IV ... MYCOTAXON 121

MYCOTAXON

VOLUME ONE HUNDRED TWENTY-ONE — TABLE OF CONTENTS

COVER SECTION

PIGGG Fs Se SLR ee BAe SOR EAs ee Oe SRR Some es 2a Oe tee es viii

TRC VFOWWCHS He EN, Sy den, 504 IM oP GAP Wiehe te Sea Re A OMNE 2 aP, 2 8 Ie ix

SUDMUSSON BIGCCUATES. Om. sey cit Neel A Rm, the Eat, Ma MGEY 2a Sue tes x

E POULAOLE CIOL, 1 ar tgd ga Pint tide. trek west eer aed ahs Ma eth tec tele dh eat xi

RESEARCH ARTICLES

Agarics of alders I - The Alnicola badia complex

Pierre-Arthur Moreau, Juliette Rochet, Enrico Bizio, Laurent Deparis,

Ursula Peintner, Beatrice Senn-Irlet, Leho Tedersoo & Monique Gardes 1

A new species of Xerocomus from Southern China

Ming Zhang, Tai-Hui Li, Tolgor Bau & Bin Song 23

Nomenclatural and taxonomic notes on Calvatia (Lycoperdaceae)

and associated genera Johannes C. Coetzee & Abraham E. Van Wyk 29

New combinations and notes in clavarioid fungi I. Olariaga & I. Salcedo 37

A new species of Stigmidium (Mycosphaerellaceae) on Aspicilia

from North America Jana Kocourkova & Kerry Knudsen 45

Notes on some Eurasian species of Anthracoidea and Entyloma

Kyrylo G. Savchenko, Vasyl P. Heluta,

Ivan S. Hirylovich, Solomon P. Wasser & Eviatar Nevo 53

Lactarius annulocystidiatus sp. nov. from India

Samidha Sharma, Munruchi Kaur & N.S. Atri 63

A new species of Lentinus from India

Gunasekaran Senthilarasu & Sanjay K. Singh 69

Two new species in the Graphidaceae (Ostropales, Ascomycota) from China

Ze-Feng Jia, Rui-Fang Wang & Jang-Chun Wei 75

Tricholomopsis in Europe -phylogeny, key, and notes on variability

Jan Holec & Miroslav Kolatik 81

Ganoderma (Basidiomycota or Agaricomycetes) in Brazil:

known species and new records Mabel Gisela Torres-Torres,

Laura Guzman- Davalos & Adriana Melo Gugliotta 93

New records of pyrenocarpous lichenized fungi from Bulgaria

Veselin V. Shivarov & Dimitar Y. Stoykov 133

Lichenological notes 5: Neotypification of Sarcogyne magnussonii

(Acarosporaceae) Kerry Knudsen & Jana Kocourkova 139

New species of Humicola and Endophragmiella from China

Yue-Ming Wu & Tian-Yu Zhang 147

JULY-SEPTEMBER 2012... V

Pholiota virescens, a new species from China En-jing Tian & Tolgor Bau 153

Conocybe hausknechtii, a new species of sect. Pilosellae

from western Caucasus, Russia Ekaterina Malysheva 159

Additions to the smut fungi of Pakistan. 1.

Cvetomir M. Denchev, Muhammad Fiaz, Teodor T. Denchev,

Habib Ahmad & Abdul Nasir Khalid 165

Two new microfungi from Portugal: Magnohelicospora iberica

gen. & sp. nov. and Phaeodactylium stadleri sp. nov.

Rafael F. Castafieda-Ruiz, Margarita Hernandez-Restrepo,

Josepa Gené, Josep Guarro, David W. Minter & Masatoshi Saikawa 171

Ellisembia karadkensis sp. nov. from southern Western Ghats, India

Kunhiraman C. Rajeshkumar, Swapnil C. Kajale,

Somnath A. Sutar & Sanjay K. Singh 181

Mycoacia angustata sp. nov. (Basidiomycota, Meruliaceae), the first

Chinese hydnoid species Hai-Sheng Yuan & Xian-Zhen Wan 187

A new species of Entoloma from Liaoning Province,

Northeast China Liang-Liang Qi, Xiao-Lan He & Yu Li 193

Beauveria lii sp. nov., isolated from Henosepilachna vigintioctopunctata

Sheng-li Zhang, Ling-ming He, Xue Chen, Zeng-zhi Li & Bo Huang 199

Kuehneola warburgiana comb. nov. (Phragmidiaceae, Pucciniales),

causing witches’ brooms on Rosa bracteata Yoshitaka Ono 207

New records of smut fungi.6 Cvetomir M. Denchev & Teodor T. Denchev 215

Suillus flavidus and its ectomycorrhizae with Pinus wallichiana

in Pakistan S. Sarwar, A.N. Khalid, M. Hanif & A.R. Niazi 225

Russulaceae of the Pakaraima Mountains of Guyana 2:

new species of Russula and Lactifluus

Steven L. Miller, M. Catherine Aime & Terry W. Henkel 233

Tuber in China: T. sinopuberulum and T. vesicoperidium spp. nov.

Li Fan, Jin-Zhong Cao & Jia Yu 255

Presence of Leucocoprinus cretaceus and L. fragilissimus

in Argentina N. Niveiro, O. Popoff & E. Alberté 265

Two new species of Lasionectria (Bionectriaceae, Hypocreales)

from Guadeloupe and Martinique (French West Indies)

Christian Lechat & Jacques Fournier 275

Steccherinum straminellum, a new record for Italy

Alessandro Saitta & Ireneia Melo 281

A new species of Inonotus (Basidiomycotina, Hymenochaetales)

from tropical Yunnan, China Hai-Jiao Li & Shuang-Hui He 285

A new species of Grammothelopsis (Polyporales, Basidiomycota)

from southern China Chang-Lin Zhao & Bao-Kai Cui 291

VI... MYCOTAXON 121

Two new species of white truffle from China Li Fan & Jin-Zhong Cao 297

Two new species of Ceriporia (Basidiomycota, Polyporales)

with a key to the accepted species in China Bi-Si Jia & Bao-Kai Cui 305

New Candelariella records for Turkey

Mehmet Gokhan Halici, Mustafa Kocakaya & Emre Kilig 313

Lepiota himalayensis (Basidiomycota, Agaricales),

a new species from Pakistan A. Razaq, A.N. Khalid & E.C. Vellinga 319

Phytophthora castaneae, the correct name for

P. katsurae nom. nov. superfl. Shaun R. Pennycook 327

Studies on Wrightoporia from China 2. A new species and

three new records from South China Jia-jia Chen & Bao-Kai Cui 333

Diplomitoporus dilutabilis belongs to Cinereomyces

(Polyporales, Basidiomycota) Otto Miettinen 345

Uncobasidium roseocremeum sp. nov. and other corticioid

basidiomycetes new to the Patagonian Andes (Argentina)

Sergio P. Gorjon, Alina G. Greslebin & Mario Rajchenberg 349

Cyathus badius and C. earlei reported from the Brazilian

Atlantic rainforest Rhudson Henrique Santos Ferreira da Cruz,

Marcos Mateus Barros Barbosa & Iuri Goulart Baseia 365

First report of Stemphylium lycopersici from Far East Russia:

a new record and new host Philipp B. Gannibal 371

Three new species of Septobasidium (Septobasidiaceae)

from southern and southwestern China Suzhen Chen & Lin Guo 375

New records of Lecanora for Bolivia Lucyna Sliwa, Karina Wilk,

Pamela Rodriguez Flakus & Adam Flakus 385

Laccariopsis, a new genus for Hydropus mediterraneus

(Basidiomycota, Agaricales)

Alfredo Vizzini, Enrico Ercole & Samuele Voyron 393

Amanita chocoana—a new species from Ecuador

Felipe Wartchow & J. Pail Gamboa-Trujillo 405

Heteroconium bannaense sp. nov. and a new record of the genus

from China _ Ji-Wen Xia, Shou-Cai Ren, Li-Guo Ma & Xiu-Guo Zhang 413

Mycobilimbia and Rinodina species new to Turkey Kenan Yazici 419

New records of smut fungi.7 | Cvetomir M. Denchev & Teodor T. Denchev 425

Type studies on four Entoloma species from South China

Xiao-Lan He, Tai-Hui Li, Zi-De Jiang & Ya-Heng Shen 435

Notes on Stereocaulon species from Bolivia

Magdalena Oset & Martin Kukwa 447

JULY-SEPTEMBER 2012... VII

Taxonomy and phylogenetic placement of the downy mildew

Peronospora saturejae-hortensis Jutta Gabler,

Gregor Hagedorn & Uwe Braun 455

Clarification of Peziza fimeti with notes on P. varia collections

on dung Gianfranco Medardi, Angela Lantieri,

Donald H. Pfister, Katherine F. LoBuglio & Gabriele Cacialli 465

New taxa of Hymenochaete (Agaricomycetes, Hymenochaetales)

with a note on H. caucasica Erast Parmastot 477

BOOK REVIEWS AND NOTICES Else C. Vellinga (Editor) 485

MYCOBIOTAS ONLINE

Checklist of the Argentine Agaricales 4. Tricholomataceae and

Polyporaceae N Niveiro & E. Alberté 499

Diversity of Colombian macrofungi (Ascomycota - Basidiomycota)

Aida Marcela Vasco-Palacios &Ana Esperanza Franco-Molano 499

A preliminary checklist of smut fungi of Croatia

Dario Ivi¢é, Zdravka Sever, Christian Scheuer & Matthias Lutz 499

A preliminary checklist of the Boletales in Pakistan

Samina Sarwar & A.N. Khalid 500

NOMENCLATURE

Nomenclatural novelties proposed in volume 121 501

PUBLICATION DATE FOR VOLUME ONE HUNDRED TWENTY

MYCOTAXON for APRIL-JUNE, VOLUME 120 (I-x1I + 1-510)

was issued on September 28, 2012

vul ... MYCOTAXON 121

ERRATA FROM PREVIOUS VOLUMES

VOLUME 105

p. 174, line 11

VOLUME 120

p-69, line 37

p-70, line 3

p-100, line 21

p- 109, line 28

p- 109, line 31

p- 110, line 18

p. 157, line 3

p-199, line 9

p-203, line 3

p-257, line 5

p-264, lines 10-11

p.289, 4" from bottom

p- 401, line 19

for: Gloeoporus niger

read: Gloeoporus dichrous vat. niger

for: with warts or a irregular covering layer of various thickness

read: with warts or an irregular covering layer of various thickness,

for: f. Asci with ascospores. g. Conidiophores and conidia.

read: f. Conidiophores and conidia. g. Asci with ascospores.

for: Shangri La

read: Shangri-La

for: lichesterinic, protolichesterinic, nephrosteranic,

read: lichesterinic, protolichesterinic, nephrosterinic,

for: except nephrosteranic,

read: except nephrosteranic,

for: the absence of lobaric and fatty acids

read: the absence of fatty acids

for: AKBAR KHODAPARAST

read: SEYED AKBAR KHODAPARAST

for: there were recorded

read: there were three records

for: Telleria

read: Telleria

for: Cleistocybe vernalis Ammirati et al. cluster sister to the

C. acromelalga/C. amoenolens pair

read: Cleistocybe vernalis Ammirati et al. clusters sister to the

C. acromelalga/ C. amoenolens pair

for: Additionally, our analyses show only a 91% pairwise

ITS sequence identity...

read: Additionally, our analyses show only a 94.4% pairwise

ITS sequence identity...

for: ...National Natural Science Foundation Program of

PR China (31160008, 31060008).

read: ... National Natural Science Foundation Program of

PR China (31160008, 31060008,31260007).

for: in July, 2009 is

read: in July 2009, is

Juty-SEPTEMBER 2012... IX

REVIEWERS — VOLUME ONE HUNDRED TWENTY

The Editors express their appreciation to the following individuals who have,

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

prepared for this volume.

Najm-u-sehar Afshan

Anibal Alves de Carvalho

Junior

Joe Ammirati

Vladimir Antonin

André Aptroot

Alan W. Archer

Eef Arnolds

Juliano M. Baltazar

Timothy J. Baroni

Annarosa Bernicchia

Joshua M. Birkebak

Peter Buchanan

Harold H. Burdsall

Marina Capelari

Lori Carris

Young-Joon Choi

J.C. Coetzee

Vagner Gularte Cortez

Bao-Kai Cui

Yu-Cheng Dai

Kanad Das

Francesco Doveri

F, Esteve-Raventés

Adam Flakus

Genevieve M. Gates

Tatiana B. Gibertoni

Ying-Lan Guo

M. Gokhan Halici

Ian R. Hall

David Hawksworth

Shuang-Hui He

Stephan Helfer

Jan Holec

Volker John

Klaus Kalb

Taiga Kasuya

Bryce Kendrick

Takahito Kobayashi

Heikki Kotiranta

Irmgard Krisai-Greilhuber

Martin Kukwa

T.K. Arun Kumar

James C. Lendemer

De-Wei Li

Jun-Feng Liang

Xiao- Yong Liu

Laszlo Lékés

Rosalind Lowen

Guo-Zhong Lit

Matthias Lutz

P. Manimohan

Eric H.C. McKenzie

Ireneia Melo

Andrew M. Minnis

Lorelei L. Norvell

Beatriz Ortiz-Santana

Shaun R. Pennycook

Ronald H. Petersen

Marcin Piatek

Orlando F. Popoff

Mykola Prydiuk

Barry M. Pryor

Mario Rajchenberg

Scott A Redhead

Steven A. Rehner

Amy Rossman

Svengunnar Ryman

Leif Ryvarden

Jean-Michel Savoie

Mark R.D. Seaward

Brij M. Sharma

Roger Graham Shivas

Gladstone Alves da Silva

Emory G. Simmons

H.J.M. Sipman

Joost Stalpers

Ave Suija

Leho Tedersoo

Erin A. Tripp

Kalman Vanky

Annemieke Verbeken

Josef Vlasak

Jan Vondrak

Long Wang

Zheng Wang

Felipe Wartchow

Martin Westberg

Hai-Sheng Yuan

Tian-Yu Zhang

Li-Wei Zhou

Wen-Ying Zhuang

x ... MYCOTAXON 121

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

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free download of distributional annotated species lists.

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

Mycotaxon publishes four 500-page volumes a year. Both open access and

subscription articles are offered.

JULY-SEPTEMBER 2012... XI

FROM THE EDITOR-IN-CHIEF

MELBOURNE CODE PUBLISHED — Authors are urged to consult the new International

Code of Nomenclature for algae, fungi, and plants (Melbourne Code) [McNeill et al.

