytospora notastroma conidiomata. A. Interspersed conidiomata of C. notastroma

(white arrows) and C. chrysosperma (red arrows). B. Erumpent conidioma; well-developed black

conceptacle tissue (yellow arrow); prominent circular grayish-white disc with two emerging black

ostioles (white arrow); black conceptacle (black arrow). C. Horizontal cross-section; rosette-like

multi-chambered locules surrounded by well developed creamish-white to olive-gray stromatic

tissues (green arrow); conceptacle (black arrow). D. Vertical section; conceptacle tissue surrounding

the disc (yellow arrow); rosette-like multi-chambered locules sharing common walls surrounded

by well developed stromatic tissues (green arrow); conceptacle delimiting the stroma (black arrow).

E. Conidiogenous cells and spores; basal cell subtending branching phialidic conidiophores

(left-most arrow); spore at apex of phialide (right-most arrow). FE. Hyaline, eguttulate, allantoid, and

aseptate conidia. Scale bars: A-D = 1.0 mm, E= 8 um, F=2 um

ADDITIONAL MATERIAL EXAMINED: USA, COLORADO: Pingree Park, on dead bole of

Populus tremuloides, 25 May 2004, collectors J.B. Kepley & EB. Reeves (NEB318542,

NEB318543).

Hosts — Populus tremuloides Michx., P. deltoides W. Bartram ex Marshall, P. xcanadensis

Moench ‘Robusta’ [P. nigra x P. deltoides], Alnus incana subsp. rugosa (Du Roi) R.T.

Clausen

DIsTRIBUTION — USA (Alaska, Colorado, Montana, Michigan)

Notes — Conidiomata of C. notastroma are more common on aspen than the

teleomorphs, although they occasionally occur in the same stroma. Conidiomata

often co-occur interspersed amongst C. chrysosperma conidiomata (PLATE 4A)

and are of similar size and external shape. However, the conidiomata of the

two species are readily distinguished, as those of C. notastroma have prominent

olive-black to black conceptacles visible on the bark surface that delimit the

stroma including the white to grayish-white disc (PLATE 4B). Entostromatic

tissue surrounding the locules is cinnamon, olive-gray to creamy-white in color,

and can be variable in terms of development. Well-developed conceptacle tissue

causes conidiomata to take on a distinctive target-like appearance (PLATE 4).

Cytospora notastroma sp. nov. (U.S.A.) ... 797

PAD

Cy Bilas sie

A*,

PLATE 5. Cytospora chrysosperma conidiomata. A. Erumpent circular conidiomata (note lack

of conceptacles and conceptacle tissues). B. Conidiomata with prominent gray, olive-gray to

olive-black circular discs, each with single black ostiole (white arrows). C. Conidioma with

labyrinthine multi-chambered locules surrounded by gray to olive-green stromatic tissues (green

arrow). D. Vertical cross section with labyrinthine multi-chambered locules sharing common

walls surrounded by dark stromatic tissues (green arrow); stroma of the conidioma is better

developed than entostroma of the ascostroma); note lack of conceptacle and conceptacle tissue.

Scale bars: A-D = 1.0 mm

In contrast, C. chrysosperma (PLATE 5) lacks conceptacles delimiting the

conidiomata and conceptacle tissues surrounding the discs; the discs are gray,

olive-gray to olive-black.

Ascostromata of C. notastroma are similar in size and shape to those of

C. chrysosperma but have a prominent dark conceptacle visible from the bark

surface delimiting the stroma and disc (PLATE 2). Such zone lines become less

prominent with increasing depth and can vary among specimens or between

ascostromata on the same specimen, ranging from well developed to somewhat

obscure. When well developed, the conceptacle tissue gives the ascostroma

a distinct target-like appearance. The zone line tends to extend less deeply

into the fruit body in comparison to similar tissues found in conidiomata.

798 ... Kepley & al.

PLATE 6. Conidiomata in vitro. Cytospora chrysosperma. A. Vertical section of conidioma of isolate

C 14 with multi-lobed locular chambers (red arrow) and layer of conidiophores (black arrow)

interspersed with long gelatinous hyphal cells lining the walls of the locular structure. B. Horizontal

cross section through conidioma of isolate C 14 showing the complex labyrinthine locular

structure. Cytospora notastroma. C. Vertical section of conidioma of isolate 3 with invaginations

lined with conidiophores (yellow arrow) and conidiophores lining the surface of the pycnidium

(black arrows). D. Horizontal cross section through conidioma of isolate K3 showing the simple

structure without an enclosed multi-lobed arrangement of locules. Scale bars: A-D = 2.0 mm

Ascostromatal discs are snowy-white to grayish-white and powdery or flaky.

The ostioles emerge through the disc and may be scattered, in rows, or in

a circular arrangement on the disc surface; as few as two or as many as 10

ostioles may be visible. Perithecia are surrounded by well-developed orangish,

cinnamon, olive-gray to creamish-white entostroma (PLATE 2c).

Ascostromata and conidiomata of C. translucens and C. nivea (including the

smaller spore form, “C. pseudonivea”) share with C. notastroma the leucostomoid

form with perithecia embedded in entostroma and with delimiting conceptacle

forming the dark zone lines. Cytospora translucens, but not C. nivea, shares the

dark zone line ring of conceptacle surrounding the disc that is visible on the

plant surface through a thin epidermis. Many other characters of C. translucens

and the small form of C. nivea overlap those of C. notastroma. On average,

C. translucens forms much smaller ascostromata with fewer perithecia and

Cytospora notastroma sp. nov. (U.S.A.) ... 799

larger ascospores. The smaller form of C. nivea also generally forms smaller

ascostromata. The conidiomata of C. translucens are smaller with less complex

labyrinthine chambers than C. notastroma, whereas those of C. nivea have

more complex labyrinthine chambers than C. notastroma.

Cytospora chrysosperma ascostromata lack the delimiting conceptacle

and discs are not snowy-white (PLATE 3) as compared with C. notastroma,

C. nivea, and C. translucens. Ascostromatal discs of C. chrysosperma are dark

and sometimes obscured by perithecial beaks. Ostioles are circinately arranged

around the margin of the disc, and beaks are often swollen and may be fused

with adjacent beaks.

In vitro, conidiomata produced by C. notastroma are simple with

invaginations (PLATE 6). In contrast, conidiomata produced by isolates of

C. chrysosperma have a complex structure comprised of multi-lobed locular

chambers enclosed within the conidioma (PLATE 6). Cultures of C. notastroma

(isolates K3, K16, K20, L1) differ distinctly from those of C. chrysosperma

(isolates C1 and C14; PLaTE 7) on common media like potato dextrose or

malt extract agars. On Leonian’ss medium C. notastroma cultures tend to be

dark with hyphae generally appressed and typically growing within the agar,

and conidiomata are often covered with white, smoke-grey to olivaceous-grey

hyphae. In contrast, C. chrysosperma cultures are light in color and zonate,

with aerial hyphae dense and quite tall; numerous pycnidia-like conidiomata,

some covered with white, buff to honey-colored hyphae, form primarily in the

darker zones. Cultures of C. notastroma do not show the scalloped concentric

rings of growth at irregular distances within the colony as described for

“C. pseudonivea” (Vasilyeva et al. 2008, as Leucostoma pseudoniveum).

Hyphal tips collected from five-day-old cultures and mounted in water

showed that isolates of C. notastroma had wide hyphae (4.0-5.5 um _ diam.)

that were bead-like and wavy in appearance. Additionally, a bursting of hyphal

tips was observed in young (ca. seven days old) cultures. Hyphae produced

by C. chrysosperma isolates are considerably narrower (1.5-2.5 um diam.) and

uniformly straight and no lysing was observed. Young (i.e., ca. 7-10 days old)

cultures of C. notastroma produced a Phialocephala-like anamorph (PLATE 7).

These synanamorphs were located in older regions of cultures just beginning to

form pigmentation where hyphae aggregated into ball-like clusters.

Discussion

A main objective of the present study was to use molecular techniques

to sort out the “species complex” associated with cytospora canker of aspen

in the Rocky Mountains and adjacent regions of the Great Plains. Based

upon evolutionary analyses of ITS and EF-la nucleotide sequence data sets,

800 ... Kepley & al.

PiaTE 7. Cultural characteristics. A. Cultures of Cytospora chrysosperma isolates (C1 and C14) are

light in color and zonate (conidiomata forming in the darker zones) unlike isolates of C. notastroma

(K3, K16, K20, T1). Cytospora notastroma. B. Phialocephala-like synanamorph produced by

isolates K16 and K20. Dichotomous branching of conidiophores (black arrow); clusters of phialidic

conidiogenous cells (yellow arrows). C. Phialocephala-like synanamorph produced by isolates

K16 and K20. Septa (black arrows); phialidic conidiogenous cells of conidiogenous apparatus

(yellow arrows). Scale bars: B—C = 5.0 um.

isolates of C. notastroma were strongly supported as a holophyletic clade with

eastern isolates from Populus deltoides (eastern cottonwood), P. xcanadensis

‘Robusta’ (Carolina poplar), and Alnus incana subsp. rugosa (speckled alder) in

Michigan. This implies that the Colorado and Michigan isolates are members

of one widespread (phylogenetic) species occurring on at least two host genera.

The other species isolated from aspen was identified as C. chrysosperma based

on morphology and homologous sequences. Cytospora chrysosperma is more

prevalent on aspen than C. notastroma and the pathogenicity of the latter has

not yet been documented.

Study of historical herbarium specimens collected in the Americas and the

opportunity to collect similar specimens from similar locations and compare

morphology and DNA homology of the ITS and EF-la sequences when

possible have led us to conclude that C. notastroma has been confused with

C. nivea in eastern and western North America. For example, our study of

herbarium specimens indicates that several determined as C. nivea by J.B. Ellis

(Ellis & Everhart 1892) and H. Kern (Kern 1957) represent C. notastroma. Ellis

& Everhart (1892) and Kern (1957) had noted that the dimensions of stromata,

perithecia, asci, ascospores, and conidia of the material from the western

hemisphere were consistently smaller than those of European material and the

type specimen of C. nivea (Vasilyeva et al. 2008).

Cytospora notastroma also shares with C. translucens the encircling

conceptacle superficially visible under the thin epidermis and bark of tree

stems and branches, and the leucostomoid character of the stroma. The fact

that the visible encircling conceptacle may be absent in collections on thicker

bark may have led to misidentification as C. nivea. Furthermore, comparison

of ITS sequence of specimens identified as C. nivea and C. translucens in

Cytospora notastroma sp. nov. (U.S.A.) ... 801

NCBI GenBank accessions from Iran and China supports our conclusions that

C. nivea and C. translucens are difficult to differentiate genetically. The collection

of an isolate from Salix sp. in Colorado that is conspecific with European

isolates identified as C. translucens by Défago (agreeing in morphology and

DNA sequence homology) may be fortuitous but is also due to our concerted

efforts toward discovering and verifying the species presence. As C. translucens

may not be present on aspen and is rarely reported on Salix spp. in the Rocky

Mountains, it may not be endemic.

We believe the smaller forms of C. nivea described by Ellis & Everhart

(1892), Gilman et al. (1957), and Kern (1957) from Colorado, Montana, Iowa,

and Michigan most likely represent C. notastroma. We assume many similar

collections will be homologous in DNA sequence to C. notastroma, but we

cannot exclude the possibility that some may have sequence homology to

C. nivea. Resolving interrelationships among a well-supported clade and

neighboring clades, unfortunately, is somewhat problematic in Cytospora

because of polytomies occurring in the phylogram may result from either much

homoplasy across the genus (Sanderson & Donoghue 1989, Farr et al. 2002)

or “very real polytomies in the tree” (Hall 2008). Farr et al. (2002) reported

problems with homoplasy across the related diaporthalean genus, Diaporthe.

Cytospora notastroma is somewhat unusual in that the anamorph occasionally

forms within the same stromatic tissues as the teleomorph; nonetheless, this does

occur in a few species such as C. cincta Sacc. and C. massariana Sacc. (Adams

et al. 2005). In both nature and culture, C. notastroma specimens are readily

distinguished from those of C. chrysosperma, which increases the usefulness

of our research for forest and plant pathologists. Cultural variation among

C. notastroma isolates is relatively negligible, although isolate K3 produced

more lobate growth, was less darkly pigmented, and did not produce pycnidia-

like conidiomata with beaks. Additionally, isoenzyme studies place isolate K3

in a different subgroup from isolates K16, K20, and L1 with a genetic similarity

(based on Jaccard’s coefficient) of 47% (Kepley 2009). Our phylogeny clusters

Colorado isolate K3 more closely with the eastern isolates from speckled alder,

eastern cottonwood, and P. xcanadensis ‘Robusta of Michigan. However, as

isolate K3 was collected from an urban landscape, the geographic origin of the

host is unknown. In this sexually reproducing species, isoenzyme profiles would

be expected to differ between populations east of the Mississippi and those of

the southern Rocky Mountain region. Obviously, further genetic, molecular,

and morphological studies should be conducted for this phylogenetic cluster of

taxa in order to resolve possible sympatric species.

Although the occurrence of two synanamorphs (albeit in culture) was quite

unexpected, two other studies have reported similar observations: Helton &

802 ... Kepley & al.

Konicek (1961) described “naked conidiophores” arising from dichotomous

branching of hyphal tips in isolates from stone fruit trees; Hildebrand (1947)

reported a similar occurrence with isolates of C. leucostoma from peach trees

(our phylogenetic analyses indicate the C. notastroma isolates as distinct but

closely related to C. leucostoma). The naked conidiophores described in the

latter studies apparently were not Phialocephala-like in morphology, and

whether naked conidiophores or a Phialocephala-like anamorph occur in the

field is not known.

With time, methods for identifying fungi and assessing their phylogeny

will continue to improve, and databases will continue to enlarge. Datasets

of 28S rDNA, B-tubulin, and histone 3A sequences are nearly complete for

100 Cytospora taxa and will, it is to be hoped, remove the polytomies in the

current phylogeny. It is important to note that phylogenetic analyses allow only

formulation of inferences and hypotheses. Hall (2008) summarizes this quite

succinctly, stating that “the right tree doesn't exist” and “all methods implicitly

acknowledge that the trees produced are only a subset of the possible trees that

are consistent with the data.” Recognizing new Cytospora species is important

when they are encountered in association with major plant epidemics. Given

their endophytic and pathogenic nature and broad host range, future studies are

needed to determine the biology, ecology, and physiology of these organisms.

Acknowledgments

We gratefully acknowledge the technical help from Professor Mursel Catal, and

we thank Professor James J. Worrall and the US Forest Service for their concern for

forest health and protection and Dr. Amy Rossman for guidance on nomenclature. We

especially appreciate the help of Professor emeritus Robert Kaul and Thomas Labedz of

the Charles E. Bessey Herbarium, Professor Alan Prather and Dr. Alan Fryday of the

Beal—Darlington Herbarium, and other herbarium curators and supporters. Professors

Ned Tisserat and James J. Worrall provided helpful manuscript reviews for which we

are grateful.

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MY COTAXON

http://dx.doi.org/10.5248/130.807

Volume 130, pp. 807-813 July-September 2015

Two new species and a new record of Hansfordia

from China

YUE-MING Wu*”", HONG-FENG WANG", JUN-JIE XU?, & TIAN-YU ZHANG?”

' Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

? Key Laboratory of Agricultural Microbiology, Shandong Province, Taian, 271018, China

* College of Life Sciences, Linyi University, Shandong Province, Linyi, 276005, China

4 Shandong Agricultural University Fertilizer Science & Technology Company Limited,

Taian, 271000, China

*CORRESPONDENCE TO: tyzhang1937@yahoo.com.cn

ABSTRACT —Two new species, Hansfordia rosea, H. ginghaiensis, and a new record of

H. caricis are described and illustrated from soil in China. A key to the 13 accepted Hansfordia

species is also provided. The specimens (dried cultures) and living cultures are deposited

in the fungal reference collections of Shandong Agricultural University, Plant Pathology

(HSAUP), and the Institute of Microbiology, Academia Sinica (HMAS).

Key Worps — dematiaceous hyphomycetes, soil fungi, taxonomy

Introduction

The genus Hansfordia was established by Hughes (1951) with H. ovalispora

as type species. The genus is characterized by solitary conidia produced in

succession on conspicuous denticles (Barron 1968). Arx (1982) regarded

this genus as a synonym of Dicyma Boulanger, but that view has not gained

acceptance (Kirk et al., 2008); currently the two genera are distinguished by

the presence of separating cells in Hansfordia and their absence in Dicyma.

IndexFungorum lists 22 taxa, of which 11 are accepted by SpeciesFungorum

(2015) while Seifert et al. (2011) estimated that the genus might contain only

7 valid species.

During a survey of soil dematiaceous hyphomycetes in China, several

unusual species of Hansfordia were collected, of which two are described and

AY.M. Wu and H.F. Wang contributed equally to this work.

808 ... Wu, Wang, & & al.

illustrated here as new and one represents a new record for China. A key to all

13 Hansfordia species is provided below.

The three species covered here are described and illustrated from cultures

grown on potato dextrose agar (PDA) according to Hughes (1951). Specimens

(dried cultures) and living cultures were deposited in the Herbarium of

Shandong Agricultural University, Plant Pathology, Tainan, China (HSAUP)

and the Herbarium of Institute of Microbiology, Academia Sinica, Beijing,

China (HMAS).

Fic. 1. Hansfordia rosea (ex holotype HSAUP II ,,2794).

Conidia, conidiophores, and conidiogenous cells. Scale. bar = 20 um.

Hansfordia spp. nov. (China) ... 809

Hansfordia rosea J.J. Xu & T.Y. Zhang, sp. nov. FIG. 1

MycoBank MB 808391

Differs from Hansfordia pulvinata by its bigger conidia.

Type: China, Zhejiang Province: Hangzhou, National Nature Reserve of Mount Tianmu,

from a forest soil, 11 Sept. 2004, J.J. Xu (Holotype, HSAUP II ,,2794; isotype, HMAS

196281; ex-type cultures, HSAUP II_,, 2794-1, HMAS 196284).

ErymMo.ocy: The epithet refers to the color of this species.

CoLonigs on PDA after 7 days at 25 °C effuse, 40-50 mm diam., velvety, pink

to pale red-brown. MyceLium mostly superficial, hyphae branched, septate,

smooth, subhyaline, 3.5-7.5 um diam. CONIDIOPHORES mononematous,

macronematous, straight or flexuous, pale brown to subhyaline, with subhyaline

branches, smooth-walled, 45 um long and 3-6 um wide. CONIDIOGENOUS

CELLS integrated, terminal, sometimes intercalary. hyaline. Conidial secession

rhexolytic by fracture of the wall of a small separating cell. Conrp1a solitary,

acrogenous, later acropleurogenous, ellipsoidal, obovate to broadly subglobose,

verrucose, 0-septate, pale brown to dark brown, 6.5-9 um diam., hilum

conspicuous.

Comments: Morphologically, Hansfordia rosea resembles H. pulvinata (Berk.

& M.A. Curtis) $. Hughes, which can be distinguished by its smaller conidia

(4-7 um; Hughes 1958).

Hansfordia qinghaiensis H.F. Wang & T-Y. Zhang, sp. nov. FIG. 2

MycoBank MB 808384

Differs from Hansfordia alba by its shorter and subhyaline conidia.

Type: China, Qinghai: Huzhu, North Mountain National Forest Park, from a forest soil,

10 Jun. 2006, H.F. Wang (Holotype, HSAUPII 5193; isotype, HMAS 196282; ex-type

cultures, HSAUP II ,.5193-1, HMAS 196285).

ETyMOLoGy: in reference to the type locality.

Cotonizs on PDA after 7 days at 25 °C effuse, 25-30 mm diam., velvety,

with distinct radial furrows, gray-black. MyceLrum superficial or immersed;

hyphae branched, septate, smooth, brown, 2-3 um diam. CONIDIOPHORES

mononematous, straight or flexuous, pale brown to brown, with subhyaline

branches, smooth-walled, <600 um long and 3-5 um diam. CONIDIOGENOUS

CELLS integrated, terminal, sometimes intercalary, hyaline. Conidial secession

rhexolytic by fracture of the wall of a small separating cell. Conip1a solitary,

acrogenous, later acropleurogenous, obovoid to fusiform, smooth, 0-septate,

subhyaline, 4-6 x 2.5-3.5 um, hilum conspicuous.

810 ... Wu, Wang, & & al.

Fic. 2. Hansfordia ginghaiensis (ex holotype HSAUP II 065193).

Conidia, conidiophores, and conidiogenous cells. Scale bar = 20 um.

ComMENTs: Morphologically, Hansfordia qinghaiensis resembles H. alba

J.A. Mey, which can be distinguished by its longer and hyaline conidia (7-9 x

2.5-3 um; Meyer 1959).

Hansfordia caricis P.M. Kirk, Trans. Br. Mycol. Soc. 86: 415. 1986. FIG. 3

CoLonigs on PDA after 7 days at 25°C effuse, 30-35 mm diam., hairy or

velvety, pale greyish brown to brown. Mycelium superficial or immersed;

hyphae branched, septate, smooth, subhyaline to pale brown, 1.5-3 um

Hansfordia spp. nov. (China) ... 811

Fic. 3. Hansfordia caricis (HSAUP II ,,3106).

Conidia, conidiophores, and conidiogenous cells. Scale bar = 20 um

wide. CONIDIOPHORES mononematous, straight or flexuous, very pale

brown to subhyaline, branched, smooth-walled, 50 um long, 2.5-4 um diam.

CONIDIOGENOUS CELLS integrated, terminal, sometimes intercalary. Conidial

secession rhexolytic by fracture of the wall of a small separating cell. CoNIDIA

solitary, acrogenous, later acropleurogenous, obovate to broadly ellipsoid,

smooth, 0-septate, pale brown to hyaline, 5-8 x 3-4.5 um, slightly protruding

basal hilum about 0.5 um diam.

SPECIMEN EXAMINED: CHINA. Macau: Seac Pai Van Park, from a forest soil, 1 Jul.

2004, J.J. Xu (HSAUP II ,,3106, HMAS 196283; cultures, HSAUP II ,,3106-1, HMAS

196286).

Hansfordia caricis is reported for the first time in China. Our specimen shares

obovate to broadly ellipsoid, 0-septate, smooth, very pale brown to hyaline

conidia with a slight hilum and measuring 5-8 x 3-4.5 um, as described

by Kirk (1986). The species is most similar in conidial shape to H. sinuosae

Wei Li & X.L. Cheng, which differs by its smaller conidia (4-6 x 3-4 um; Cheng

et al. 2011).

812 ... Wu, Wang, & & al.

Key to species of Hansfordia

4b -Conidial surface verruculose orechinulate nt; 3 nit: nh-ts nbs. nod tapos ap-utznee 2.

re Conidialigui face smo ouir 5 5g ches 5, ot eey ge cob eae: «neha ceco-m ahs. mib on gan6 gh ancogdeele mogteree nek 5

2. Conidia all hyaline, globose or ellipsoid .....................0004. H. triumfettae

2eGonidia viotall iva ners. 05.0). o tae iedes ers Baba satlentea Bake: ephRdey apbatahe come ades are des 3

3. Conidia pale brown to dark brown, ellipsoid, obovate to broadly subglobose H. rosea

3. Conidia hyaline, very pale brown, or pale brown ................ 0. eee ee eee eee 4

4. Conidia subspherical to spherical, hyaline to pale brown ............ H. pulvinata

4. Conidia obovoid to broadly ellipsoid, very pale brown ................. H. caricis

SeConidia subhiyaline Or Wyalime oo owe « Pecwies< bcm o Naruto Neceisese Necessities et ecw os ewie 6

Sawonidianotsubnyaline ot hyaline sci. 8 kote e beat g Secs) eal d watts oats s oath 9

Ge COnidis OCOVGIA Or MUSTONI Fee he bon pct tency OF ge ees sen ncy UF anahe ae agen ee yaaa 7

6. Conidia narrowly obovoid, ellipsoid or globose ............ 0... cece eee eee eee 8

7eConidiatusitorin, 79% 25S Sei PS OF, Rel ake tee i BEL EAE hig H. alba

7. Conidia obovoid to fusiform, 4-6 x 2.5-3.5 UM ....... 2. ee eee eee H. ginghaiensis

8. Conidia narrowly obovoid to ellipsoid, 6-10 x 2.5-4 um ............ H. parasitica

8. Conidia ellipsoid to globose, 8-13.5 x 5.5-7.5 UM ....... eee eee eee eee H. pallens

9. Conidia very pale brown, spherical or oval .................. 00000. H. canescens

9. Conidia pale brown, brown or dark brown ............. see eee cece eee eee 10

10. Conidia ovoid, pale brown to dark brown .............. 2. sees eee eee H. indica

10. Conidia obovoid, turbinate, ellipsoid, subglobose to globose,

PalebtownrordRo ware .8. 6 2oe. FF hs elas eel at A hark, Mel bet cea teh east aah ee bad 11

11. Conidia broadly obovoid, turbinate or sometimes ellipsoid, brown ... H. catalonica

11. Conidia ellipsoid, broadly subglobose to globose, pale brown ................ 12

12. Conidia globose or ellipsoid, diam. 9.5-10 um ................04. H. arborescens

12. Conidia ellipsoid to broadly subglobose, 4-6 x 3-4 um ............. H. sinuosae

Acknowledgments

The authors are grateful for pre-submission comments and suggestions provided

by Dr. R.E. Castafieda-Ruiz, Dr. David W. Minter, and Dr. Yong Wang, and Dr. Shaun

Pennycook. This project was supported by the National Science Foundation of China

(no. 30970011 & 30499340).

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marine brown alga, Colpomenia sinuosa. Mycotaxon 116: 431-436.

http://dx.doi.org/10.5248/116.431

Hansfordia spp. nov. (China) ... 813

Hughes SJ. 1951. Studies on micro-fungi. IX. Calcarisporium, Verticicladium, and Hansfordia

(gen. nov.). Mycological Papers 43. 25 p.

Hughes SJ. 1958. Revisiones Hyphomycetum aliquot cum appendice de nominibus rejiciendis.

Canadian Journal of Botany 36: 727-836. http://dx.doi.org/10.1139/b58-067

Index Fungorum. 2015. http://www.indexfungorum.org/Names/Names.asp; accessed 29 Aug.

2015.

Kirk PM. 1986. New or interesting microfungi XV. Miscellaneous hyphomycetes from the British

Isles. Transactions of the British Mycological Society 86: 409-428.

http://dx.doi.org/10.1016/S0007-1536(86)80185-1

Kirk PM, Cannon PF, Minter DW & Stalpers J. 2008. Dictionary of the Fungi. Edn 10. CAB

International, Wallingford, UK. 771 pp.

Meyer JA. 1959. Moisissures du sol et des litieres de la région de Yangambi (Congo Belge).

Publications de l'Institut National pour Etude Agronomique de Congo Belge, Série Scientifique

75.211 p.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p. http://dx.doi.org/10.3767/003158511X617435

Species Fungorum. 2015. http://www.speciesfungorum.org/Names/Names.asp (accessed 29 Aug.

2015).

MY COTAXON

http://dx.doi.org/10.5248/130.815

Volume 130, pp. 815-820 July-September 2015

Marthamyces chinensis sp. nov. on Fissistigma from China

Hat-LIn Gu’, YING-REN LIN?*, FAN PENG’, QING LI’, & SHI-JUAN WANG?

" School of Life Science &? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

ABSTRACT — A new species, Marthamyces chinensis, collected from fallen leaves of Fissistigma

sp. in tropical rain forest of Hainan Province, is described and illustrated. The type collection

is deposited in the Reference Collection of Forest Fungi of Anhui Agricultural University,

China (AAUF).

Key worps — Annonaceae, morphology, Rhytismataceae, Rhytismatales, taxonomy

Introduction

Marthamyces Minter, typified by M. emarginatus (Minter 2003), is a small

genus in the Rhytismataceae (Rhytismatales, Leotiomycetes, Ascomycota).

Members of this genus are usually leaf-inhabiting and are characterized by

apothecial circular or angular immersed ascomata not associated with an

anamorph, bleached spots and zone lines, a persistent covering layer that

often contains crystals, a rather thin subhymenium, cylindric to sub-saccate

asci, colorless filiform ascospores that are usually transversely septate, and

continuous or septate paraphyses often branched or flexuous at the tips (Minter

2003, Johnston 2006).

The genus was proposed to accommodate nine leaf-inhabiting fungi

with filiform ascospores that had previously been treated in Propolis sensu

Sherwood (1977) and subsequent authors. Johnston (2006) added three new

species and a new combination to Marthamyces, so that thirteen species are

currently accepted in the genus. No Marthamyces species has yet been reported

from China (Lin et al. 2012).

Materials & methods

The ascomata were observed with a stereomicroscope at 10-50x magnification.

After rehydration in water for ca. 10 min, 10-15 um thick sections of fruitbodies were

made using a YD-1508-III freezing microtome (Jinhua, China). Material mounted in

816... Gu & al.

water, KOH solution, Melzer’s reagent, cotton blue in water, or lactophenol-glycerin

was examined microscopically. The color of internal ascomatal structures was observed

in water. Gelatinous caps and sheaths of ascospores were examined in 0.1% (w/v)

lactophenol-cotton blue. Measurements were made using material in 5% KOH and

from more than 30 paraphyses, asci, and ascospores for each specimen. Point and line

integrated illustrations of external status and internal structures of ascomata were drawn

using a Panasonic XSJ-2 microscope drawing device (Osaka, Japan). Photographs were

taken using a Sony DSC-TX7C camera (Tokyo, Japan), and digital cameras connected

to an Olympus CX31RTSF microscope and Olympus SZ2-ILST dissecting microscope

(Tokyo, Japan). The type collection is deposited in the Reference Collection of Forest

Fungi of Anhui Agricultural University, Hefei, China (AAUF).

Taxonomy

Marthamyces chinensis Y.R. Lin & H.L. Gu, sp. nov. FIGs 1, 2

MycoBank MB811489

Differs from Marthamyces emarginatus by its covering layer composed of basically intact

epidermal and mesophyllous cells containing randomly arranged brown to blackbrown

angular cells and unevenly sized dense crystals mostly in the inside part of the upper

1/3-2/3, its smaller asci, and its narrower ascospores tapered gradually towards the

subacute or rounded base.

Type: China, Hainan, Jianfengling National Forest Park, Rainforest Valley, alt. ca 700 m,

on fallen leaves of Fissistigma sp. (Annonaceae), 17 June 2014, Y.R. Lin, S.J. Wang, HY.

Liu & Y.E. Xu 2765 (Holotype, AAUF 68873).

EryMo_oey: chinensis, referring to the country where the specimen was collected.

ASCOMATA not associated with bleached spots, zone lines, or an anamorph,

developing on both sides of leaves, principally on the upper side of the leaf,

scattered or rarely clustered.

In surface view, ascomata 350-530 um diam., triangular to pentagonal,

occasionally elliptical, without a clearly marked outline, initially appearing

as patches concolorous with substratum surface, patches becoming raised

as ascomata mature, eventually opening by (1-)3-5 somewhat radiate slits,

the covering host and fungal tissue folding back as several more or less

triangular, grey-pruinose lobes to widely expose the white-pruinose surface of

a hymenium. Lips absent.

Fic. 1. Marthamyces chinensis (holotype AAUF 68873) on Fissistigma sp.: A. Habit on leaf (most

of the bleached areas are colonised by Lophodermium agathidis). B. Enlargement of ascomata

(note immature, mature, and over-mature ascomata and association with L. agathidis on bottom

right). C. Ascoma in median vertical section. D. Portion of ascoma in median vertical section.

E. Detail of apical covering layer. F. Hymenium and lower wall (note crystals in upper hymenium).

G. Asci and paraphyses. H. Ascospores. Bars: A =1 cm; B = 500 um; C, D = 50 um; E = 30 um;

F = 20 um; G, H= 10 um.

817

Marthamyces chinensis sp. nov. (China) ...

818 ... Gu & al.

Fic. 2. Drawing of Marthamyces chinensis (holotype AAUF 68873) on Fissistigma sp.: A. Portion

of ascoma in median vertical section. B. Asci and paraphyses. C. Ascospores. Bars: A = 50 um;

B, C = 10 um.

In median vertical section, ascomata deeply embedded with host cells

becoming filled with fungal tissue as the ascoma continues development.

CovERING LAYER 70-130 um thick, slightly thinning towards the edge,

connecting to the lower wall, composed of basically intact epidermal and

mesophyllous cells containing sparse to dense, brown, grey-brown or black-

brown, thick-walled angular cells 3-7 um diam., and a group of colorless or

pale, refractive crystals of uneven size (3-14 um diam.) in the inside part of

the upper 1/3-2/3. LowER WALL somewhat poorly developed, 8-14 um thick,

consisting of black-brown textura angularis-globulosa with thick-walled cells

4-8 um diam., but a small number of dark fungal cells embedding degraded

mesophyll tissue beneath the lower wall. SUBHYMENIUM nearly flattened,

10-15 wm thick, composed of colorless textura angularis and intricata.

HyMENIvM colorless or sometimes pale grayish brown in the upper 1/4-1/3,

usually intermixed with sparse crystals in the upper layer. PARAPHYSES 72-80

Marthamyces chinensis sp. nov. (China) ... 819

x 1.6-2 um, filiform, straight or somewhat tortuous, 1-2-septate at the base,

slightly tapered towards the upper, with several short, narrow, irregular

branches near the tip, with no gelatinous matrix. Asci ripening sequentially,

65-76 x 5.2-6.5 um, cylindrical, apex subacute, rounded or subtruncate,

smooth, thin-walled, without a circumapical thickening, nearly sessile, J-,

8-spored. ASCOsPoRES arranged in a fascicle, 60-72 x 1.2-1.6 um, filiform,

colorless, tapered gradually towards the rounded or subacute base, slightly

curved or sigmoid, 0-1-septate, with apical and basal gelatinous caps and an

inconspicuous thin gelatinous sheath.

HOST SPECIES, HABITAT, & DISTRIBUTION: Producing ascomata on fallen

leaves of Fissistigma sp. Known only from the type locality in Hainan Province,

China.

COMMENTS — ‘The new species is very similar to the Marthamyces type

species, M. emarginatus (Cooke & Massee) Minter, in shape, size, and opening

form of ascomata. However, M. emarginatus has covering layer composed of

remains of broken down epidermal cells, upper wall of ascoma and sparse large

crystals, subhymenium comprising angular to cuboidal cells with walls slightly

gelatinized, much larger asci ((75-)85-95(-100) x (7-)7.5-8.5(-9.5) um or

75-100 x 6-8 um) (Johnston 2006; Cannon & Minter 1986), and wider

ascospores (2(-2.5) um) tapering slightly to both rounded ends. Besides, the

upper wall of ascoma is composed of two layers, with the outer layer comprising

angular to cylindric cells oriented roughly parallel to host surface and the

inner layer comprising rows of short-cylindric cells oriented more or less

perpendicular to host surface, with poorly developed periphysoids (Johnston

2006).

Marthamyces quadrifidus (Lév.) Minter is easily distinguished from this

new taxon by its triangular or quadrangular, subepidermal ascomata, thinner

covering layer (50-65 um), upper wall composed of two layers (the outer layer

very poorly developed, consisting of colorless cylindric cells oriented parallel to

host surface, and the inner layer comprising hyaline thin-walled periphysoid-

like elements), a subhymenium consisting of angular to prismatic cells, and

much larger asci (75-95(-100) x 6.5-9.5 um) often widest near the base

(Johnston 2006).

Marthamyces chinensis is often accompanied by Lophodermium agathidis

Minter & Hettige, sometimes growing on the same leaves. From our

observations, it seems likely that this new species is a weak parasite or a saprobe.

Acknowledgments

We are grateful to Prof C.L. Hou (Capital Normal University, China) and Prof M. Ye

(Hefei University of Technology, China) for serving as pre-submission reviewers and to

820 ... Gu & al.

Ms H.Y. Liu and Mr Y.F. Xu for the field investigations. This study was supported by the

National Natural Science Foundation of China (No. 31270065, 31500019).

Literature cited

Cannon PF, Minter DW. 1986. The Rhytismataceae of the Indian subcontinent. Mycological Papers

155. 123 p.

Johnston PR. 2006. Rhytismatales of Australia: the genus Marthamyces. Australian Systematic

Botany 19: 135-146. http://dx.doi.org/10.1071/SB05010

Lin YR, Liu HY, Hou CL, Wang SJ, Ye M, Huang CL, Xiang Y, Yu SM. 2012. Flora fungorum

sinicorum, vol. 40, Rhytismatales [in Chinese]. Science Press. Beijing. 261 p.

Minter DW. 2003. Propolis and Marthamyces gen. nov. Mycotaxon 87: 43-52.

Sherwood MA. 1977. Taxonomic studies in the Phacidiales: Propolis and Propolomyces. Mycotaxon

5: 320-330.

MY COTAXON

http://dx.doi.org/10.5248/130.821

Volume 130, pp. 821-825 July-September 2015

Blastophragma chonggingense sp. nov. and a new record of

Bahusutrabeeja angularis from southern China

JIAN- MEI GAO, JI-WEN XIA, YING-RuI MA, ZHUANG LI, & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: sdau613@163.com, liz552@126.com

ABsTRACT — A new species, Blastophragma chongqingense is described and illustrated

from specimens collected on dead branches in Chongqing Province. It differs in conidial

morphology from the two known species of Blastophragma. In addition, Bahusutrabeeja

angularis is newly recorded from China.

Key worps —hyphomycetes, taxonomy

Introduction

The genus Blastophragma, containing two species, was described by

Subramanian (1993), but was invalid because there was no indication of

where the type was conserved. Subsequently, Subramanian (1995) validated

the genus, which is characterized by single conidiophores with an integrated,

apical, polyblastic conidiogenous cell that shows sympodial extension and

produces solitary, acrogenous, distoseptate blastoconidia. The type species,

B. subulatum Subram., was found on dead twigs of an unidentified plant from

Malaysia, while the second species, B. rostratum Subram., was found on dead

twigs of Antidesma cuspidatum Mull. Arg. (Phyllanthaceae).

Bahusutrabeeja dwaya Subram. & Bhat (Subramanian & Bhat 1977),

the type species of Bahusutrabeeja Subram. & Bhat, has distinct individual

conidiophores with integrated, terminal, cylindrical conidiogenous cells that

produce conidia in succession by percurrent extension froma single fertile locus.

The conidia are hyaline, smooth, thick-walled, aseptate, spherical, rounded-

cubical or obpyriform to obclavate, with several to many slender appendages

distributed over the surface; they accumulate in a slimy mass at the apex of

the conidiogenous cell. Six species have been accepted in Bahusutrabeeja (Bhat

1994, Bhat & Kendrick 1993, Li et al. 2014, McKenzie 1997, Rao & Hoog 1986,

Subramanian & Bhat 1977).

822 ... Gao & al.

A species with the morphological characteristics of Blastophragma was

collected on decaying twigs and dead stems in the tropical forests of Chongqing

Province; it is proposed here as a new species. In addition, Bahusutrabeeja

angularis was collected and is a new record for China.

Fic. 1. Blastophragma chonggingense (holotype, HSAUP H9611). A-C. Conidiophores,

conidiogenous cells, and conidia. D. Conidiophore and conidiogenous cell. E-G. Conidia.

Blastophragma chonggqingense J.M. Gao & X.G. Zhang, sp. nov. FIG. 1

MycoBAnk MB 814400

Differs from Blastophragma subulatum by its larger fusiform conidia and from

B. rostratum by its fusiform 5-11-distoseptate conidia.

Type: China. Chongqing Province: Mount Xiannv, on dead stems of unidentified broad-

leaved tree, 20 Oct. 2014, J.M. Gao (holotype, HSAUP H9611; isotype, HMAS 243458).

Erymo ocy: referring to the province where the type was found.

Blastophragma chonggingense sp. nov. (China) ... 823

Colonies on natural substrate effuse, pale brown to brown, hairy. Mycelium

partly superficial, partly immersed in the substratum, composed of septate,

pale brown, smooth, 2-4 um wide hyphae. Conidiophores distinct, single,

caespitose, unbranched, erect, straight or slightly flexuous, dark brown,

slightly paler in the distal part, geniculate, cylindrical, smooth, thick-walled,

0-6-septate, 50-175 x 4.8-9.6 um. Conidiogenous cells polyblastic, integrated,

terminal, with sympodial extension, subhyaline to pale brown. Conidia

acrogenous, blastic, solitary, dry, subhyaline to pale brown, fusiform with a flat,

dark basal scar, widening then narrowing above, smooth, rounded at the apex,

5-11-distoseptate, 36-44 x 12-16 um; conidial wall brown; basal scar 3 um

wide.

Comments -Blastophragma chonggingense differs from the two previously

described species of Blastophragma in conidial shape, size, and number of

septa. Blastophragma subulatum differs by its smaller subulate conidia (15-25

x 4.5-6.0 um; Subramanian 1993), and B. rostratum differs by its subobclavate

to subfusiform conidia with fewer septa (40-50 x 7-10 um, 4-septate;

Subramanian 1993).

Bahusutrabeeja angularis V. Rao & de Hoog, Stud. Mycol. 28: 67 (1986) Fic. 2

Colonies on natural substrate effuse, farinose, dark brown. Mycelium

scanty, immersed, composed of septate, pale brown, smooth-walled hyphae.

Conidiophores distinct, single, erect, straight or slightly flexuous, unbranched,

arising from minute, semi-immersed sclerotial bodies, cylindrical, smooth,

thick-walled, 145-290 x 4.8-7.2 um, dark reddish brown near the base, light

brown in the apical part. Conidiogenous cells polyphialidic, often extending

percurrently after breakage, terminating with a phialide. Phialides integrated,

cylindrical with conical apex, 4.8-6.5 um wide at the base, 2 um wide just below

the terminal collarette; collarettes terminal, sometimes also lateral, pale brown,

funnel-shaped, 2 um long, 4 um wide at the tip. Occasionally the collarette

extends and develops a new phialide opening. Conidia hyaline, aseptate,

smooth, thick-walled, rounded-cubical to polygonal, 7.8-10.2 um diam., one

of the angles being the basal scar, each corner provided with a thin, hyaline

appendage; conidia aggregating in slimy masses at the apex of the conidiophore.

SPECIMEN EXAMINED: CHINA. HAINAN PROVINCE, tropical forest of Jianfengling, on

dead stems of unidentified broad-leaved tree, 18 Apr. 2014, J.M. Gao (HSAUP H9599;

HMAS 243459).

Comments -Bahusutrabeeja angularis, which was described from India, is

reported for the first time from China. The Chinese specimen closely matched

the protologue description (Rao & Hoog 1986), except that the Indian type

specimen had longer conidiophores (200-500 x 5-7 um) and slightly smaller

conidia (7-8 um diam.).

824 ... Gao & al.

Fic. 2. Bahusutrabeeja angularis (HSAUP H9599). A. Conidiophores and conidiogenous cells.

B, D. Conidiophores, conidiogenous cells, and conidia. C. Conidia.

Blastophragma chonggqingense sp. nov. (China) ... 825

Acknowledgments

The authors express gratitude to Dr. Bryce Kendrick and Dr. Eric H.C. McKenzie for

serving as pre-submission reviewers and for their valuable comments and suggestions.

This project was supported by the National Natural Science Foundation of China

(Nos. 31093440, 31230001) and the Ministry of Science and Technology of the People’s

Republic of China (Nos. 2006FY120100).

Literature cited

Bhat DJ. 1994. Two undescribed species of conidial fungi from forests of western Ghats in southern

India. Indian Journal of Forestry 17: 129-133.

Bhat DJ, Kendrick WB. 1993. Twenty-five new conidial fungi from the Western Ghats and the

Andaman Islands (India). Mycotaxon 49: 19-90.

Li XX, Xia JW, Ma LG, Castaneda Ruiz RF, Zhang XG. 2014. A new species of Bahusutrabeeja from

Guangxi, China. Mycotaxon 126: 227-230. http://dx.doi.org/10.5248/126.227

McKenzie EHC. 1997. Bahusutrabeeja bunyensis sp. nov. (Hyphomycetes) from Queensland,

Australia, and a new name for Chalara australis McKenzie. Mycotaxon 61: 303-306.

Rao V, Hoog GS de. 1986. New or critical hyphomycetes from India. Studies in Mycology 28. 84 p.

Subramanian CV. 1993. Blastophragma gen. nov. for two interesting hyphomycetes from southeast

Asia. Cryptogamie, Mycologie 14: 39-44.

Subramanian CV. 1995 [“1992-1993”]. Validation of names of some taxa (hyphomycetes). Kavaka

20/21: 57-58.

Subramanian CV, Bhat DJ. 1977. Bahusutrabeeja, a new genus of the hyphomycetes. Canadian

Journal of Botany 55: 2202-2206. http://dx.doi.org/10.1139/b77-249

MY COTAXON

http://dx.doi.org/10.5248/130.827

Volume 130, pp. 827-833 July-September 2015

Sporidesmiopsis malloti sp. nov.

and new records from southern China

JI-WEN X1A, YING-RuI Ma, JIAN-MEI GAo,

ZHUANG LI, & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: sdau613@163.com, liz552@126.com

ABSTRACT — Sporidesmiopsis malloti is described and illustrated from specimens collected

from dead branches of Mallotus hookerianus in Hainan Province, China. The new fungus is

characterized by macronematous conidiophores with branches at the apex and conidiogenous

cells arising on both stipe and branches with 3-7-euseptate conidia. A key to Sporidesmiopsis

species is provided. Gangliostilbe malabarica and Vamsapriya indica are newly recorded from

China.

KEY worps — conidial fungi, taxonomy

Introduction

Subramanian & Bhat (1989) erected the genus Sporidesmiopsis with

S. malabarica Subram. & Bhat as type species. The genus is distinguished from

Sporidesmium Link by its mononematous conidiophores with apical branches

and conidiogenous cells that occur on both stipe and branches (Wongsawas

et al. 2008). Four other species have been included in the genus: S. dennisii

(J.L. Crane & Dumont) Bhat et al., S. goanensis Bhat & W.B. Kendr., S. guangxiensis

J.W. Xia & X.G. Zhang, and S. zhejiangensis Wongs. et al. (Bhat & Kendrick

1993, Wongsawas et al. 2008, Xia et al. 2014). However, Bhat & Kendrick

(1993) synonymized S. malabarica under S. dennisii, and with the transfer of

S. zhejiangensis to Ellisembiopsis based on its distoseptate conidia (Santa Izabel

et al. 2013), only three species are currently accepted in Sporidesmiopsis.

We propose a new Sporidesmiopsis species, collected on dead branches from

Hainan Province, China. We also note two other hyphomycete species on dead

branches from Guangdong Province as newly recorded from China.

828 ... Xia & al.

Fic. 1. Sporidesmiopsis malloti. (ex HSAUP H6426): A. Conidiophores, conidiogenous cells,

and conidia. B, C. Conidiogenous cells and conidia. D-F. Conidia.

Sporidesmiopsis malloti sp. nov. (China) ... 829

Sporidesmiopsis malloti J.W. Xia & X.G. Zhang, sp. nov. FIG. 1

MycoBAnk MB 814399

Differs from Sporidesmiopsis goanensis and S. guangxiensis by its larger conidia and from

S. dennisii by its smaller conidia with fewer septa.

Type: China, Hainan Province: Jianfengling, on dead stems of Mallotus hookerianus

(Seem.) Mull. Arg. (Euphorbiaceae), 22 Apr. 2014, J.W. Xia (Holotype, HSAUP H6426;

isotype, HMAS 245586).

ETryMoOLoGey: in reference to the host genus Mallotus.

CoLonigs on the natural substrate effuse, brown to dark brown, hairy.

Mycelium superficial and partly immersed, composed of septate, pale brown,

smooth hyphae, 1-2 um diam. Conip1opHorEs distinct, single, branched,

erect, straight or slightly flexuous, cylindrical, smooth, thick-walled, brown

to dark brown, 7-12-septate, 200-380 x 6-9.5 um. CONIDIOGENOUS CELLS

monoblastic, integrated and discrete, narrowly doliiform, truncate at the apex

after conidium secession, 20-40 x 4.5-8 um. Conip1a solitary, dry, obclavate to

fusiform, base truncate, apex rounded, pale brown to brown, 35-60 x 8.5-11.5

um, 3—7-euseptate.

ComMENTs - Sporidesmiopsis malloti bears some resemblance to S. goanensis

and S. guangxiensis in conidial shape. However, the conidia of S. goanensis

are smaller (20-30 x 5-7 um, 3-4-euseptate; Bhat & Kendrick 1993), and the

conidia of S. guangxiensis are smaller (32-40 x 6-7.5 um, 5-6-euseptate) and

have a subhyaline to hyaline apical cell (Xia et al. 2014).

Key to species of Sporidesmiopsis

1. Conidia >70 um long, 10-15-euseptate 1.0.0.0... eee eee ee eee S. dennisii

tiConidia:<70 pip lones-<10-SUSE plate 4 2. gece we geen ee gt gun ue pt in BB nh BAG ove SEA ote 2

Px SE Onidea SSO IMA ONG ee ey ie ce oS dee eke hha ete gree AMS nee au: Rite teat 3

2. Conidia 20-30 x 5-7 um, 3-4-euseptate ........ 0. eee ee ee eee ee S. goanensis

3. Conidia 32-40 x 6-7.5 um, 5-6-euseptate ........... ee eee eee S. guangxiensis

3. Conidia 35-60 x 8.5-11.5 um, 3-7-euseptate ..... eee eee eee eee S. malloti

Gangliostilbe malabarica Subram. & Bhat, Kavaka 15: 54, 1989 [“1987”]. Fic. 2

COLONIES on natural substrate effuse, dark brown, hairy. Mycelium mostly

immersed in the substratum. SYNNEMATA arising singly, rarely in pairs, on

the surface of the substratum, erect, with the stipes comprising compact

aggregations of parallel hyphae, terminating in fertile heads, with stipes

dark brown and fertile heads brown, 200-550 x 20-40 um. CONIDIOPHORES

branched, percurrently extending at the tip up to 4 times, septate, smooth,

pale brown, divergent in the apical part of the synnema forming a loose clavate

head up to 70 um wide, and 25-35 um high. CONIDIOGENOUS CELLS producing

conidia singly at the tip, integrated, subhyaline, smooth, 8-12 x 2.5-3.5 um.

830 ... Xia & al.

miton ACL,

wngz

CELPELS The. > CEE PEE,

Thipttcb> HOME> can “ooo

Fic. 2. Gangliostilbe malabarica. (ex HSAUP H6398):

B. Synnema. C. Apex of synnema. D. Conidia.

Sporidesmiopsis malloti sp. nov. (China) ... 831

Conip1A pale brown, obclavate to fusiform, elongated, rounded at the tip,

apical cell sometimes surrounded by a globose mucilaginous tunica, truncate

at the base, broader in the middle, 6-11-euseptate, 30-55 x 6-8 um.

SPECIMEN EXAMINED: CHINA, GUANGDONG PROVINCE: Baiyun Mountain, on dead

stems of unidentified broadleaf tree, 28 May 2013, J.W. Xia (HSAUP H6398; HMAS

245587).

Comments - The genus Gangliostilbe Subram. & Vittal, with G. indica Subram.

& Vittal as the type species, was proposed in Vittal (1976) and characterized

by solitary, obovoid, ovoid, or fusiform conidia borne on integrated,

terminal, monoblastic conidiogenous cells on synnematous conidiomata.

Five species have been described in this genus (Vittal 1976, Bhat & Sutton

1985, Subramanian & Bhat 1989, Mercado-Sierra et al. 1997, Ma et al. 2014).

Bactrodesmium longisporum is similar to G. malabarica but is distinguished by

its sporodochia that are scattered, punctiform, and black and a conidial apex

that is often enveloped by a thin spherical mucilaginous tunica (Ellis 1976).

Gangliostilbe malabarica is reported here for the first time from China. The

Chinese specimen fits well with the protologue description, except that the

conidia of the Indian type collection have more septa (up to 17; Subramanian

& Bhat 1989).

Vamsapriya indica Gawas & Bhat, Mycotaxon 94: 150, 2006 [“2005”]. FIG. 3

COLONIES on natural substrate effuse, dark brown to black. Mycelium

mostly immersed in the substratum, composed of subhyaline, septate,

branched, smooth hyphae, 2-3 um wide. SYNNEMATA erect, rigid, dark

brown, composed of compact parallel conidiophores, 400-600 x 15-30 um.

CONIDIOPHORES distinct, dark brown, smooth, septate, branched, 3-4.5 um

wide. CONIDIOGENOUS CELLS monotretic, non-cicatrized, integrated or

discrete, terminal, clavate, slightly curved toward the exterior, 15-20 x 3-4.5

um. CONIDIA dry, catenate, acrogenous, brown, smooth, simple, cylindrical,

vermiform, 2-16-euseptate, constricted at the septa, 15-110 x 4-6 um,

developing in acropetal chains; terminal conidia rounded at the apex, slightly

truncate at the base; other conidia slightly truncate at both ends.

SPECIMEN EXAMINED: CHINA, GUANGDONG PROVINCE: Baiyun Mountain, on dead

stems of unidentified broadleaf tree, 28 May 2013, J.W. Xia (HSAUP H6386; HMAS

245588).

ComMENTs - There are only two accepted taxa of Vamsapriya, V. indica and

V. mahabaleshwarensis Pratibha & Bhat (Gawas & Bhat 2006, Pratibha & Bhat

2008). Vamsapriya indica is reported here for the first time from China. The

Chinese specimen is similar to the protologue description, but the Indian type

collection has longer synnemata (700-870 um) and a slightly lower range of

conidial lengths (10-80 um; Gawas & Bhat 1989).

832 ... Xia & al.

He TL

Choi ST TF.

{ pul Gee:

SSA ewVenn es =

Fic. 3. Vamsapriya indica. (ex HSAUP H6386):

A, B. Synnema. C. Conidiogenous cells and conidia. D. Conidia.

Sporidesmiopsis malloti sp. nov. (China) ... 833

Acknowledgments

The authors express gratitude to Dr. Bryce Kendrick and Dr. Rafael F. Castafeda-

Ruiz for serving as pre-submission reviewers and for their valuable comments and

suggestions. This project was supported by the National Natural Science Foundation of

China (Nos. 31093440, 31230001) and the Ministry of Science and Technology of the

People’s Republic of China (Nos. 2006FY120100).

Literature cited

Bhat DJ, Kendrick B. 1993. Twenty-five new conidial fungi from the Western Ghats and the

Andaman Islands (India). Mycotaxon 49: 19-90.

Bhat DJ, Sutton BC. 1985. New and interesting hyphomycetes from Ethiopia. Transactions of the

British Mycological Society 85: 107-122. http://dx.doi.org/10.1016/S0007-1536(85)80160-1

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute,

Kew, UK.

Gawas P, Bhat DJ. 2006 [“2005”]. Vamsapriya indica gen. et sp. nov., a bambusicolous, synnematous

fungus from India. Mycotaxon 94: 149-154.

Ma LG, Xia JW, Ma YR, Castafieda-Ruiz RF, Zhang XG. 2014. Two new species of

Spadicoides and Gangliostilbe from southern China. Mycological Progress 13: 547-552.

http://dx.doi.org/10.1007/s11557-013-0937-z

Mercado-Sierra A, Gené J, Guarro J. 1997. Some Costa Rican hyphomycetes. I. Nova Hedwigia

64: 455-465.

Pratibha J, Bhat DJ. 2008. New and unusual hyphomycetes from Mahabaleshwar, India. Mycotaxon

105: 423-431.

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 on

Sporidesmium complex. Mycosphere 4: 156-163. http://dx.doi.org/10.5943/mycosphere/4/2/1

Subramanian CV, Bhat DJ. 1989 [“1987”]. Hyphomycetes from South India I. Some new taxa.

Kavaka 15: 41-74.

Vittal BPR. 1976 [“1975”]. Gangliostilbe indica, a new synnematous hyphomycetes from India.

Kavaka 3: 69-71.

Wongsawas M, Wang HK, Hyde KD, Lin FC. 2008. New and rare lignicolous hyphomycetes

from Zhejiang Province, China. Journal of Zhejiang University 9: 797-801.

http://dx.doi.org/10.1631/jzus.B0860008

Xia JW, Ma LG, Castafeda-Ruiz RE, Zhang XG. 2014. A new species of Sporidesmiopsis and three

new records of other dematiaceous hyphomycetes from southern China. Nova Hedwigia

98: 103-111. http://dx.doi.org/10.1127/0029-5035/2013/0145

MY COTAXON

http://dx.doi.org/10.5248/130.835

Volume 130, pp. 835-841 July-September 2015

Codinaea jianfenglingensis sp. nov.

and new records from southern China

JI-WEN X1A, YING-RuI Ma, JIAN-MEI GAo,

ZHUANG LI & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: sdau613@163.com, liz552@126.com

ABSTRACT — Codinaea jianfenglingensis is described and illustrated from specimens

collected on dead branches in Hainan Province. The new fungus is characterized by distinct

conidiophores and single apical monophialidic conidiogenous cells that produce fusiform to

lunate conidia with a single unbranched setula at each end. Craspedodidymum hyalosporum

and Ellisembiopsis brasiliensis are newly recorded from China.