2012]. Fortunately for web-users, the Articles, Notes, and Recommendations are freely

available online at <http://www.iapt-taxon.org/nomen/main.php>. Official hard copy

may be ordered from Koeltz Scientific Books [Regnum Vegetabile 154. A.R.G. Gantner

Verlag KG. ISBN 978-3-87429-425-6].

MycoBANnk, INDEXFUNGORUM & FUNGAL NAME ARE THE 2013. OFFICIAL

NOMENCLATURAL REPOSITORIES — The MELBOURNE CODE requires that an identifier

issued by a recognized repository be cited in the protologue for valid publication of a

new taxon in 2013. MycoBank <http://www.mycobank.org>, established in 2005 as the

first fungal nomenclatural repository and now overseen by the International Mycological

Association, was joined by INDEx FuNGoRuM <http://www.indexfungorum.org> in

2009 and FUNGAL NaME <http://fungalinfo.im.ac.cn/fungalname/fungalname.html> in

2010. MycoTaxon initially required a MycoBank identifier seven years ago; recently,

however, we have also accepted numbers from INDEXFUNGORUM and FUNGAL NAME

and will continue to do so.

This past November (2012) representatives from the repositories noted above

signed a Memorandum of Cooperation agreeing to work together under the guidance

of the Nomenclature Committee for Fungi (NFC). In December the NCF voted by a

71% majority to accept all three registries beginning 1 January 2013, with the NCF

recommendation to be ratified by the 2014 Bangkok International Mycological Congress,

provided the IMC finds multiple registries workable and effective.

For more information, see Redhead & Norvell (2012; IMA Fungus 3(2): 44-45).

MyYCOTAXON 121 FINALLY COMPLETED! — We greatly regret our very late ‘July-

September’ volume and appreciate even more everyone's patience during the recent

difficult period. Although 2012 brought health challenges that both editors would

prefer not to have encountered, we are now conquering our manuscript backlog and

going forward. Fortunately, the 55 papers in this volume offer MycorTaxon’s usual

cornucopia of fungal delicacies and insights, and there are at least another 55 fascinating

submissions awaiting review for Mycotaxon 122, now scheduled for February.

We take this opportunity to thank our authors and experts for their hard work and

to wish everyone health, happiness, peace, progress, and health in 2013. (Health is

mentioned twice because we are now convinced it is the most important wish of all!)

Warm regards,

Lorelei Norvell (Editor-in-Chief)

1 January 2013

MYCOTAXON

http://dx.doi.org/10.5248/121.1

Volume 121, pp. 1-22 July-September 2012

Agarics of alders 1 - the Alnicola badia complex

PIERRE-ARTHUR MOREAU? , JULIETTE ROCHET’, ENRICO BIZzI03,

LAURENT DEPARIS‘*, URSULA PEINTNER®, BEATRICE SENN-IRLET‘,

LEHO TEDERSOO’ & MONIQUE GARDES?

'Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France,

BP83. 3 rue du Pr Laguesse, F-59006 Lille cedex, France

?Laboratoire Evolution et Diversité Biologique, Université de Toulouse,

3 Paul Sabatier CNRS, UMR 5174, F-31062 Toulouse Cedex 4, France

*Societa Veneziana di Micologia, c/o Museo di Storia Naturale,

1730 S. Croce. I-30135 Venezia, Italy

4305, rue des Ecoles, F-74930 Reignier, France

°Universitat Innsbruck, Institut fiir Mikrobiologie,

Technikerstrafse 25, A-6020 Innsbruck, Austria

° Swiss Federal Institute for Forest, Snow and Landscape Research WSL,

Ziircherstrafe, 111. CH- 8903 Birmensdorf, Switzerland

“Institute of Ecology and Earth Sciences and Natural History Museum, University of Tartu,

40 Lai Street, EST-51005 Tartu, Estonia

* CORRESPONDENCE TO *: pierre-arthur.moreau@univ-lille2.fr

ABSTRACT — Three closely related species in Alnicola sect. Alnicola, Alnicola, are described,

illustrated, and compared. Alnicola badia is revised based on the type collection and recent

collections made under Alnus alnobetula. Two species (A. longicystis, A. xanthophylla)

associated with Alnus glutinosa and A. incana and locally frequent in western and central

Europe are described as new.

Key worps — Agaricales, Basidiomycota, Hymenogastraceae, Naucoria, taxonomy

Introduction

The agaric genus Alnicola Kiihner (Naucoria s. auct.) was defined by Kthner

(1926) as strictly an Alnus-associated genus with small-sized basidiomata

and rough basidiospores. Four species were cited by Kithner: three of them

(A. badia, A. luteolofibrillosa and A. submelinoides) were new to science, all

discovered by Kithner under Alnus alnobetula (Ehrh.) K. Koch (Moreau 2005).

This paper is dedicated to the memory of Denise Dailly-Lamoure (26 January 1928 - 26 September

2012), French mycologist, disciple of and successor to Robert Kithner at the University of Lyon.

2 ... Moreau & al.

Although a number of agaricologists studied Alnicola species (e.g. Orton 1960,

Singer 1978, Reid 1984, Runge 1990, Bon 1992, Ludwig 2000, 2001), each of

them presented their own nomenclature and made taxonomy of the genus

more and more complex and confusing.

Recent molecular work (Moreau et al. 2006) suggested that species

hypotheses in Alnicola could be resolved by using several gene sequences to

create a robust phylogeny. A four-gene analysis by Rochet et al. (2011) showed

that at least 16 phylogenetic Alnicola species could be identified in Alnus-

associated ectomycorrhizal communities in Europe. Most of them could be

named according to traditional taxonomy, but some were new to science and

given provisional names. Moreover, a complex infrageneric structure of Alnicola

was suggested in the same study, but no details were given on morphological

interpretations of the taxa defined there.

As a first step towards a general taxonomic revision of the genus Alnicola in

Europe, this article treats the “/badia clade”; a small clade identified by Rochet

et al. (2011: fig. 2), which currently encompasses three well-defined species in

Europe. Morphological descriptions based on numerous observations of fresh

collections and microscopical studies are presented for these three species. One

of them, only collected under Alnus alnobetula so far, is identified as Alnicola

badia, the two others, found with A. incana and A. glutinosa, could not be

assigned to any previously described species and are given new names.

Material & methods

Taxonomy and nomenclature

Generic and infrageneric nomenclature follows Moreau (2005). All new names

proposed are deposited in MycoBank; holotypes are deposited in herb. LIP (Lille,

France), isotypes in herb. IB (Innsbruck, Austria) and ZT (Ziirich, Switzerland).

Basidiomata sampling

Basidiomata were collected in Austria (Tirol), France (Alps), Italy (Trentino) and

Switzerland (Graubiinden). Photographs and morphological descriptions were made

from specimens soon after sampling; all specimens were cut and air-dried. Additional

collections (exsiccata, notes, and pictures) were provided by collaborators from France

and various parts of Europe. Also included are collections of R. Kithner (G); D. Dailly-

Lamoure, personal herbarium; M. Moser (IB); U. Peintner (IB); L. Tedersoo (TU); and

those in the general herbarium of the Kew Botanical Garden (K). Because intraspecific

variability is often important in these species, the following descriptions are based on

the holotype; species variability is discussed after each description on the basis of all

collections studied.

Descriptions

Morphological descriptions are adapted from notes taken on fresh specimens;

colour references (when quoted on fresh material) correspond to Munsell (1975; = M)

TABLE 1: Basidiospores in representative Alnicola collections.

Alnicola spp. nov. in the A. badia complex ...

COLLECTION # SPORES L (um) W (um) 0

A. badia

oe (ays B 88189

abi n oggeate ORL OUI

pi ci cy rr a Ce ECS

A. longicystis

nee Se rt CC

en so OMG1as- — EP 30-228)

eee po OSS OES 8a(-2.08)

anar or BR BB OT) haL-2 10)

A. xanthophylla

aepennal wo Oayessras- Poy “Ng 308)

re ORO Re OO “aaie237)

on RO pt a 7) OT

pits ORAS Beery aca)

Basidiospores measured from spore deposits on stipe in mature, old*, and young** sporocarps;

dimensions presented as (minimum value) 1st decile-average value-9th decile(-maximum value).

L= Length, W = Width, Q = Quotient (L/W).

or Cailleux & Taylor (1963; = C). Microscopical descriptions are based on observations

made at the Laboratory of Botany (Lille), with a light microscope Nachet Andromede

018 at x100, x400 and x1000 magnifications on hand-sectioned mounts in Melzer’s

reagent, 5% KOH, and Congo red (1 mg in 10 ml NH,OH) after reviving in 10% KOH

for a few minutes. For all collections at least 30 basidiospores have been measured from

spore prints or from natural deposits on stipe or pileus surfaces. Statistical treatments

and notations follow Fannechére (2005) and measurements were made using the

software Mycomeéetre 2.02 (Fannechére 2011). Estimations of spore dimensions for each

collection are given in detail separately in TaBLE 1. In the corresponding description

they are re-calculated for all measurements made on collections attributed to the species.

In the following descriptions basidiospore dimensions are given as follows: (minimum

value—)1st decile-average value-9th decile(-maximum value), and calculated on

all basidiospores measured for each species. Pileipellis structures are described and

illustrated from radial sections, additional observations on suprapellis structure are

made on tangential cuts (scalps). Descriptive terminology for pileus structures s adapted

4 ... Moreau & al.

from Heilmann-Clausen et al. (1997) - proposed for Lactarius but perfectly adapted for

the description of structures of Alnicola and most agaricales. Stipitipellis structures are

described from the upper part of stipe, 1-3 mm below the lamellae.

TABLE 2: Main distinguishing features of Alnicola badia, A. longicystis,

and A. xanthophylla.

CHARACTER

PILEUS

Young color

Striation : To centre even when young Faintly with age

Surface Minutely scurfy, Smooth, Furfuraceous,

i then glabrous i then fibrillose-silky i then glabrous, + cracking

aR A rere eer nee ees 36-30 (1-3); pale gray to Prrerrery 33-34 (3), PERT ETere reer ere rereere reer eres 30-36 (3-3), POPEeeeTee rer ererrerereerrerrreys

i pale yellow-ochre i yellowish ochre i butter yellow

mn gieraerernerrenneire a ae ee ae a a iaiagogar i ae ge ae

Color Pale grey + darker at apex, Purplish black at apex, Dull ochre-yellow, early

i darkening with age i darkening with age i darkening from base

Apex + sheathed by Coarsely floccose Densely pruinose-floccose

i caulohymenium,

i not pruinose

oy SMELL eee reer errr ree rer reees . Weak "Yesinous, ‘cold Serre rer rere rer fungoid to ‘taphanoid Pererrererer ey Strongly raphanoid rere rere reer errr

i tobacco, cedar oil

SEE DT Mild; ¥ astringent eee eeeceeerereeereesen

“CHEILOCYSTIDIA "38-50 x 45-8 x 2-25 um, “45-55 x 6.5-8 x 15-2 um, - “28-36 x 8-9.5 x 2 pum, neck ~

i neck 9-26 um long, i neck 18-38 um long, : 10-15 um long,

i with yellow walls i partly with yellow walls: with bright yellow walls

“PLEUROCYSTIDIA Y Scatiéted towards edge" Abundant towards edge" Scattered towards edge"

a CAULOCYSTIDIA a vaneeee Scattered at apex, ba onevnerccnenne ee nadant ab ‘apex dave nevernonenanne : in ‘dence Ghasters ‘at apex, pe eeenene

i intermixed with clavate i intermixed with clavate-

i elements : catenulate elements

pre cri aeetrit POSE Tena FSO MERE aan aS ‘i ML rc

Morphology Oedotrichoderm Hyphoepithelium Hyphoepithelium

Subpellis : 40-50 pm thick : 35-50 um thick : 50-70 um thick

Hyphal Smooth to punctuate Smooth Incrusted in subpellis,

pigments : smooth in suprapellis

“BASIDIOSPORES penenenes 75-105 X 46-65 jim pe enenecneee SB TT3 X56 8 iim pe neerenenes OS-135 <S6274 im se neeeeceneene

ALNUS HOST;

SUBSTRATE

A. badia

Bicoloured:

i pale straw ochre margin,

: date-brown disc

Late in age

i Ovo-amygdaliform not

i papillate

: A. alnobetula,

i mainly on mineral soil

A. longicystis

Bicoloured:

dirty yellowish margin,

dark brown/+ olivaceous

disc

i Amygdaliform partly

i subpapillate

: A. glutinosa, A. incana,

! on humus-rich or acidic

: soil

A. xanthophylla

Uniformly

i dull yellowish ochre,

i aging to + brownish at disc

: Amygdaliform + elongated,

i not papillate

: mainly on bare,

i + mineral soil

Alnicola spp. nov. in the A. badia complex ... 5

Taxonomy

Alnicola badia Kihner, Botaniste 17: 176. 1926. FIGs 1-2

= Naucoria phaea Maire & Kiihner, in Maire, Bull. Inst. Bot. Barcelona 3(4): 101. 1937.

= Alnicola phaea (Maire & Kihner) Romagn., Bull. Soc.

Mycol. France 58: 126. 1942, nom. superfl.

?= Naucoria cedriolens Bresinsky & Schmid-Heckel, in Schmid-Heckel,

Forschungsber. Nationalpark Berchtesgaden 8: 163. 1985.

= Alnicola cedriolens (Bresinsky & Schmid-Heckel) Bon, Doc. Mycol. 21(83): 37. 1991.

“Alnicola phaeodisca Kithner in sched. [R. Kithner, unpublished notes, (G)].

ILLUSTRATED REFERENCE — Moreau et al. (2011: 33).

DESCRIPTION OF HOLOTYPE (adapted from handwritten notes of R. Kithner

on the holotype n° 9.100, G) — “Pileus 1.5-2.5 cm, convex-flat + umbonate,

opaque or hardly striolate on one specimen, reddish bistre (margin paler), under

whitish pruinose flakes (furfuraceous aspect), then red-brown and pruinose,

hygrophanous. Taste mild, odour none. Lamellae 22-23 at stipe, 3-7 lamellules

per lamella, almost spaced, cinnamon brownish. Stipe 3.5-4.5 x 0.17-0.25 cm,

equal, flexuose, base + coated with a white cotton-like mycelium; fulvous-brown,

glabrescent, slightly pale-fibrillose, with slightly fluffy or minutely pruinose

apex, narrowly fistulose; context brown. Spore amygdaliform, 9.5-10.0 x 5.5

(10.5 x 6.0) um, faintly rugose-punctate, cheilocystidia cuspidate. Cellular

cuticle distinctly deviating.’ Trama regular, made of short hyphae. Basidia

4-spored, 28-29 x 8-9 um; paraphyses shorter, cylindro-clavate. Subhymenium

narrow, cellular-branched.”

DESCRIPTION OF RECENT COLLECTIONS — Pileus 0.8-2.5(-3) cm, early

flattened without distinct umbo or lowly convex-flattened, completely scurfy-

squamulose at first with pale ochre to whitish squamules, especially towards

margin, becoming glabrous towards centre by erosion, never cracked; distinctly

bicoloured at first: margin pale straw-ochre (2.5YR2/4, M), centre date brown

to dark red-brown (2.5YR5/6), not or hardly striate (only when old and wet),

hygrophanous, quickly fading from disc to yellowish ochre slightly darker at

centre. Lamellae crowded, 20-30 reaching stipe, 1-2 series of lamellulae, narrow,

adnate-subdecurrent to + emarginate, pale grey-ochre to slightly yellowish

ochre, when mature cinnamon ochre (7.5YR6/4, M); edge white pruinose.

Stipe 2.5-6(-9) x 0.1-0.2 cm, not pruinose at apex, somewhat sheathed by

caulohymenium under lamellae, fibrillose-twisted below, pale greyish, on some

collections slightly darker gray at apex, then greying, blackish brown at the

"Kithner frequently used this term for describing pileus structures in naucorioid groups (Kihner

1931, 1942, 1980). A “deviating” structure refers to a basically hymenodermioid structure,

more or less “degraded”, i.e. with inflate terminal cells of more or less radial orientation and

adpressed (transitions to hyphotrichoderm).

6 ... Moreau & al.

base, with white + strigose mycelium. Smell weak, + resin-like, reminding of

cedar oil, cooking oil or cold tobacco; taste mild, insipid to weakly fungoid-

raphanoid.

Pileipellis an oedotrichoderm (Heilmann-Clausen et al. 2007) 60-90 um

thick; suprapellis made of 3-5 layers of hyphae, each a long chain of up to 15

catenulate cells, more or less fasciculate and erected, prostrate with age, cells

cylindrical to inflate, subterminal cells often constricted, 9-20 um wide, usually

of thin- (<0.2 um thick) yellowish walls, but some (about 10%) with thicker

yellow to brownish walls or thickened only at septa, smooth to locally encrusted;

terminal cells frequent, cylindrical to fusiform, 45-70 x 7-15 um, smooth

and thick-walled; rarely with superficial very slender hyphae 2-2.5 um wide,

observed on surface (scalp, sections) with yellow thickened wall and coarsely

incrusting yellow pigment; subpellis hardly differentiated from suprapellis,

40-50 um thick, made of 4-6 layers of short cylindrical to polygonal cells 7-15

um wide, with yellow walls up to 0.5 um thick, minutely punctuate to locally

distinctly incrusted, not stained by KOH. Pileus trama pale, with somewhat

brownish zones, composed of variously shaped hyphae, slender cylindrical

hyphae 3-5 um wide, and cylindrical to ampullaceous hyphae 8-25 um wide,

mostly smooth, some incrusted or with light yellow walls especially at septa.

Stipitipellis with sparse clusters of caulocystidia at apex, 45-90 x 6-14 um,

some repent, cylindro-clavate more or less curved, with or without lateral neck,

mixed with more typical ampullo-fusiform cystidia, all with yellow thickened

walls. Superficial hyphae 4-6 um wide, wall yellow and coarsely incrusted. Stipe

context composed of cylindrical hyphae 5-14 um wide, with smooth strongly

thickened walls up to 1.5 um thick, yellowish towards cortex, paler in centre.

Basidiospores (5 collections, 196 measurements) (6.7—)7.5-9.02-10.5

(-12.2) x (4.2-)4.6-5.48-6.5(-7.2) um, Q = 1.51-1.65-1.79, ovoid to slightly

amygdaliform with obtuse apex, never subpapillate, ochre-yellow in KOH, not

dextrinoid, uniguttulate in KOH, ornamentation of low warts less than 0.2

um long, punctiform-rounded, not confluent, weakly contrasted; episporium

not distinct. Basidia 22-26(-32) x 6.5-8 um, cylindro-clavate, 4-spored, base

variously elongated, before maturity with granular content, sometimes weakly

brownish; necrobasidia abundant, deep ochre yellow in KOH. Subhymenium

12-15 um thick, ramose with short cylindrical articles 3-4.5 um wide, hyaline.

Hymenial trama regular, mediostratum almost colourless to pale brownish

zoned, made of short cylindrical hyphae 15-45 x 4-14 um, with yellowish

wall locally thickened (up to 1 um thick) and incrusted. Lamella edge sterile;

cheilocystidia 38-50 x 4.5-8 um (base), x 2-2.5 um (apex), ampullaceous at

base, with distinct attenuate neck 9-26 um long, sometimes obtuse to + inflate

at apex (e.g. on holotype, Fic. 1), all with thickened yellowish wall up to 0.5 um

thick, most colourless, scattered ones with deep yellow content. Pleurocystidia

Alnicola spp. nov. in the A. badia complex ... 7

Figure 1: Alnicola badia (09082204, LIP). a: basidiospores; b: portion of hymenium with basidia

and subhymenium;; c: cheilocystidia; d: pleurocystidia; e: pileipellis, radial section; b: stipitipellis at

stipe apex (longitudinal section.). Scale bar = 10 um.

scattered but not unfrequent towards edge, much rarer 300 um in from the edge,

36-56 x 6-6.5 um (base), x 2 um (neck), fusiform, neck 22-40 um long and

attenuate, occasionally abruptly inflated at apex, walls thickened and yellowish

up to 0.3 um thick, originating from deep in subhymenium. Gloeoplerous

hyphae absent to very rare, pale yellow, slender, cylindrical, in stipe and pileus

context. Clamp connections present at all septa.

8 ... Moreau & al.

FiGuRE 2: Alnicola badia (Holotype, R. Kiihner 9.100, G). a: basidiospores; b: cheilocystidia;

c: pileipellis, radial section. Scale bar = 10 um.

SPECIMENS EXAMINED — AUSTRIA, Trrot, Obergurgl, Alnus alnobetula, 25 August

2005, P.-A. Moreau n° 05082717 (LIP); 28 August 2005, P.-A. Moreau 05082808b (LIP).

FRANCE, Corse bu Sup, Bocca Palmente, Alnus alnobetula subsp. suaveolens along a

stream, leg. F Richard & P.-A. Moreau, 12 October 2004, P.-A. Moreau 04101209 and

04101206 (LIP); Bastelica, val d’Ese, Alnus alnobetula subsp. suaveolens in sand along

a stream, 7 September 2007, P.-A. Moreau 07090822 (LIP); 8 September 2007, P.-A.

Moreau 07090905 (LIP); 3 September 2008, P.-A. Moreau 08090340 (LIP; illustrated in

Moreau et al. 2011: 33); HAUTE-CoRSE, Vizzavona, Monte d’Oro, under Alnus alnobetula

subsp. suaveolens along a stream, leg. F Richard & P.-A. Moreau, 11 October 2004, P.-

A. Moreau 04101101 (LIP); Haute-Savoie, Samoéns, Collet d’Anterne, 6 September

1958, R. Kithner Sa-58-58 (G, “Naucoria subescharoides” in herb., “Alnicola phaeodisca”

in sched.); Savor, La Perriére, Praz Joseph, 31 August [see note 1], under alders on

ground, R. Kihner 9.100 (G, holotype); la Rosiére, left side of the road to La Jairaz below

the dam, under Alnus alnobetula, 8 September 1942, R. Kithner (G, “Naucoria (Alnicola)

Joniss A”); under La Rosiére, 10 September 1942, under Alnus alnobetula, R. Kithner (G,

“Alnicola [Nauc. pannosa °] Joniss A’); above La Rosiére, Alnus alnobetula, 18 September

1961, R. Kiihner (G, “sp.1961”); Bourg-Saint-Maurice, Arc 1800, above the golf area, in

old Alnus alnobetula thickets along a stream on acidic soil, 1870 m, 7 September 1993,

Alnicola spp. nov. in the A. badia complex ... 9

P.-A. Moreau n° 93090702 (LIP); ruisseau du Villard, 19 August 2002, J.-C. Deiana, P.-A.

Moreau JCD19-8-02; 21 August 2003, P.-A. Moreau 03082102 (LIP); Peisey-Nancroix,

Nancroix, les Lanches, under Alnus alnobetula, calcareous watercourse, P. Saviuc, 21

October 2003, P.-A. Moreau 03082110 (LIP); Landry, Barmont, under Alnus alnobetula,

leg. M. Broussal, 23 August 2003, P.-A. Moreau 03082301 (LIP); 26 October 2007,

P.-A. Moreau 07082637 (LIP); Bessans, Ecot de Bonneval, under Alnus alnobetula, 1

October 2008, leg. M. Durand, M. Durand 08100119 and 08100121 (LIP); Crest-Voland,

APPB Tourbiére des Saisies, along the Nant Pareu stream, acidic peaty soil under Alnus

alnobetula subsp. alnobetula amongst liverworts (Pellia spp.), 22 October 2009, P.-A.

Moreau 09082204 (LIP); Beaufort, Cormet d’Aréches above the Lac des Fées, under Alnus

alnobetula on calciferous schists, 22 October 2008, P.-A. Moreau 08082201 (LIP); leg. C.

Hugouvieux, 27 October 2009, P.-A. Moreau 09082703 (LIP); locality unknown (Savoie

or Haute-Savoie), 1964, R. Kithner n° 64-20 (G). SWITZERLAND, GRAUBUNDEN,

Davos, leg. H. Gamper, under Alnus alnobetula isolated bush in a pasture, 1 August

2003, P.-A. Moreau 03080101 (LIP); Bergiin, above the Palpuogna lake, under Alnus

alnobetula on acidic ground along a stream, 14 August 2003, P.-A. Moreau 03081403

(LIP); 2 September 2006, P.-A. Moreau 06090202 (LIP); Lenzerheide, Parpan, under a

thicket of Alnus alnobetula, 31 August 2006, P.-A. Moreau 06083102 (LIP).

Note 1 — The year of collection of the holotype is neither mentioned in the

notes nor in any of Kihner’s publications, but the notes (kind of paper, handwritten

indications and style) are identical to those on A. submelinoides and A. luteolofibrillosa.

The collection of A. submelinoides was dated 1924 by Kithner (1931) himself, therefore

31 August 1924 should be accepted as the date of collection of the original material of

A. badia.

GENBANK ACCESSION NUMBERS (Moreau et al. 2006, Rochet et al. 2011) — ITS1-5.8S-

ITS2: AY900031, AY900032, AY900033, AY900036, AY900037, AY900039, HQ714656,

HQ714657, HQ714658, HQ714664, HQ714670, HQ714676, HQ714755, HQ714756,

HQ714781, HQ714787. GPD: HQ714896, HQ714597, HQ714598, HQ714599,

HQ714603, HQ714608, HQ714614. RPB2: HQ714796, HQ714797, HQ714799,

HQ714800, HQ714811, HQ714818. V9: HQ714500, HQ714508.

ECOLOGY AND DISTRIBUTION — Only known with Alnus alnobetula, usually

on acidic substrate, on wet humus amongst liverworts, more rarely on schists;

known from subalpine altitudes of Western and Central Alps (Austria, France,

Switzerland) and Corsica. Widespread but usually not abundant, often

scattered.

OBSERVATIONS — Kihner (1926, 1931) found his very first collections of

A. badia and A. submelinoides (a species with club-shaped cystidia) together

in 1924 at the same locality of Praz Joseph, La Perrie¢re (near Bozel, Savoie). In

fact both species often grow together and are easy to mistake in the field. Dry

A. badia can be detected by darker colours, and especially blackish stipe on

mature specimens, while dry basidiomata of A. submelinoides remain pale.

It cannot be proved that Heim’s (1931) representation of the pileipellis

of A. badia was based on Kihner’s holotype, but we are inclined to think so

because: 1) Ktihner contributed to Heim’s work; 2) Heim does not seem to

have collected any agaric himself under Alnus alnobetula in the Alps; 3) pileus

10 ... Moreau & al.

structure excludes all species known to us from lowland species of Alnus; and

4) at this date Kihner only knew A. badia from his type collection. However

Heim’s figure is a representative - although somewhat stylized - illustration

of the oedotrichodermial structure observed on our typical collections of

A. badia; it is reproduced by Clémengon (1997: 58, fig. 2.61) who interprets it

as “moniliform physalohyphae’.

VARIABILITY OF THE SPECIES —The pileus colour of A. badia is variable,

making identification uncertain in the field. Also, basidiomata are often

scattered, intermixed with other species, and young and old specimens can

be easily confused with other species without microscopic observations.

However, a pale pileus margin with light ochre tones, distinctly bicoloured on

darker specimens, and pale browning stipe are apparently constant features

distinguishing it from other Alnicola species found with Alnus alnobetula. The

presence of pleurocystidia is usually confined to the vicinity of lamella edge,

and sometimes not conspicuous (very scarce on PAM07090905). A spectacular

variation is found in spore size within the same otherwise homogenous collection

(see TABLE 1), ranging from rather narrow (3.8-4.8 um wide in average) to

widely ovo-amygdaliform (6.0-6.6 um wide in average), partly attributed to

age of specimens; but they are rather well characterized by low, thin punctuate

ornamentation and obtuse, neither attenuate nor mucronate apex.

EXTRALIMITAL COLLECTIONS — 1) A few collections from mineral

substrates in Corsica (riverbed) and Savoy (calciferous schists) show a more

distinct cedar-like smell; such collections are described in the diagnosis of

A. cedriolens (Schmid-Heckel 1985). There is no morphological or molecular

evidence that this character supports a distinct taxon. The same cedroid smell

has also been reported on collections of A. luteolofibrillosa (Moreau 2005;

E. Campo, pers. com.); 2) two rather poor collections also from Savoy and

Corsica are morphologically conform to A. badia but have a slight aromatic

smell reminiscent of Inocybe bongardii (Weinm.) Quél. (or Alnicola suavis

(Bres.) Kithner); basidiospores are at the larger size limit for A. badia (suggesting

senescent or damaged specimens) and no sequence could be generated for

these collections.