KEY worps — conidial fungi, taxonomy

Introduction

During our ongoing survey of microfungi associated with woody debris in

tropical and subtropical forests of Hainan and Guangdong provinces, three

species with morphological characteristics of the genera Codinaea Maire,

Craspedodidymum Hol.-Jech., and Ellisembiopsis T.S. Santa Izabel & Gusmao

(Maire 1937, Holubova-Jechova 1972, Santa Izabel et al. 2013) were collected

on decaying twigs and dead stems. One of these is an undescribed species

of Codinaea, while the other two species are new records for China. The

specimens are deposited in the Herbarium of Department of Plant Pathology,

Shandong Agricultural University, Taian, China (HSAUP) and the Mycological

Herbarium, Institute of Microbiology, Chinese Academy of Sciences, Beijing,

China (HMAS).

Codinaea jianfenglingensis J.W. Xia & X.G. Zhang, sp. nov. FIG. 1

MycoBAnk MB 814404

Differs from Dictyochaeta caatingae, D. macrospora, D. matsushimae, and D. triseptata

by its monophialidic conidiogenous cells that produce fusiform to lunate conidia with a

filiform setula at each end.

836 ... Xia & al.

Fic. 1. Codinaea jianfenglingensis (ex holotype, HSAUP H6456): A. Conidiophores,

conidiogenous cells, and conidia. B. Conidiophores and conidiogenous cells with

collarettes. C. Setae. D. Conidia.

Type: China, Hainan Province: Jianfengling, on dead stems of unidentified broadleaf

tree, 22 Apr. 2014, J.W. Xia (Holotype, HSAUP H6456; isotype, HMAS 245589).

EryMo_ocy: in reference to the type locality.

COLONIES on natural substrate effuse, brown, hairy. Mycelium partly superficial,

partly immersed in the substratum, composed of septate, pale brown, smooth

hyphae, 1-2 um wide. Setae solitary, erect, straight or flexuous, 3-5-septate,

Codinaea jianfenglingensis sp. nov. (China) ... 837

smooth, brown, 100-130 x 3.4-4.5 um. CoNIDIOPHORES distinct, single,

erect, straight or slightly flexuous, cylindrical, smooth, thick-walled, brown,

4-7-septate, 120-200 x 4.5-5 um. CONIDIOGENOUS CELLS monophialidic,

integrated, cylindrical, subhyaline to pale brown, 28-50 x 4.5-5 um; collarette,

3.5-4.5 x 2.5-3.5 um. Conrpia fusiform to lunate, 3-septate, smooth,

subhyaline, 22-30 x 4-6 um, with a filiform setula, 7.5-11.5 um long, at each

end.

ComMMENTS - ‘The genus Codinaea was proposed by Maire (1937), with

C. aristata Maire as the type species. Gamundi et al. (1977) rediscovered and

redescribed Dictyochaeta fuegiana Speg., the type species of Dictyochaeta

(Spegazzini 1923) and proposed Codinaea as a synonym. However, molecular

studies by Réblova & Winka (2000) suggest that species with setulate conidia

(Codinaea) are distinct from those lacking conidial setulae (Dictyochaeta).

Seifert et al. (2011) and Li et al. (2012) supported segregation of Codinaea and

Dictyochaeta as separate genera delineated by the presence/absence of conidial

setulae.

Codinaea jianfenglingensis is morphologically similar to Dictyochaeta

caatingae A.C. Cruz & Gusmao, D. macrospora Kuthub. & Nawawi,

D. matsushimae (Hewings & J.L. Crane) Whitton et al., and D. triseptata

(Matsush.) R.F. Castafieda in having 3-septate conidia. However, D. caatingae,

D. macrospora, D. matsushimae, and D. triseptata can be easily separated by

their polyphialidic conidiogenous cells. (Castafteda-Ruiz 1986, Kuthubutheen

& Nawawi 1991, Whitton et al. 2000, Cruz et al. 2008 ).

Craspedodidymum hyalosporum Bhat & W.B. Kendr., Mycotaxon 49: 35, 1993. Fic. 2

COLONIES on natural substrate effuse, dark brown to black, hairy. Mycelium

partly superficial, partly immersed in the substratum. CONIDIOPHORES

distinct, single, erect, straight or slightly flexuous, smooth, thick-walled,

brown to dark brown, unbranched, often percurrently regenerating at cut

ends, 8-12-septate, 150-270 x 5.5-8 um. CONIDIOGENOUS CELLS terminal,

integrated, monophialidic, pale brown, 25-38 x 5-6 um, with a prominent,

thin-walled, cup-shaped and flared collarette. Conip1A pale brown, ellipsoidal

to cylindrical, rounded at both ends, smooth, 0-1-septate, slightly constricted

at the septum, 6.5-11 x 3.5-5 um, accumulating in colourless slimy masses at

the conidiophore apex.

SPECIMEN EXAMINED: CHINA, GUANGDONG PROVINCE: Baiyun Mountain, on dead

stems of unidentified broadleaf tree, 28 May 2013, J.W. Xia (HSAUP H6358, HMAS

245590).

ComMENTs - The genus Craspedodidymum was erected by Holubova-Jechova

(1972) with C. elatum Hol.-Jech. as the type species. It is characterized by

distinct, single, unbranched, or branched conidiophores bearing integrated,

838 ... Xia & al.

Fic. 2. Craspedodidymum hyalosporum (ex HSAUP H6358): A. Conidiophores,

conidiogenous cells, and conidia. B. Conidiophores and conidiogenous cells.

C. Conidiogenous cell with collarette and conidium. D. Conidia.

terminal, monophialidic, cylindrical, apically swollen conidiogenous cells with

a large and distinct funnel-shaped terminal collarette (Holubova-Jechova 1972,

Ellis 1976, Yanna et al. 2000).

Craspedodidymum hyalosporum is reported for the first time from China.

The Chinese specimen is similar to the protologue description, with almost

the same conidial size as the Indian type collection (8-12.5 x 4-6 um; Bhat &

Kendrick 1993).

Codinaea jianfenglingensis sp. nov. (China) ... 839

Ellisembiopsis brasiliensis T.S. Santa Izabel & Gusmao, Mycosphere 4: 158, 2013. Fic. 3

COLONIES on natural substrate effuse, brown to dark brown. Mycelium

partly superficial, partly immersed in the substratum, composed of subhyaline,

septate, branched, smooth hyphae, 1.5-2.5 um wide. CONIDIOPHORES distinct,

single, erect, straight or flexuous, branched at the apical region, 7—9-septate,

Fic. 3. Ellisembiopsis brasiliensis (ex HSAUP H6442): A. Colonies on natural substratum.

B. Conidiophore, conidiogenous cells, and conidia. C. Conidiophore and conidiogenous cells.

D. Conidia. E. Conidiogenous cells and conidia.

840 ... Xia & al.

smooth, brown to dark brown, 150-200 x 5.5-6.5 um. CONIDIOGENOUS CELLS

monoblastic, integrated, lageniform to doliiform, 12-14 x 4.5-5.5 um. Conidial

secession schizolytic. Conrp1 solitary, obclavate to fusiform, 5-6-distoseptate,

smooth, brown, 30-50 x 7.5-8.5 um.

SPECIMEN EXAMINED: CHINA, HAINAN PROVINCE: Jianfengling, on dead stems of

unidentified broadleaf tree, 28 May 2013, J.W. Xia (HSAUP H6442, HMAS 245591).

Comments - There are only two accepted taxa of Ellisembiopsis, E. brasiliensis

and E. zhejiangensis (Wongs. et al.) T.S. Santa Izabel & Gusmao. Ellisembiopsis

brasiliensis is reported for the first time from China. The Chinese specimen is

similar to the protologue description, with the Brazilian type collection having

somewhat larger conidia (35-80 x 7.5-13.5 um; Santa Izabel et al. 2013).

Acknowledgments

The authors express gratitude to Dr. Bryce Kendrick and Dr. Rafael F. Castafeda-

Ruiz for serving as pre-submission reviewers and for their valuable comments and

suggestions. This project was supported by the National Natural Science Foundation of

China (Nos. 31093440, 31230001) and the Ministry of Science and Technology of the

People’s Republic of China (Nos. 2006FY120100).

Literature cited

Bhat DJ, Kendrick B. 1993. Twenty-five new conidial fungi from the Western Ghats and the

Andaman Islands (India). Mycotaxon 49: 19-90.

Castaneda-Ruiz RF. 1986. Fungi Cubenses. Havana, Instituto de Investigaciones Fundamentales en

Agricultura Tropical “Alejandro de Humboldt” (INIFAT), Havana.

Cruz ACR, Ledo-Ferreira SM, Barbosa ER, Gusmao LFP. 2008. Conidial fungi from semi-arid

Caatinga biome of Brazil. New and interesting Dictyochaeta species. Mycotaxon 106: 15-27.

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew,

Gamundi IJ, Arambarri AM, Giaiotti AL. 1977. Microflora de la hojarasca de Nothofagus dombeyi.

Darwiniana 21: 81-114.

Holubova-Jechova V. 1972. Craspedodidymum, new genus of phialosporus hyphomycetes. Ceska

Mykologie 26: 70-73.

Kuthubutheen AJ, Nawawi A. 1991. Dictyochaeta macrospora sp. nov.: a litter-inhabiting

hyphomycete from Malaysia. Mycological Research 95: 248-250.

http://dx.doi.org/10.1016/S0953-7562(09)81022-8

Li DW, Kendrick B, Chen JY. 2012. Two new hyphomycetes: Codinaea sinensis sp. nov. and

Parapleurotheciopsis quercicola sp. nov., and two new records from Quercus phillyraeoides leaf

litter. Mycological Progress 11: 899-905. http://dx.doi.org/10.1007/s11557-011-0805-7

Maire R. 1937. Fungi Catalaunici: series altera. Contributions a étude de la flore mycologique de la

Catalogne. Publicacions del Instituto Botanico, Barcelona. 3(4). 128 p.

Réblova M, Winka K. 2000. Phylogeny of Chaetosphaeria and its anamorphs based on morphological

and molecular data. Mycologia 92: 939-954. http://dx.doi.org/10.2307/3761589

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 on

Sporidesmium complex. Mycosphere 4: 156-163. http://dx.doi.org/10.5943/mycosphere/4/2/1

Codinaea jianfenglingensis sp. nov. (China) ... 841

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p. http://dx.doi.org/10.3767/003158511X617435

Spegazzini C. 1923. Algunos hongos de Tierra del Fuego. Physis 7: 7-23.

Whitton SR. McKenzie EHC, Hyde KD. 2000. Dictyochaeta and Dictyochaetopsis species from the

Pandanaceae. Fungal Diversity 4: 133-158.

Yanna, Ho WH, Goh TK, Hyde KD. 2000. Craspedodidymum nigroseptatum sp. nov., a new

hyphomycete on palms from Brunei Darussalam. Mycological Research 104: 1146-1151.

http://dx.doi.org/10.1017/S0953756299002178

MY COTAXON

http://dx.doi.org/10.5248/130.843

Volume 130, pp. 843-855 July-September 2015

Fuscoporia atlantica sp. nov., a new polypore

from the Brazilian Atlantic Rainforest

RICARDO MATHEUS PIRES”, VIVIANA MOTATO- VASQUEZ,

& ADRIANA DE MELLO GUGLIOTTA

Instituto de Botanica, Nucleo de Pesquisa em Micologia,

Av. Miguel Stéfano 3687, 04301-902, Sdo Paulo, SP, Brazil

* CORRESPONDENCE TO: sals.bio@gmail.com

ABSTRACT — Fuscoporia atlantica (Basidiomycota, Hymenochaetaceae) is described and

illustrated based on specimens collected in Atlantic Rainforest areas of Sao Paulo state, Brazil,

and its inferred phylogenetic relationships are based on sequence data from the ribosomal

ITS and LSU regions. The new species is characterized by an effused-reflexed to pileate

basidioma, golden-yellow-ferruginous pore surface, tiny pores, dimitic hyphal system with

thin-walled hyphae encrusted at dissepiment edges, mostly uncinate or hooked hymenial

setae, and broadly ellipsoid to ellipsoid basidiospores. An identification key to the Fuscoporia

species recorded from Brazil is also provided.

Key worps — basidiomycete, Hymenochaetales, neotropics, phylogeny, taxonomy

Introduction

Species of the genus Fuscoporia Murrill traditionally have been included

in Phellinus s.l., a morphologically heterogeneous and phylogenetically

polyphyletic group (Fiasson & Niemela 1984; Dai 1999; Wagner & Fischer

2001, 2002b; Larsson et al. 2006). However, molecular phylogenetic analyses

have shown that Fuscoporia species form a clade independent from Phellinus

s.s. (Wagner & Fischer 2002b, Larsson et al. 2006).

Fuscoporia was described to accommodate species with resupinate to

pileate basidiomes occurring principally on coniferous and deciduous trees

characterized by monomitic to dimitic hyphal system, encrusted generative

hyphae at the dissepiments (considered diagnostic within Phellinus s.1.),

hymenial setae, and cylindric to ellipsoid, hyaline, inamyloid, nondextrinoid,

thin-walled, smooth basidiospores (Murrill 1907; Fiasson & Niemela 1984; Dai

1999, 2010; Niemela et al. 2001; Wagner & Fischer 2001).

844 ... Pires, Motato- Vasquez, & Gugliotta

TABLE 1. ITS dataset of Fuscoporia and Phellinidium spp. used in the phylogenetic

analyses.

SPECIES

Fuscoporia atlantica

E callimorpha

E contigua

E ferrea

E ferruginosa

E gilva

E rufitincta (Cooke) Murrill

FE senex

E torulosa (Pers.) T. Wagner & M. Fisch.

E viticola (Schwein.) Murrill

Phellinidium sulphurascens

P. weirii

VOUCHER SPECIMENS

SP445618 (holotype)

SP465829 (paratype)

JV090487

Jvo40914J

JV1007/3

JV0907/2°

JV0309/118

JV0309/171

CBS 444.48

D4-3B-b

DLL2009-080

SP5108

JV0202/2

JV8909/74

JV1109/24

KCTC 6653

xsd08128

SACCR 11076

ASIS24343

PRM915961

JV0610/14PK

CBS 442.76

KUC20110922-13

Q32

Pt134

Pto

Pt7

CBS 381.82

JV0709/163

Type 10

Type 11

CFS504

CFS586

GENBANK ACCESSION

NUMBERS

KP058515 {m}

KP058514 {m}

JF692190 {I}

JF692191 {I}

JF692193 {I}

JQ794546 {I}

JQ794547 {I}

JQ794548 {I}

JQ794550 {I}

AY558617 {f}

DQ516525 {i}

JQ673178 {b}

KJ677120 {k}

J794572 {I}

JQ794573 {I}

JQ794577 {I}

AY558620 {f}

FJ481039 {e}

JX427049 {d}

KF692068 {j}

GU594160 {j}

JQ794579 {I}

JQ794580 {j}

AY558647 {f}

JX463658 {e}

KC414230 {f}

EF068139 {c}

EF068238 {h}

EF068239 {h}

AY558653 {f}

JQ358814 {a}

JQ794583 {I}

EF527215 {g}

EF527216 {g}

AY829341 {f}

AY829342 {f}

Sources for published sequences: {a}= Alfredsen et al. (GenBank, unpublished); {b}= Brazee et al.

(2012); {c}= Campanile et al. (GenBank, unpublished); {d}= Cantrell et al. (2013); {e}= Jang

et al. (2012); {f} = Jeong et al. (GenBank, unpublished); {g} = Lim et al. (2005); {h} = Lim et

al. (2008); {i} = Lim et al. (GenBank, unpublished); {j} = Seok et al. (GenBank, unpublished);

{k} = Spirin et al. (GenBank, unpublished); {l} = Vlasak et al. (GenBank, unpublished); and

{m}= new sequences provided by this study.

Fuscoporia atlantica sp. nov. (Brazil) ... 845

Of the approximately 50 species worldwide currently accepted in Fuscoporia

(Mycobank 2014), 13 have been reported in Brazil, mostly from the Atlantic

Rainforest (Baltazar et al. 2009): EF. bifurcata Baltazar et al., FE callimorpha (Lév.)

Groposoetal., E contigua (Pers.) G.Cunn., F chrysea (Lév.) Baltazar & Gibertoni,

F. ferrea (Pers.) G. Cunn., F. ferruginosa (Schrad.) Murrill, E flavomarginata

(Murrill) Groposo et al., E gilva (Schwein.) T. Wagner & M. Fisch., FE palmicola

(Berk. & M.A. Curtis) Bondartseva & S. Herrera, E punctatiformis (Murrill)

Zmitr. et al., FE rhabarbarina (Berk.) Groposo et al., F senex (Nees & Mont.)

Ghob.-Nejh., and FE. wahlbergii (Fr.) T. Wagner & M. Fisch.

During the study of wood-inhabiting fungi in Atlantic Rainforest areas of

Sao Paulo state, an unknown Fuscoporia species was found. The phylogeny

inferred from ribosomal ITS and LSU sequence analyses of the unknown

fungus support it in a monophyletic terminal clade, distinct from all the other

Fuscoporia species for which DNA sequence data are available, and we describe

species here as F. atlantica.

Materials & methods

Morphological study

Specimens collected during 2012-2013 in Atlantic Rainforest areas in the Sao Paulo

state, Brazil, were examined according to Gilbertson & Ryvarden (1986) and Nunez

& Ryvarden (2001). Basidioma colors are described according to Kippers (2002).

For microscopic analysis, free hand sections of the basidiomata were mounted on

microscope slides with a drop of 3% KOH solution and 1% aqueous phloxine solution.

Amyloid and dextrinoid reactions were observed in Melzer’s reagent. Drawings of the

microstructures were made with the aid of camera lucida. Thirty basidiospores from

each specimen were measured. Abbreviations and codes used for the measurements are:

Dm = diameter means, L x W = length mean x width mean, Q = range of length/width

ratios, Qm = length/width mean, and n = x/y (x = number of measurements of a given

number (y) of specimens) (Coelho 2005). The specimens were deposited in Herbario,

Instituto de Botanica, Sao Paulo, Brazil (SP).

Sequencing

Total DNA was extracted from dried basidiomata. Extractions were carried out using

the Sigma-Aldrich Gen Elute™ Plant Genomic DNA Miniprep Kit and later amplified

with Sigma-Aldrich ReadyMix™ Taq PCR P4600 (Sigma-Aldrich Corporation,

St. Louis, MO, USA), following the manufacturer’s recommendations. Primers pairs

used were ITS1 and ITS4 for the ITS region (including ITS1, 5.88 and ITS2) and LROR

and LR7 for nLSU (White et al. 1990). Reactions were amplified in a thermal cycler

(C1000 Touch™ Thermal Cycler Bio-Rad) using the following parameters: 1 cycle at

94°C for 2 min; 5 cycles at 94°C for 45 s, 60°C for 50 s and 72°C for 1 min and 20 s

where the annealing temperature decreases 1°C each cycle until it reaches 56°C under

the touch-down technique (Korbie & Mattick 2008); 30 cycles at 95°C for 45 s, 55°C for

50 s, 72°C for 1 min and 20 s; and 72°C for 10 min. PCR products were checked on 2%

agarose gel before sequencing and then sequenced in both directions using the same

846 ... Pires, Motato- Vasquez, & Gugliotta

TABLE 2. LSU dataset of Fuscoporia and related genera used in the phylogenetic

analyses.

SPECIES

Coltricia cinnamomea (Jacq.) Murrill

Coltricia montagnei (Fr.) Murrill

Coltricia perennis (L.) Murrill

Coltriciella baoshanensis Y.C. Dai & B.K. Cui

Coltriciella dependens (Berk. & M.A. Curtis)

Murrill

Coltriciella oblectabilis (Lloyd) Kotl. et al.

Fomitiporella umbrinella (Bres.) Murrill

Fomitiporia mediterranea M. Fisch.

Fomitiporia punctata (P. Karst.) Murrill

Fomitiporia robusta (P. Karst.) Fiasson & Niemela

Fulvifomes fastuosus (Lév.) Bondartseva &

S. Herrera

Fulvifomes robiniae (Murrill) Murrill

Fuscoporia atlantica

Fuscoporia contigua

Fuscoporia ferrea

Fuscoporia ferruginosa

Fuscoporia gilva

Fuscoporia wahlbergii

Hymenochaete cyclolamellata T. Wagner &

M. Fisch.

Hymenochaete innexa G. Cunn.

Hymenochaete tropica S.H. He & Y.C. Dai

Inocutis dryophila (Berk.) Fiasson & Niemela

Inocutis jamaicensis (Murrill) A.M. Gottlieb

et al.

Inocutis tamaricis (Pat.) Fiasson & Niemela

Inonotus cuticularis (Bull.) P. Karst.

Inonotus hispidus (Bull.) P. Karst.

Inonotus linteus (Berk. & M.A. Curtis) Teixeira

Inonotus obliquus (Ach. ex Pers.) Pilat

Inonotus tropicalis (M.J. Larsen & Lombard)

T. Wagner & M. Fisch.

Inonotus vaninii (Ljub.) T. Wagner & M. Fisch.

Mensularia crocitincta (Berk & M.A. Curtis)

T. Wagner & M. Fisch.

Mensularia hastifera (Pouzar) T. Wagner &

M. Fisch.

Mensularia radiata (Sowerby) Lazaro Ibiza

Onnia tomentosa (Fr.) P. Karst.

Onnia triquetra (Pers.) Imazeki

Phellinidium ferrugineofuscum (P. Karst.) Fiasson

& Niemela

VOUCHER SPECIMENS

2464 REG

MF 96-96 REG

MF 92-96 REG

Dai 13075 BJFC

LR 39016 O

RC 11835 O

CBS 303.66

FMG3

MF 85-74 REG

TW 242 REG

CBS 213.36

CBS 211.36

SP465829 (paratype)

SP445618 (holotype)

TW 699 REG

ME 87-8 REG

MF 82-930 REG

MEF 91-42e REG

MEF 89-922

Cui 8548

He 555

He 493

MF 87-918 REG

Gilb. 14740 O

MF 96-415 REG

MEF 97-97 REG

MF 92-829 REG

TAA 84-12 TAA

TW 705 REG

CBS 617.89

Dai 1980 REG

LR 41826 O

MF 84-1023a REG

TW 704 REG

TW 445 REG

TW 411 REG

TN 6121 REG

GENBANK ACCESSION

NUMBERS

AF311003{s}

AY039683{t}

AF311004{s}

KC857267{o}

AY059059{u}

AY059061{u}

AY059036{u}

AY621000{r}

AF311007{s}

AF311008{s}

AY059057{u}

AY059038{u}

KP058516{m}

KP058517{m}

AF311029{s}

AF311030{s}

AF311032{s}

AY059025{u}

AF311045{s}

JQ279630{n}

JQ279674{n}

JQ279677{n}

AF311012{s}

AY059048{u}

AF311021{s}

AF311010{s}

AF311014{s}

AY059018{u}

AF311017{s}

AY059037{u}

AY059056{u}

AY059043{u}

AF311013{s}

AF311018{s}

AF311023{s}

AF311024{s}

AF311031{s}

Fuscoporia atlantica sp. nov. (Brazil) ... 847

Phellinidium fragrans CBS 202.90 AY059027{u}

Phellinus arctostaphyli (Long) Niemela MF 91-329a REG AY059026{u}

Phellinus cinereus (Niemela) M. Fisch. MF 85-917 REG AF311027{s}

Phellinus igniarius (L.) Quél. MF 83-1110a REG AF311033{s}

Phellinus laevigatus (P. Karst.) Bourdot & Galzin TN 3260 REG AF311034{s}

Phellinus overholtsii Ginns CBS 169.55 AY059019{u}

Phellinus spiculosus (W.A. Campb. & R.W. CBS 345.63 AY059055{u}

Davidson) Niemela

Phylloporia chrysites (Berk.) Ryvarden N.W. Legon O AF411821{v}

Phylloporia pectinata (Klotzsch) Ryvarden R. Coveny 113 O AF411823{v}

Phylloporia spathulata (Hook.) Ryvarden Chay 456 O AF411822{v}

Bondarzewia montana SARs.n. 258 AF042646{q}

Russula violacea AF218559{p}

Stereum hirsutum TW 235 REG AF385165{u}

Sources for published sequences: {n} = He & Dai (2012); {o} = Dai & Cui (GenBank, unpublished);

{p} = Miller et al. (2001); {q} = Moncalvo et al. (2000); {r} = Pilotti et al. (2005); {s} = Wagner &

Fischer (2001); {t} = Wagner & Fischer (2002a); {u} = Wagner & Fischer (2002b); {v} = Wagner

et al. (2002); and {m} = new sequences provided by this study.

primers as the amplification. Nucleotide sequences were determined using an Applied

Biosystems 3730xl DNA Analyser (Macrogen, Korea). Edited sequences have been

deposited in GenBank (TABLEs 1, 2).

Phylogenetic analyses

All sequences were matched against the entire GenBank database using BLAST

(http://blast.ncbi.nlm.nih.gov/), and their pairwise identity was recorded. Sequences

retrieved from GenBank (NCBI) included in the study were selected based on the

quality, identity, length, and alignment test to build the ingroup. The selected outgroup

for ITS analyses was Phellinidium fragrans (M.J. Larsen & Lombard) M. Fisch., P. weirii

(Murrill) Y.C. Dai, and P. sulphurascens (Pilat) Y.C. Dai, based on the results of the LSU

region that places Phellinidium as a sister group of Fuscoporia; the outgroup for LSU

analyses was Bondarzewia montana (Quél.) Singer, Russula violacea Quél., and Stereum

hirsutum (Willd.) Pers. (based on Wagner & Fischer 2002b). Vouchers and sequences

used in this study are provided in TaBLEs 1-2.

The LSU dataset comprising sequences of species in Hymenochaetaceae was selected

to assess the relative position of the new taxon within the family, while an ITS dataset

was assembled to verify placement of the new species among closely related taxa

present in GenBank. Nucleotide sequences were aligned automatically with MAFFT v.7

(http://mafft.cbrc.jp/aligment/server/), and then optimized manually with BioEdit 7.2.0

(Hall 1999). Best models of evolution were estimated by using JModeltest 2.c1.4 (Darriba

et al. 2012). Substitution models were TrN+I+G in the LSU and TVM+G for the ITS

dataset. Bayesian analysis was performed based on independent Markov chain Monte

Carlo (MCMC). The run consisted of five million generations, two independent runs,

and four independent chains. The sampling frequency was every 500 generations. The

first ten percent of the analysis (burn-in phase) was discarded. Posterior probabilities

(PP) were determined in MrBayes 3.2.2 (Ronquist et al. 2012) to test the support of

the branches’ nodes. PP values 20.95 were considered statistically significant, and all

848 ... Pires, Motato- Vasquez, & Gugliotta

branches supported below this value were disregarded. Maximum likelihood (ML)

analysis was performed using RaxML 7.5.4-standard (Stamatakis 2006) with 1000 rapid

bootstrap replicates. Trees from ML analysis were compared with those resulting from

the Bayesian analysis and bootstrap values >50% were used to support the respective

nodes compatible between two trees.

Results & discussion

Taxonomy

Fuscoporia atlantica Motato-Vasquez, R.M. Pires & Gugliotta, sp. nov. Fics 1-9

MycoBAnk MB 810911

Differs from Fuscoporia gilva by its dark brown crust at the pileus base, its golden-

yellow-ferruginous pore surface, its hooked setae, and its larger basidiospores.

TYPE: Brazil. So Paulo state: Parque Estadual da Cantareira, 23°32’36”S 46°37'59”W,

27 June 2012, V. Motato-Vasquez, M.C. Westphalen & A.C. Bolafos 230 (Holotype,

SP445618; GenBank KP058515, KP058517).

Erymo ocy: atlantica - refers to the Brazilian Atlantic Rainforest biome, the only

known habitat of this species.

BASIDIOMA annual, pileate, sessile to effuse-reflexed, flexible when fresh to

leathery when dry, solitary to imbricate, without odor or taste. PILEUS broadly

attached, semicircular, applanate, <(5.0-)8.0-9.0 x 4.0-6.0 x 0.1-0.4 cm; upper

surface concentrically zonate, radially wrinkled, glabrous to tomentose, brown

(N,,A,,.M,,) to yellowish-brown (N,,A,,M.,,), with a matte dark brown crust

(N,,A,.™M,,) at the base. MARGIN thin, obtuse, entire, not involute when dry,

50° 99

sterile, up to 0.2 cm, yellowish-brown (N,,A,,M.,) to black brown (N,,.M,,C,,).

CONTEXT homogeneous, dense, azonate, golden-yellow-ferruginous

(N,,A,.™M,,). PORE SURFACE golden-yellow-ferruginous (N,A,,M,,), pores

rounded, invisible to the naked eye, 7-9 per mm, tubes concolor with the pore

surface, <0.2 cm deep, dissepiments entire to slightly lacerated, thin to slightly

thick. HypHAL sysTEM dimitic, generative hypha simple-septate, hyaline,

golden yellow to rusty brown, thin to thick-walled, straight, sparsely branched,

3.0-6.5 um diam., rarely branched, thin-walled hyphae at dissepiments edges

encrusted; skeletal hyphae thick-walled, straight, unbranched, dark-brown,

3.0-6.0 um. HYMENIAL SETAE abundant, subulate to ventricose, mostly uncinate

or hooked, dark brown and thick-walled, 20-45 x 7-10 um. CysTIDIOLES

abundant. Basrp1a clavate, hyaline, 4-sterigmated, thin-walled, 9-11 x 5-7 um.