Alnicola longicystis P.-A. Moreau, Bizio & Deparis, sp. nov. FIG. 3

MycoBank MB 563643

Similar to Alnicola badia and A. xanthophylla by cheilocystidia with yellow thickened

walls, amygdaliform punctate basidiospores, pleurocystidia, and mild taste. Diagnosed

by purplish-black stipe apex when young. Differs from Alnicola badia by association

with Alnus glutinosa and A. incana, pileus striate from early stages, and frequent

pleurocystidia.

TyPE: Italy, Trentino, Balsega di Pine, Alnus incana, 25.VIII.2005, leg. A. Aiardi & E.

Bizio, herb. P-A. Moreau n° 05082501 (Holotype, LIP; isotypes, IB 20050690, ZT).

Alnicola spp. nov. in the A. badia complex... 11

FiGurRE 3: Alnicola longicystis (Holotype, 05082501, LIP). a: basidiospores; b: portion of hymenium

with basidia, pleurocystidia and subhymenium; c: cheilocystidia; d: pleurocystidia; e: pileipellis,

radial section; e: stipitipellis at stipe apex, longitudinal section. Scale bar = 10 um.

Erymo oey: “with long cystidia’, referring to the distinctive lanceolate cheilocystidia

with elongated neck.

Pileus 0.8-2.5 cm, convex then soon flattened, striate to centre, at first pale

yellowish ochre with subpapillate dark brown centre, this dark colour expanding

12 ... Moreau & al.

with age, when old gradually brownish-grey at centre with somewhat olivaceous

tones (H63-H64, C) to dirty yellowish at margin (B72), hygrophanous, fading

uniformly to pale ochraceous (A84), when fresh almost glabrous and smooth,

becoming fibrillose-silky from margin when drying. Lamellae crowded,

22-24 reaching the stipe, 2 series of lamellulae, adnexed-horizontal to slightly

emarginated, at first yellowish ochre to pale lemon-ochre (A84), darkening

slightly with age to dull ochre (C74-D74), rusty ochre when old; edge white,

minutely pruinose. Stipe 3-8 x 0.1-0.3 cm, fistulose, slightly enlarged at base,

initially dull ochre brown with purplish-black apex, coarsely floccose at apex,

below whitish-fibrillose, fading to pale yellowish in lower part with age, apex

remaining darker, base darkening on older specimens. Context greyish in pileus,

dark grey-brown at apex, yellowish in stipe cortex. Smell strongly fungoid or

raphanoid. Taste raphanoid or fungoid, mild.

Basidiospores (3 collections, 292 measurements): 8.8-10.48-12.3 x 5.5-5.97-

6.8 um, Q = 1.60-1.79-2.10, amygdaliform, dimorphic with a variable

proportion of large basidiospores with more elongate to subpapillate apex,

rather bright yellow in KOH, not dextrinoid, ornamentation when mature

warty-punctate to echinulate, up to 0.3 um high, warts isolated, irregular in

size but rarely confluent, before maturity often as low patches (especially when

observed on hymenium); content homogenous, not distinctly guttulate, or with

a single irregular drop; episporium not loosening. Basidia all with 4 sterigmata,

strongly heteromorphic: 21-36 x 6.5-8.5 um, cylindrical, slightly strangulate

under apex when mature, with variously elongated base, the longest generating

larger basidiospores; necrobasidia with yellowish brown content frequent,

collapsed, but also some basidioles and young basidia with yellow cytoplasmic

content present, otherwise colourless; most mature basidia with dense colourless

granular content. Subhymenium compact and weakly developed, 10-12 um

wide, composed of small short cells forming a thin pseudoparenchymatic

tissue. Mediostratum of lamellae regular, pale yellow-brown in KOH, composed

of slender and wide hyphae intermixed, 3-14 um wide, with smooth slightly

thickened walls. Lamella edge sterile; cheilocystidia (38-)45-55 x 6.5-8 um

(base), x 1.5-2 um (neck), all lanceolate with an exceptionally long and slender

neck (18-38 um long), slightly thick-walled (up to 0.3 um thick), some with

the neck filled with yellow pigment towards apex; often with small basidioles

intermixed. Pleurocystidia relatively frequent up to 200 um from the lamella

edge, larger and slenderer than cheilocystidia, 55-62 x 5.5 um (base), x 1.5-2.5

tum (neck), with a long neck often difformed: capitate, inflated, ramose or bifid,

often yellow at apex, arising distinctly from hymenium, disappearing early

from edge to sinus.

Pileipellis a thick hyphoepithelium (Heilmann-Clausen et al., 2007);

suprapellis 12-20 um thick, distinctly yellowish coloured, made of 2-4 layers

Alnicola spp. nov. in the A. badia complex ... 13

of cylindrical-filamentous hyphae 2.5-7 um wide, walls yellow, up to 0.5(-1)

uum thick, smooth; terminal cells cylindrical with rounded to slightly inflated

apex. Subpellis 35-50 um thick, very distinct, formed of inflated to subglobose

cells 25-75 x 6-28 um, with pale yellow smooth walls, moderately thick-walled

except towards septa (up to 1 m thick). Pileus trama of short, pale and smooth

walled hyphae, 3-8 um wide. Stipitipellis covered by a velipellis of cylindrical,

incrusted hyphae 5-6.5 um diam, with scattered erected clavate cells; at apex

intermixed with large clusters of caulocystidia 40-70 x 5.5-11 um, lanceolate

with long neck, most with yellow apex, some collapsed with yellow-brown

necropigment. Stipe trama made of parallel hyphae, 3-12 um diam, with

yellowish slightly incrusted wall, often thickened and more brownish at septa;

gloeoplerous hyphae very rare, long, sinuous and slender (3-4 um diam.), with

yellow-brown thick walls in KOH. Clamp connections present at all septa.

ADDITIONAL SPECIMENS EXAMINED — ESTONIA, VILIANDI, Songa, 58°18'N 25°22’E,

Alnus glutinosa, 12 September 2009, L. Tedersoo 110256 and 110257 (TU); LAANE,

Vormsi, Fallarna, 59.001°N 23.202°E, A. glutinosa, 17 September 2010, L. Tedersoo

110285 (TU); Hullo, 59.008°N 23.2275°E, A. glutinosa, 17 September 2010, L. Tedersoo

110300 (TU); Sdderby, 59.0049°N 23.3520°E, 18 September 2010, L. Tedersoo 110304

(TU); POLvA, Jarvselja tirgkvartal, 58°16'N 27°19’E, A. glutinosa, 30 September 2010,

L. Tedersoo 110343 (TU); Tartu, Haage, 58.363°N 26.63°E, 26 September 2010,

L. Tedersoo 110321 (TU); Maksa, 58°22'N 27°03’E, A. incana, 30 September 2010,

L. Tedersoo 110344 (TU); Porijée, 58°23'N 26°43’E, A. incana, 6 October 2010, L.

Tedersoo 110358 (TU). FRANCE, HauTE-Savoteg, les Houches, under A. incana and

A. alnobetula intermixed, 22 September 2004, P.-A. Moreau 04092201 (LIP); Argentiére,

under A. incana on acidic peaty ground, 24 September 2004, leg. L. Deparis & P.-A.

Moreau, P.-A. Moreau 04092401 (LIP); LANDEs, Yrieu, edge of brook with A. glutinosa,

29 avril 2000, leg. J. Guinberteau & P.-A. Moreau, P.-A. Moreau 100 and 101 (LIP); Pas-

DE-CALAIS, Boulogne-sur-Mer, domanial forest, oceanic deciduous forest on clay soil,

under A. glutinosa along a rivulet, 10 September 2009, P.-A. Moreau 09091001 (LIP);

SAVOIE, Séez, along the Iséere river, under A. incana in a mixed riparian forest, leg.

M. Gardes, J. Rochet & P.-A. Moreau, 28 August 2008, P.-A. Moreau 08082805 (LIP);

Bozel, les Perriéres, under A. incana along a rivulet in a Fagus forest, 780 m, 26 August

2010, P.-A. Moreau 10082611 (LIP); Beaufort, Aréches, le Perthuis, wet mesotropic

riparian forest with A. incana, 22 August 2011, P-A. Moreau 11082203 (LIP). SPAIN,

CANTABRIA, Ruente, Parque natural, under A. glutinosa on acidic peaty soil, 29 October

2011, PR-A. Moreau 11102901 (LIP).

GENBANK ACCESSION NUMBERS (Moreau et al. 2006, Rochet et al. 2011, Tedersoo et al.

2009) — ITS1-5.8S-ITS2: AY900038, AY900075, FM993128, FM993140, FM993256,

FM993273, HQ714667, HQ7 14757, HQ714795, HQ714668: holotype; GPD: HQ714605,

HQ714606: holotype, HQ714641; RPB2: HQ714808, HQ714809: holotype, HQ714885;

V9: HQ714576: holotype.

ECOLOGY AND DISTRIBUTION— With Alnus glutinosa and A. incana, most

often on humus-rich substrates, typically in wet mixed Alnus—Corylus thickets

on acidic ground (often with Polytrichum or Sphagnum spp.), but also in

neutrophilic to slightly basophilic river sides with A. xanthophylla, known from

14 ... Moreau & al.

lowland to mountainous altitudes (1000 m); known from siliceous mountains

of Western and Southern Europe (Pyrenees, Cantabrian Mountains, Alps; also

isolated from mycorrhizae in Corsica, unpublished data) and scattered localities

in coastal forests (Aquitaine, North of France), as well as Estonia. Probably

widespread in Europe, gregarious but rarely abundant.

OBSERVATIONS — This is one of the most frequent species of Alnicola associated

with Alnus incana in the mountains. It is found on river sides (with Gyrodon

lividus and Paxillus filamentosus) as well as in acidic peaty alder stands amongst

Sphagnum. It is also present but rare and scattered in coastal peaty forests

under Alnus glutinosa. In alpine regions it can be found together with Alnicola

xanthophylla, which differs macroscopically on the field by distinctly striate

pileus, yellow lamellae, and the absence of purplish black spot at stipe apex.

Because colours evolve strongly with age, old or atypical specimens can be

difficult to interpret in the field, but can be recognized easily by long-necked

cheilocystidia and pleurocystidia (e.g., most cheilocystidia measuring 45-55 x

6.5-8 um in A. longicystis, 28-36 x 8-9.5 in A. xanthophylla). Alnicola badia,

which can show the same characteristic dark tones at stipe apex, differing

especially by the host tree and a pileus becoming striate only with age. Alnicola

umbrina (Maire) Kithner (Kithner 1931: 241) differs by more foxy to brown

colours and the fusiform spores and cystidia (Moreau 2005).

Alnicola xanthophylla P.-A. Moreau, Peintner & Senn-Irlet, sp. nov. FIG 4

MycoBank MB 563644

Differs from Alnicola longicystis by a pileus that is striate only in age, uniform yellowish

colors, and shorter cheilocystidia with bright yellow walls and necks < 15 um long.

Type: SWITZERLAND, GRAUBUNDEN, Rothenbrunnen, leg. P.-A. Moreau & B. Senn-

Irlet, 1.1X.2006, herb. P.-A. Moreau 06090101 (Holotype, LIP; isotypes, IB20060540,

Zit),

EryMo_oey: “with yellow gills’, referring to the characteristic butter-yellow tinges of

lamellae.

Pileus 1-2.5 cm diam, soon flattened to slightly obtuse, slightly striate when

mature, entirely furfuraceous, glabrescent, often cracked-squamulose when old,

dull yellowish ochre (C74-C66, C), subpapillate, brownish at centre, gradually

pale yellowish towards margin, when old dark grey, strongly hygrophanous

fading to uniformly dull greyish ochre to somewhat more rusty at centre (C72

or a bit darker), slightly fibrillose under a lens, without trace of veil. Lamellae

very crowded, 20-26 reaching the stipe, 2(-3) series of lamellulae, adnexed to

almost free, initially butter yellow (D68-C68), becoming dull greyish ochre

when old, at the end tobacco brown; edge pale yellow, smooth to pruinose-

pubescent. Stipe 2-5 x 0.1-0.2 cm, equal or slightly enlarged at base, when

very young dark grey-brown at apex, then uniformly dull ochre-yellow (C74),

Alnicola spp. nov. in the A. badia complex... 15

FiGurE 4: Alnicola xanthophylla (Holotype, 06090101, LIP). a: basidiospores; b: portion of

hymenium with basidia and subhymenium; c: cheilocystidia; d: pleurocystidia; e: pileipellis, radial

section; f: stipitipellis at stipe apex, longitudinal section. Scale bar = 10 um.

soon browning from base, at maturity dark grey-brown with yellowish tones at

apex, densely pruinose-floccose 2-3 mm under the lamellae, slightly covered

16 ... Moreau & al.

by fibrils more conspicuous on old darkened specimens, without trace of veil;

mycelium white hardly visible. Context grey when fresh then whitish, amber-

yellow in cortex, when old dirty grey in stipe base. Smell strongly raphanoid.

Taste mild, raphanoid, more or less astringent.

Basidiospores (4 collections, 149 measurements): 9.2-11.20-13.5 x 5.6-6.46-

7.4 um, Q = 1.55-1.76-2.21, ovo-amygdaliform, rather polymorphic and

partly elongated with acute apex but not distinctly subpapillate (a majority of

abnormally elongate basidiospores on coll. PAM06090104), bright yellow in

KOH, not dextrinoid, ornamentation when mature warty-punctate, up to 0.3

um high, warts low and obtuse, irregular in size, punctiform to slightly cristate;

content with a large irregular droplet and often granular cytoplasm; episporium

not or only slightly loosening. Basidia all with 4 sterigmata, cylindro-clavate

with more or less elongate base, 22-32 x 7.5-9.5 um; necrobasidia abundant,

with amber-yellow content, usually collapsed, young and mature basidia

usually colourless. Subhymenium thin, filamentous, 5-8 um, made of slender

hyphae 2-4 um wide, with small, short cells forming a weakly developed

pseudoparenchymatic structure. Lamella mediostratum regular, 80-100 um

wide, pale to almost colourless in KOH, made of shortly cylindrical to polygonal

or strongly inflated cells 7-35 um wide, pale yellow, with smooth, slightly

thickened walls. Lamella edge sterile; cheilocystidia 28-36(-40) x 8-9.5 um

(base), x 2 um (neck), shortly fusi-lageniform with strongly inflated base and

rather short attenuate neck (10-15 um long), apex obtuse to slightly rounded,

walls bright yellow, especially thickened, up to 0.5 um at apex, without basidia

found intermixed. Pleurocystidia scattered, only frequent very close to the

edge but sometimes up to 200-300 um from the edge, 32-40 x 6-8 um (base),

x 2 um (neck), fusiform with longer neck than cheilocystidia (15 um long),

sometimes tortuous, bright yellow, arising from subhymenium, disappearing

towards sinus.

Pileipellis a hyphoepithelium; suprapellis 20-30 um thick made of 1-2(-3)

layers of cylindrical to catenulate hyphae of cells 18-45 x 4.5-11(-16) um, +

erected towards centre, terminal cells cylindrical-attenuate; walls yellowish

to brownish on some cells, smooth; subpellis 50-70 um thick, very distinct,

reddish ochre in KOH, cellular, with slightly radial-ascendant orientation,

made of cylindrical to polygonal or subglobose isodiametric cells, 15-90 x

8-30 um, with yellow incrusted walls, especially thickened and brownish at

septa. Pileus trama reddish brown in KOH, under subpellis gradually paler,

composed of cylindrical hyphae 4-16 um wide, more inflated towards surface,

walls minutely incrusted. Stipitipellis at apex with sparse short catenulate

hyphae 5-8 um wide, dark yellowish, with thickened strongly incrusted walls,

intermixed towards apex with dense clusters of caulocystidia 22-28 x 10-18

um (base) x 2-2.5 um (neck), strongly inflated with attenuate neck 12-25 um

Alnicola spp. nov. in the A. badia complex ... 17

long, only slightly thick-walled (some with thickened yellow neck). Stipe trama

made of parallel hyphae, 3-12 um wide, with yellowish incrusted walls in

cortex, colourless in depth; gloeoplerous hyphae sparse, more frequent towards

base, short and rather wide (5-8 um), dark reddish brown in KOH. Clamp

connections present at all septa.

ADDITIONAL SPECIMENS EXAMINED — CZECH REPUBLIC, BOHEMIA, Prague-Radotin,

under Alnus glutinosa, J. Borovicka n° JB2010-2515; Prague, Reporyje, herb. J. Borovicka

BORE2008-9 (part in LIP). ESTONIA, Tartu, Janese matkarada/Kvissentali, Alnus

incana, 24 September 2010, L. Tedersoo 110319 (TU); Liilli, 58°27.5'N 26°49'E, Alnus

incana, 2 October 2010, L. Tedersoo 110349 (TU). FRANCE, AIN, Vanchy, calcareous

sand pit, under a small Alnus glutinosa in a watercourse, R. Fillion & P.-A. Moreau,

1 May 1999, P.-A. Moreau 99050110 (LIP, as “A. pseudoscolecina”); NORD, Trélon, A.

glutinosa, 25 April 1999, J.-J. Wuilbaut 99042503 (LIP); Zuydcoote, fixed dune, under

a single Alnus glutinosa along a sandy path, 9 November 2008, herb. P.-A. Moreau

08110901 (LIP); Pas-pE-CatLats, Evin-Malmaison, under Alnus incana planted in a

periurban park, on wet clay-calcareous soil along a ditch, 30 October 2007, leg. A. Hayet

& B.-A. Moreau, P.-A. Moreau 07103002 (LIP); Savoig, la Perriére, under Alnus incana

along a rivulet in a Fagus forest, 780 m, 26 August 2010, P.-A. Moreau 10082610 (LIP);

Beaufort, la Turnaz, with young Alnus incana in a mineral river bed, 1015 m, 28 August

2010, P.-A. Moreau 10082802 (LIP). SLOVENIA, Alnus incana, L. Tedersoo L7054 (TU).

SWEDEN, JAMTLAND, Ostersund, Rannason, Alnus incana on calcareous gravel, 26

September 2007, leg. H. Lindstrém, H. Lindstrém 06.373 (part in LIP); SKANE, Ivo, Iv6

klack, 56.139766°N 14.403568°E, under Alnus incana, Salix and Betula on calcareous

clay, poor in organic matter, 22 October 2010, K. Gillen, K. Hansen, L. Orstadius and

I. Olariaga, F-176866 (S, as “Alnicola cf. escharoides”. SWITZERLAND, GRAUBUNDEN,

Filisur, leg. U. Peintner, 18 September 1998, IB1998/0446 (IB, as “Alnicola escharoides”);

Rothenbrunnen, under Alnus incana on black alluviums, alt. 600 m, leg. P.-A. Moreau

& B. Senn-Irlet, 1 September 2006, P.-A. Moreau 06090104; Alvaneu-Bad, under Alnus

incana, 600 m, 1 September 2006, leg. V. Queloz, P.-A. Moreau 04090104 (LIP); BERN,

Kandersteg, Filfalle, 1200 m, Alnus incana on calcareous soil, 30 September 2003, leg. H.

Woltsche, P.-A. Moreau (LIP); idem, A. incana along a stream, on black alluvions, 22 July

2004, leg. B. Senn-Irlet & P.-A. Moreau, P.-A. Moreau 04072201 (LIP).

GENBANK ACCESSION NUMBERS (Moreau et al. 2006, Rochet et al. 2011) — ITS1-5.8S-

ITS2: AF325630, HQ714705, HQ714709, HQ714765; GPD: HQ714627, HQ714628,

HQ714648; RPB2: HQ714839, HQ714844, HQ714889; V9: HQ714530, HQ714534,

HQ714581.

ECOLOGY AND DISTRIBUTION— With Alnus incana, more rarely A. glutinosa,

usually on mineral-rich substrates, typically on black alluviums on river banks

in alpine valleys (Alps), but also on coastal calcariferous sands (Northern

France) or chalk in disturbed continental sites; known from lowland to montane

altitudes (1200 m), from France to Estonia. Probably strictly basophilic,

widespread in Europe and locally abundant.

OBSERVATIONS — ‘This species is easily recognizable in the field by its general

yellow colour when young, persistent in lamellae with age, non-striate pileus

(when young), and a preference for basic mineral-rich sedimentary substrates.

18 ... Moreau & al.

It is typically found under Alnus incana on river banks in alpine regions, where

it seems frequent and locally dominant, but also in plantations on calcareous

soil, and a single collection from calcareous sand dunes under Alnus glutinosa

in Northern France, what suggests an oro-septentrional distribution. It has

probably been misidentified as Alnicola escharoides by various mycologists,

despite the mild taste, and shorter basidiospores and cystidia that are usually

described for this variously interpreted species (Moreau 2005; Fernandez Sasia

& Moreau 2011).

The closest species is Alnicola longicystis, which has the same biogeographical

distribution but seems to prefer humus-rich or more acidic soils. Macroscopically

it can be possible to misidentify old or single specimens of the two species;

microscopically cystidia shape and size make a clear distinction between them.

Analysis of ITS sequences clearly separate these species (Rochet et al. 2011).

Discussion

The identity of Alnicola badia

Alnicola badia is one of the most common agarics associated with Alnus

alnobetula and one of the first (with A. luteolofibrillosa and A. submelinoides)

described under this plant, but paradoxically one of the less documented in

the difficult genus Alnicola. Kiihner (1926) described A. badia from a single

collection, a provision he wrote later (1931: 239): “... récolte unique sur laquelle

nous avions cru pouvoir baser une espéce indépendante de N. escharoides’,

with a detailed description of this unique material. “Praz Joseph,” now called

“Praz Juget,’ is a locality of the village of La Perriere (Gros 1973), close to Bozel

(Savoie), at low subalpine level (1890 m), with a northern exposure; the only

alder species found there is Alnus alnobetula. In this locality Kihner also found

two species exclusively associated with Alnus alnobetula (Basso 1999): Lactarius

alpinus Peck (illustrated by Ktihner 1929a as “L. subalpinus R. Kithner” [nom.

illegit.], also known as Lactarius kuehneri Joss) and L. lepidotus Hesler & A.H.

Sm. (illustrated by Kiihner 1929b as “Lactarius griseus Peck”).

Unfortunately, this single collection did not give the young Kuhner a clear

concept of his Alnicola badia, and at that time he was not experienced in

lowland alnicolous fungi. By contrast, his friend Marcel Josserand, an active

mycologist familiar with wet lowland forests around Lyon, was experienced in

a common species identified by René Maire as Tubaria umbrina (Maire 1930),

found abundantly under Alnus glutinosa. Josserand was convinced that Alnicola

badia and Tubaria umbrina were synonyms, and did his best to convince Maire

and Kihner of his opinion (unpublished correspondence, coll. R. Kihner (G),

and coll. R. Maire (MPU)). Kithner (1931: 240) hesitated to synonymize these

taxa, but finally accepted Josserand’s point of view (Kiihner 1942: 1). He also

rejected his own genus name Alnicola under the influence of Maire (Maire

Alnicola spp. nov. in the A. badia complex... 19

1937), and together they formalized the synonymy between A. badia and

A. umbrina under a new name, Naucoria phaea.

Despite his affirmations, Kithner’s numerous unpublished notes and

collections prove that he had never been really convinced with this synonymy,

but failed to define the limits of his A. badia. The unpublished names

“A. phaeodisca,” “A. subescharoides,” “A. alnobetulae,” “A. phaea from 1961, etc.

in his abundant notes on collections made after 1954 under Alnus alnobetula,

show his repeated attempts to compare his collections and to separate them

from the common lowland species A. umbrina, to which he had become

familiar after 1942. But he never completed this challenge (Kiihner 1981) and

never published any definitive decision about A. badia after 1931.

The epithet name badia is in fact quite deceptive for a species typically

bicoloured when fresh, only turning red brown with age or on drying. We

formerly (Moreau 2006a; Moreau et al. 2006) interpreted it erroneously as a dark-

coloured species (provisionally named “A. badiofusca” in Rochet et al. 2011),

before understanding the importance of pileus structure and pigmentation for

the distinction of these species. Before clearly separating these two species,

we had already intuitively separated them in the field by their general colours

and pileus striations under the respective field names “A. badia pale [light]” or

“A. badia bicolore” (A. badia) and “A. badia sombre [dark]” or “A. badia rouge

[red]” (= “A. badiofusca”). The type collection of A. badia (herb. R. Kiihner, G)

is small, only made of two fragmented pilei, but fortunately we could precisely

observe pileipellis (typically catenulate with mainly smooth pigmentation) and

basidiospores, and establish identity with our own collections.

Morphology and ecology in the Alnicola badia complex

Alnicola badia, although strictly associated with Alnus alnobetula, is

morphologically not a very distinctive species. With the two other species

described here, both associated with Alnus glutinosa and A. incana, it forms a

small group of mainly orophilic species, sharing the following characteristics:

1) lack of reddish, ferruginous or fulvous tinges, 2) absence of any trace of

partial veil, 3) mild taste, 4) abundant necrobasidia, 5) cheilocystidia with

thickened wall and neck partly filled with yellow content, 6) pleurocystidia

around lamella edge, and 7) medium-sized amygdaliform basidiospores

with low punctuate ornamentation. Species in the Alnicola umbrina group

(‘A. badiofusca” ad int., A. sphagneti, A. subconspersa, A. umbrina; Rochet et

al. 2011) differ by the presence of red-brown colours in lamellae and/or pileus

even when young, and spores frequently fusiform. Alnicola citrinella P.-A.

Moreau & A. de Haan (= A. escharoides sensu auct.; de Haan & Moreau 2012),

the most common Alnicola species under Alnus glutinosa and A. incana, differs

by absence of pleurocystidia, bitter taste, and non-striate pileus.

20 ... Moreau & al.

Alnicola longicystis is macroscopically closest to A. badia. Its distinctive

features include the distinctly bicoloured pileus (especially on young

specimens) and distinct yellow tones in young lamellae, contrasting strongly

with the typical grey-brown apex of stipe. Its distribution is apparently wide

but irregular. It especially frequent under Alnus incana in the Alps but also

found occasionally under Alnus glutinosa on peaty ground in lowlands, and

also occurs at montane levels in the Pyrenees where Alnus glutinosa fills the

natural niche of A. incana as known from the Alps. When Alnus alnobetula

and A. incana grow intermixed, as in some localities in the Alps, the grayish

to pale cream-yellow colour of lamellae and the very weakly striate pileus of

A. badia are good field characters as compared to the clearly yellowish lamellae

and distinctly striate pileus of young specimens of A. longicystis. Old or too wet

specimens are almost impossible to recognize without a rigorous microscopic

analysis.

Alnicola xanthophylla is also a very distinct species, easily separated from

A. longicystis (with which it can grow closely intermixed in wet peaty

Alnus—Picea forests on rich ground, as observed in several sites in Savoy)

by the uniformly yellow colours of fresh basidiomata, slightly striate pileus,

and absence of gray colour on the stipe apex even on youngest stages. Both

species should be confused with Alnicola striatula (P.D. Orton) Romagn., a

species with pale colours (not bright yellow; Orton 1960: 322), striate pileus,

“amygdaliform-limoniform” and verrucose basidiospores and_ lanceolate

cystidia, which belongs to the Alnicola umbrina complex (see Moreau, 2004,

2006). An updated identification key including the new taxa described here will

be the object of a further publication.

Acknowledgments

We are grateful to all curators and technicians of herbaria who kindly gave us access

to the European collections and documents cited above: G (P. Clerc), IB (R. Kihnert-

Finkernagel), LIP (R. Courtecuisse, C. Lécuru), M (D. Triebel), PC (B. Buyck), and to

Denise Dailly-Lamouret+ and Egon Horak for having put at our disposal their personal

notes and collections and for bibliographic help. We are indebted to all mycologists who

submitted us their personal collections cited here, helped us in field excursions, and

provided us information or bibliography, especially: Andrea Aiardi, Didier Borgarino,

Jan Borovicka, Micheline Broussal, Emanuele Campo, Gilles Corriol, Jean-Claude

Déianat, Maurice Durand, Hannes Gamper, Jacques Guinberteau, Audrey Hayet,

Chantal Hugouvieux, Patricia Jargeat, Hakan Lindstrém, Ibai Olariaga Ibarguren,

Valentin Queloz, Heinz Woltsche and Jean-Jacques Wuilbaut. We also thank Andreas

Bresinsky (Munich) for confirmation of the loss of original material of Naucoria

cedriolens, Francois Chassagne, Patricia Jargeat and Sophie Manzi for technical

assistance in molecular work in Toulouse, Laetitia Hugot (Conservatoire Botanique de

la Corse, Corte), Jean Alesandri, Myriam Coulom, Claude Lavoise (Société Mycologique

d’Ajaccio) and Franck Richard (CEFE-CNRS, Montpellier) for field assistance in Corsica,

Alnicola spp. nov. in the A. badia complex... 21

and Ita Paz Conde (Sociedad Micoldgica Cantabra) for her invitation to field excursions

in Cantabria, Spain. Lastly the authors thank Régis Courtecuisse for his corrections and

comments on the text, and Joseph F Ammirati (Seattle) and Fernando Esteve-Raventdés

(Alcala de Henares) for their pre-submission review of the manuscript.