BASIDIOSPORES abundant, broadly ellipsoid to ellipsoid, hyaline to pale yellow,

smooth, thin-walled, inamyloid, 4-4.5 x (2-)3-3.5 um (Xm = 4.2 x 2.8 um),

n = 60/2, Q = (1.1-)1.3-2, Qm = 1.5.

ECOLOGY & DISTRIBUTION: growing on dead branches and trunks of

angiosperms. Known only from Brazilian Atlantic Rainforest, Sao Paulo state.

Fuscoporia atlantica sp. nov. (Brazil) ... 849

Figures 1-4. Fuscoporia atlantica (Holotype, SP445618): 1. Basidioma. 2. Margin when dry.

3. Crust on upper surface. 4. Hymenial surface. Scale bars: 1 = 5 cm; 2, 3 = 0.1 cm; 4 = 0.05 cm.

ADDITIONAL SPECIMENS EXAMINED —Fuscoporia atlantica: BRAZIL. SAo PAULO

STATE: Parque Estadual da Serra do Mar, Nucleo Santa Virginia, 11 May 2013, R.M.

Pires 8 (SP465829; GenBank KP058514, KPKP058516).

Fuscoporia gilva: BRAZIL. SAo PAULO sTaTE: Parque Estadual da Cantareira, 21

October 2011, V. Motato- Vasquez & A.M. Gugliotta 22 (SP417971); 16 February 2012,

M. Capelari & J.J.S. Oliveira 4683 (SP445350).

Fuscoporia rhabarbarina: BRAZIL. SANTA CATARINA STATE: 26 December 1988,

F. Furlani & C.L. Leite 186 (FLOR 10929). CANADA. on wood, Dr. Richardson 4

(K187843, lectotype).

ComMENTs — Fuscoporia atlantica is a distinct species that produces effused-

reflexed/pileate basidiomata with a radially wrinkled and tomentose pileus

and matte dark brown crust at the base, a margin that is yellowish-brown

when fresh to black brown after drying, tiny pores, hooked hymenial setae,

and broadly ellipsoid to ellipsoid basidiospores. Fuscoporia atlantica is very

similar to E gilva, which differs microscopically by its straight setae and

smaller basidiospores and macroscopically by its crust-less pileus base and

850 ... Pires, Motato- Vasquez, & Gugliotta

dark purplish brown pore surface (Ryvarden & Johansen 1980, Loguercio-Leite

& Wright 1995, Dai 1999). Fuscoporia wahlbergii, another effuse-reflexed to

pileate species with hooked hymenial setae that occurs in Brazil, can be easily

distinguished macroscopically by its woody basidiomata with a reddish-brown

to umber upper surface that is narrowly banded in concentric sulcate to flat

zones and rusty to chestnut brown pore surface. Fuscoporia atlantica is also

morphologically similar to FE rhabarbarina, which can be easily distinguished

macroscopically by a glabrous pileus with sulcate zones and distinctive black

crust covering the entire pileus surface and its yellowish brown context and

microscopically by the smaller basidiospores (3-4 x 2-3 um) and straight

ventricose hymenial setae (Groposo et al. 2007).

Key to species of Fuscoporia from Brazil

l“Basidiomarcompletely tesupitiate: 2-5 rh ret eh ian el aa ey ek het die ber keee bee 2

I, Basidioma eftused-retlexedto pileate= 040. nse aw et new ene ee ge e EE Hee hE ewe’ 5

2, Itamal sétae present; spores ellipsoid:.\ b..2.. 5004-054 40.0n4 pn dna gone eae ae be ne 3

2: Famal.setaeabsent, spores cylindtical: a8 8.8 neat Meet laa Mceel ae dt ae 4

3. Pores 2-3 per mm, hymenial setae subulate......................00.. E contigua

3. Pores 5-6 per mm, hymenial setae ventricose ..................06. E ferruginosa

4. Basidioma dark reddish-brown, hymenial setae 18.0-36.0 x 5.0-8.0 um,

basidiospores cylindric, 4.0-7.0 x 2.0-3.0 Um .......... eee eee eee E ferrea

4, Basidioma pale yellow to brown, hymenial setae 10.0-15.0 x 5.0-7.5 um,

basidiospores ellipsoid, 4.5-7.0 x 2-2.5um...............0.. E punctatiformis

SAPs With CiStnCtDIACKACEUS be ies Fo nd Bl toy stan ween tne at E rhabarbarina

Ss ULeLio WALNO ied DIACKGHIGE gel c's fat oc.'2, 0M AON BRON tate tat RON Rt ee DR, hee 6

Gy menial setae: bituncateOn-Spinyy =. eagkilie soneenste ances Uapenstcaten ogee atlveee E bifurcata

6. Hymenial setae nor bifurcate neither spiny (all setae apically regular) ........... 7

7. Hymenial setae hooked to straight, both present .................... 0. eee eee 8

% Hymenial setae only strarcht ews 2 Secece < Stewie < Werwicese Ubcmiese Mcwie« NEcwitare Wes os deme # 9

8. Basidioma woody, upper surface without a crust,

basidiospores subglobose, 4.0-5.0 x 3.5-4.5 um... 2.2... eee eee E. wahlbergii

8. Basidioma leathery, upper surface with a dark brown crust,

basidiospores ellipsoid to broadly ellipsoid, 4.0-4.5 x 2.0-3.5 um.... F atlantica

OEP OTESH ES PCAN 4.m, Tutus ths aah eee tad cial tok eg Sek eer ee E palmicola

9. Porestlateer, SA: per amin iy 468s ode y ooh eg ee Ey ee ey yeh eee eee eigen 10

EOS ROLES 9 CRIN ee sc. ares pdb sMidbityh pill dip LV iat dipe kV aldiye bVo-uldipm bye whdtpe prot em byte | 11

11. Context reddish-brown, basidiospores 2.0-2.5 um wide ............... EF. chrysea

11. Context yellowish brown, basidiospores 3.0-3.5 um wide .................00. 12

Fuscoporia atlantica sp. nov. (Brazil) ... 851

B88" Dd

— 8

FiGuRES 5-9. Fuscoporia atlantica (from the holotype): 5. Basidiospores. 6. Hymenial setae. 7.

Generative hyphae. 8. Skeletal hyphae. 9. Hyphal tips at dissepiment edges. Scale bar = 5 um.

12. Basidiospores ellipsoid to ovoid, 4.0-5.0 x 3.0-3.5 um,

hymenial setae 20250) 55 —G py ew Sache hades tee ale ee Pade ee Balle reall E gilva

12. Basidiospores ellipsoid, 3.0-4.5 x 2.5-3.0 um,

hymenial setae 15-30 X 4.5-7.5 um ........ eee eee eee eee E flavomarginata

13. Basidiospores ellipsoid to subcylindrical, 3.5-4.5 x 2-3 um ........ E callimorpha

13. Basidiospores broadly ellipsoid, 4.5-6 x 3.5-5 um ...... eee eee eee EF senex

Phylogenetic analyses

The final LSU matrix comprised 51 sequences and 838 characters, of which

534 (63.72%) were parsimony-informative. Bayesian analysis produced an

average standard deviation of split frequencies = 0.003721. The phylogeny

852 ... Pires, Motato- Vasquez, & Gugliotta

Fomitiporella umbrinella AYO059036

Inocutis dryophilus AF311012

Inocutis tamaricis AF311021

Inocutis jamaicensis AY059048

ie Fulvifomes fastuosus AY059057

Fulvifomes robiniae AY059038

4/100 Phylloporia chrysites AF411821

97/85 [| Phylloporia pectinata AF411823

Phylloporia spathulata AF411822

Mensularia hastifera AF311013

pc radiata AF311018

Inonotus tropicalis AYO59037

Inonotus linteus AY059018

Inonotus vaninii AYO59056

i Phellinus cinereus AF311027

Phellinus laevigatus AF311034

96/71 Phellinus igniarius AF311033

feo Phellinus arctostaphyli AYO059026

Phellinus spiculosus AYO59055

Phellinus overholtsii AYO59019

1/100 Fomitiporia punctata AF311007

Fomitiporia robusta AF311008

Fomitiporia mediterranea AY621000

Hymenochaete cyclolamellata JQ279630

99/91 Hymenochaete innexa JQ279674

'__ Hymenochaete tropica JQ279677

Onnia tomentosa AF311023

L— Onnia triquetra AF311024

1 99/88 Coltricia cinnamomea AF311003

—\H44 97/81 Coltricia montagnei AY039683

1/100 Coltricia perennis AF311004

Coltriciella dependens AY059059

LL Coltriciella oblectabilis AYO59061

Coltriciella baoshanensis KC857267

.97/89__

Fuscoporia contigua AF311029

Fuscoporia wahlbergii AF311045

r— Fuscoporia gilva AY059025

1/100;— Fuscoporia atlantica KP058516

99/95 Fuscoporia atlantica KP058517 holotype

Fuscoporia ferruginosa AF311032

99/96 = Fuscoporia ferrea AF311030

ee | Phellinidium ferrugineofuscum AF311031

=| Phellinidium fragrans AY059027

-—___________ Stereum hirsutum AF385165

Bondarzewia montana AF042646

.96/-

99/61 1/59

99/95

Russula violacea AF218559 0.05

FiGuRE 10. Phylogenetic placement of nLSU sequences from Fuscoporia atlantica and related

species and genera, inferred from Bayesian analysis and ML. The new species is presented in bold

font. Support values consist of the Bayesian posterior probability (PP) >0.95, followed by ML

bootstrap (BS) values >50%.

derived from LSU sequences of different taxa of Hymenochaetaceae from

GenBank (Fic. 10) clusters the new species within Fuscoporia genus with full

support.

The ITS analysis included 37 sequences and 585 characters, of which 302

(51.62%) were parsimony-informative. Bayesian analysis produced an average

standard deviation of split frequencies = 0.005049. The ITS-derived tree

(Frc. 11) recovered the newly described taxon among a limited number of the

most closely related taxa within Fuscoporia and supported separation of the

new species, F atlantica (PP = 1; bootstrap = 100%). Our morphological and

phylogenetic analyses indicated a close relationship between E atlantica and

EF gilva.

Fuscoporia atlantica sp. nov. (Brazil) ... 853

1/100 Fuscoporia viticola AY558653

ete Fuscoporia viticola JQ794583

Fuscoporia viticola JQ358814

-—— Fuscoporia ferruginosa JQ794577

1/100 _- Fuscoporia ferruginosa JQ794573

“_ Fuscoporia ferruginosa JQ794572

1/100 Fuscoporia ferrea JQ673178

ef Fuscoporia ferrea KJ677120

1/100 Fuscoporia ferrea AY558617

1/99'_ Fuscoporia ferrea DQ516525

1/100

- Fuscoporia ruftincta GU594160

Fuscoporia rufitincta JQ794580

1/100 We taenne rufitincta JQ794579

1/94 Fuscoporia contigua JQ794550

Fuscoporia contigua JQ794546

1/- Fuscoporia contigua JQ794548

Fuscoporia contigua JQ794547

Fuscoporia torulosa EF068139

Fuscoporia torulosa EF068239

L Fuscoporia torulosa EF068238

1/100 Fuscoporia senex JX463658

7 1/90 Fuscoporia senex AY558647

-/74, '— Fuscoporia senex KC414230

Fuscoporia callimorpha JF692193

.98/100

1/100

Fuscoporia callimorpha JF692190

Fuscoporia callimorpha JF692191

-— Fuscoporia gilva FJ481039

27100 ; Fuscoporia gilva AY558620

1/100 1/100 Fuscoporia gilva Kr692068

Fuscoporia gilva JX427049

1/100 Fuscoporia atlantica KP058514

Fuscoporia atlantica KP058515 holotype

Phellinidium fragans AY558619

1/100 Phellinidium weiri AY829341

1/100 Phellinidium weirii AY829342

Phellinidium sulphurascens EF527215

Phellinidium sulphurascens EF527216 0.08

FicureE 11. Phylogenetic analysis of ITS sequences from Fuscoporia atlantica and related species,

based on Bayesian analysis and ML. The new species is presented in bold font. Support values

consist of Bayesian posterior probability (PP) values >0.95, followed by ML bootstrap (BS) values

>50%.

Our introduction of a new taxon from the Brazilian Atlantic Rainforest,

Sao Paulo, Brazil, offers another example of how poorly known is the fungal

diversity in the area.

Acknowledgments

The authors are grateful to the curators of the herbaria FLOR and K for the loan of

specimens, especially those of type material. VMV gratefully acknowledges the financial

support received from the Programa Estudantes-Convénio de Pés-Graduacao — PEC-

PG, from CAPES/CNPq - Brazil, and RMP gratefully acknowledges the financial support

received from Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP

(12/25493-7). We also extend our thanks to Dr. Juliano M. Baltazar (Universidade

Estadual de Maringa, Brazil) and Dr. Michal TomSovsky (Mendel University in Brno,

Czech Republic), who kindly reviewed the manuscript.

854 ... Pires, Motato- Vasquez, & Gugliotta

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Hymenochaetales (Basidiomycota) supported by nLSU rDNA sequence data. Mycol. Res.

105: 773-782.

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Phellinus s.l. and Inonotus s.1., and phylogenetic relationships of allied genera. Mycologia

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(Hymenochaetales). Mycol. Progr. 1: 105-116.

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methods and applications. San Diego, Academic Press.

MY COTAXON

http://dx.doi.org/10.5248/130.857

Volume 130, pp. 857-865 July-September 2015

Volvariella rava sp. nov. from southern China

JIANG Xu”?, Tat-Hur Lr’*, YA-HENG SHEN’, & MING ZHANG”?

' School of Bioscience & Bioengineering, South China University of Technology,

Guangzhou 510006, China,

’ State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key

Laboratory of Microbial Culture Collection and Application, Guangdong Institute of

Microbiology, Guangzhou 510070, China

*CORRESPONDENCE TO: mycolab@263.net

ABSTRACT— Volvariella rava, a new grey species collected from Guangzhou City in southern

China, is described, illustrated, and compared with closely related taxa of Volvariella. The new

species is characterized by a medium-sized basidiome and greyish brown appressed-fibrillose

pileus; it closely resembles V. volvacea but differs by its smaller spores. Molecular analysis

based on nrITS sequences confirmed that V. rava is distinct from other morphologically

similar Volvariella species.

Key worps—Agaricales, Basidiomycota, fungal diversity, phylogeny, taxonomy

Introduction

Volvariella Speg. (traditionally referred to Pluteaceae, but see Justo et al.

2011) is characterized by a small to large agaricoid basidiome having a slender

to fleshy stipe with a volva but no ring, crowded and free (often remote)

lamellae, and pink basidiospores in mass (Orton 1986). The genus has been

reported from tropical, subtropical, and temperate regions of both the eastern

and the western hemispheres (Shaffer 1957). A review of the bibliographical

and herbarium records (Mycological Herbarium of Microbiology Institute

the Chinese Academy of Science (HMAS), Herbarium of Cryptogams,

Kunming Institute of Botany Academy of Science (HKAS), Mycological

Herbarium of Mycological Institute of Jilin Agriculture University (HMJAU),

and Mycological Herbarium of Microbiology Institute Guangdong Academy

of Science (GDGM)) revealed that a total of 22 Volvariella species have been

described and reported from China (Teng 1963, Tai 1979, He & Feng 1987, Bi

et al. 1994, Ying & Zang 1994, Shao & Xiang 1997, Mao 2000, Li et al. 2009, Bao

858 ... Xu & al.

& Wang 2013). A grey species of Volvariella discovered from Tianlu Lake Forest

Park of Guangzhou City, southern China, is described here as a new species

based on morphological and molecular characters.

TABLE 1. Sequences of Volvariella (V.) and outgroup species used in the

phylogenetic analyses.

SPECIES VOUCHER No. GENBENK No.

Volvariella bombycina AJ244 HM562212.1

SP393635 EU920673.1

V. caesiotincta MA54717 HM562211.1

V. dunensis SCM3513 JF415140.1

V. hypopithys AV137 HM246492.1

V. nivea GDGM25489 FJ749127.1

V. nullicystidiata SP393639 EU920671.1

V. pusilla AJ51(LOU) JF415137.1

AV 139 HM246494.1

V. rava GDGM41955 KP784686

V. sathei AMH 9436 JN792550.1

V. cf. sathei Hamal93 KF926663.1

Hama390 KF926666.1

Hamal94 KF926664.1

Hama204 KF926665.1

V. strangulata AV141 HM246493.1

V. surrecta AJ55 HM562213.1

V. taylorii AJ54 HM562210.1

V. terrea LUG11010 JF415141.1

V. volvacea AV 143 HM246500.1

ATCC MYA-4696* HQ999973.1

Volvariella sp. LOU18924 JF415139.1

Volvopluteus earlei (as V. acystidiata) HG1973 HM246499.1

Volvopluteus earlei (as V. media) HG2001 HM246498.1

Volvopluteus earlei (as V. cookei) AV133 HM246496.1

Volvopluteus earlei (as V. earlei) AV134 HM246497.1

Schizophyllum commune IFM 56967* AB566277.1

* Strain no.

Material & methods

The macroscopic descriptions were based on the fresh basidiomata (Vellinga 1988).

The microscopical characters were observed using an Olympus DP25 microscope.

Thin handmade sections were made from dried specimens and revived in 5% KOH.

Volvariella rava sp. nov. (China) ... 859

The pileipellis and stipitipellis were examined in KOH and basidiospores, basidia

and cystidia were observed in 1% Congo red. Basidiospore dimensions are based on

the random measurements of 30 basidiospores. The abbreviation [n/m/p] represents

measurements made on n basidiospores in m basidiomata from p collections.

Basidiospores dimensions follow the form (a—)b-c(—d) where b-c represents 95% of the

measured values and extreme values shown in parentheses. The following abbreviations

are used in the descriptions: avl for average length, avw for average width, Q for quotient

of length/width, avQ for average quotient. Colour designations follow Kornerup &

Wanscher (1978). The type specimen is deposited at the Herbarium of Guangdong

Institute of Microbiology, Guangzhou, China (GDGM).

Genomic DNA of the new species was extracted from herbarium materials using

the Sangon Fungus Genomic DNA Extraction kit (Sangon Biotech Co., Ltd., Shanghai,

China) according to the manufacturer’s instructions. The ITS region was amplified by

PCR with universal primers ITS4 and ITS5 (White et al. 1990). The amplified strands

were sequenced with an ABI 3730 DNA analyzer (Invitrogen Life Technologies Co., Ltd,

Shanghai). The newly generated sequences were submitted to GenBank. For phylogenetic

analysis, additional related sequences of Volvariella obtained from NCBI (TaBLE 1) were

edited and aligned by Clustal W (Thompson et al. 1994) and manually checked and

adjusted. The outgroup comprised Schizophyllum commune Fr. and Volvopluteus earlei

(Murrill) Vizzini et al. A phylogenetic tree was generated with MEGA v.6.0 (Tamura et

al. 2013). All characters were treated as unordered and equal weight. Gaps were treated

as missing data. Bootstrap values were calculated for 1000 replicates.

Taxonomy

Volvariella rava T.H. Li & Jiang Xu, sp. nov. PxaTEs 1, 2

MycoBank MB 811615

Differs from Volvariella volvacea by its smaller basidiospores, glossy pileus with

appressed fibrils, and irregularly lobed volva.

Type: China, Guangdong Province, Guangzhou City, Tianlu Lake Forest Park,

23°13.38’N 113°25.05’E, alt. 50-100 m asl, 14.09.2012, Zhang M, Zhou SH (Holotype,

GDGM41955; GenBank KP784686).

ErymMo oey: The epithet ‘rava’ refers to the tawny grey to grayish brown colour of the

pileus.

BASIDIOMATA medium. PILEus 25-60 mm diam., ovoid when young, becoming

subconical with an obtuse apex, convex at maturity; surface tawny grey to

greyish brown (5E3-5F3), darker at the disc, subviscid to viscid when wet,

nearly glabrous to innately appressed-fibrillose; margin estriate, often radially

rimulose on drying, eroded. CONTEXT white, unchanging on bruising, 5-6 mm

thick near stipe. LAMELLAE free, remote, white when young, becoming sordid

and pinkish, <6 mm wide, densely crowded, with lamellulae of different

lengths. StrpE 70-80 x 5-10 mm, cylindrical, compressed, slightly attenuated

860 ... Xu & al.

towards apex, white, glabrous, solid. VoLva free, concolorous with the pileus

above, whitish (4A2) below, irregularly lobed, relatively thick, <12 mm high

and 15 mm wide. Opour and TasTE mild.

BASIDIOSPORES (60/2/2) (4-)5-6.5 x 4-5 um, (avl x avw = 5.47 x 4.28),

Q = 1.1-1.5, avQ = 1.28, ovoid to ellipsoid, smooth, stramineous to salmon-

coloured, pinkish at thickened wall. Basrp1a 24-30 x 7-10 um, cylindric-

clavate to clavate, 4-spored, rarely 2-spored; sterigmata short (<2 um), slender.

LAMELLAR EDGE heteromorphous. CHEILOCysTIDIA abundant, 55-80 x

15-25 um, fusoid to ventricose, or subclavate with blunt end, thin-walled, easily

collapsing. PLEUROCYSTIDIA scattered, 35-71 x 11-30 um, cylindric-clavate,

mucronate with a broadly rounded apex, thin-walled, similar to cheilocystidia.

LAMELLAR TRAMA convergent, composed of hyaline hyphae, 4-7 um diam.,

thin-walled. PILEIPELLIS a regular cutis consisting of procumbent hyphae,

12-23 um diam., gelatinous, thin-walled; terminal elements blunt or slightly

acute, 54-86 x 10-18 um, hyaline. Sripitipetuis hyphae cylindric, rarely

septate, 7-25 um diam., with rounded or slightly acute apex. VoLva made up

of filamentous hyphae, with inflated cells (32-50 x 10-15 um) at outer and

inner surfaces, often containing brownish vacuolar pigments. CAULOCYSTIDIA

absent. CLAMP CONNECTIONS absent in all hyphae.

Hasitat & DiIsTRIBUTION—Solitary on ground of bamboo grove in a

subtropical evergreen broad-leaved forest. Known only from the type locality.

Additional specimen examined: CHINA, GUANGDONG PROVINCE, Guangzhou City,

Tianlu Lake Forest Park, 12.05.2011, Zhang M, Xu J (GDGM41548).

Phylogenetic analyses

The 788 bp nrITS sequence from the new species (GDGM41955) differs

from all other known Volvariella sequences deposited in the NCBI-GenBank

database (http://www.ncbi.nlm.nih.gov/).

An initial BLASTn search comparing the nrITS sequence from V. rava with

the NCBI nrITS sequence database showed maximum 98% similarity with

V. nivea T.H. Li & Xiang L. Chen (FJ749127) followed by 96% similarity with

V. strangulata (Romagn.) Vizzini & Contu (HM246493) and V. dunensis (Vila

et al.) Justo & M.L. Castro (JF415140), 90% similarity with V. surrecta (Knapp)

Singer (HM562213), and 84% similarity with V. caesiotincta P.D. Orton

(HM562211).

Twenty-six additional sequences obtained from NCBI, representing

15 Volvariella spp. and an outgroup of Volvopluteus earlei (deposited as

Volvariella) and Schizophyllum commune, were included in the analyses. In

the phylogenetic tree (PLATE 3), all Volvariella species were grouped in two

main clades I and II. Volvariella rava and V. nivea clustered in subclade Ib with

Volvariella rava sp. nov. (China) ... 861

PiatTE 1. Volvariella rava: basidiomata in the natural habitat. a. Surface and gill views (holotype,

GDGM41955); b. young basidiomata (GDGM41548). Scale bars = 1cm.

PiaTE 2. Volvariella rava (holotype, GDGM41955). a. Basidiospores; b. basidia; c. cheilocystidia;

d. pleurocystidia; e. pileipellis elements. Scale bars: a = 5 um; b = 10 um; c, d = 20 um; e = 40 um.

862 ... Xu &al.

84% boot strap support, but the different branch lengths imply considerable

sequence divergence between these two species.

In this study, a high degree of variability (including considerable

misalignment and several indels) among the Volvariella species was notable

throughout the ITS 1 and ITS 2 regions, although the 5.8S region had few

changes and remained conserved, as indicated by Li et al. (2009), Vizzini et al.

(2011), and Senthilarasu et al. (2012) for V. nivea and V. sathei Senthil. et al.

Discussion

Volvariella rava can be placed in stirps Volvacea sect. Volvariella based

on its medium-sized pileus with appressed fibrils and small (<12 um long)

basidiospores (Singer 1986, Justo et al. 2011). Volvariella rava closely resembles

other species in stirps Volvacea: V. volvacea (Bull.) Singer, V. bakeri (Murrill)

Shaffer, V. esculenta (Massee) Singer, and V. diplasia (Berk. & Broome) Singer.

However, V. volvacea is distinguished from V. rava by its white to dark erect

pileus fibrils and much larger basidiospores (7-10 x 4.5-7 um; Shaffer 1957,

Pegler 1977, 1986, Priest & Conde 2006). Volvariella bakeri has a dark-

fuliginous pileus with concolorous volva and larger basidiospores (6.9-9.3 x

4.6-6.9 um; Shaffer 1957). Volvariella esculenta differs from V. rava in having

a glabrous grayish violet pileus with striate margin and elongate basidiospores

(6-7 x 4-5 um; Massee 1908, Singer 1949). Volvariella diplasia has a white

to golden yellow silky-floccose pileus and larger basidiospores (7.5-10.5 x

4.7-6.5 um; Singer 1949, Pegler 1986).

Volvariella rava somewhat resembles V. apalotricha (Berk. & Broome)

Pegler, V. pseudovolvacea (Berk. & Broome) Singer, and V. terastia (Berk. &

Broome) Singer in its medium-sized basdiomata, grayish to dark pileus,

and <7 um long basidiospores. However, V. apalotricha has an ash-grey to

greyish brown pileus with blackish radial fibrils, a small white membranous

volva, narrower basidiospores (3.5-4.5 um diam.), and ventricoso-clavate

cheilocystidia and pleurocystidia (Pegler 1986, Pradeep et al. 1998, Priest &

Conde 2006). Volvariella pseudovolvacea has dense black fibrillose striae on

the pileus, slightly smaller basidiospores (4.2-5.2 x 2.5-3.5 um), and fusoid

ventricose to lageniform cheilocystidia (Pegler 1986, Bi et al. 1994, Pradeep

et al. 1998). Volvariella terastia differs from V. rava in its larger basidiome

and pileus (60-110 mm diam.) covered with dark brown fibrils, thicker stipe

(<20 mm diam., <30 mm thick at the base), larger volva (<80 mm high), and

smaller cheilocystidia (32-36 x 10-13 um; Pegler 1986).

Volvariella aethiops A. Favre & Vialard, V. caesiotincta, V. lepiotospora

Singer, and V. taylorii (Berk. & Broome) Singer are also somewhat similar to

V. rava in their greyish brown or darker pileus colour. However, V. aethiops has

Volvariella rava sp. nov. (China) ... 863

41 -— Volvariella pusilla AV139

qF Volvariella pusilla AJ51(LOU)

Volvariella hypopithys AV137

31) g7l Volariella sp. LOU18924

64 Volvariella surrecta AJ55

88 Volvariella strangulata AV141

Volvariella terrea LUG11010

Volvariella taylorii AJ54

Volvariella dunensis SCM3513 la

oF Volvariella nullicystidiata SP 393639

Volvariella caesiotincta MA54717

JN792550 Volvariella sathei. AMH 9436

70 Volvariella cf. sathei Hama193

100)! Volvariella cf. sathei Hama390

64! Volvariella cf. sathei Hama194

36 Volvariella cf. sathei Hama204

Volvariella volvacea AV143

q1 Volvariella rava GDGM41955

B4 Volvariella nivea GDGM25489

Volvariella volvacea strain ATCC MYA-4696

100 Volvariella bombycina AJ244 II

100 _ Volvariella bombycina SP393635

KF679517 Schizophyllum commune

Volvopluteus earlei HG1973

Volvopluteus earlei HG2001

Volvopluteus earlei AV133

Volvopluteus earlei AV134

0.05

PLATE 3. Phylogenetic tree of Volvariella sequences generated from ITS dataset using NJ method.

Bootstrap values 250% are shown on the branches. Scale bar indicates nucleotide substitutions

per site.

a distinctly shaggy fibrillose pileus and longer basidiospores (6.2-8.6 x 4-4.7

um; Favre & Vialard 2007, Kibby 2011). Volvariella caesiotincta is lignicolous

and has greyish white to greyish brown to pale bluish pileus, a membranous

and often 3-lobed volva, and narrow basidiospores (5.5-7.5 x 3.3-4 um;

Orton 1974, 1986). Volvariella lepiotospora differs from V. rava in its smaller

(32 mm diam.) pileus covered by brownish black to black fibrils and narrower

basidiospores (4.7-6(-7) x 3-3.7 um; avQ = 1.67; Singer 1955, Shaffer 1957).