This study has been partly supported by the ETH Ziirich (post-doctoral project of

P.-A. Moreau, 2003-2004), the Office de Environnement de la Corse (Corte), and the

University Toulouse 3-Paul Sabatier, CNRS, and Region Midi-Pyrenees (Toulouse, PhD

thesis program of J. Rochet).

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MYCOTAXON

http://dx.doi.org/10.5248/121.23

Volume 121, pp. 23-27 July-September 2012

A new species of Xerocomus from Southern China

MING ZHANG »?, Ta1-Hut Li", TOLGOR BAU” & BIN SONG?

‘Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application,

Guangdong Open Laboratory of Applied Microbiology & State Key Laboratory of

Applied Microbiology (Ministry—Guangdong Province Jointly Breeding Base),

South China, Guangdong Institute of Microbiology, Guangzhou 510070, China

Jilin Agricultural University, Changchun 130118, China

“CORRESPONDENCE TO: *' mycolab@263.net, “junwusuo@126.com

ABSTRACT — Xerocomus puniceiporus, discovered from Heishiding National Nature Reserve

in Guangdong province of China, is described as new. The new species is morphologically

distinctive because of its small basidioma, greyish ruby to purplish red pileus hairs or scales,

reddish purple stipe, and yellow tubes with reddish purple pores. Its rDNA ITS sequence

differs from any known sequence of Xerocomus and Boletus. The holotype is deposited in the

Fungal Herbarium of Guangdong Institute of Microbiology (GDGM).

KEY worbD — taxonomy, Boletales

Introduction

Xerocomus, which is considered to have a broad concept, is a heterogeneous

mixture of several groups of species (Singer 1986, Ladurner & Simonini 2003)

and is not monophyletic (Sutara 2008). Some mycologists have incorporated

Xerocomus into Boletus, whereas others still treat it as an independent genus

(Watling & Hills 2005, Hills 2008). At least 45 Xerocomus species have been

reported in China (Bi et al. 1993, 1997; Chiu 1948, 1957; Li & Song 2000, 2003;

Teng 1939, 1963; Zang 1996, 2006; Zang et al. 2001, 2002; Zang & Petersen

2004; Zhang et al. 2001). Recently, when tidying the bolete specimens collected

from Heishiding National Nature Reserve in Guangdong province of China,

we found a species different from any other bolete known from China. It is

consistent with the definition of Xerocomus following the opinions of Watling

& Hills (2005) and Sutara (2008). Further study revealed that as a new species,

which we formally describe below. The holotype is deposited in the Fungal

Herbarium of Guangdong Institute of Microbiology (GDGM).

24 ... Zhang & al.

20mm

PLATE 1. Xerocomus puniceiporus.

a. Basidiomata. Bar = 20 mm. b. Basidiospores under scanning electron microscope

Materials & methods

Specimens were annotated and photographed in the field, and dried in an electric

drier. Colour description was according to Kornerup & Wanscher (1978). Tissue

sections were mounted in 5% KOH for microscopic examination. At least 30 basidio-

spores were measured. Basidiospore dimensions follow the notation (a—)b-c(-d), with

b-c representing 90% or more of the measured values and with extreme values “a”

and “d” shown in parentheses. Q = length/width ratio of an individual basidiospore,

Q = average Q of all basidiospores + sample standard deviation, and n = number of

spores measured per specimen.

Genomic DNA was isolated from dried specimens and the ITS1-5.8S-ITS4 segment

from the ribosomal DNA (rDNA) was amplified with primer sets ITS1 (5-cTT GCG TTG

ATT ACG-3 ) and ITS4 (5-TCc TCC GCT TAT TGA TAT GC-3’) by polymerase chain reaction

(PCR) techniques. Amplified products were examined with agarose gel electrophoresis

using a 2kb DNA marker. The amplified PCR product was directly sequenced and the

datum is deposited in GenBank.

Taxonomy

Xerocomus puniceiporus T.H. Li, Ming Zhang & T. Bau, sp. nov. PLATE1, FIG. 1

MycoBAnk MB 563347

Differs from Boletus aokii by its purplish red pores, nearly unchanging reaction when

injured, and small basidiospores.

Type: China, Guangdong Province, Fengkai County, Heishiding National Nature

Reserve, 1 July 2010, TaiHui Li & ChunYing Deng (Holotype, GDGM 27443; GenBank

JF514559).

ETYMOLOGY: puniceiporus refers to the purplish red pore-surface or pore-edge.

PiLEus 20-25 mm broad, fleshy, convex to expanded, greyish ruby (12E5) to

greyish Magenta (13E5) in the mass, mixed with greyish red (11D5), tomentose

to fibrillose or villose, soon becoming areolate, and with tufts of greyish ruby

(12E5), greyish Magenta (13E5) to purplish red (13B7) hairs or villose scales on

Xerocomus puniceiporus sp. nov. (China) ... 25

ERS

Fic. 1. Microscopic features of Xerocomus puniceiporus.

a. Basidia; b. Basidiospores; c. Pleurocystidia; d. Pileipellis. Bars = 10 um.

surface, not viscid; with a narrow purplish red (13B7) zone and slightly incurved

at margin. Exposed background or context nearly white, pinkish white, yellowish

white (1A1), pinkish (1A2) to light purplish pink (14A2). FLesH 2.5-4 mm thick

at center, soft, white (1A1), nearly unchanging to slightly blueing (21A2, 22A2)

when exposed. TuBEs 1.5-2.5 mm deep, decurrent, light yellow (2A5), lemon

yellow (2B5) to greenish yellow (2C6), PoREs 1-2 mm wide, angular, somewhat

radially elongated or lamellate close to stipe, with a very narrow purplish red

(13A7) pore-surface or edge. STIPE 20-27 x 2-3 mm, central, cylindrical, equal

to slightly enlarged downwards, purplish red to magenta (13B7, 13C7), with

purplish red furfuraceous scurfs, slightly or obscurely striate, solid, brittle, with

white mycelium at base; STIPE CONTEXT white (1A1), nearly unchanging to

slightly blueing (21A2, 22A2) when exposed.

BASIDIOSPORES (7.2—)7.8-9.8(-10.6) x (3.5-)3.9-4.3(-4.8) um, Q = (1.88-)

2.0-2.33(-2.51),Q_ = 2.20 £0.15, n = 30, ellipsoid to ellipsoid-fusiform, smooth,

pale yellowish brown to olive yellow, thin-walled. BAsip1a 4-spored 27.0-35.0

x 7.5-11.0 um, clavate, hyaline to yellowish white; sTERIGMATA 1.4-2.7 um

long. PLEUROCySTIDIA 48.0-70.0 x 12.0-15.0 um, fusoid ventricose to fusoid,

thin-walled, with hyaline to yellowish. HYMENOPHORAL TRAMA subparallel to

nearly bilateral type with hyphae of the lateral strata loosely arranged, hyaline

to yellowish, 7.8-15 um broad. PILEIPELLIS composed of tufts of erect hyphal

elements 9.8-13.2 um broad, but initially a characteristic palisadoderm, with

brownish yellow to yellowish. CLAMP CONNECTIONS absent in all tissues.

26 ... Zhang & al.

ECOLOGY AND DISTRIBUTION — Solitary or scattered on soil with humus

and debris under conifers mixed with other broadleaf trees; June to July. Only

known from the type locality in South China (Guangdong).

ComMENTS — According to Sutara (2008), Xerocomus s.l. can be divided into

five genera as Xerocomus s. str., Phylloporus, Xerocomellus, Hemileccinum, and

Pseudoboletus. Following this classification, our new species would be placed

in Sutara’s Xerocomellus based on the smooth as observed under the scanning

electron microscope (PLATE 1B), the hymenophoral trama comprises an

intermediate structure between the boletoid and phylloporoid types with hyphae

of the lateral strata not touching each other, and the pileipellis at the initial stage

is composed of a characteristic palisadoderm (Sutara 2005, 2008). However,

when compared with the other species within Xerocomellus group possessing

a similar cracking pileus and large angular pores, Xerocomus puniceiporus can

be distinguished from other species by the combination of small basidiomes

with greyish ruby to purplish red hairs or scales on the pileus, yellow tubes with

purplish red pores, nearly unchanging to slightly blueing context, and purplish

red scurfs on the stipe. Among the other bolete groups, Boletus aokii Hongo

macroscopically resembles the new species to some extent in size, pileus colour,

yellow tubes, and angular pores, but it has longer basidiospores (9-13.5 x 4-5

um), yellow pores, and a noticeable blueing reaction on tubes and flesh when

injured (Hongo 1984, Zang 2006).

The nrDNA-ITS sequence (660 bp) of X. puniceiporus (JF514559) differs

from any known sequences of Xerocomus and Boletus. A Blast search of

GenBank produced as the closest matches several unknown Xerocomus species

(AY372287, AY372285, DQ066413, DQ066412, DQ066378), with only 87%

maximal identities. Therefore, we distinguish Xerocomus puniceiporus as a new

species based on both morphological and molecular characters.

Acknowledgments

The authors are very grateful to Dr. Beatriz Ortiz-Santana (US-Forest Service,

Northern Research Station, Center for Forest Mycology Research) and Dr. Genevieve M.

Gates (University of Tasmania) who reviewed the manuscript. The study is supported by

the National Natural Science Foundation of China (No. 30970023, 31070024, 31170026),

the Natural Science Foundation of Guangdong (Nos. E05202480), and the Foundation

of Guangdong Academy of Sciences, China for 2008 Outstanding Young Science and

echnology Talents. We also thank Ms. Chun-Ying Deng, Xiao-Lan He, Mr. Ting Li and

Cheng-Shu Qiu who have participated in the study.

Literature cited

Bi ZS, Zheng GY, Li TH. 1993. The macrofungus flora of Chinas Guangdong Province. The Chinese

University Press: Hong Kong. (in Chinese)

Bi ZS, Li TH, Zhang WM, Song B. 1997. A preliminary agaric flora of Hainan Province. Guangdong

Higher Education Press. (in Chinese)

Xerocomus puniceiporus sp. nov. (China) ... 27

Chiu WE. 1948. The boletes of Yunnan. Mycologia 40: 199-231. http://dx.doi.org/10.2307/3755085

Chiu WE. 1957. Atlas of the Yunnan boletes. Science Press, Beijing. (in Chinese)

Hills AE. 2008. The genus Xerocomus. A personal view, with a key to the British species. Field

Mycology 9: 77-96. http://dx.doi.org/10.1016/S1468-1641(10)60416-1

Hongo T. 1984. Materials for the fungus flora of Japan (35). Transactions of the Mycological Society

of Japan 25: 281-284.

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

Ladurner H, Simonini G. 2003. Xerocomus s. 1. Fungi Europaei, vol. 8. Edizioni Candusso, Alassio.

Li TH, Song B. 2000. Chinese boletes: a comparison of boreal and tropical elements. 4, in: AJS

Walley (ed.). Tropical mycology (The millennium meeting on tropical mycology, main meeting

2000).

Li TH, Song B. 2003. Bolete species known from China. Guizhou Science 21: 78-86.

Singer R. 1986. The Agaricales in modern taxonomy, 4th edition. Koeltz Scientific Books,

Koenigstein, Germany.

Sutara J. 2005. Central European genera of the Boletaceae and Suillaceae, with notes on their

anatomical characters. Czech Mycology 57: 1-50.

Sutara J. 2008. Xerocomus s. |. in the light of the present state of knowledge. Czech Mycology

60: 29-62.

Teng SC. 1939. Contribution to our knowledge of higher fungi of China. National Institute of

Zoology & Botany, Academia Sinica.

Teng SC. 1963. Fungi of China. Science Press. (in Chinese)

Watling R, Hills AE. 2005. Boletes and their allies. Boletaceae: Strobilomycetaceae: Gyroporaceae:

Paxillaceae: Coniophoraceae: Gomphidiaceae (revised and enlarged edition). British Fungus

Flora. Agarics and Boleti 1. 173 p.

Zang M. 1996. A contribution to the taxonomy and distribution of the genus Xerocomus from

China. Fungal Science 11: 1-15.

Zang M. 2006. Flora Fungorum Sinicorum vol.22. Boletaceae (1). Science Press, Beijing. 215 p.

(in Chinese)

Zang M, Petersen RH. 2004. Notes on tropical boletes from Asia. Acta Botanica Yunnanica 26:

619-627.

Zang M, Li TH, Petersen RH. 2001. Five new species of Boletaceae from China. Mycotaxon 80:

481-487.

Zang M, Lin JT, Huang NL. 2002. Xerocomus tengii, a new Xerocomus species from China.

Mycosystema 21: 480-482.

Zhang WM, Li TH, Song B. 2001. A survey on the macrofungi in Guangdong Province. Ecologic

Science 20: 48-58. (in Chinese)

MY COTAXON

http://dx.doi.org/10.5248/121.29

Volume 121, pp. 29-36 July-September 2012

Nomenclatural and taxonomic notes on

Calvatia (Lycoperdaceae) and associated genera

JOHANNES C. COETZEE? & ABRAHAM E. VAN Wyk?

"Department of Horticultural Sciences, Cape Peninsula University of Technology,

PO. Box 1906, Bellville 7535 Republic of South Africa

?H.G. WJ. Schweickerdt Herbarium, Department of Plant Science, University of Pretoria, Pretoria

0002 Republic of South Africa

* CORRESPONDENCE TO: coetzeej@cput.ac.za

ABSTRACT — Various nomenclatural aspects pertaining to author citations, orthography and

the validity of names in the genus Calvatia, and, in one instance, Bovista, are discussed. The

name Calbovista subsculpta var. fumosa is validated and the biogeographic status of Calvatia

gigantea in southern Africa is discussed. The name Calvatiella lioui is lectotypified.

Key worps — Bovista cacao, Calvatia sericella, Calvatia sinclairii, Calvatia versipora,

Calvatiella lioui

Introduction

In the course of our ongoing study of the genus Calvatia Fr. in southern

Africa, we are continually encountering noteworthy facts and odd snippets

of information, some of which—especially those pertaining to non-southern

African taxa—are unlikely to find mention elsewhere in our papers on the

southern African Lycoperdaceae. Individually, some items might perhaps be

regarded as rather trivial, but collectively it all adds up to a substantial body

of information that might be of value to other workers in the field as well as,

in a few cases, even serve as stimulus for further investigation. A selection of

those (often purely nomenclatural) items of information is presented in this

miscellany of short notes. All references to the International Code of Botanical

Nomenclature (ICBN) pertain to the “Vienna Code’ (McNeill et al. 2006).

1. Validation of the name Calbovista subsculpta var. fumosa

Calbovista subsculpta var. fumosa A.H. Sm. ex J.C. Coetzee & A.E. van Wyk,

var. Nov. MycoBANnk 513113

“Calbovista subsculpta var. fumosa” A.H. Sm., Mycopathol. Mycol. Appl. 26: 396. 1965,

nom. inval.

30 ... Coetzee & Van Wyk

DESCRIPTION (Latin and English) — Smith (1965).

TyPE — United States, Idaho, Bonner Co., Dickensheet Campground, Priest River,

Kaniksu National Forest, gregarious under Pinus contorta, A.H. Smith 71347, 21 October

1964. [HoLoTyPE, MICH].

KEY CHARACTERS — See Smith (1965).

ComMMENTS — When Morse (1935) erected the monotypic genus Calbovista,

no Latin diagnosis or reference to any previously published Latin diagnosis was

provided, making the names “Calbovista” Morse and “Calbovista subsculpta”

Morse invalid according to ICBN Art. 36.1. Consequently, the varietal name

“Calbovista subsculpta var. fumosa” (Smith 1965) was also invalid in terms of

ICBN Art. 43.1, which rules that the name of a taxon below the rank of genus

is not validly published unless the name of the genus or species to which it is

assigned has been validly published. Although Seidl (1995) validated Morse’s

invalid generic and specific names, no consideration was given to Smith's

varietal name. That situation is here rectified by formally validating the name.

2. The names Calvatia umbrina and Bovista cacao revisited

Although the fungus that has gone under the above two names was properly

described in Lloyd (1904a: 2), it is evident from Lloyd’s discussion that he

regarded Calvatia umbrina as merely provisional, thereby making “Calvatia

umbrina” Lloyd invalid according to ICBN Art. 34.1(b). Nonetheless, despite

this and despite Stevenson & Cash's (1936) exclusion of C. umbrina from their

catalogue of new fungus names proposed by Lloyd, Zeller & Smith (1964)

continued to use Lloyd's invalid name.

This error was perpetuated by Ponce de Leén (1975) when he transferred

C. umbrina to the genus Bovista. Ponce de Leén accepted Lloyd's provisional

name but, realising that the epithet umbrina was unavailable in Bovista (having

been used previously by Bottomley (1948) for a different fungus), proposed the

new name Bovista cacao P. Ponce de Leén. However, since C. umbrina is invalid

and thus has no status under the ICBN, Ponce de Leon should not have treated

Bovista cacao as a nomen novum, but as a species novum which, in the absence

of a Latin diagnosis, must be rejected as invalid according to ICBN Art. 36.1.

A re-examination of the original (and apparently only) collection of this

fungus [U.S.A., California, Mountain View, on the ground, Erhorn s.n. (Lloyd

Myc. Coll. 20679, BPI)] is required and, if found to be a good species, the name

needs to be validated. Kreisel (1992, 1994) did not include this fungus in his

survey and key to the genus Calvatia.

3. On the identity of Calvatia sericella

Calvatia sericella (Berk.) Lloyd ex J.A. Stev. & E.K. Cash, Bull. Lloyd Libr. Bot. (35):

173. 1936.

= Lycoperdon sericellum Berk., Hooker's J. Bot. Kew Gard. Misc. 3: 171. 1851.

Ho.ortyPe — India, Darjeeling, 7000 feet, on the ground, Hook fil. 32. [K!]

Notes on Calvatia ... 31

As far as could be ascertained, the combination Calvatia sericella was first

validly published in Stevenson & Cash (1936). They ascribed the combination

to Lloyd (1904b: 2), but as Lloyd did not definitely associate the epithet with the

genus name, he failed to create a combination according to ICBN Art. 33.1 +

Ex. 2. Demoulin (1971) believed that C. sericella probably was conspecific with

Calvatia excipuliformis (Scop. : Pers.) Perdeck, although Lloyd (1904b) and Van

Overeem (1927) suggested a relationship with Calvatia gardneri (Berk.) Lloyd

[= Calvatia pyriformis (Lév.) Kreisel]. However, after having examined the type

material at K (consisting of three specimens in the Lycoperdon sericellum type

folder but filed under C. excipuliformis), we are convinced that it represents

neither C. excipuliformis nor C. pyriformis and—based on macroscopic

similarity, spore morphology, and the presence of conspicuous, widely

gaping perforations in the branched, septate capillitium—strongly suspect

conspecificity with the well-known Calvatia rugosa (Berk. & M.A. Curtis) D.A.

Reid. Should that be the case, the name C. sericella will enjoy priority over

C. rugosa, since its basionym (L. sericellum) antedates the basionym of C. rugosa

(= Lycoperdon rugosum Berk. & M.A. Curtis, J. Linn. Soc., Bot. 10: 345. 1868) by

17 years. Molecular analysis of the relevant type specimens should throw more

light on this intriguing question.

4. A matter of orthography: ‘versispora’ or ‘versipora’?

The fungus originally described by Lloyd (1915: 7-8) as Calvatia versipora

does not belong to the Lycoperdaceae at all. It is, in fact, the anamorph of

Laetiporus sulphureus (Bull. : Fr.) Murrill (Kreisel et al. 1983, Stalpers 1984,

Reid 1985), which Stalpers (1984) transferred to the genus Sporotrichum Link :

Fr. [as Sporotrichum versisporum (Lloyd) Stalpers].

According to Lloyd (1916), the name C. versipora was an “...original blunder...’

and he therefore suggested a change to Calvatia versispora since that ‘..would

better indicate the idea...? According to ICBN Art. 51.1 and 60.1, however,

changes such as this are not permitted. Nonetheless, various later authors such

as Stevenson & Cash (1936), Kreisel et al. (1983), Stalpers (1984), and Reid

(1985), adopted Lloyd’s proposed ‘correction’ Stalpers (1984) defends this as

follows: “The code allows the correction of printing errors and thus the epithet

‘versispora is used.... Perusal of Lloyd’s (1915) original publication revealed,

however, that the name of this fungus appears in print not only once, but at

least three times (once on p. 4 and twice on p. 7) and, in all cases, as ‘versipora.

As a result of this consistent use by Lloyd (1915), we are much more inclined to

believe that the use of the original ‘versipora was intentional, and that it should

not merely be dismissed as a correctable typographic error. We have little doubt

that the blunder referred to by Lloyd (1916) represents an error of semantics

and not of typography. Consequently we believe that Lloyd's original spelling

should also be retained in later combinations derived from the original name.

32 ... Coetzee & Van Wyk

5. Correct author citation for the superfluous combination

Calvatia bovista (Pers. : Pers.) Kambly & Lee

T.C.E. Fries’s (1921) transfer of Lycoperdon bovista Pers. : Pers. to the genus

Calvatia [as C. bovista (Pers. : Pers.) T.C.E. Fr., an illegitimate later homonym of

C. bovista (L.) T. Macbr.] went unnoticed by many authors, including Kambly

& Lee (1936) who, in their paper on the gasteromycetes of Iowa, also made

the transfer and have since then often been credited with the combination

Calvatia bovista [basionym: L. bovista Pers. : Pers. {= Calvatia utriformis

(Bull. : Pers.) Jaap}], not to be confused with Calvatia bovista (L.) T. Macbr.

[= Lycoperdon bovista L. {= Calvatia gigantea (Batsch : Pers.) Lloyd}]. Kambly &

Lee’s combination is, however, illegitimate on two counts: (1) it is superfluous

and (2) (like the Fries combination) it is a later homonym of C. bovista (L.) T.

Macbr.

Establishing the correct author citation for Paul Kambly and Robert E.

Lee’s isonym proved to be an unexpected and time-consuming irritation.

Throughout the literature, the name has merely been cited as “Calvatia bovista

(Pers. : Pers.) Kambly & Lee.” With Brummitt & Powell (1992) listing no

fewer than 29 Lees as authors of plant names, the abbreviation requires some

refinement. Unfortunately, however, Brummitt & Powell (1992) list not one, but

two authors as Robert E.... Lee, neither of whom is cited as an author of fungal

names. Robert Edward Lee is listed for his algal names, while Robert Edwin Lee

is remembered as an author of spermatophyte names. Who then collaborated

with Kambly on the 1936 paper on the gasteromycetes of Iowa? Was it Robert

Edward, Robert Edwin, or someone entirely different? The literature available

to us yielded no clues.

The problem was eventually solved after repeated enquiries revealed the

existence of a 1932 MS thesis on gasteromycetes by one ‘Robert Edward Lee’

in the University of Iowa library. This ‘discovery’ left us with little doubt that

Robert Edward was our man and that the correct author citation for Kambly &

Lee’s illegitimate combination should be Calvatia bovista (Pers. : Pers.) Kambly

& R.Ed. Lee.

6. The status of Calvatia gigantea in South Africa

Bottomley’s (1948) inclusion of C. gigantea in her list of South African

gasteromycetes was based on a single, late 18th century, Thunberg collection

of a specimen(s?) found in June in hilly terrain outside Cape Town (Thunberg

1800, 1823). Thunberg’s designation of his material as Lycoperdon bovista sensu

Linnaeus (= C. gigantea) was an error, but understandable for the time. It is

inexplicable however, how this error could have been perpetuated in Bottomley

(1948), which has led to other authors (e.g. Kreisel 1992, 1994) also citing C.

gigantea as occurring in South Africa. Thunberg (1800, 1823) described his

material as follows: “..globosum, lacero-dehiscens, stipite validissimo, clavato-

Notes on Calvatia ... 33

ventricoso, carne alba, seminis atris’ The reference to ‘white flesh’ indicates

an immature specimen, which would have made identification difficult. The

distinct stipe, however, clearly excludes Thunberg’s material from the stipeless

C. gigantea, which is, apart from Thunberg’s record, not known to occur in

the Cape Town area or anywhere else in South Africa. Thunberg’s description

is, however, very reminiscent of an immature Calvatia cyathiformis (Bosc)

Morgan, which is, from our own experience, the most common large puffball

occurring in the Cape Peninsula area. We would be surprised if the Thunberg

specimen did not represent an immature C. cyathiformis specimen.

Thunberg’s collection does not seem to have survived. In the Thunberg

herbarium at UPS there is only a single sheet (no. 27495, with no locality

indicated) filed under Lycoperdon bovista. A photograph of the sheet clearly

shows the specimens mounted on it to be globose, stalkless, and dehiscing by

means of a relatively wide ostiole. These are characteristics of Bovista, and the

sheet also bears the inscription “Bovista nigrescens.’ In addition to the missing

locality information on the sheet, the stalkless nature of the specimens at

UPS clearly separate them from Thunberg’s ‘L. bovista’ specimen(s?) from the

Cape.

In July 1985 a huge puffball, unfortunately still immature but macroscopically

very reminiscent of C. gigantea, was collected in a garden in Durban (T.D.

Steinke s.n.) and sent to PREM for identification (PREM 48248). A small piece

of the carpophore was later sent to Kew, on which Dr. Derek Reid commented

in a letter to the PREM correspondent (copy of letter filed in the Langermannia

sp. nov. file at K!): ‘It is not L. gigantea..’ because the spores ‘.. are smaller and

much less ornamented... [and] ...the capillitium is also perforate’? He concluded:

‘.. Lam inclined to agree with you that the specimen may well represent a new

species of Langermannia but I feel it would be unwise to describe it on this

abnormal specimen:

No other southern African material of C. gigantea could be found in any

South African herbarium, and a search at K also failed to locate anything

from the African continent. A specimen from Kenya in a box at K, labeled

Langermannia gigantea, represents Calvatia argentea (Berk) Kreisel. The

US. National Fungus Collections also contain no record of this fungus from

Africa (Farr & Rossman 2006). Therefore, in the absence of any substantiating

evidence, the occurrence of C. gigantea in South Africa remains unconfirmed.

7. Identity of Lycoperdon sinclairii confirmed

Calvatia sinclairii (Berk. ex Massee) Lloyd, Lycoperd. Australia: 37. 1905.

= Lycoperdon sinclairii Berk. ex Massee, J. Roy. Microscop. Soc. London 1887: 716. 1887.

Ho.LotyPe — New Zealand, Nelson 1500 ft., on the ground, Sinclair s.n. [K!]

Calvatia sinclairii has in the past been cited as a synonym of C. utriformis

(Kreisel 1962, Zeller & Smith 1964) and, according to Demoulin (1971),

34 ... Coetzee & Van Wyk

it is close to C. utriformis. Kreisel (1989), however, regarded it as a doubtful

synonym of that species. After having had the opportunity to examine the type

specimen at K, however, we are confident that C. sinclairii is conspecific with,

and should indeed be regarded as a later synonym of, C. utriformis.

8. On the status of Calvatia purpurea

“Calvatia purpurea” (O.F. Mill. : Fr.) LR. Hall, PK. Buchanan, Y. Wang & A.LJ.

Cole, Edible and poisonous mushrooms: 174. 1998, comb. inval.

= Clavaria purpurea O.F. Mill. Fl.dan. 14: t.837 f.2. 1780 : Fr., Syst. Mycol. 1: 480. 1821.

This combination first appeared in a list of fungi published in Hall et al.

(1998), and both MycoBank (http://www.mycobank.org, 30-3-2011) and Index

Fungorum (www.indexfungorum.org, 30-3-2011) accepted it as a good name,

citing it as “Calvatia purpurea (Lloyd) I.R. Hall, PK. Buchanan, Wang{?} &

Cole” with Bovista purpurea Lloyd as basionym. ‘That, however, is not correct.

The combination “Calvatia purpurea” was created unintentionally and is an

error to be ignored. A mistake was made in compiling the list in Hall et al.

(1998); the entry should have read Clavaria purpurea O.F. Mill. : Fr., not

“Calvatia purpurea” (Dr. I.R. Hall, pers. comm.). Since the combination was

published without indicating the basionym and as it is also not accepted by the

authors who inadvertently created it, the name is invalid according to ICBN

Art. 33.3 and 34.1(a).