Volvariella taylorii is separated by its silky fibrillose pileus, smaller stipe (35-55

x 2-4 mm), and smaller lageniform pleurocystidia (32-48 x 8-12 um), and

cheilocystidia (25-45 x 9-13 um; Pegler 1983, Pradeep et al. 1998).

864 ... Xu &al.

Volvariella nivea was described from the Baiyun Mountain Forest Park (very

close to Tianlu Lake Forest Park) growing on the ground of a bamboo grove.

It thus has a similar habitat to V. rava and shares similar spore dimensions

(6-7 x 4.5-5.5 um), but clearly differs in its snow-white basidiomata with the

distinctly free-ended fibrils on the pileus (Li et al. 2009).

Acknowledgments

We would like to thank Dr. Alfredo Justo (Universidad Nacional Aut6noma de

México, Mexico) and Dr. Gunasekaran Senthilarasu (SRM Research Institute, SRM

University, Tamil Nadu, India) for their pre-submission reviews. This study was

supported by the National Natural Science Foundation of China (Nos. 31093440,

31170026, 31370072) and the early-stage basic research key projects of National Basic

Research Program of China (2014CB460613).

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MY COTAXON

http://dx.doi.org/10.5248/130.867

Volume 130, pp. 867-874 July-September 2015

First report of Leptographium pini-densiflorae

causing sapstain on Pinus densiflora in Korea

Joo-HyuUN HONG’, YEONGSEON JANG’, SEOKYOON JANG’,

MIHEE MIN’, & JAE-JIN KIm'*

‘Division of Environmental Science & Ecological Engineering,

College of Life Sciences & Biotechnology, Korea University,

145, Anam-ro, Seongbuk-gu, Seoul 02841, Korea

Division of Wood Chemistry & Microbiology, Korea Forest Research Institute,

57, Hoegi-ro, Dongdaemum-gu, Seoul 02455, Korea

*Department of Converging Technology e& Industry Strategy,

Bureau of Health Industry Policy, Korea Health Industry Development Institute,

187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si,

Chungcheongbuk-do 28159, Korea

*CORRESPONDENCE TO: jae-jinkim@korea.ac.kr

AsBstRAcT —Morphological observation and phylogenetic analysis revealed that the

dominant sapstain fungus isolated during a recent survey of sapstain of Japanese red pine

logs in Korea represents Leptographium pini-densiflorae, a new record for Korea. A detailed

description of L. pini-densiflorae accompanies illustrations of cultural and microscopical

characters.

KEY worps — ascomycetes, blue-stain fungi, taxonomy

Introduction

Species of Leptographium Lagerb. & Melin are well known as agents of

sapstain in conifers and have been reported worldwide (Seifert et al. 2013).

Some Leptographium spp. are known to have a symbiotic relationship with bark

beetles such as red turpentine beetle (Dendroctonus valens), pine shoot beetle

(Tomicus piniperda), and European spruce bark beetle (Ips typographus) (Jacobs

& Wingfield 2001, Wingfield et al. 1993, Masuya et al. 2003, 2009, Seifert et al.

2013, Solheim et al. 1993, Zhou et al. 2002). Associated insects provide a means

of mobility for Leptographium spp.

868 ... Hong & al.

Among the insects associated with Leptographium species, some

Dendroctonus bark beetles cause devastation in Canada and China (Lee et al.

2006, Lu et al. 2009). Lodgepole pine (Pinus contorta) forest in British Columbia

has suffered from considerable mortality due to the combination of mountain

pine beetle (D. ponderosae) and Leptographium longiclavatum S.W. Lee et al.

(Lee et al. 2006). A similar phenomenon happened in northern China, where

extensive tree death occurred in forests over large areas. Lu et al. (2009) noted

that transportation of unprocessed logs was a dispersal means for the rapid

spread of exotic insects and Leptographium spp.

Research on sapstain fungi has focused on commercial logs and boards in

Korea, where Grosmannia koreana (Masuya et al.) Q. Lu et al., Ophiostoma

floccosum Math.-Kaarik, O. ips (Rumbold) Nannf., O. piceae (Miinch) Syd. &

P. Syd., O. piliferum (Fr.) Syd. & P. Syd., and O. quercus (Georgev.) Nannf. have

been reported (Kim et al. 2005, 2007, 2010, 2011). Among the ophiostomatoid

fungi, Grosmannia koreana [= Leptographium koreanum J.J. Kim & G.H.

Kim,] was the most dominant sapstain fungal species isolated from conifer

logs and wood products in Korea (Masuya et al. 2005, Kim et al. 2005, 2007).

However, during recent surveys of log yards, another Leptographium sp. was

found dominant in Japanese red pine (Pinus densiflora Siebold & Zucc).

Morphological and phylogenetic analyses revealed that the isolates represent

Leptographium pini-densiflorae, which has not been reported previously in

Korea. Detailed description and figures for this species are provided.

Materials & methods

Morphological examination

Leptographium isolates were collected from the sapwood of Japanese red pine, at Mt.

Chiak and the Bonghwa log yard in Korea. The fungus was isolated by inoculating 2%

malt extract agar (MEA, malt extract 20 g, agar 15 g, distilled water 1000 ml) containing

cycloheximide with a small piece of sapwood (Masuya et al. 2000). The pure isolate

was subcultured on 2% MEA and incubated at 20°C for 14 days for morphological

investigation. For microscopic characteristics, fungal structures were mounted in 85%

lactic acid and observed at <1000x using an Olympus BX51 light microscope. Fifty

measurements were made for each character. Isolates were deposited in the Korean

Agricultural Culture Collection, National Academy of Agricultural Science, Jeonju-si,

Korea (KACC) and the Korea University Culture Collections, Korea University, Seoul,

South Korea (KUC).

Phylogenetic analysis

Genomic DNA was extracted according to Jang et al. (2013a). PCR reactions were

performed for the: [1] internal transcribed spacer and part of the large subunit (ITS2-

28S), [2] translation elongation factor-la (TEF), and [3] B-tubulin (TUB) region using

the primers [1] ITS3 / LR3 (White et al. 1990), [2] EFIF / EF2R (Jacobs et al. 2004),

Leptographium pini-densiflorae sapstain (Korea) ... 869

TABLE 1. Fungal species and GenBank accession numbers of sequences

used in this study.

Be aac Oy Par er GENBANK ACCESSION No.

ITS2-28S TUB TEF

Grosmannia serpens CMW 304, CBS 141.36 JN135314 JN135334 JN135307

Leptographium bhutanense CMW 18650 EU650186 EU650190 EU650194

CMW 18649 EU650187 EU650191 EU650195

L. gracile CMW 12316 HQ406842 HQ406890 HQ406866

CMW 12398 HQ406840 HQ406888 HQ406864

L. latens CMW 12319 HQ406844 HQ406892 HQ406868

CMW 12438 HQ406845 HQ406893 HQ406869

L. manifestum CMW 12461 HQ406838 HQ406886 HQ406862

CMW 12436 HQ406839 HQ406887 HQ406863

L. pini-densiflorae CMW 25611 EU785398 EU785371 EU785427

CMW 25600 EU785397 EU785370 EU785426

CMW 5158 AY707200 KM491367 KM491467

CMW 5157 AY707199 KM491366 KM491475

KACC 47825 KP264113 KM058085 KP264112

L. procerum MUCL46357 EU502795 EU502806 EU502821

MUCL47244 EU502796 EU502807 EU502822

L. profanum CMW 10550 DQ354943 KM491375 KM491484

CMW 10552 DQ354944 KM491376 KM491485

L. sibiricum CMW 4482 KM491425 KM491379 KM491488

CMW 4481 KM491424 KM491378 KM491487

L. sinoprocerum MUCL 46331 EU296772 EU296778 EU296785

MUCL 47246 EU296774 EU296780 EU296787

L. terebrantis ATCC 58098 JF798476 JF798460 JF798471

CBS 337.7 EU296777 EU296784 EU296791

“CMW: Culture Collection of the Forestry and Agricultural Biotechnology Institute (FABI), University

of Pretoria, Pretoria, South Africa; CBS: Centraalbureau voor Schimmelcultures, Utrecht, The

Netherland; MUCL: Mycothéque de l'Université catholique de Louvain, Lou-vain-la-Neuve, Belgium;

ATCC: American Type Culture Collection, Manassas, Virginia, USA; KACC: Korean Agriculture

Culture Collection, Suwon, Korea.

and [3] T10 / BT2b (O'Donnell & Cigelnik 1997, Glass & Donaldson 1995). The PCR

conditions followed Jacobs et al. (2004) for ITS2-28S and TEF and Masuya et al. (2000)

for TUB. The products were sequenced by Macrogen (Korea) and the obtained sequences

were deposited in GenBank (TABLE 1). The sequences were proofread and compared

with reference sequences in GenBank. The phylogenetic tree was inferred according to

Jang et al. (2013b). Closely related Leptographium sequences (TABLE 1) were retrieved

from GenBank. The sequences were aligned with MAFFT 7.130 (Katoh & Toh 2008)

870 ... Hong & al.

with L-INS-i method and then manually edited using MacClade 4.08 (Maddison &

Maddison 2005). The best-fit model was searched by MrModeltest 2.3 (Nylander 2004)

under AIC criterion and majority rule consensus tree was generated using MrBayes

3.2.2 (Ronquist et al. 2012). Nodal support was assessed with posterior probabilities.

The tree was visualized using FigTree 1.4.0 (http://tree.bio.ed.ac.uk/software/figtree/).

Taxonomy

Leptographium pini-densiflorae Masuya & M.J. Wingf.,

Mycoscience 41: 428. 2000. Fic. 1

Conidiophores macronematous, mononematous with rhizoids at the

bases, single or occasionally in groups. Stipes erect, olivaceous, cylindrical,

1-6-septate, 43-160(-245) um (mean = 102.9 um) long and 3.7-8.7(-11.3) um

(mean = 5.9 um) wide at base. Conidiogenous apparatus 30-84 um long (mean

= 51 um) excluding conidial mass, 3-5 (mostly 4) series of branches. Primary

branches, 2-4 (mostly 2), without larger central branch, light olivaceous to

hyaline, smooth, 8-24.4 x 2.2-6 um (mean = 12.7 x 3.9 um). Conidiogenous

cells discrete, cylindrical, tapering slightly at the apex, 7-16.6 x 1-2.3 um

(mean = 10.8 x 1.5 um). Conidia oblong to ellipsoid, sub-truncate at base,

widely rounded at apex, 2.7-8.5 x 1.2-3 um (mean = 4.6 x 1.9 um).

Colonies with optimal growth at 25°C on 2% MEA, reaching 36 mm in

diameter in 6 days. No growth below 5°C or above 35°C. Cycloheximide tolerant

with a 65% reduction in growth on 5 g/l cycloheximide after 6 days at 25°C in

the dark. Colonies hyaline to light olivaceous. Colony margin smooth. Hyphae

submerged in the medium or aerial, light olivaceous to hyaline, verrucose, not

constricted at the septa, 1.3-11.6 um wide.

SPECIMEN EXAMINED: KOREA, Gangwon-do, Mt. Chiak, 37°24’10”N 128°2’57’”E, on

log of Japanese red pine, 26 July 2012, Joo-Hyun Hong (KACC47825 = KUC20013).

DISTRIBUTION — China, Japan, Thailand, and Korea.

REMARKS — The characteristics of Leptographium pini-densiflorae KUC20013

from Korea are consistent with the data from Masuya et al. (2000). The species

was first reported by Masuya et al. (2000) from infected Japanese red pine in

association with Tomicus piniperda in Japan, and later with T. minor, Cryphalus

fulvus, and Orthotomicus angulatus. This species was also reported from Pinus

kesiya Royle ex Gordon in Thailand and two years later from invasive pine-

infesting Dendroctonus valens in China (Yamaoka et al. 2007, Lu et al. 2009).

After L. pini-densiflorae was first discovered in Korea in 2012, it became the

major sapstain fungus detected in surveys of Japanese red pine in Korea, and

the previously dominant species, G. koreana, was no longer found. At present,

L. pini-densiflorae appears restricted to east and southeast Asia.

Leptographium pini-densiflorae sapstain (Korea) ... 871

Fic. 1. Leptographium pini-densiflorae (KUC20013). A, growth on MEA in 9 cm diam Petri dishes

after 14 d at 20°C. B, conidiophore. C, conidiogenous apparatus. D, conidiogenous cells. E, conidia.

Scale bar = 10 um.

Some Leptographium species are closely associated with bark beetles, and

the species composition of sapstainers can be directly related to the beetle

populations. The bark beetles associated with L. pini-densiflorae in Korea

should be examined.

Phylogeny

Amplification generated sequences of 954 bp from TIS2-28S, 504 bp from

TEF, and 442 bp from TUB. ‘The aligned multi-locus dataset comprised 24

isolates and 1365 characters including gaps. The best-fit models for Bayesian

phylogenetic analysis were GTR+G for ITS2-288, HKY+I for TEF, and SYM+G

for TUB. Grosmannia serpens Goid. (CMW304) was used as outgroup. The

50% majority rule consensus tree using Bayesian analysis is shown in Fie. 2.

872 ... Hong & al.

Leptographium latens CMW12319

Leptographium latens CMW12438

Leptographium gracile CMW12316

0.83

Leptographium gracile CMW12398

Leptographium sinoprocerum MUCL46331

Leptographium sinoprocerum MUCL47246

Leptographium bhutanense CMW18650

Leptographium procerum MUCL46357

Leptographium procerum MUCL47244

Leptographium profanum CMW10550

Leptographium profanum CMW10552

Leptographium pini-densiflorae CMW5158

Leptographium pini-densiflorae CMW5157

9.95 |t L eptographium pini-densiflorae CMW25600

Leptographium pini-densiflorae CMW25611

Leptographium pini-densiflorae KACC47825

Leptographium sibiricum CMW4482

Leptographium sibiricum CMW4481

Leptographium manifestum CMW12461

Leptographium manifestum CMW12436

Leptographium terebrantis ATCC58098

Leptographium terebrantis CBS337.70

Grosmannia serpens CMW304

0.1

Fic. 2. 50% majority-rule consensus tree of 600 trees resulting from a bayesian analysis of 24

ITS2-28S, TEE, and TUB sequences. Bayesian posterior probabilities >50% are shown.

The Leptographium pini-densiflorae KUC20013 was monophyletic with the

L. pini-densiflorae isolates from China (CMW25600 and CMW25611) and

Japan (CMW5157 and CMW5158) with high posterior probability (PP = 1);

L. sibiricum K. Jacobs & M.J. Wingf. was shown as a sister taxon (PP = 1).

These two species again clustered with six Leptographium spp. (PP = 0.99):

L. bhutanense X.D. Zhou et al., L. gracile Paciura et al., L. latens Paciura et

al., L. procerum (W.B. Kendr.) M.J. Wingf., L. profanum K. Jacobs et al., and

L. sinoprocerum Q. Lu et al., among which four (L. bhutanense, L. gracile,

L. latens, L. sinoprocerum) are also Asian species (Paciura et al. 2010). Unlike

L. pini-densiflorae, L. sibiricum lacks rhizoids, and although L. bhutanense,

L. gracile, L. latens, and L. sinoprocerum produce rhizoids, their colony

morphologies, conidiophore lengths, and conidial sizes (Paciura et al. 2010) all

differ from L. pini-densiflorae.

Leptographium pini-densiflorae sapstain (Korea) ... 873

Acknowledgments

This work was supported by National Research Foundation of Korea (NRF) Grant

funded by the Korean Government (Fostering Core Leaders of the Future Basic Science

Program, 2012H1A8003235). Support for this study was also provided by the Indigenous

Species Survey and Investigation project from the National Institute of Biological

Resources (NIBR) under the Ministry of Environmental, Republic of Korea. We are

grateful to Dr. Hayato Masuya and Dr. Xudong Zhou for their valuable suggestions on

the manuscript.

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basidiomycete in East Asia. Mycotaxon 124: 209-217. http://dx.doi.org/10.5248/124.209

Jang Y, Lee YM, Kim GH, Kim JJ. 2013b. Two species of Cladosporium associated with wood

discoloration in Korea. Mycotaxon 124: 21-29. http://dx.doi.org/10.5248/124.21

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

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

Kim JJ, Lim YW, Breuil C, Wingfield MJ, Zhou XD, Kim GH. 2005. A new Leptographium

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Kim GH, Kim JJ, Breuil C. 2007. Sap-staining fungi from logs and boards of two commercially

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Kim JJ, Suh DY, Kim SH. 2010. Sapstain of Japanese black pine caused by Ophiostoma piceae in

Korea. Plant Pathology Journal 26: 427-427. http://dx.doi.org/10.5423/PPJ.2010.26.4.427

Kim JJ, Suh DY, Shin SC. 2011. Disease reports: Ophiostoma ips isolated from reddish brown

stained Japanese red pine wood. Plant Pathology Journal 27: 397-397. h

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Lee S, Kim JJ, Breuil C. 2006. Pathogenicity of Leptographium longiclavatum associated with

Dendroctonus ponderosae to Pinus contorta. Canadian Journal of Forest Research 36:

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Lu Q, Decock C, Zhang XY, Maraite H. 2009. Ophiostomatoid fungi (Ascomycota) associated with

Pinus tabuliformis infested by Dendroctonus valens (Coleoptera) in northern China and an

assessment of their pathogenicity on mature trees. Antonie van Leeuwenhoek 96: 275-293.

http://dx.doi.org/10.1007/s10482-009-9343-6

Maddison D, Maddison W. 2005. MacClade 4: Analysis of phylogeny and character evolution.

Version 4.08. Sinauer Associates, Sunderland, MA, USA.

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Masuya H, Kim JJ, Wingfield MJ, Yamaoka Y, Kaneko S, Breuil C, Kim GH. 2005. Discovery and

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Masuya H, Kaneko S, Yamaura Y, Yamaoka Y. 2009. Ophiostomatoid fungi isolated from

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MY COTAXON

http://dx.doi.org/10.5248/130.875

Volume 130, pp. 875-878 July-September 2015

Xylomyces acerosisporus sp. nov.

from submerged leaves from Brazil

Mayra S. OLIVEIRA’, ELAINE MALOSSO?*, MARCELA A. BARBOSA’,

MarInéA A.G. ARAUJO’, & RAFAEL FE. CASTANEDA-RUIZ?

‘Programa de Pés-Graduagao em Biologia de Fungos, Universidade Federal de Pernambuco,

Avenida da Engenharia, s/n. Cidade Universitaria, Recife, PE, 50.740-600, Brazil

*Centro de Ciéncias Biolégicas, Departamento de Micologia/ Laboratorio de Micorrizas,

Universidade Federal de Pernambuco, Avenida da Engenharia, s/n.

Cidade Universitaria, Recife, PE, 50.740-600, Brazil

*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

* CORRESPONDENCE TO: elainemalosso@yahoo.com.br

ABSTRACT — A new species, Xylomyces acerosisporus, collected on submerged decaying

leaves in a river in the Brazilian Atlantic Forest, is described and illustrated. It is distinguished

by filiform to acerose, 7-15-septate, brown, catenate, smooth chlamydospores.

Key worps — freshwater fungi, asexual fungi, taxonomy

Introduction

Xylomyces Goos et al. (Goos et al. 1977, Castaheda-Ruiz & Kendrick 1990,

Goh et al 1997, Kohlmeyer & Volkmann-Kohlmeyer 1998, Hyde & Goh 1999)

is distinguished by multiseptate, cylindrical to fusiform, solitary or catenate,

intercalary or terminal, brown to dark brown chlamydospores that resemble

phragmoconidia but in which the conidial ontogeny is neither blastic nor

thallic-arthric (Kendrick 1971). A fungus found on decaying leaves submerged

in a river in Brazil is described here as a new species of Xylomyces.

Materials & methods

During an expedition in November 2014 through “Rio Formoso, Reserva Biologica

de Saltinho,’ Pernambuco State, in the Atlantic Forest of Brazil, samples of submerged

decaying leaves were stored in sterile glass jars with water from the site, taken to the

876 ... Oliveira & al.

Fic.1. Xylomyces acerosisporus (ex holotype, URM 87587).

Colonies and chlamydospores on the natural substrate.

Xylomyces acerosisporus sp. nov. (Brazil) ... 877

laboratory and treated according to Ingold (1975) and Schoenlein-Crusius & Milanez

(1989). The colonized leaves were sectioned by hand with ophthalmic micro-surgical

blades (Shreeji Micro Systems Inc., Indiamar Company), washed with 4% KOH for

5-10 min., then washed twice with sterile distilled water. Mounts were prepared in PVL

(polyvinyl alcohol and lactic acid) and measurements made at a magnification of x1000.

Photomicrographs were obtained with a Nikon microscope with bright field optics. The

holotype was deposited in the Herbarium of Universidade Federal de Pernambuco,

Recife, Brazil (URM).

Taxonomy

Xylomyces acerosisporus M.S. Oliveira, Malosso & R.E. Castafieda, sp.nov. _—‘Fia. 1

INDEX FUNGORUM IF550946

Differs from Xylomyces giganteus and X. rhizophorae by its smaller chlamydospores with

fewer septa and from X. foliicola by its cylindrical to acerose (occasionally narrowly

fusiform) chlamydospores that tend to have more septa.

Type: Brazil, Pernambuco, Rio Formoso, Reserva Biologica de Saltinho, 8°43’S 35°11’W,

on submerged decaying leaves of an unidentified plant in the Formoso River, 13

November 2014, coll. M.S. Oliveira (Holotype: URM87587).

EryMo oey: Latin, acerosus (acerose; needle-like, with tapering ends) + sporus (spore),

referring to the chlamydospore shape..

CoLonigs on the natural substrate spreading, amphigenous, funiculose,

brown. Mycelium superficial, composed of septate, branched, brown to dark

brown, smooth hyphae, 2-3.5um diam. CHLAMyDosPoRES filiform to acerose,

occasionally narrowly fusiform straight, intercalary and terminal, catenate,

rarely solitary, unbranched, 7-15-septate, brown, dark brown at the septa,

95-180 x 8-10 um, smooth.

Note: Xylomyces acerosisporus is superficially similar to X. foliicola W.B. Kendr.

& R.F. Castafieda, X. giganteus Goh et al., and X. rhizophorae Kohlm. & Volkm.-

Kohlm. However, X. foliicola has intercalary, fusiform, 4-11-septate (mostly

11-) concolorous brown chlamydospores, 126-152 x 7-9 um (Castafeda-Ruiz

& Kendrick 1990); X. giganteus has 6-26-septate, yellowish brown to medium

brown straight or curved chlamydospores (constricted at the septa and with

irregular longitudinal striations), 140-575 x 25-50 um (Goh et al 1997); and

X. rhizophorae has straight or curved, concolorous dark brown chlamydospores

that are 11-43(-64)-septate (rarely with longitudinal or oblique septa, mostly

widest at the tips and tapered toward the base, constricted at some septa),

95-370(-500) x 9-16 um (Kohlmeyer &Volkmann-Kohlmeyer 1998).

Acknowledgments

The authors express their sincere gratitude to Prof Dr. Bryce Kendrick and Dr. De-Wei Li

for their critical review of the manuscript. The authors are deeply grateful to Dr. T.K. Goh

878 ... Oliveira & al.

for his commentaries and opinion. The authors are grateful to the “Coordenacao de

Aperfeicoamento de Pessoal de Nivel Superior (CAPES)” for financial support through

project 88881.062172/2014-01 and the “Programa Ciéncia sem Fronteiras”.. RFCR

is grateful to the Cuban Ministry of Agriculture and “Programa de Salud Animal y

Vegetal’, project P131LH003033 for facilities. We acknowledge the facilities provided

by Dr. P.M. Kirk and Drs. V. Robert and A. Decock through the Index Fungorum and

MycoBank websites. Dr. Lorelei L. Norvell’s editorial review and Dr. Shaun Pennycook’s

nomenclature review are greatly appreciated.

Literature cited

Castaneda Ruiz RE, Kendrick B. 1990. Conidial fungi from Cuba: II. University of Waterloo Biology

Series 33. 61 p.

Goh TK, Ho WH, Hyde KD, Tsui KM. 1997. Four new species of Xylomyces from submerged wood.

Mycological Research 101: 1323-1328. http://dx.doi.org/10.1017/S0953756297004164

Goos RD, Brooks RD, Lamora BJ. 1977. An undescribed hyphomycete from wood submerged in a

Rhode Island stream. Mycologia 67: 280-286. http://dx.doi.org/10.2307/3758653

Hyde KD, Goh TK. 1999. Fungi on submerged wood from the River Coln, England. Mycological

Research 103: 1561-1574. http://dx.doi.org/10.1017/S0953756299008989

Ingold CT. 1975. An illustrated guide of aquatic and water borne hyphomycetes (fungi imperfecti)

with notes on their biology. Freshwater Biological Association, Scientific Publication 30. 96 p.

Kendrick WB. 1971. Taxonomy of fungi imperfecti. University of Toronto Press, Canada.

Kohlmeyer J, Volkmann-Kohlmeyer B. 1998. A new marine Xylomyces on Rhizophora from the

Caribbean and Hawaii. Fungal Diversity 1: 159-164.

Schoenlein-Crusius IH, Milanez AI. 1989. Sucessao fingica em folhas de Ficus microcarpa L. f.

submerses no Lago Frontal situado no Parque Estadual das Fontes do Ipiranga, Sao Paulo.

Revista de Microbiologia, Sao Paulo 20:95-101.

MY COTAXON

http://dx.doi.org/10.5248/130.879

Volume 130, pp. 879-881 July-September 2015

Tubulicrinis indicus, a new corticioid species from India

JyoT1 SHARMA”, AVNEET P. SINGH, & G.S. DHINGRA

Department of Botany, Punjabi University, Patiala 147002, India

*CORRESPONDENCE TO: jyotisharma.botany@gmail.com

AsBsTRACT — Tubulicrinis indicus sp. nov. is described and illustrated from Jammu and

Kashmir, India.

Key worps — Basidiomycota, Agaricomycetes, Himalaya

While conducting fungal forays in the Duggi area of district Doda, Jammu and

Kashmir, India, Jyoti Sharma collected a corticioid fungus on decaying wood

of Cedrus deodara. After comparing macroscopic and microscopic features

(Rattan 1977, Hjortstam et al. 1988, Bernicchia & Gorjén 2010, Mycobank

2015), we identified it as a new species of Tubulicrinis, close to T. hirtellus and

T. effugiens. It is characterized by awl-shaped lyocystidia with a narrow capillary

lumen that extends up to the apex and large basidiospores. The type specimen is

conserved in the Herbarium, Botany Department, Punjabi University, Patiala,

India (PUN). Dr. Nils Hallenberg examined the specimen and supported the

description of a new species.

Tubulicrinis indicus Jyoti Sharma, Avneet P. Singh & Dhingra, sp. nov. PLatEs 1, 2

MycoBank 812015

Differs from Tubulicrinis hirtellus and T. effugiens by having lyocystidia with a narrow

capillary lumen that extends up to the apex and larger basidiospores.

Type: India, Jammu and Kashmir: Doda, Bhadarwah, Duggi, on bark of log of Cedrus

deodara (D. Don) G. Don, 28 September 2014, Jyoti Sharma 7087 (PUN, holotype).

ErymMo.oey: The epithet refers to the country of collection, India.

Basidiocarp resupinate, adnate, effused, <150 um thick in section; hymenial

surface smooth to porulose under lens, creamish white to pale yellow when

fresh, not changing much on drying; margins thinning, concolorous to

indeterminate. Hyphal system monomitic. Generative hyphae <4 um wide,

branched, septate, clamped, thin-walled. Cystidia 110-133 x 4.5-5.5 um,

880 ... Sharma, Singh, & Dhingra

PiaTE 1. Tubulicrinis indicus (holotype, PUN). Basidiocarp showing hymenial surface.

cylindrical, somewhat flexuous, with a subulate and mucronate apex, with

rooting base, grayish, thick-walled, narrow capillary lumen extending up to the

apex, weakly amyloid; projecting up to 50 um out of hymenium. Basidia 25-40

x 4.5-5.8 um, subclavate to subcylindrical, 4—sterigmate, with basal clamp;

sterigmata <5.5 um long. Basidiospores 7.8-11 x 2.5-3.5 um, subcylindrical,

apiculate, thin-walled, smooth, inamyloid, acyanophilous, with oily contents.

ReMARKS— Tubulicrinis hirtellus (Bourdot & Galzin) J. Erikss. differs from

T. indicus by its lyocystidia with a thin-walled apex, and T. effugiens (Bourdot

& Galzin) Oberw. is distinguished by its capillary lumen ending gradually in a

thin-walled apex; both these species also differ in having smaller basidiospores.

Acknowledgements

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

providing research facilities, UGC-BSR for providing financial assistance, Dr. Nils

Hallenberg (Professor Emeritus, Denmark) for expert comments and peer review,

Prof. B.M. Sharma (Department of Plant Pathology, COA, CSKHPAU, Palampur, H.P.,

India) for peer review, and Mycotaxon, Ltd. for underwriting publication charges.

Literature cited

Bernicchia A, Gorjon SP. 2010. Corticiaceae s.1. Fungi Europaei 12. Edizioni Candusso. Alassio.

Italia. 1008 p.

Hjortstam K, Larsson KH, Ryvarden L. 1988. The Corticiaceae of North Europe - VIII. Oslo. pp.

1450-1614.

MycoBank. 2015. Fungal databases. Nomenclature and species banks.

http://www.mycobank.org/ [Accessed: 20/03/2015].

Rattan SS. 1977. The resupinate Aphyllophorales of the North Western Himalayas. Bibliotheca

Mycologica 60. 427 p.

MY COTAXON

http://dx.doi.org/10.5248/130.883

Volume 130, pp. 883-891 July-September 2015

Four Rhizocarpon species new to China

WEI-CHENG WANG, ZUN-TIAN ZHAO, & LU-LU ZHANG"

College of Life Science, Shandong Normal University, Jinan, 250014, P. R. China

* CORRESPONDENCE TO: lichenzll@aliyun.com

ABSTRACT — Four new lichen records are reported from China — Rhizocarpon lavatum,

R. reductum, R. saurinum, and R. umense. Detailed taxonomic descriptions with photos and

comments are provided.

Key worps — Rhizocarpaceae, Asia, taxonomy

Introduction

The genus Rhizocarpon was established by De Candolle in 1805 and is

usually recognized by a crustose thallus, a usually distinct prothallus, black

lecideine apothecia, Rhizocarpon-type asci, and hyaline to brown, halonate

ascospores that can be transversely septate or submuriform to muriform

(Fletcher et al. 2009, McCarthy & Elix 2014). Rhizocarpon species frequently

occur on siliceous rocks (some on basic substrata) in extra-tropical regions,

while other species are parasitic on other lichens (Matwiejuk 2008).

Rhizocarpon includes about 200 species worldwide, of which 31 species have

been found in China (Aptroot 2002, Aptroot & Sparrius 2003, Guo 2005, Zhao

et al. 2013). During our study of the lichen flora of China, we identified four

Rhizocarpon species new to China, which are reported here. Two species are

rather widespread and frequently reported, but the other two are rare lichens

previously known only from restricted areas, one from northern Sweden and

the second from North America and Iran.

Materials & methods

The examined specimens are preserved in the Lichen Section of Botanical

Herbarium, Shandong Normal University, Jinan, China (SDNU) and the lichen

herbarium of the Kunming Institute of Botany, Chinese Academy of Sciences, Kunming,

China (KUN-L). Their morphological and anatomical characters were examined under

a stereomicroscope (Olympus SZ) and a polarizing microscope (Olympus CX21). Both

thallus and medulla were tested with K (10% aqueous solution of potassium hydroxide),

884 ... Wang, Zhao, & Zhang

C (saturated solution of aqueous sodium hypochlorite), I (10% aqueous solution of

aqueous potassium iodide), and P (saturated solution of p-phenylenediamine in 95%

ethyl alcohol) for identification. The lichen substances were identified using standardized

thin layer chromatography techniques (TLC) with system C (Orange et al. 2010). Photos

of these lichens were taken under Olympus SZX16 and BX61 with DP72.

Taxonomic descriptions

Rhizocarpon lavatum (Ach.) Hazsl., Magyar Birodalom Zuzm6-Fléraja:

206 (1884) FIG. 1

Saxicolous. THALLUS rimose to areolate, <1.0 cm diam.; medulla I-;

prothallus black, distinct; areoles grayish white, grey or brown, contiguous or

scattered, thin or medium thick, 0.20-0.40 mm diam.. APOTHECIA sessile or

partly immersed, black, round, 0.60-1.50 mm diam., dispersed, mostly single,

disc flat to slightly convex, epruinose, margin distinct, thick, <125 um wide;

exciple thick, 85-125 um wide, brown in inner part, rim brownish black,

K-, no crystals; epihymenium pale brown, or brown and olive-green

intermixed, K-, no crystals; hymenium hyaline, 85-150 um tall; hypothecium

brown, K-; paraphyses branched and anastomosing. Asci 8-spored; ascospores

persistently colorless, eumuriform, with 15-24 cells in optical view, oblong-

ellipsoid, halonate, 31-40 x 12.5-13.5 um.

CHEMISTRY — Medulla K-, C-, P-; no substance detected by TLC.

SPECIMENS EXAMINED: CHINA. JILIN, Baishan Co., western side of Mt. Changbai, alt.

2300 m, on rock, 22 Jul. 2014, W.C. Wang 20141093-A, 20141088, 20141123, 20141094,

20141098, 20141072, 20141059, 20141117, 20141118, 20141079, 20141080, 20141087

(SDNU)); alt. 2400 m, on rock, 22 Jul. 2014, W.C. Wang 20141207 (SDNU); northern

side of Mt. Changbai, alt. 1850 m, on rock, 20 Jul. 2014, W.C. Wang 20141213, 20141214

(SDNU)); alt. 1950 m, on rock, 20 Jul. 2014, W.C. Wang 20141168, 20141169, 20141171,

20141172, 20141174, 20141221 (SDNU). YUNNAN, Lijiang City, Mt. Laojun, alt. 4000 m,

on rock, 7 Nov. 2009, Y.L. Cheng 20100689 (SDNU); alt. 3950 m, on rock, 25 Aug. 2015,

W.C. Wang 20150101 (SDNU). SHANNXI, Baoji City, Mt. Taibai, Doumugong, alt. 2850

m, on rock, 25 Aug. 2014, X.Y. Wang, X. Ye 14-44711 (KUN-L).

DISTRIBUTION — Rhizocarpon lavatum has been reported from South Korea,

Europe, North America, Australia, and New Zealand (McCarthy & Elix 2014,

Ovstedal et al. 2009, Joshi et al. 2010). New to China.

ComMENTs — Rhizocarpon lavatum was found in shaded or damp habitats,

such as rocks in forests or near the river. This species is usually recognized

by its numerous, large, sessile, and flat apothecia with thick margins, long

narrow ascospores with many cells (15-24), and a rimose thin thallus lacking

secondary substances that separates it from other Rhizocarpon species.

Rhizocarpon reductum is similar to R. lavatum but has smaller ascospores

(23-31 x 10-13.5 um), narrower exciple (<33 um wide), and contains stictic

acid. Rhizocarpon timdalii is also close to R. lavatum but has strongly convex

Rhizocarpon spp. new for China... 885

Fic. 1. Rhizocarpon lavatum (Wang 20141173, SDNU). A: thallus; B: apothecia; C: apothecium

section; D: K reaction; E: no crystals in epihymenium and exciple; F: eight spores per ascus;

G: ascospores. Scale bars: A = 1 mm; B = 500 um; C-E = 50 um; F, G = 10 um.

apothecia and areoles; R. sublavatum is distinguished by smaller apothecia

with a much thinner margin and broad ellipsoid ascospores; R. anaperum has a

brown granular thallus and a brown epihymenium (Fletcher et al. 2009).

Rhizocarpon reductum Th. Fr., Lich. Scand. 1(2): 633 (1874) Fic. 2

Saxicolous. THALLUS areolate, <1.0 cm diam.; medulla I-; prothallus black,

present at the margin; areoles dark grey to brown, dull, usually containing

886 ... Wang, Zhao, & Zhang

10 ym |

Fic. 2. Rhizocarpon reductum (Wang 20141075, SDNU). A: thallus; B: apothecia; C: apothecium

section; D: K reaction; E: crystals in epihymenium and exciple; F: eight spores per ascus;

G: ascospores. Scale bars: A = 1 mm; B = 200 um; C-E = 50 um; F, G = 10 um.

granules. APOTHECIA black, distributed between the areolae, 0.50-0.75 mm

diam., disc flat or slightly convex, rough, epruinose; exciple thin, <33 um wide,

brown in inner part, rim greenish black, K-, containing crystals dissolving in

K; epihymenium olive-green, intermixed with brown, K-, containing crystals

dissolving in K; hymenium hyaline, 108-125 um tall; hypothecium dark

brown, K-; paraphyses branched and anastomosing. Asci 8-spores; ascospores

persistently colorless, muriform, with 8-18 cells in optical view, ellipsoid,

halonate, 23-31 x 10-13.5 um.

Rhizocarpon spp. new for China... 887

CHEMISTRY — Medulla K+ yellow, C-, P+ orange; stictic acid detected by

Tee

SPECIMENS EXAMINED: CHINA. SHAANXI, Baoji City, Mt. Taibai, alt. 3700 m, on rock,

5 Aug. 2005, C.L. Wang & FE Yang TBW099 (SDNU). Jrzin, Baishan Co., western side

of Mt. Changbai, alt. 2300 m, on rock, 22 Jul. 2014, W.C. Wang 20141132, 20141065,

20141075, 20141125, 20141102 (SDNU). X1njIANG, Urumai City, Mt. Tianshan, Glacier

No. 1, alt. 2700 m, on rock, 28 Aug. 2011, L.L. Zhang 20126751 (SDNU); Nanshan

xiaoqvzi, alt. 2100 m, on rock, 28 Aug. 2011, L.L. Zhang 20126750 (SDNU); Yili City,

Qiongbola National Forest Park, on rock, 12 Jun. 2014, PM. Wang 20140109 (SDNU);

Changji City, Augest First Forest Farm, alt. 2100 m, on rock, 10 Aug. 2012, X. Zhao

20129234 (SDNU). ZHEJIANG, Hangzhou City, Mt. Tianmu, Tianchi, alt. 1100 m, on

rock, 20 Oct. 2010, H.Y. Wang 20104183 (SDNU).

DISTRIBUTION — Rhizocarpon reductum has been reported from Europe, Asia,

North America, Africa, Australia, and New Zealand (Feuerer 1991, Fryday

2000, Ihlen 2004, McCarthy & Elix 2014). New to China.

ComMENTS — Rhizocarpon reductum is recognized mainly by the presence

of stictic acid, smaller muriform ascospores, and narrower exciple. Other

Rhizocarpon species that also contain stictic acid and have hyaline and

muriform ascospores include R. furfurosum, R. distinctum, R. petraeum, and

R. umbilicatum. Rhizocarpon furfurosum differs from R. reductum in having

an isidiate thallus, while R. distinctum is distinguished by its I+ blue medulla;

R. petraeum and R. umbilicatum both have a pruinose thallus. Rhizocarpon

timdalii is close to R. reductum but can be separated by its blue-green

epihymenium, small convex areoles, lack of stictic acid, and ascospores that

are longer and broader and with more cells (Ihlen 2004). For the differences

between R. reductum and R. lavatum, see COMMENTS under R. lavatum.

Rhizocarpon saurinum (W.A. Weber) Bungartz, Bryologist 107: 77 (2004) Fic. 3

Saxicolous. THALLuS dispersed or contiguous, <1.5 cm diam., granular

verrucose to areolate; medulla I-; prothallus absent; areoles pale yellow, up to

1 mm diam., convex, angular or orbicular, dull. AporHecta black, round, 0.4-

1.0 mm diam., convex, immersed in thalli, rough, epruinose; exciple brown-

black, narrow, <40 um wide, K+ violet; epihymenium brown-black, K+ violet,

no crystals; hymenium hyaline and intermixed with brown, 100-125 um tall,

containing crystals, K+ violet; hypothecium brown, K-; paraphyses branched

and anastomosing. Asc 8-spores; ascospores ellipsoid to globose, with 2-4-(6)

cells in optical view, brown-black, halonate, 15-20 x 10-12.5 um.

CHEMISTRY — Medulla K-, C-, P-. rhizocarpic acid detected by TLC.

SPECIMENS EXAMINED: CHINA. XINJIANG, Yili City, Zhaosu Co., Kuopiertigou, alt.

2436 m, on rock, 18 Jun. 2014, PM. Wang 20140276, 20140283, 20140285 (SDNU).

XIZANG, Basu Co., Sharao village, alt. 3867 m, on rock, 18 Sep. 2014, L.S. Wang et al.

14-46374 (KUN-L).

888 ... Wang, Zhao, & Zhang

oo a vate

Fic. 3. Rhizocarpon saurinum (Wang 20140276, SDNU). A: thallus; B: apothecia; C: apothecium

section; D: K reaction; E: crystals in hymenium; F: eight spores per ascus; G: ascospores. Scale bars:

A = 1mm; B= 500 um; C-E = 50 um; 5 G = 10 um.

DISTRIBUTION — Rhizocarpon saurinum has been reported from North

America and Iran (Bungartz & Fryday 2004, Moniri et al. 2009). New to China.

ComMENTs — Rhizocarpon saurinum is recognized by its I- medulla, small

2-6 celled ascospores, brown-black K+ violet epihymenium, and containing

rhizocarpic acid only (Bungartz & Fryday 2004). The similar Rhizocarpon

viridiatrum is distinguished by its larger ascospores (15-28 x 7-14 um) and

lichenicolous (at least initially) habit.

Rhizocarpon spp. new for China... 889

. > - G 2

F ‘a 10 um |

Fic. 4. Rhizocarpon umense (Zhang 20126751, SDNU). A: thallus; B: apothecia; C: apothecium

section; D: K reaction; E: no crystals in epihymenium and exciple; F: eight spores per ascus;

G: ascospores. Scale bars: A = 500 um; B = 200 um; C-E = 20 um; F, G = 10 um.

Rhizocarpon umense (H. Magn.) A. Nordin, Graphis Scripta 17(2): 37 (2005) Fie. 4

Saxicolous. THALLUS areolate, small, <0.5 cm diam.; medulla I+ blue;

prothallus black, distinct at the margin; areoles brown, 0.30-0.55 mm diam.,

medium thick, flat to slightly convex, angular, contiguous in the central

portion but thin and scattered at the margin. APOTHECIA black, round to

angular, multi-shaped, 0.25-0.55 mm diam., dispersed, distributed between

the areolae, immersed, disc persistently flat, rough, epruinose; margin present,

890 ... Wang, Zhao, & Zhang

thin, completely separate from areoles; exciple and epihymenium black,

K+ red; hymenium hyaline below, upper part pale red, 60-90 um tall,

I+ blue; hypothecium brown, K-; paraphyses branched and anastomosing, no

crystals in the apothecia. Asci 8-spores; ascospores brown, 1-septate, ellipsoid,

occasionally slightly bent, halonate, 12.5-17.5 x 6-7 um.

CHEMISTRY — Medulla K-, C-, P-. no substance detected by TLC.

SPECIMENS EXAMINED: CHINA. XINJIANG, Urumgi City, Mt. Tianshan, Glacier No. 1,

alt. 2700 m, on rock, 28 Aug. 2011, L.L. Zhang 20126751, 20126752 (SDNU).

DISTRIBUTION — Rhizocarpon umense has previously been reported only from

Sweden (Nordin 2005). New to China.

ComMMENTS — Rhizocarpon umense is characterized by its small 1-septate

brown ascospores, I+ blue medulla, K+ red epihymenium, and absence of

secondary substances. The similar Rhizocarpon alaxense can be distinguished

by its I- medulla, K- epihymenium, and rusty-ochraceous thallus.

Acknowledgements

We thank Dr. A. Aptroot (ABL Herbarium, Soest, the Netherlands) and Prof. Shou-Yu

Guo (Institute of Microbiology, Chinese Academy of Sciences, Beijing, China) for

presubmission reviews; we also thank A. Nordin (Sweden) for confirmation on

Rhizocarpon umense and E. Timdal (Oslo) for helpful suggestions during our research.

This study was supported by the National Natural Science Foundation of China

(31170187, 31400015, 31570017), and the Scientific Research Innovation Foundation of

Shandong Normal University (SCX1525).

Literature cited

Aptroot A. 2002. Corticolous and saxicolous lichens from Xishuangbanna, Southern Yunnan,

China. http://www.nhm.uio.no/botanisk/lav/Yunnan

Aptroot A, Sparrius LB. 2003. New microlichens from Taiwan. Fungal Diversity 14: 1-50.

Bungartz F, Fryday A. 2004. Buellia saurina belongs to the genus Rhizocarpon (Rhizocarpaceae,

lichenized Ascomycetes). Bryologist 107(1): 75-78.

http://dx.doi.org/10.1639/0007-2745(2004)107[75:BSBT TG]2.0.CO;2

Feuerer T. 1991. Revision der europaischen Arten der Flechtengattung Rhizocarpon mit

nichtgelbem Lager und veilzelligen Sporen. Bibliotheca Lichenologica 39. 218 p.

Fletcher A, Gilbert OL, Clayden S, Fryday AM. 2009. Rhizocarpon. 792-808, in: CW Smith et al.

(eds). The lichens of Great Britain and Ireland. London: British Lichen Society.

Fryday A. 2000. On Rhizocarpon obscuratum (Ach.) Massal., with notes on some related species in

the British Isles. Lichenologist 32(3): 207-224. http://dx.doi.org/10.1006/lich.2000.0269

Guo SY. 2005. Lichens. 31-82, in: WY Zhuang (ed.). Fungi of northwestern China. Mycotaxon Ltd.,

Ithaca, New York.

Ihlen PG. 2004. Taxonomy of the non-yellow species of Rhizocarpon in the Nordic countries,

with hyaline and muriform ascospores. Mycological Research 108(5): 533-570.

http://dx.doi.org/10.1017/S0953756204009803

Joshi Y, Koh YJ, Hur JS. 2010. Three new records of lichen genus Rhizocarpon from South Korea.

Mycobiology 38(3): 219-221. http://dx.doi.org/10.4489/MYCO.2010.38.3.219

Rhizocarpon spp. new for China... 891

Matwiejuk A. 2008. Noteworthy species of the genus Rhizocarpon Ramond ex DC. (Rhizocarpaceae,

lichenized Ascomycota) in the LBL herbarium. Annales UMCS, Biologia 63(1): 79-92.

http://dx.doi.org/10.2478/v10067-008-0006-1

McCarthy PM, Elix JA. 2014. The lichen genus Rhizocarpon in mainland Australia. Telopea

16: 195-211. http://dx.doi-org/10.7751/telopea20148124

Moniri MH, Kamyabi S, Fryday AM. 2009. Rhizocarpon saurinum new to Asia, and other reports

of Rhizocarpon species from Razavi Khorasan Province, Iran. Mycologica Balcanica 6: 89-92.

Nordin A. 2005. Rhizocarpon umense, a distinct but originally misplaced species. Graphis Scripta

17: 37-38.

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: 1-393.

http://dx.doi.org/10.2478/v10208-011-0013-5

Zhao ZT, Li C, Zhao X, Zhang LL. 2013. New records of Rhizocarpon from China. Mycotaxon

125: 217-226. http://dx.doi.org/10.5248/125.217

MY COTAXON

http://dx.doi.org/10.5248/130.893

Volume 130, pp. 893-898 July-September 2015

Terriera transversa sp. nov. from Hainan, China

QING LI", SHI-JUAN WANG”, YU-XIA CHEN’,

YAN-PING TANG?, & YING-REN LIN”

' School of Life Science &? School of Forestry & Landscape Architecture,

Anhui Agricultural University,

West Changjiang Road 130, Hefei, Anhui 230036, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

ABSTRACT — A new Species, Terriera transversa, is described, illustrated, and compared

with similar taxa. This fungus was collected from dead leaves of Pandanus sp. in Jianfengling

National Forest Park of Hainan Province, China. It is distinguished by deeply embedded

ascomata perpendicular to the host veins and asci with truncate apices. The type specimen

is deposited in the Reference Collection of Forest Fungi of Anhui Agricultural University,

China (AAUF).

Key worps — morphological character, Pandanaceae, Rhytismataceae, Rhytismatales,

taxonomy

Introduction

During a visit to Hainan, South China, in June 2014, a week was devoted

to collecting fungal specimens. Jianfengling National Forest Park, the largest,

richest, and best-preserved tropical rainforest in China, was chosen as the

destination due to its unique natural environment. Amongst the collections

obtained, a new Terriera species was found from dead leaves of Pandanus sp. in

Tianchi alpine lake at an altitude of 880 m.

The genus Terriera was proposed by Eriksson (1970) with T. cladophila (Lév.)

B. Erikss. as its only species; subsequently an additional 25 species and one

variety have been included in the genus (IndexFungorum 2015). For China, ten

Terriera species have been recorded (none of them from tropical rainforests):

T: angularis, T. brevis, T: camelliae, T. coacervata, T: huangshanensis, T. illiciicola,

T: mangiferae, T: petrakii, T. rotundata, and T. simplex (Sivanesan & Hsieh 1989,

* Qinc Li and SHI-JUAN WANG contributed equally to this work.

894 ... Li, Wang, & al.

Frohlich & Hyde 2000, Yang et al. 2011, Chen et al. 2012, Zheng et al. 2012,

Song et al. 2012, Gao et al. 2012, Zhou et al. 2013, Li et al. 2014).

Our new species from Hainan, described here as Terriera transversa, is the

first report of Terriera from tropical rainforests of China.

Materials & methods

The study was based on a collection made in 2014 from Hainan, China. Specimens

were rehydrated in water for 15 min. and ascomata were sectioned vertically at a

thickness of 10-15 um using a YD-1508-III freezing microtome (Jinhua, China). The

outlines of fruitbodies were observed in 0.1% (w/v) cotton blue in lactophenol-glycerin.

The color of internal fruitbody structures was observed in water. Asci, ascospores and

paraphyses were measured from squash mounts in 5% KOH solution. Line and point

integrated drawings of internal structures of ascomata were prepared using a Panasonic

XSJ-2 microscope drawing device (Osaka, Japan). Photographs were taken using a

Sony DSC-TX7C digital camera (Tokyo, Japan), and a digital camera connected to an

Olympus CX31RTSF microscope (Tokyo, Japan). The type specimen is deposited in the

Reference Collection of Forest Fungi of Anhui Agricultural University, Hefei, China

(AAUF).

Taxonomy

Terriera transversa Y.R. Lin & Qing Li, sp. nov. FIGs 1, 2

MycoBank MB 811488

Differs from Terriera cladophila by its deeply embedded ascomata perpendicular

to the host veins and not associated with conidiomata, smaller asci with truncate or

subtruncate apices, and ascospores tapering to both ends.

Type: China, Hainan, Jianfengling National Park, Tianchi, alt. 880 m, on dead leaves of

Pandanus sp. (Pandanaceae), 21 June 2014, Y.R. Lin, S.J. Wang, & Q. Li 2794 (Holotype,

AAUF 68902).

ETYMOLOGy: transversa (Latin = transverse), referring to the arrangement of the

ascomata, which are mostly perpendicular to the host veins.

Cotontgs on both sides of leaves, forming subcircular or irregular dull brown

to grey-yellow bleached spots 2-5-(7) mm diam. that sometimes coalesce into

larger irregular shapes.

ZONE LINES infrequent or sometimes absent, pale brown, wide, diffused,

entirely or partly surrounding the paler areas without an obvious edge.

CONIDIOMATA not observed.

AscoMaTA developing on both sides of dead leaves, predominantly

hypophyllous, mostly perpendicular to the host veins, sometimes confluent in

groups of two or three. In surface view, ascomata 470-1150 x 250-330 um,

elliptical or oblong-elliptical in outline, ends slightly acute to obtuse. Whole

surface of ascomata black, matt or slightly glossy, with a clearly marked outline,

moderately raising the substratum surface, opening by a single longitudinal

895

Terriera transversa sp. nov. (China) ...

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PRET I =e

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Fic. 1. Terriera transversa (holotype AAUF 68902) on Pandanus sp.: A. Ascomata on a leaf.

B. Detail of ascomata. C. Ascoma in median vertical section. D. Portion of ascoma in median

vertical section. E. Hymenium and basal stroma. F. Asci and paraphyses. G. Ascospores. Bars:

20 um; E-G = 10 um.

10 mm; B = 1 mm; C = 50 um; D =

896 ... Li, Wang, & al.

Woy cag ss

CHALE

SAO

Fic. 2. Terriera transversa (holotype AAUF 68902): A. Portion of ascoma in median vertical section

B. Asci and paraphyses. C. Ascospores. Bars: A = 20 um; B, C= 10 um.

split nearly extend to the edge of the ascomata. Lips absent. In median vertical

section, ascomata deeply embedded, with host cells becoming filled with fungal

tissue as the ascoma continues development. COVERING STROMA 30-45 um

thick, composed of textura angularis and globulosa with blackish brown to

black thick-walled cells 2—5.5 um diam., but with a group of nearly colourless,

thin-walled, pale cells in the inside part of the upper half; a short, 12-20 um

thick extension comprising strongly carbonized tissue with no obvious cellular

structure and adjacent to the top of the covering stroma. ExcipULUM well

developed, 10-18 um thick, arising from the inner layer of the basal stroma,

septate, consisting of colorless textura porrecta-intricata with hyphae 1.2-1.8

um diam., sometimes forming a loose reticular structure in some local

Terriera transversa sp. nov. (China) ... 897

parts. BASAL sTROMA 8-18 um thick, dark brown or black-brown composed

of angular to globose, thick-walled cells 2.5—4 um diam. A 35-60 um thick

textura prismatica with somewhat thin-walled, nearly colorless to grey-brown

cells existing between the covering stroma and basal stroma. SUBHYMENIUM

12-22 um thick, consisting of hyaline textura angularis and intricata.

PARAPHYSES 85-105 x 1.5-2 um, filiform, hyaline, sparsely septate, often

branching 1-2 times and gradually irregularly swollen to 2.8-4 um near the

apex, with an inconspicuous thin gelatinous matrix forming a light grey-brown

15-20 um thick epithecium above the asci. Asci ripening sequentially, 70-86

x 5-6 um, cylindrical, somewhat short-stalked, thin-walled, apex truncate

or subtruncate, without circumapical thickening, J-, 8-spored. Ascospores

arranged in a fascicle, 45-68 x 1.0—1.2 um, filiform, gradually tapered towards

the both ends, hyaline, aseptate, smooth-walled, with a gelatinous sheath ca 0.5

um thick.

HOsT SPECIES, HABITAT, & DISTRIBUTION: Producing ascomata on dead

leaves of a Pandanus sp. Known only from the type locality, Hainan Province,

China.

ComMMENTs — ‘The new species resembles the type species of Terriera,

T. cladophila (Eriksson 1970), which produces brown diffuse zone lines, small

bleached spots, and a well-developed subhymenium of textura angularis and

intricata. However, T. cladophila irregularly scatters on dead or living twigs

and has circular to elliptical ascomata associated with conidiomata, hyphal

bridges at the base of adjacent paraphyses, larger asci (80-125 x 6—9 um) with

rather rounded apices, and non-tapered ascospores without a mucous sheath

(Minter 1996).

Terriera simplex Y.L. Lin et al., which is very similar to T. transversa in the

truncate or subtruncate apices of its asci, differs in the presence of conidiomata,

subepidermal ascomata not associated with zone lines, a poorly developed

excipulum (7-12 um), synchronously maturing asci, and thinner paraphyses

(1.2—1.5 um wide) that are apically unbranched and do not form an epithecium

(Gao et al. 2012).

Terriera minor (Tehon) P.R. Johnst. is easily distinguished from the new

species by its smaller ascomata (400-800 x 200-300 um) with rounded or

obtuse ends, aseptate paraphyses branching 2-3 times in the upper 30-40 um,

and larger ascospores (70-100 x 1.5—2 um) with 0-1 septa (Johnston 1988,

1989).

Acknowledgements

The authors are grateful to Dr C.L. Hou (Capital Normal University, China) and

Dr M. Ye (Hefei University of Technology, China) for serving as pre-submission

898 ... Li, Wang, & al.

reviewers and to Mr Y.F. Xu for the field investigations. This study was supported by the

National Natural Science Foundation of China (No. 31270065, 31500019).

Literature cited

Chen JL, Lin YR, Hou CL, Wang SJ.2012 [“2011”]. Species of Rhytismataceae on Camellia spp. from

the Chinese mainland. Mycotaxon 118: 219-230. http://dx.doi.org/10.5248/118.219

Eriksson B. 1970. On Ascomycetes on Diapensiales and Ericales in Fennoscandia. I. Discomycetes.

Symbolae Botanicae Upsalienses 19(4): 1-71.

Frohlich J, Hyde KD. 2000. Palm microfungi. Fungal Diversity Press. Hong Kong. 364 p.

Gao XM, Zheng CT, Lin YR. 2012. Terriera simplex, a new species of Rhytismatales from China.

Mycotaxon 120: 209-213. http://dx.doi.org/10.5248/120.209

IndexFungorum. 2015. www.indexfungorum.org (viewed online on 10 August 2015).

Johnston PR. 1988. An undescribed pattern of ascocarp development in some non-coniferous

Lophodermium species. Mycotaxon 31: 383-394.

Johnston PR. 1989. Lophodermium (Rhytismataceae) on Clusia. Sydowia 41: 170-179.

Li ZJ, Cao N, Chen HE, Taylor JE, Hou CL. 2014. New species and new records of Rhytismataceae

from Japan. Mycological. Progress 13: 951-958. http://dx.doi.org/10.1007/s11557-014-0979-x

Minter DW. 1996. Terriera cladophila. IMI Descriptions of Fungi & Bacteria, no. 1296.

Sivanesan A, Hsieh WH. 1989. New species and records of ascomycetes from Taiwan. Mycol. Res.

93: 340-351. http://dx.doi.org/ 10.1016/S0953-7562(89)80161-3

Song JF, Liu L, Li YY, Hou CL. 2012. Two new species of Terriera from Yunnan Province, China.

Mycotaxon 119: 329-335. http://dx.doi.org/10.5248/119.329

Yang ZZ, Lin YR, Hou CL. 2011. A new species of Terriera (Rhytismatales, Ascomycota) from

China. Mycotaxon 117: 367-371. http://dx.doi.org/10.5248/117.367

Zheng Q, Lin YR, Yu SM, Chen L. 2012 [“2011”]. Species of Rhytismataceae on Lithocarpus spp.

from Mt Huangshan, China. Mycotaxon 118: 311-323. http://dx.doi.org/10.5248/118.311

Zhou F, Wang XY, Zhang L, Lin YR. 2013 [“2012”]. Terriera angularis sp. nov. on Illicium simonsii

from China. Mycotaxon 122: 355-359. http://dx.doi.org/10.5248/122.355

MY COTAXON

http://dx.doi.org/10.5248/130.899

Volume 130, pp. 899-905 July-September 2015

New records of Clauzadea and Immersaria from China

Lu-Lu ZHANG*, LING Hu*, XIANG-XIANG ZHAO, & ZUN-TIAN ZHAO*

Key Laboratory of Plant Stress Research, College of Life Sciences,

Shandong Normal University, Jinan, 250014, PR. China

* CORRESPONDENCE TO: ztzhao@sohu.com

ABSTRACT —'Three lichen species in the Lecideaceae (Clauzadea monticola, Immersaria

iranica, I. usbekica) are reported for the first time from China.

Key worps —Lecideales, Asia, taxonomy

Introduction

Lecideaceae Chevall. (Chevallier 1826, as ‘Lecideae’) originally included

all crustose lecideoid lichen genera but now consists of relatively few genera

with (crypo-)lecideine or (crypo-)lecanorine apothecia, mostly simple hyaline

ascospores, and a Lecidea-type or Porpidia-type ascus structure. Currently the

family contains about 23 genera and 547 species, of which the largest genus is

Lecidea Ach., containing about 427 species (Kirk et al. 2008, Fryday & Hertel

2014). Two species of Immersaria Rambold & Pietschm. have been reported

from China (Hertel 1977, Wei 1991; Aptroot & Sparrius 2003; Obermayer

2004) but no species of Clauzadea Hafellner & Bellem.

Clauzadea and Immersaria are closely related and are both characterized

by a Porpidia-type ascus, + halonate ascospores, and branched paraphyses.

Immersaria can best be distinguished by its thick brown areolate thallus with

a distinct epinecral layer, its immersed lecideine or lecanorine apothecia, and

its siliceous or calcareous habitat, while Clauzadea has only lecideine apothecia

and is restricted to calcareous rock (Rambold 1989, Thomson 1997, Calatayud

& Rambold 1998, Smith et al. 2009, Valadbeigi et al. 2011).

During our research on the lecideoid taxa in China, we identified three

species new to the country: Clauzadea monticola, Immersaria iranica, and

I. usbekica.

*Lu-Lu Zhang & Ling Hu contributed equally.

900 ... Zhang, Hu, & al.

Materials & methods

The studied specimens are preserved in the Lichen Section of the Botanical

Herbarium, Shandong Normal University, Jinan, China (SDNU). Morphological and

anatomical characters were examined under a stereo-microscope (COIC XTL7045B2)

and a polarizing microscope (OLympus CX41). The thalli and medullae were tested

for identification with K (10% aqueous KOH solution), C (saturated aqueous NaClO

solution), I (10% aqueous IKI solution), and P (saturated p-phenylenediamine solution

in 95% ethyl alcohol). The lichen substances were identified using standardized thin

layer chromatography techniques (TLC) with solvent system C (Orange et al. 2010).

These lichens were photographed under OLympus SZX16 and BX61 with DP72.

Taxonomic descriptions

Clauzadea monticola (Ach.) Hafellner & Bellem., Beih. Nova Hedwigia

79: 319 (1984) Fic. 1

MorpHoLocy — THALLUs granular to almost rimose and superficial, or +

immersed, brownish grey to ochre; medulla I-; prothallus absent. APOTHECIA

sessile, constricted at the base, not forming pits; disc (0.3-)0.4-0.7(-0.9)

mm diam., flat to convex, dark brown to black, becoming brownish when

wet, not pruinose; margin slightly raised, generally persistent but sometimes

finally excluded. ExcrpLte black-brown, 40-50 um wide, without crystals;

epihymenium reddish brown, without crystals; hymenium hyaline, 65-80 um

tall; hypothecium deep red brown, without crystals; paraphyses 2-3.5 um diam.,

branched and occasionally anastomosing, the apices capitate and widening to

5.0 um diam. Asci clavate, Porpidia-type; ascospores hyaline, simple, ellipsoid,

9-12(-13) x 4-5.5 um, halonate when young, the perispore <1.5 um thick.

PyYcNIDIA not observed.

CHEMISTRY — Thallus and medulla K-, C-, KC-, P-. No lichen substances

were detected by TLC.

SPECIMENS EXAMINED: CHINA. XINJIANG, Chabuer, Mt. Wusongshan, alt. 2215 m, on

rock, 10 Jun. 2014, P. M. Wang 20140049, 20140022, 20140012 (SDNU).

DISTRIBUTION —Clauzadea monticola has been reported from Asia, Africa,

Europe, Macaronesia, North and South America, and New Zealand (Thomson

1997, Smith et al. 2009). New from China.

COMMENTS — Clauzadea monticola can be most easily distinguished from

other Clauzadea species by its generally superficial thallus and sessile, epruinose

apothecia. It is morphologically similar to Lecidella stigmatea and Farnoldia

jurana, both of which occur in similar habitats. Lecidella stigmatea can be

separated by its Lecanora-type ascus, larger non-halonate ascospores (11-17x

6-9 um), and hyaline hypothecium (Thomson 1997), while F. jurana differs

in its blue-green epihymenium, non-capitate paraphyses, larger apothecia

(0.5-1.5 mm), and larger ascospores (13-28 x 7-14 um; Smith et al. 2009).

Clauzadea & Immersaria spp. new for China... 901

: : ri

' ozs f Gxt ae

10 pm _ 10 um | Ra 10um .

= 7X + : ~

Fic. 1. Clauzadea monticola (Wang 20140049, SDNU). A: thallus; B: apothecium section; C: ascus

and ascospores; D: amyloid reaction of ascus; E: ascospores; F: paraphyses.

Immersaria iranica Valadb., Sipman & Rambold, Lichenologist 43: 204 (2011) Fic. 2

MorpPHOLocy — THALLUS crustose, areolate, 0.3-0.4 mm thick; areoles

brown to slightly reddish brown, flat to slightly convex, often with irregular,

whitish rims; epinecral layer 15-23 um high, (pheno-)cortical layer 30-40

um thick; medulla I+; hypothallus black to grey, frequently visible between

the areoles. APOTHECIA immersed, 0.3-0.6 mm diam., usually one per areole,

round or angular, sometimes surrounded by a white rim, with crypto-lecanorine

margin; disc black-brown, plane to weakly convex, epruinose. ExcrpPLe thin

to absent; epihymenium dark brown, 15-20 um; hymenium hyaline, 120-150

um tall; hypothecium hyaline; paraphyses 1.5-3.5 um diam., anastomosing

and apically branched, apical cells widening slightly to 4.5 um. Asc clavate,

Porpidia-type; ascospores hyaline, simple, ellipsoid to subglobose, 9-13(-15) x

5.5-7 um, halonate. Pycnip1a immersed, opening by + stellate winding cracks

<0.5 mm long; conidia clavate to occasionally pyriform, 3-5 x c. 1.5 um.

CHEMISTRY — Thallus and medulla K-, C-, KC-, P-. No lichen substances

were detected by TLC.

SPECIMENS EXAMINED: CHINA. XINJIANG, Urumdi, Mt. Tianshan-glacier No.1, alt.

3800 m, on rock, 27 Aug. 2011, Z.L. Huang 20126106, 20129049 (SDNU).

902 ... Zhang, Hu, & al.

‘ a 10 um 10 um ~10ym 10 ym

Fic. 2. Immersaria iranica (Huang 20126106 SDNU). A: thallus; B: pycnidia; C: thallus section;

D: apothecium section; E: amyloid reaction of ascus; F: paraphyses; G: ascospores; H: conidia.

DISTRIBUTION — Immersaria iranica has been reported only from Iran

(Valadbeigi et al. 2011). New from China.

COMMENTS —Immersaria iranica is characterized by the crypto-lecanorine

apothecia, the I+ medulla, the pycnidia opening by + stellate winding cracks,

the round or lobed apothecia, and possession of 2’-O-methylsuperphyllinic

acid (Valadbeigi 2011). The Chinese specimens differ in their non-lobed

apothecia and lack of secondary substances.

Clauzadea & Immersaria spp. new for China... 903

Immersaria usbekica (Hertel) M. Barbero, Nav.-Ros. & Cl. Roux, Bull. Soc. Linn.

Provence 41: 140 (1990) Fic. 3

MorPHOLOGY — THALLUS crustose, cracked-areolate, 0.15-0.5(-0.7) mm

thick; areoles yellow-brown, flat to slightly convex; epinecral layer 14-20 um

high, (pheno-)cortical layer 25-35 um thick; medulla I+; hypothallus black,

more or less distinct between the areoles. APOTHECIA immersed, 0.2-0.5

mm diam., usually one or two per areole, with crypto-lecanorine margin;

disc black-brown, plane to weakly concave, epruinose. EXCIPLE usually very

Fic. 3. Immersaria usbekica (Li 2013814, SDNU). A: thallus; B: pycnidia; C: thallus section;

D: apothecium section; E: amyloid reaction of ascus; F: paraphyses; G: ascospores; H: conidia.

904 ... Zhang, Hu, & al.

reduced where the apothecia are attached to the areoles and better developed

where the apothecia reach the areole margins; epihymenium olive-brown to

brown, 17-25 um, hymenium hyaline, 125-140 um tall; hypothecium hyaline;

paraphyses 2.5-4 um diam., anastomosing and apically branched, apical cells

widening to 5.5 um. Asci clavate, Porpidia-type; ascospores hyaline, simple,

ellipsoid, (15-)18-23 x 9-13 um, halonate. Pycnip1A immersed, opening by

simple or graphidoid cracks up to 0.25 mm long; conidia short bacilliform,

4.5-6.5 x c. 1.5 um.

CuEmMIistTry — Thallus and medulla K-, C+ pale red, KC+ pale red, P-.

Gyrophoric acid and a confluentic acid syndrome were detected by TLC.

SPECIMEN EXAMINED: CHINA. XINJIANG, Urumai, Xishuigou, alt. 1950 m, on rock, 7

Sep. 2013, S.X. Li 2013814 (SDNU).

DISTRIBUTION — Immersaria usbekica has been reported from Asia, Europe,

and North Africa (Barbero et al. 1990; Hertel 1977, 2001; Valadbeigi et al.

2011). New to China.

ComMENTS —Immersaria usbekica is morphologically similar to I. iranica but

differs by the presence gyrophoric and confluentic acids, bacilliform conidia,

and calcareous habitat.

Acknowledgements

The authors thank Dr. H. J. M. Sipman (Botanical Garden and Museum, Berlin,

Germany) and Dr. Shou-Yu Guo (Chinese Academy of Sciences, Beijing, China) for

presubmission reviews. We also thank Dr. Tahereh Valadbeigi for helping us identify

Immersaria iranica specimens. This study was supported by the Program for Scientific

Research Innovation Team in Colleges and Universities of Shandong Province, the

National Natural Science Foundation of China (31170187, 31400015), the Foundation

of the Key Laboratory, CAS (KLBB-201306), and the National Training Programs of

Innovation and Entrepreneurship for Undergraduates (201410445093).

Literature cited

Aptroot A, Sparrius LB. 2003. New microlichens from Taiwan. Fungal Diversity 14: 1-50.

Barbero M, Navarro-Rosinés P, Roux C. 1990. Immersaria usbekica (Hertel) Barbero, Nav.-Ros. et

Roux comb. nov. [= Amygdalaria tellensis Esnault et Roux] nove trovita en Europo. Bulletin de

la Société Linnéenne de Provence 41: 139-142.

Calatayud V, Rambold G. 1998. Two new species of the lichen genus Immersaria (Porpidiaceae).

Lichenologist 30: 231-244.

Chevallier FF. 1826. Flore générale des environs de Paris. Paris: Ferra Jeune.

Fryday AM, Hertel H. 2014. A contribution to the family Lecideaceae s. lat. (Lecanoromycetidae

inc. sed., lichenized Ascomycota) in the southern subpolar region; including eight

new species and some revised generic circumscriptions. Lichenologist 46: 389-412.

http://dx.doi.org/10.1017/S0024282913000704

Hertel H. 1977. Gesteinsbewohnende Arten der Sammelgattung Lecidea (Lichenes) aus Zentral-,

Ost- und Siidasien. Khumbu Himal, Ergebnisse des Forschungsunternehmens Nepal Himalaya

6: 145-378.

Clauzadea & Immersaria spp. new for China... 905

Hertel H. 2001. Floristic and taxonomic notes on saxicolous lecideoid lichens. Sendtnera 7: 93-136.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th edition. CABI

Bioscience: CAB International. 771 p.

Obermayer W. 2004. Additions to the lichen flora of the Tibetan region. Bibliotheca Lichenologica

88: 479-526.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

Rambold G. 1989. A monograph of the saxicolous lecideoid lichens of Australia (excl. Tasmania).

Bibliotheca Lichenologica 34:1-345.

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.

Thomson JW. 1997. American Arctic lichens, vol. II. University of Wisconsin Press.

Valadbeigi T, Sipman H, Rambold G. 2011. The genus Immersaria (Lecideaceae) in Iran, including

I. iranica sp. nov. Lichenologist 43: 203-208.

Wei JC. 1991. An enumeration of lichens in China. International Academic Publishers, Beijing.

MY COTAXON

http://dx.doi.org/10.5248/130.907

Volume 130, pp. 907-909 July-September 2015

Phlebiopsis punjabensis sp. nov. from India

GURPREET KAUR, AVNEET P. SINGH *, & G.S. DHINGRA

Department of Botany, Punjabi University, Patiala, 147002, India

*CORRESPONDENCE TO: avneetbot@gmail.com

ABSTRACT — A new corticioid species, Phlebiopsis punjabensis, is described from Punjab, India.

Key worps — Basidiomycota, Agaricomycetes, Phanerochaetaceae, Ropar

During September 2012, Gurpreet and Avneet collected an unknown corticioid

fungus growing on the trunk of Psidium guajava from the Ropar Boat Club,

Ropar district, Punjab, India. After comparing macroscopic and microscopic

characters (Rattan 1977, Eriksson et al. 1978 1981, Dhingra 1987, Bernicchia

& Gorjon 2010, Priyanka et al. 2011, Mycobank 2015), we identified the

specimen as representing a new species of Phlebiopsis, distinguished from

P. mussooriensis Priyanka et al. and P. peniophoroides Gilb. & Adask. by brown-

pigmented basal hyphae. A portion of the basidiocarp was also sent to Dr. Nils

Hallenberg (Denmark), who confirmed the findings. The holotype specimen is

conserved in the Herbarium, Botany Department, Punjabi University, Patiala,

India (PUN).

Phlebiopsis punjabensis G. Kaur, Avneet P. Singh & Dhingra, sp. nov. — PLaTes 1, 2

MycoBank MB 812265

Differs from Phlebiopsis mussooriensis by its basidiocarp not cracking on drying, from

P. peniophoroides by its larger basidiospores, and from both these species by its brown-

pigmented basal hyphae.

Type: India, Punjab, Ropar boat club, on trunk of Psidium guajava L. (Myrtaceae),

5 September 2012, Gurpreet & Avneet 7150 (PUN, holotype).

ErymMo.ocy: The epithet refers to the state of collection, Punjab, India

Basidiocarp resupinate, adnate, effused, <75 um thick in section, crustaceous,

hymenial surface smooth, whitish when fresh, not changing much on drying;

margins thinning, paler concolorous or indeterminate. Hyphal system

monomitic. Generative hyphae simple-septate, densely packed and difficult to

discern; basal hyphae <4.6 um wide, agglutinated and making up a thin layer

908 ... Kaur, Singh, & Dhingra

PiatE 1. Phlebiopsis punjabensis (holotype, PUN). Basidiocarp showing hymenial surface.

1. fresh; 2. dry.

of hyphae parallel to the substrate, thin- to somewhat thick-walled; context

and subhymenial hyphae <3.3 um wide, vertically oriented, densely united and

richly branched, thin-walled. Basal hyphae and lower part of context hyphae

light, but distinctly brown-pigmented. Cystidia 20-36 x 7-9.8 um, subfusiform,

thick-walled, without basal clamp, encrusted with crystalline matter. Basidia

14-26 x 4-4.5 um, clavate to subcylindrical, 4-sterigmate, without basal

clamp; sterigmata <4.6 um long. Basidiospores 5.3-8.5 x 2.5-4 um, ellipsoid to

subcylindrical, smooth, inamyloid, acyanophilous.

REMARKS— P. mussooriensis differs in having a basidiocarp that cracks after

drying, a grayish yellow hymenial surface, more narrow basal hyphae, and

larger cystidia (Priyanka et al. 2011), whereas P peniophoroides differs in having

smaller basidiospores (Gilbertson & Adaskaveg 1993).

Acknowledgements

The authors thank SERB, DST, Government of India for financial assistance, Head,

Department of Botany, Punjabi University, Patiala, for providing research facilities,

Dr. Leif Ryvarden (Oslo, Norway) for expert comments, Dr. Nils Hallenberg (Sweden)

for expert comments and peer review, Prof. B.M. Sharma (Department of Plant

Pathology, COA, CSKHPAU, Palampur, H.P., India) for peer review, and Mycotaxon

Ltd., for underwriting publication charges.

Literature cited

Bernicchia A, Gorjon SP. 2010. Corticiaceae s.1. Fungi Europaei 12. Edizioni Candusso. Alassio.

Italia. 1008 p.

Dhingra GS. 1987. The genus Phlebiopsis in the eastern Himalayas. Nova Hedwigia 44: 221-227.

Eriksson J, Hjortstam K, Ryvarden L. 1978. The Corticiaceae of North Europe. Vol. 5. Mycoaciella -

Phanerochaete. Fungiflora, Oslo. pp. 889-1047.

Eriksson J, Hjortstam K, Ryvarden L. 1981. The Corticiaceae of North Europe. Vol. 6. Phlebia -

Sarcodontia. Fungiflora, Oslo. pp. 1049-1276.

Gilbertson RL, Adaskaveg JE. 1993. Studies on wood-rotting basidiomycetes of Hawaii. Mycotaxon

49: 369-397.

MycoBank. 2015. Fungal databases. Nomenclature and species banks. [Accessed: 26/03/2015].

Phlebiopsis punjabensis sp. nov. (India) ... 909

Priyanka, Dhingra GS, Kaur N. 2011. Phlebiopsis mussooriensis (Agaricomycetes), a new corticioid

species from India. Mycotaxon 115: 255-258. http://dx.doi.org/10.5248/115.255

Rattan SS. 1977. The resupinate Aphyllophorales of the north western Himalayas. Bibliotheca

Mycologica 60. 427 p.

ALK

PLATE 2. Phlebiopsis punjabensis (holotype, PUN).

1. basidiospores; 2. vertical section through basidiocarp.

MY COTAXON

http://dx.doi.org/10.5248/130.911

Volume 130, pp. 911-919 July-September 2015

Ocellularia lumbschii and O. saxicola spp. nov. from Vietnam

SANTOSH JOSHI’, DALIP KUMAR UPRETI’, EIMY RIVAS PLATA’,

Tur THuy NGUYEN, ANH DZUNG NGUYEN’, SOON-OK OH* & JAE-SEOUN HuR**

'Lichenology laboratory, CSIR-National Botanical Research Institute,

Rana Pratap Marg, Lucknow (UP)-226001, India

?Integrative Research Center, Science and Education, The Field Museum,

1400 South Lake Shore Drive, Chicago, IL 60605, USA

*Biotechnology Center, Tay Nguyen University, 567 Le Duan, Buon Ma Thuot City, Vietnam

‘Korean Lichen Research Institute, Sunchon National University, Suncheon-540 950, Korea

CORRESPONDENCE TO *: jshur1@sunchon.ac.kr

ABSTRACT — Two new species of thelotremoid Graphidaceae, Ocellularia lumbschii and

O. saxicola, are described from the Central Highlands of Vietnam. Ocellularia lumbschii has

an off-white to creamish shiny smooth thallus, apically carbonized excipulum and columella,

transversely septate ascospores (20-27 x 7-8 um), and isonotatic and norisonotatic acid.

Ocellularia saxicola is characterized by a grey-white to silver-white thallus, hyaline to

pale brown proper exciple, slightly carbonized columella developing in mature ascomata,

submuriform ascospores (22-27 x 7-9 um), and psoromic acid. Two other species, O. fumosa

and O. minutula, represent new records for Vietnam. A working key to the Ocellularia from

Vietnam is presented.

KEY worps — Chu Yang Sin National Park, lichens, taxonomy, Thelotremataceae, tropics

Introduction

During the recent investigations on the lichen biota of Vietnam, several

rich elements of tropical lichenized ascomycetes have been found (Joshi et al.

2013a, b). The most diverse group of crustose lichens in Vietnam is the order

Ostropales, in which the Graphidaceae form the largest component of tropical

species and comprise more than 2500 species worldwide (Licking et al. 2009;

Rivas Plata et al. 2012a; Kraichak et al. 2014). In Vietnam, approximately

50 species of Graphidaceae have so far been recorded (Aptroot & Sparrius

2006; Joshi et al. 2013a, b), including those previously placed separately in

the Thelotremataceae. Aptroot & Sparrius (2006) reported ten species of

thelotremoid lichens from Vietnam in the genera Chroodiscus, Myriotrema,

Thelotrema, and Ocellularia. However, following the recent modification of

912 ... Joshi & al.

taxonomic and systemic concepts in this group, some of these species are now

accommodated in Chapsa, Leucodecton, and Rhabdodiscus.

Ocellularia G. Mey. comprises more than 300 species world-wide (Frisch

et. al. 2006; Mangold et al. 2009; Rivas Plata et al. 2012b Kraichak et al.

2014; Pelaez et al. 2014) and accommodates species with usually round and

porinoid apothecia, a + carbonized proper exciple lacking lateral paraphyses,

and columellate structures. The genus has not been thoroughly investigated

in Vietnam where only seven species have been reported (Aptroot & Sparrius

2006; Joshi et al. 2013a): Ocellularia allosporoides (Nyl.) Patw. & C.R. Kulk.,

O. asiatica (Vain.) Hale [= Rhabdodiscus asiaticus (Vain.) Rivas Plata et al.],

O. dolichotata (Nyl.) Zahlbr., O. papillata (Leight.) Zahlbr., O. perforata (Leight.)

Mull. Arg., O. tenuis (Hale) Hale, and O. thelotremoides (Leight.) Zahlbr. Here

we describe two new Ocellularia species, O. lumbschii and O. saxicola, recently

collected from the Central Highlands of Vietnam. We also report two new

Vietnamese records (O. fumosa, O. minutula) and provide an artificial key to

the Ocellularia species recorded from Vietnam.

Materials & methods

A lichenological expedition at Chu Yang Sin National Park in the Central Highland

area in Vietnam was organized in 2013 by Prof. A.D. Ngyuen, Tay Nguyen University,

Vietnam. The specimens were deposited in the lichen herbarium of the Korean

Lichen Research Institute, Suncheon, Korea (KoLRI), and were studied to identify the

thelotremoid species; duplicate specimens were preserved in the Biotechnology Center

of Tay Nguyen University, Buon Ma Thuot City, Vietnam (BCTNU). The morphology

was examined under a dissecting microscope (SMZ 168), while anatomical observations

of thin hand-cut sections of thallus and ascomata were observed in tap water and

measured using a compound microscope (Olympus BX50). Colour spot reaction tests

and thin-layer chromatography (TLC) were adapted from Orange et al. (2001). Thin-

layer chromatography was performed in solvent systems A [toluene (180): 1, 4-dioxane

(45): acetic acid (5)] and C [toluene (170): acetic acid (30)]. Lugol’s solution (I) was

used to check the amyloidity of the asci, interascal filaments, and ascospores. The

measurements were based on 20-25 sections of the thallus and ascomata. Relevant

literature on thelotremoid lichens (Hale 1981; Nagarkar & Hale 1989; Awasthi 1991;

Nagarkar et al. 1988; Homchantara & Coppins 2002; Frisch et al. 2006; Mangold et al.

2009; Rivas Plata et al. 2010) was consulted for species identifications. Images were taken

using Zeiss Scope Al (AX10) and Axio cam ERc 5s, and illustrations were prepared

using Coral Draw (version 12).

Taxonomy

Ocellularia lumbschii S. Joshi & Hur, sp. nov. PL.1

MycoBANnk MB812262

Differs from Ocellularia chonestoma by its off-white thallus and its apically carbonized

excipulum and columella

Ocellularia spp. nov. (Vietnam) ... 913

PLATE 1. Ocellularia lumbschii (holotype, KoLRI). A. habit; B. cross section of ascoma;

C. transversely septate ascospores (note I+ blue reaction). Scale bars: A = 1 mm; B = 150 um;

C= 25 um.

Type: Vietnam, Dak Lak province, Chu Yang Sin National Park, 12°27’57”N

108°20'34.9’E, alt. 780 m, on bark, 21 April 2012, Hur & Oh VN120024 (Holotype,

KoLRI; isotype, BCTNU).

EryMoLoGy: The new species is named in honour of the prominent lichenologist,

Dr. H.T. Lumbsch.

Thallus corticolous, epiperidermal, off-white to creamish, + shiny, smooth,

uneven due to the substrate, 200-250 um thick; cortex continuous, made up

of periclinal hyphae, 20-25 um thick; algal layer continuous, inspersed with

crystals, 100-130 um thick; medulla white, crystalline, indistinct, mostly

endoperidermal; prothallus whitish.

914 ... Joshi & al.

Ascomata apothecioid, porinoid with round margins, immersed to slightly

emergent, solitary, dispersed, 0.4-0.6 mm in diam.; pore rounded, lined by

whitish thalline rim, filled by the tip of columella, 0.15-0.17 mm in diam.; disc

not seen from above, flesh coloured; thalline margin slightly raised, 100-200 um

thick; proper exciple apically carbonized, fused, surrounded by periderm layer

40-70 um thick; epihymenium greyish, crystalline, <15 um high; hymenium

hyaline, clear, 80-130 um high; paraphyses lax, simple, unbranched, 1-2 um

thick; subhymenium hyaline, 20-30 um high; columella simple, cylindrical,

apically carbonized, 200-250 um in width; asci narrowly clavate, uni- to

biseriate, 8-spored, 80-100 x 15-17 um, I-; ascospores hyaline, transversely

septate, 7-9-loculate, oblong to ellipsoidal, with rounded ends, 20-27 x 7-8 um,

I+ blue, halo not seen.

CHEMISTRY: K+ yellow, PD-, C-, KC-; isonotatic and norisonotatic acids

detected in TLC.

DISTRIBUTION & ECOLOGY: The new taxon is known only from its type

locality in Vietnam, where it was growing on thin bark of trees in evergreen

forests, at an altitude between 600-700 m. The associated species growing in

the same forest climate were Rhabdodiscus asiaticus, Ocellularia allosporoides,

O. papillata, O. thelotremoides, and members of lirellate graphioid taxa.

ADDITIONAL SPECIMEN EXAMINED: VIETNAM. Dak LAK PROVINCE: Chu Yang Sin

National Park, 12°28’04.4”N 108°20’39’E, alt. 622 m, on tree, 20 April 2012, Hur & Oh

VN120060 (KoLRI, BCTNU).

REMARKS: Ocellularia lumbschii is well characterized by its off-white

epiperidermal thallus, immersed to slightly emergent ascomata, apically

carbonized proper exciple and columella, transversely 6-8-septate ascospores,

and the presence of unknown compounds. Ocellularia chonestoma (Leight.)

Zahlbr., which also produces transversely septate ascospores of 18-30 x 5-6 um

and isonotatic and norisonotatic acids, differs from O. lumbschii chiefly by its

olivaceous thallus and fully carbonized excipulum and columella. Ocellularia

roseotecta Homchant. & Coppins is similar to the new taxon in having immersed

to + emergent ascomata, an apically carbonized excipulum and columella, and

transversely septate ascospores, but its pigmented medulla, slightly smaller

(15-25 x 6-8 um) ascospores, and unidentified compound (reacting PD+

orange) separate O. roseotecta from O. lumbschii. Ocellularia pyrenuloides

Zahlbr., which shares transversely septate ascospores (<30 x 5-8 um),

an apically carbonized excipulum, and a simple, carbonized columella with

the new species, differs in producing the stictic acid chemosyndrome. Other

similar species with transversely septate ascospores and the same chemistry

as O. lumbschii are O. allosporoides, which has fully carbonized excipula and

columellae, and larger ascospores, 50-120(-130) x 10-18 um (Mangold et

Ocellularia spp. nov. (Vietnam) ... 915

al. 2009), and O. baileyi Mull. Arg., which has fully carbonized excipula and

columellae and a pink medulla (Mangold et al. 2009).

Ocellularia saxicola S. Joshi & Hur, sp. nov. PL. 2

MycoBank MB812263

Differs from Ocellularia planaria by its small ascomata, distinctly submuriform and

rather large ascospores, and saxicolous habitat.

Type: Vietnam, Dak Lak province, Chu Yang Sin National Park, 12°27’57”N

108°20'34.9”E, alt. 780 m, on rock, 21 April 2012, Hur & Oh VN120129 (Holotype,

KoLRI; isotype, BCTNU).

EryMmotocy: The specific epithet refers to the rock habitat of the new species.

PLATE 2. Ocellularia saxicola (holotype, KoLRI). A. habit; B. ascomata; C. cross section of ascoma

(note the developing columella and brown excipulum); D. ascospores. Scales: A= 1 mm; B=0.5 mm;

C = 100 um; D = 50 um.

916... Joshi & al.

Thallus saxicolous, epilithic, off-white, grey-white to silver-white, corticate,

shiny, smooth, coarsely cracked due to the substratum, 150-200 um thick;

cortex continuous, made up of periclinal hyphae, thin, 15-20 um; algal layer

continuous, inspersed with crystals, 90-100 um; medulla white, crystalline,

epi- to endolithic, 70-90 um; prothallus not seen.

Ascomata apothecioid, porinoid with round margins, immersed, solitary to

rarely fused in aggregates of 2 or 3, 0.1-0.25 mm in diam.; pore 0.05-0.1 mm

in diam., lined by entire, pale brown to creamish thalline margin; disc mostly

visible, flesh coloured, thinly covered by white pruina; thalline margin

inconspicuous, whitish than the thallus, rarely erumpent, up to 80 um thick;

proper exciple fused, brownish to + hyaline, becoming carbonized apically in

mature ascomata, 20-30 um thick; epihymenium greyish, granular, crystalline,

indistinct to 15 um high; hymenium hyaline, clear, 120-150 um high; paraphyses

rather lax, simple, unbranched, 1-2 um thick; subhymenium hyaline, 30-40 um

high; columella seen only in mature and fused ascomata, apically well

developed, brown to slightly carbonized in late maturity 80-100(-120) um in

width; asci narrowly clavate, uni- to biseriate, 8-spored, 100-135 x 10-18 um,

I-; ascospores hyaline, submuriform, 8 x 1-2- loculate, subglobular to oblong,

20-27 x 7-9 um, I+ violet-blue, halonate, halo up to 5 um thick.

CHEMISTRY: K-, PD+ golden yellow, C-, KC-; psoromic acid detected in

TLC.

DISTRIBUTION & ECOLOGY: The new species is known only from its type

locality in Vietnam, where it was spreading on uneven rock surfaces along

with other porinoid members of the thelotremoid group at an altitude between

700-800 m.

ADDITIONAL SPECIMENS EXAMINED: VIETNAM. Dak LAK PROVINCE: Chu Yang Sin

National Park, 12°28’04.4”N 108°20’39’E, alt. 622 m, on rock, 20 April 2012, Hur & Oh

VN120036 (KoLRI, BCTNU); 12°28’12.3”N 108°20’59.9’E, alt. 763 m, on rock, 20 April

2012, Hur & Oh VN120082 (KoLRI, BCTNU).

REMARKS: Ocellularia saxicola is characterized by its grey- to silver-white,

shiny, and continuous to + cracked thallus, a pale brown to apically carbonized

excipulum, a poorly developed, brownish to carbonized, thin columella usually

seen in mature or fused ascomata, submuriform ascospores, and its psoromic

acid chemistry. The new species closely resembles O. planaria (Hale) Hale

in having a grey-white thallus producing psoromic acid and ecolumellate to

columellate flush ascomata. However, O. planaria differs in its somewhat larger

(0.4-0.8 mm) ascomata, transversely septate ascospores of 15-20 x 9-10 um,

and corticolous habit. Rhabdodiscus asiaticus is close to O. saxicola in having

ascospores <30 x 7-10 um and a thallus producing psoromic acid, but differs

in its greyish to pale greenish or olive verrucose thallus, emergent and larger

Ocellularia spp. nov. (Vietnam) ... 917

(<1.4 mm wide) hemispherical to urceolate ascomata, a carbonized excipulum,

a well developed columella up to 800 um thick with distinct carbonization,

and corticolous habit. Two other psoromic acid containing species, O. minutula

and O. terebrata, also have similar ascospore dimensions (O. minutula, 20-30

x 6-8 um; O. terebrata, 17-32 x 7-10 um) and a weakly developed columella

similar to the new species, but their + warty thallus, transversely septate and

non-halonate ascospores, and corticolous habit clearly distinguish them from

O. saxicola.

Ocellularia minutula Hale, Smithsonian Contr. Bot. 38: 24, 1978.

This corticolous species is characterized by a green to olive-green, smooth,

dull to + glossy, continuous thallus; inconspicuous, immersed, solitary to fused,

round, porinoid apothecia, 0.1-0.2 mm wide; the pore filled by columella at

maturity; a disc not visible from above; a brown to slightly carbonized proper

exciple; a thick, brownish to slightly carbonized, weakly developed entire

columella, 100-120 um thick; a hyaline, indistinct epihymenium; hyaline, clear

hymenium, 140-150 um; 8-spored asci; hyaline, transversely septate, oblong to

ellipsoidal, distinctly amyloid ascospores, 18-22 x 6-8 um, and the presence

of psoromic acid in the thallus. The material examined closely resembles

O. terebrata in apothecial anatomy and chemistry but differs in having a

thinner thallus and mostly immersed apothecia with small pores. The species

has a pantropical and southern temperate distribution (Mangold et. al. 2009).

SPECIMEN EXAMINED: VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National Park,

12°27'57”N 108°20'34.9’E, alt. 936 m, on tree, 21 April 2012, Hur & Oh VN120269

(KoLRI, BCTNU).

Ocellularia fumosa (Ach.) Mill. Arg., Mém. Soc. Phys. Genéve 29(8): 7, 1887.

This corticolous species is characterized by a pale-green to olive-green,

+ verrucose, dull to + glossy, continuous thallus; conspicuous, round, porinoid

apothecia, up to 0.8 mm, which are immersed to prominent at maturity, solitary

or in aggregates of two or three; a greyish disc; pores filled by columella at

maturity; a brown to slightly carbonized proper exciple; brownish to apically

carbonized, entire columella distinct in mature apothecia and 100-120

um thick; a hyaline, indistinct epihymenium; a hyaline, densely inspersed

hymenium, 120-150 um thick; 8-spored asci; hyaline, transversely septate,

oblong to ellipsoidal, distinctly amyloid ascospores, 30-45 x 6-9 um, and a

thallus producing hirtifructic and conhirtifructic acids as major compounds,

and cinchonarum unknown and other accessory compounds in traces. It differs

from O. amplior (Nyl.) Redinger, another species with inspersed hymenium,

in having a columella and in its chemistry. The species has a pantropical

distribution (Mangold et al. 2009).

918 ... Joshi & al.

SPECIMEN EXAMINED: VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National Park,

12°28'04.4”"N 108°20’39”E, alt. 622 m, on tree, 20 April 2012, Hur & Oh VN120297

(KoLRI, BCTNU).

Key to Ocellularia species recorded from Vietnam

Data for O. dolichotata and O. tenuis from Awasthi (1991) and Mangold et al. (2009)

1. Asci 2-spored; thallus ashy-white, grainy verruculose; ascomata emergent;

columella carbonized; ascospores transversely septate, 120-200 x 15-20 um;

nolichem substances: present. «2 + ow. d « Nae e < Peewee, Narnjore Nacwene Biwi O. dolichotata

AUS CLR SSP OTE he's Ak cis AM tes Le Reg Red eR be RB Be ES Rie lA Suite 2

2. | Hymenium inspersed; ascospores transversely septate, 30-45 x 6-9 um;

Zo. (AyAMenilitM Ot Insp CeseU cog! bez. cout Bas eu Hap sence pang pert ee panne de eee pero oe EB 3

S22 _ Lichen sulstanicess presen aot Ck a ae int Mack inet Mae as Anat 8 Asie Me 2 4

Syme Lac WensSMOStAN CES ADSCI Ty tel estates ge Steer Eior SI Ree Nor STP Eg Tae pr SMES cers AAS oi 9

AT alt ES OTOMMUC ACI DRES OM a a. Ata. cen Seen aaa ee arn ace we ae ate hbo hates Pe fe)

4° Psoromicacid absent: ). oss texneaens cing ey ely es eet yeh ote ne eM ae Melnge » 6

5. Ascospores submuriform, 20-27 x 7-9 um, halonate;

proper exciple mostly hyaline to pale brown;

thallus off white to grey, smooth, saxicolous...................004. O. saxicola

5. Ascospores transversely septate, 18-22 x 6-8 um, non-halonate;

proper exciple brown to slightly carbonized;

thallus in shades of green, + warty, corticolous ................... O. minutula

6. sonotatic and norisonotatic.acids:presents....-2.2% 2.0 od eek) ected oped. 7

6; “Protoeetraticacit, present’... 2424458 6445.5 oe end Fe hts yee eee eee es ee 8

7. Thallus green to olive-green; ascomata usually emergent; proper exciple

and columella mostly fully carbonized; ascospores transversely septate,

hyaline, GO=7O XBT ooh SFL bite eu, cto bannato heme fin O. allosporoides

7. Thallus off-white; ascomata immersed to slightly emergent; proper exciple

and columella apically carbonized; ascospores transversely septate, hyaline,

20D 7S SB (IIS Penta gee he a Mart es sitet tis Sletten sete te: Shag ras autteet os sighs O. lumbschii

8. Ascospores submuriform, 24-26 x 12-13 um; proper exciple rarely marginally

carbonized; columella entire, brownish to carbonized ........ O. thelotremoides

8. Ascospores transversely septate, 20-32 x 6-9 um; proper exciple brown, rarely

marginally carbonized; columella entire, brownish to carbonized . . . O. perforata

9. Ascospores transversely septate, 19-20 x 6-7 um; ascomata conspicuous,

immersed to moderately emergent; proper exciple apically to marginally

carbonized; columella developed at maturity, brown to carbonized. ... O. papillata

2. Ascospores transversely septate to submuriform, 10-20 x 5-8 um;

ascomata inconspicuous, immersed; proper exciple marginally carbonized;

columella developed at maturity, carbonized....................000. O. tenuis

Ocellularia spp. nov. (Vietnam) ... 919

Acknowledgments

This work was supported by a grant from the National Research Foundation of

Korea (#2011-0031494) and the Korea National Research Resource Center Program.

The authors are grateful to Dr. Robert Liicking and Dr. Ze-Feng Jia for their valuable

comments on the manuscript.

Literature cited

Aptroot A, Sparrius LB. 2006. Addition to the lichen flora of Vietnam, with an annotated checklist

and bibliography. Bryologist 109: 358-371. http://dx.doi.org/10.1639/0007-2745

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

40. 337 p.

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

Thelotremataceae. Bibliotheca Lichenologica 92. 556 p.

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

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

Homchantara N, Coppins BJ. 2002. New species of the lichen family Thelotremataceae in SE Asia.

Lichenologist 34: 113-140. http://dx.doi.org/10.1006/lich.2002.0382

Joshi S, Jayalal U, Oh SO, Koh YJ., Nguyen TT, Dzung NA, Hur JS. 2013a. New species and new

records in the family Graphidaceae (Ascomycota: Ostropales) from Vietnam. Lichenologist 45:

599-609. http://dx.doi.org/10.1017/S002428291300025X

Joshi S, Nguyen TT, Dzung NA, Jayalal U, Oh SO, Hur JS. 2013b. New records of corticolous lichens

from Vietnam. Mycotaxon 123: 479-489. http://dx.doi.org/10.5248/123.479

Kraichak E, Parnmen §, Liicking R, Rivas Plata E, Aptroot A, Caceres MES, Ertz D, Mangold A,

Mercado-Diaz JA, Papong K, Broeck DVD, Weerakoon G, Lumbsch HT. 2014. Revisiting

the phylogeny of Ocellularieae, the second largest tribe within Graphidaceae (lichenized

Ascomycota: Ostropales). Phytotaxa 189: 52-81. http://dx.doi.org/10.11646/phytotaxa.189.1.6

Liicking R. 2009. The taxonomy of the genus Graphis sensu Staiger (Ascomycota: Ostropales:

Graphidaceae). Lichenologist 41: 319-362. http://dx.doi-org/10.1017/S0024282909008305

Mangold A, Elix JA, Lumbsch HT. 2009. Thelotremataceae. Flora of Australia 57: 195-420.

Nagarkar MB, Hale ME Jr. 1989. New species in the lichen family Thelotremataceae from Asia

(Ascomycotina). Mycotaxon 35: 437-447.

Nagarkar MB, Sethy PK, Patwardhan PG. 1988. Lichen genus Ocellularia (Family Thelotremataceae)

from India. Biovigyanam 14: 24-43.

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society. 101 p.

Pelaez RN, Moncada B, Licking R. 2014. High diversity of Ocellularia (Ascomycota: Graphidaceae)

in the Colombian Llanos, including two species new to science. Phytotaxa 189: 245-254.

http://dx.doi.org/10.11646/phytotaxa.189.1.17

Rivas Plata E, Licking R, Sipman HJM, Mangold A, Kalb K, Lumbsch HT. 2010. A world-wide

key to the thelotremoid Graphidaceae, excluding the Ocellularia~-Myriotrema-Stegobolus clade.

Lichenologist 42: 139-185. http://dx.doi.org/10.1017/S0024282909990491

Rivas Plata E, Liicking R, Lumbsch HT. 2012a. A new classification for the family

Graphidaceae (Ascomycota: Lecanoromycetes: Ostropales). Fungal Diversity 52: 107-121.

http://dx.doi.org/10.1007/s13225-011-0135-8

Rivas Plata E, Licking R, Lumbsch HT. 2012b. Molecular phylogeny and systematics of the

Ocellularia clade (Ascomycota: Ostropales: Graphidaceae). Taxon 61: 1161-1179.

MYCOTAXON

http://dx.doi.org/10.5248/130.921

Volume 130, pp. 921-924 July-September 2015

Book reviews and notices

ELSE C. VELLINGA (GUEST BOOK REVIEW EDITOR)

"861 Keeler Avenue, Berkeley CA 94708 U.S.A.

" CORRESPONDENCE TO: bookreviews@mycotaxon.com

ABSTRACT— —Books reviewed include: GENERAL—California mushrooms.

The comprehensive identification guide (Desjardin, Wood, & Stevens 2015.);

BASIDIOMYCETES—Atlas of the Chinese species of Amanitaceae (Yang 2015).

GENERAL

California mushrooms. The comprehensive identification guide. By

D.E. Desjardin, M.G. Wood, & EA. Stevens. 2015. Timber Press Inc., 133 S.W.

Second Avenue, Suite 450, Portland OR 97204-3527, <timberpress.com>. ISBN

978-1-60469-353-9. 560 p., over 700 colour photos. Price $60.

The state of California in the western United States of America is well known

for its wide range of habitats and its rich biodiversity. Fungi in the state are

not as well investigated as plants, and only a few field guides to the California

mushrooms exist (Biek 1984; Arora 1986, 1991; Davis et al. 2012), this in

contrast to a huge number of regional and statewide floras. This new book,

California mushrooms, is a welcome up-to-date addition, and the authors

should be congratulated on producing this beautiful well-executed book.

The contents of the book follow the familiar set-up of guides with an

introduction in which the topic is introduced, an explanation of the way the

book has been made, very useful introductions to taxonomy and nomenclature

(why do names change?), mushroom ecology, and biology, and a few pages

on edible and toxic mushrooms. Chapters on ‘How to identify mushrooms’

and “How to use the book are followed by the main part, “Ihe mushrooms.

* Book reviews or books for consideration for coverage in this column should be sent to the

address above OR to the Editor-in-Chief at editor@mycotaxon.com or 6720 NW Skyline,

Portland OR 97229 USA.

922 ... MYCOTAXON 130

650 species of fungi are well illustrated (each with one picture), described, and

compared with species and other taxa that are or are not illustrated in the book.

Keys to morphological groups, followed by more in-depth keys per group to

species are, of course, present as well. The glossary of used terms, a list of plant

names, references, a list of California mushroom clubs, photo credits, an index,

and biographies of the authors follow the main part.

There are several things that make this book stand out among similar books

in the USA.

Its large format (pages are 21 x 28 cm) means that the photos are also large,

which is fantastic.

The descriptions are based on observations by the authors on material from

California and surrounding areas; microscopic data were recorded by the

first author mainly from material present in the Harry D. Thiers Herbarium

or from recent publications from California. However, it is not clear to which

vouchered specimen each photograph is linked. Most of the photos were taken

in California.

The nomenclature is current, and a list of updates since the book’s text was

finished is posted on www.californiamushrooms.us/errata.html. In some cases

the authors give provisional new combinations to indicate to which genus a

species belongs according to the newest phylogenetic insights, and a number

of as-yet unpublished new species names are also presented (e.g., in Agaricus

where a monograph is expected to be issued soon). Indeed it is a challenge

to write a book on California mushrooms, as it is very clear that many of the

names generally used are not applicable in the state, but new names have not

been proposed yet.

This is a beautiful book that introduces the most common and most

commonly encountered species of northern California. The emphasis of the

book is on the areas the authors have visited most: the San Francisco Bay

region, Mendocino and the Sequoia sempervirens and Picea sitchensis forests

in the northwest of the state, and the central part of the Sierra Nevada. The

introduction is very helpful and informative, but is written for an audience with

some background in biology. The emphasis in the keys and in the descriptions

is on macroscopic characters, but in some cases microscopic characters

are mentioned; unfortunately not all terms are included in the glossary (for

instance, an explanation of the term “broom cells’ is lacking). For some groups,

adding a microscopic character (or illustration thereof) in the key would

make identification so much easier. The difference between Laccaria and the

Hygrophoraceae is always hard for beginners, but with the addition of the spore

ornamentation of Laccaria spores, distinguishing the two groups becomes a

piece of cake. The same applies to the pink-spored gilled mushrooms where the

Book Reviews ... 923

different spore shapes of the Entolomataceae are mentioned in the key, but not

illustrated. The species are arranged by general shape, not by their phylogenetic

affiliations, so the gilled boletes stand in the middle of gilled mushroom species,

and Auriscalpium finds its place with other species with a spiny hymenophore

and not with its closest relatives in the Russulales. Only species that are fully

treated in the book are keyed out, which limits the usefulness of the book.

And although the number of species covered in the book is not small, many

species are left out; the undoubtedly most species rich genus, Cortinarius, is

represented by only 25 species, and Pluteus only by P leoninus and three species

in P. sect. Pluteus. The one real mistake I came across is in the placement of

Neolentinus, which does not belong to the Polyporaceae, but is in the middle

of the Gloeophyllales (Garcia-Sandoval et al. 2011). Here, the authors followed

Index Fungorum, which also has not been updated to reflect the newest insights.

The photos are of good to high quality showing the mushrooms well. Of

course some are not as typical as I would like them to appear (e.g., Mycena

haematopus lacks the characteristic hanging pileus margin).

As I said, this is indeed a beautiful, well-illustrated, and informative book

that definitely will be used widely, both in and outside California. Because of its

size and weight (more than 2 kg), it will not fit in a pocket or backpack. But just

leafing through it at home, or in the car, will be pleasure enough!

Arora D, 1986. Mushrooms demystified, 2‘ Ed. Ten Speed Press, Berkeley.

Arora, D., 1991. All that the rain promises, and more. Ten Speed Press,

Berkeley.

Biek, D., 1984. The mushrooms of northern California. Spore prints, Redding.

Davis RM, Sommer R, Menge JA. 2012. Field guide to mushrooms of western

North America. California Natural History Guides 106. University of

California Press, Berkely, Los Angeles, London.

Garcia-Sandoval R, Wang Z, Binder M, Hibbett 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

BASIDIOMYCETES

Atlas of the Chinese species of Amanitaceae. By Z.-L. Yang. 2015. Science Press,

Beijing China. <www.sciencep.com>; ISBN 978-7-03-043627-6; 213 pages, many

colour photos. Price US$ 53.00 [in Chinese with short introduction in English and

Latin names].

China is a vast country with many different habitats and a very rich funga; in

recent years it has become especially clear how rich the funga is. This book

924 ... MYCOTAXON 130

reflects the present knowledge of one mushroom family, the Amanitaceae, in

China. 130 taxa representing the genera Amanita and Limacella are illustrated

and described, with many photos per species to illustrate various developmental

stages. The taxa are organized by genus and by section. An introduction to the

important morphological characters and keys to all taxa treated in the book are

given as well. Very importantly, a list of voucher specimens is provided, which

enables verification if needed. A list of references is present as well.

This book is in the first place aimed at a Chinese audience to make clear

which species are edible and which are deadly poisonous. For this reason the

photos of taxa in the latter category are provided with a red warning sign.

Research of this family showed that many taxa occur in a white form besides

the normally coloured specimens. Generalizations about species’ distribution

patterns are also shown to be often wrong; some European species, such as

Amanita spissa, do occur in China, others, including A. phalloides, have not

yet been found. And lastly, some species turned out to represent a complex of

various taxa, as in the stunningly beautiful orange-capped A. hemibapha.

Some as-yet unnamed taxa are tagged with a provisional name, such as A.

pseudopantherina, or number, e.g., Amanita sp. 10.

Although the text is in Chinese and most likely not accessible to most people

outside China, the illustrations can easily be understood, and the diversity,

beauty, and variability of the species is visible for all. This book gives a very

good first introduction to the family in China. We are looking forward to more

such publications, and to a follow-up of the present one.

MYCOTAXON

http://dx.doi.org/10.5248/130.925

Volume 130, pp. 925-926 July-September 2015

Regional annotated mycobiotas new to the Mycotaxon website

ABSTRACT — MycoTaxon is pleased to announce two new species distribution lists

to our ‘web-list’ page covering jelly fungi in Brazil (by Alvarenga & Xavier-Santos) and

the lichens of Burdur Province in Turkey (by Yazici & al.). The revised mycobiota on

Fusarium in Turkey by Ahmet Asan has also been uploaded. A new feature, MycoTaxon

now provides a global database (in Excel format) on coprophilous fungi developed

by Michael Richardson during a lifetime of study. This substantial offering brings to

116 the number of free access mycobiotas now available on the MycoTaxon website:

http://www.mycotaxon.com/resources/weblists.html

GLOBAL

MIcHAEL J. RICHARDSON. Records of coprophilous fungi - a data set. 5 p. +

>12,300 records with ~1380 dates/localities/substrates available in Excel

format.

ABSTRACT — Many coprophilous fungi are adapted to their habitat by

having specialised spore dispersal mechanisms to improve the chance of the

spores being consumed, since they require passage through an animal's gut

to encourage their germination. Analysis of coprophilous fungus records

allows a study of regional, substrate, and seasonal differences, and has

demonstrated that fungi increase in biodiversity with decreasing latitude.

1386 samples of dung, mainly of herbivorous mammals and birds, were

collected, mostly between 1994 and 2014, from the wild from various parts

of the world. Most (88%) were from north temperate areas, with 7% from

south temperate areas, 4% from the tropics, and 0.3% from the Arctic. Over

12,300 records of coprophilous fungi were obtained from these samples on

incubation in damp chambers. Details of the fungi found and their origin

are presented as files that provide a data set that triples the amount of data

used in a 2001 analysis. The data set includes collection details for the

samples (locality, country, latitude/longitude coordinates, elevation), date

of collection, dung type, species recorded from each sample, location of

herbarium deposits, and citations to references where the records have been

published. Notes of observations made on each sample during incubation

[one pdf for each sample] are also in the on-line checklist, with ppFs of

publications in which the records are cited.

2 ... New regional mycobiotas online

SOUTH AMERICA

Brazil

RENATO LUCIO MENDES ALVARENGA & SOLANGE XAVIER-SANTOS. A checklist

of Jelly Fungi (Agaricomycotina: Basidiomycota) recorded in Brazil. 14 p.

ABSTRACT — Based on an intensive search of literature records on jelly

fungi (Agaricomycotina: Basidiomycota) in Brazil, a list of 88 species

was compiled. These are distributed into four orders (Auriculariales,

Dacrymycetales, Sebacinales, and Tremellales) and seven families

(Auriculariaceae, Dacrymycetaceae, Hyaloriaceae, Phragmoxenidiaceae,

Sebacinaceae, Sirobasidiaceae, and Tremellaceae), with the most frequent

taxa being Auricularia nigricans, A. fuscosuccinea, and A. delicata sensu lato.

Among the 16 Brazilian states with occurrence records, the most frequent

are Rio Grande do Sul, Sao Paulo and Parana. The facts that in 40% of the

states there is no occurrence and that most species are represented by a

single record reinforce the need for taxonomic studies about the group

MIp-EAST

Turkey

KENAN YAZICI, ANDRE APTROOT, ALI ASLAN, HARRIE SIPMAN, & MICHELE D.

PreRcEY-NormokeE. The lichen biota of Burdur province (Turkey). 30 p.

ABsTRACT — As a result of lichenological exploration in the Burdur

province (Turkey), a total of 397 lichenized fungi, representing 126 genera

in the Ascomycota, including 3 subspecies and 6 varieties, were determined

from 185 different localities. Aspicilia uxoris, Cladonia merochlorophaea,

Cladonia pulvinella, Hypocenomyce caradocensis, Lecidella pulveracea,

Rinodina freyi, and R. griseosoralifera are new to Turkey, and R. freyi and

R. griseosoralifera are also new to Asia. Collection localities and substrata

are also presented.

MY COTAXON

http://dx.doi.org/10.5248/130.927

Volume 130, pp. 927-928 July-September 2015

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 130(3)

Anthostomella saltensis Sir, Hladki & A.I. Romero, p. 722

Antrodiella indica G. Kaur, Avneet P. Singh & Dhingra, p. 625

Artomyces nothofagi M.E. Sm. & Kneal, p. 656

Blastophragma chonggingense J.M. Gao & X.G. Zhang, p. 822

Codinaea jianfenglingensis J.W. Xia & X.G. Zhang, p. 841

Cytospora notastroma Kepley & EB. Reeves, p. 793

Elmerina fragilis F. Wu & Hai J. Li, p. 683

Fuscoporia atlantica Motato- Vasquez, R.M. Pires & Gugliotta, p. 848

Hansfordia qinghaiensis H.F. Wang & TY. Zhang, p. 809

Hansfordia rosea J.J. Xu & T.Y. Zhang, p. 809

Inonotus griseus L.W. Zhou, p. 664

Marthamyces chinensis Y.R. Lin & H.L. Gu, p. 816

Mirandina inaequalis Y.R. Ma & X.G. Zhang, p. 777

Ocellularia lumbschii S. Joshi & Hur, p. 912

Ocellularia saxicola S. Joshi & Hur, p. 915

Pachyphloeus depressus L. Fan, p. 615

Pertusaria alticola Q. Ren, p. 690

Phaeomonilia guangxiensis Y.R. Ma & X.G. Zhang, p. 776

Phaeostilbelloides Armando, Z.M. Chaves & Dianese, p.258

Phaeostilbelloides velloziae Armando, Z.M. Chaves & Dianese, p. 258

Phlebiopsis punjabensis G. Kaur, Avneet P. Singh & Dhingra, p. 907

Selenodriella amoena M.A.G Aratijo, M.A. Barbosa, Malosso & R.E Castaneda, p. 622

Selenosporella minima Fiuza, Gusmao & R.F. Castafieda, p. 602

Sporidesmiopsis malloti J.W. Xia & X.G. Zhang, p. 829

Terriera transversa Y.R. Lin & Qing Li, p. 894

Tubulicrinis indicus Jyoti Sharma, Avneet P. Singh & Dhingra, p. 879