9. Lectotypification of Calvatiella lioui

Calvatiella lioui C.H. Chow, Bull. Fan Mem. Inst. Biol., Bot. 7: 93. 1936.

LECTOTYPE (designated here) — plate III of Chow (1936).

Calvatiella lioui was originally described from a single specimen found in the

herbarium of the National University of Peking (Chow 1936). This specimen

disappeared during World War II, however (Kreisel & Calonge 1993). In view

of this holotype loss, the only remaining original material associated with the

name is here formally designated as lectotype of Calvatiella lioui according to

ICBN Art. 7.10, 7.11, 9.9 and 9.10. Based on the description and illustrations

in Chow (1936), Kreisel & Calonge’s (1993) relegation of Calvatiella lioui to

synonymy under Calvatia utriformis certainly seems probable but, in the

absence of the holotype, cannot be corroborated.

Acknowledgments

We are indebted to Prof. Lois Tiffany of the Department of Ecology, Evolution and

Organismal Biology at Iowa State University for the information on Robert Edward

Lee and to Dr. Ian Hall, Dunedin, New Zealand, for the information on Calvatia

purpurea. The curators of K and PREM are thanked for entrusting us with and allowing

examination of specimens in their custody. The hospitality extended to the senior

author by Dr. David Pegler, Dr. Brian Spooner, and the rest of the staff during study

Notes on Calvatia ... 35

visits to the Mycological Herbarium at Kew is acknowledged with appreciation. Prof.

Vagner G. Cortez of Universidade Federal do Parana (Brazil) and Dr. Taiga Kasuya of

the University of Tsukuba (Japan) are thanked for valuable comments and critically

reviewing an earlier draft of this paper.

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Brummitt RK, Powell CE. 1992. Authors of plant names. Kew, Royal Botanic Gardens.

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Demoulin V. 1971. Le genre Lycoperdon en Europe et en Amérique du Nord. Etude taxonomique et

phytogéographique. Doctoral Thesis, Université de Liége.

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Hall IR, Buchanan PK, Wang Y, Cole ALJ. 1998. Edible and poisonous mushrooms: an introduction.

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Kambly PE, Lee RE. 1936. The Gasteromycetes of Iowa. Studies in Natural History, lowa University

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Kreisel H. 1962. Die Lycoperdaceae der Deutschen Demokratischen Republik. Feddes Repertorium

64: 89-201.

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Kreisel H. 1994. Studies in the Calvatia complex (Basidiomycetes) 2. Feddes Repertorium

105: 369-376. http://dx.doi.org/10.1002/fedr.19941050516

Kreisel H, Calonge FD. 1993. Calvatiella Chow, a synonym for Bovistella Morgan. Mycotaxon

48: 13-25.

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Basidiomycetes) from India. Feddes Repertorium 94: 543-547.

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Lloyd CG. 1904b. Puff ball letters, no. 2. Cincinnati. 2p.

Lloyd CG. 1915. Letter no. 56. Cincinnati. 12p.

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Prado J, Silva PC, Skog JE, Wiersema JH, Turland NJ. 2006. International Code of Botanical

Nomenclature (Vienna Code). (Regnum Vegetabile 146). Liechtenstein, A.R.G. Gantner Verlag

KG.

Morse EE. 1935. A new puffball. Mycologia 27: 96-101. http://dx.doi.org/10.2307/3754047

Ponce de Leon P. 1975. Notes on Calvatia (Lycoperdaceae), I. Fieldiana: Botany 38: 1-3.

Reid DA. 1985. An annotated list of some fungi from the Channel Islands, mostly from Jersey.

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http://dx.doi.org/10.1016/S0007-1536(85)80128-5

Seidl MT. 1995. Validation of the puffball genus Calbovista. Mycotaxon 54: 389-392.

Smith AH. 1965. New and unusual Basidiomycetes with comments on hyphal and spore wall

reactions with Melzer’s solution. Mycopathologia et Mycologia Applicata 26: 385-402.

http://dx.doi.org/10.1007/BF02049566

36 ... Coetzee & Van Wyk

Stalpers JA. 1984. A revision of the genus Sporotrichum. Studies in mycology 24: 1-105.

Stevenson JA, Cash EK. 1936. The new fungus names proposed by C.G. Lloyd. Bulletin of the Lloyd

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Edman.

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MY COTAXON

http://dx.doi.org/10.5248/121.37

Volume 121, pp. 37-44 July-September 2012

New combinations and notes in clavarioid fungi

I. OLARIAGA' & I. SALCEDO?

' The Swedish Museum of National History. Dpt. Cryptogamic Botany,

O. Box 50007, Svante Arrhenius vdg 7 SE-104 05 Stockholm, Sweden

? Dpt. Plant Biology and Ecology (Botany), University of the Basque Country, UPV/EHU,

Apdo. 644, 48080 Bilbao, Spain

* CORRESPONDENCE TO: ibai.olariaga@nrm.se

ABSTRACT — ‘The revision of several families of clavarioid fungi in the Iberian Peninsula

necessitated new combinations and names to be used for the Iberian fungal flora. Eight new

combinations are proposed in Clavulina, Ramariopsis, and Typhula, and nomenclatural and

taxonomic comments are provided in each case. Lectotypes are designated for Clavaria

contorta, Clavaria fistulosa (type species of Macrotyphula), and Clavaria phacorrhiza (type

species of Typhula). The genus Macrotyphula is reduced to synonymy under Typhula.

Key worps — Clavariaceae, Clavulinaceae, Typhulaceae, nomenclature, taxonomy

Introduction

Clavarioid fungi form an artificial assemblage of fungi, since clavarioid

fruitbodies have evolved in several lineages (Parmasto 1965, Pine et al.

1999, Dentinger & McLaughlin 2006, Hibbett 2007). However, for practical

reasons, clavarioid fungi have often been the target group for monographs

and smaller papers (Corner 1950, Petersen 1988, Dentinger & McLaughlin

2006). During the last years, a monograph that deals with some families of

clavarioid fungi (Clavariaceae, Clavulinaceae, Pterulaceae, and Typhulaceae)

has been completed as a result of a PhD (Olariaga 2009), following the

framework of Flora Mycologica Iberica (Telleria & Melo 1995). The taxonomic

and nomenclatural revision of many names and type collections in the light of

current phylogenetic knowledge has necessitated the creation of new names

to be used in the context of the Iberian fungal flora. Thus, the purpose of this

work is to effectively publish nomenclatural novelties required as a result of the

mentioned monograph (Olariaga 2009).

38 ... Olariaga & Salcedo

Materials & methods

Herbaria abbreviations follow Holmgren & Holmgren (1998). The collections

examined in this study are deposited in BIO, G, MA, PORTU (Mycological Society

of Portugalete, Biscay), S, SEST (Natural Science Society of Sestao, Biscay), and UPS

herbaria. Abbreviations of author names, periodicals, and non-periodical publications

follow Kirk & Ansell (2003), Bridson & Smith (1991), and TL2 (Stafleu & Cowan 1976,

1979), respectively. All validly published homotypic synonyms are given under each

new combination.

Taxonomy

Clavulina incarnata (Corner) Olariaga, comb. et stat. nov.

MycoBANnk 563526

= Clavulina cristata var. incarnata Corner, Ann. Bot. Mem. 1: 692. 1950.

Of the few Clavulina species known to have cystidia (Corner 1970), C. cristata

var. incarnata is the only taxon that has been recorded in Europe (Corner 1950,

Pilat 1958). Corner (1950) described the variety for two British collections that

developed a pink to grayish drab colour and, above all, possessed cystidia. He

granted the taxon the rank of variety, not without noting that it might deserve

the species rank. This taxon would differ from C. cristata (Holmsk. : Fr.) J. Schrot.

as treated by Olariaga et al. (2009) in the early development of pink to grey

tones, presence of cystidia, and the lack of profusely cristate apices. Two recent

Swedish collections that match Corner’s description have enabled the senior

author to confirm these differences and thus to propose a new combination to

accommodate the only cystidiate Clavulina species known in Europe.

SPECIMENS EXAMINED—SWEDEN. UPpPLAND: Sollentuna parish, Hansta naturreservat,

33VXF64579094, under Quercus robur, Corylus avellana, Populus tremula, and

Fraxinus excelsior on rich ground, 08/IX/2010, Hansen, Gillen & Olariaga, S-F197088;

33VXF6462690942, 14/IX/2010, Olariaga, S-F197089. Stockholm, Hagaparken, under

Pinus and Betula, on acidic ground, 24/X/2011, Zamora & Olariaga, BIO-Fungi 16400.

Clavulina reae Olariaga, nom. et stat. nov. [non Clavulina gracilis Corner 1950].

MycoBAnk 516723

= Clavaria cinerea var. gracilis Rea, Trans. Brit. Mycol. Soc. 6: 62. 1918 [“1917”].

= Clavulina cinerea var. gracilis (Rea) Corner, Ann. Bot. Mem. 1: 309. 1950.

Morphological and phylogenetic studies among the European Clavulina taxa

indicate that more species than are generally accepted are present in Europe.

Olariaga et al. (2009) showed that specimens identified as C. cinerea var. gracilis

that conform to the original description by Rea (1918) form a well-supported

clade that should be assigned to a species. Because the epithet gracilis is not

available, we propose a substitute with a new epithet honouring the memory of

Carleton Rea. No type material is extant at K (Begofa Aguirre-Hudson, pers.

comm.).

Clavulina, Ramariopsis & Typhula combs. & noms. nov. (Iberia) ... 39

Ramariopsis bispora (Schild) Olariaga, comb. et stat. nov.

MycoBAnk 516724

= Ramariopsis kunzei var. bispora Schild, Westfalische Pilzbriefe 8: 30. 1970.

The type material deposited in Schild’s personal herbarium could not be traced.

However, after examining the comprehensive original description and further

Iberian specimens, we consider that Ramariopsis kunzei var. bispora deserves

to be recognized at the species level. In addition to the 2-spored basidia and

the absence of clamps (attributed to parthenogenesis as a reproduction mode),

R. kunzei var. bispora differs macroscopically from R. kunzei (Fr. : Fr.) Corner in

having smaller basidiomata and lacking pinkish patches in age. Microscopically,

although bi-pyramidal crystals are present among the context hyphae of the

branches in all the material examined, as Schild (1970) depicted, we have

never observed crystals in the typical variety of R. kunzei. The narrower basal

tomentum hyphae (2.5-3 um) of R. kunzei var. bispora appear to be a good

taxonomic character to separate the variety from R. kunzei, with hyphae 3-4

(5.5) um diam.

SPECIMENS EXAMINED—SPAIN. Bizxka1a: Larrinagatxu, Izurtza, 30TWN2878, 280 m,

under Chamaecyparis lawsoniana along a slope, 01/X/2003, Iglesias, Arauzo & Olariaga,

BIO-Fungi 12383; Sopuerta, barrio Castano Viejo, 30T WN2464, 600 m, forest with Fagus

sylvatica var. purpurea and Chamaecyparis, 01/VII/2004, Fernandez-Vicente, PORTU

7100704 [as Ramariopsis kunzei], BIO-Fungi 12563. BuRGos: Pantano de Ordunte,

under Chamaecyparis lawsoniana on the soil, 07/X/2007, Pérez-Butron, SEST 7100705

[as Ramariopsis sp.], BIO-Fungi 12567. LEON: Riafio, Puerto del Pontén, 30TUN3677,

1000 m, slope along the road, 01/X/2003, Olariaga, BIO-Fungi 9922. AsTuRIAs: Proaza,

29TQH4192, 280 m, under Quercus ilex on calcareous soil, 27/X/2002, Olariaga, BIO-

Fungi 9705.

Ramariopsis luteonana (Schild) Olariaga, comb. nov.

MycoBAnk 516725

= Clavulinopsis luteonana Schild, Fungorum Rar. Icon. Color. 5: 28. 1971.

The type material could not be traced. The complete original description by

Schild (1971) deals with a fungus with simple to forked fruitbodies, small

smooth spores, and short basidia. This short basidia and the small spores make

Clavulinopsis luteonana closer to Ramariopsis kunzei [type of Ramariopsis

(Donk) Corner] than to Clavulinopsis miniata (Berk.) Corner (= Clavulinopsis

sulcata Overeem, the type of Clavulinopsis Overeem). Furthermore, the Iberian

material matching Schild’s fungus in basidioma shape, presence of bi-pyramidal

crystals, short basidia, and small spores also shows a very faint ornamentation

in some spores when viewed through the light microscope. Hence, we believe

that the species is better placed in Ramariopsis.

SPECIMENS EXAMINED—SPAIN. BizxKata: Korostadui, Iurreta, 30TWN2981, 140

m, under Chamaecyparis lawsoniana and Pseudotsuga menziesii, 17/X/2007, Iglesias,

Arauzo & Olariaga, BIO-Fungi 12374.

AO ... Olariaga & Salcedo

Ramariopsis subumbrinella (S. Imai) Olariaga, comb. nov.

MycoBANnk 516726

= Clavaria subumbrinella S. Imai, Trans. Sapporo Nat. Hist. Soc. 13(4): 386. 1934.

= Clavulinopsis subumbrinella (S. Imai) Corner, Ann. Bot. Mem. 1: 392. 1950.

The type material could not be located. The original description by Imai (1934)

noting small spores, small basidia, and branched basidiomata suggests a species

of Ramariopsis. The Iberian material that matches in several aspects the original

description shows a very low ornamentation in some spores through the light

microscope, also typical of many Ramariopsis species.

SPECIMENS EXAMINED—SPAIN. CAp1z: Parque Natural de los Alcornocales, Arroyo

Carlos el Tiradero, 30SF61, 150 m, under Pistacia lentiscus, Olea europaea and Quercus

suber, 26/X1/2003, Pérez-Daniéls & Olariaga, BIO-Fungi 10015. CORDOBA: Cordoba,

Arroyo Pedroches, 30SUG4398, 200 m, on bare soil under Myrtus communis, 5/XII/2003,

Pérez Daniéls & Olariaga, BIO-Fungi 10224. SeGovia: Pradena, El Nido del Cuervo,

30TVL4351, 1350 m, bajo Ilex aquifolium, 20/X1/1997, Pérez Daniéls, MA-Fungi 40223

[as Clavulinopsis subtilis], MA-Fungi 40225 [as Clavulinopsis sp.].

Typhula contorta (Holmsk. : Fr.) Olariaga, comb. nov.

MycoBAnk 516730

= Clavaria contorta Holmsk., Beata Ruris 1: 29. 1790: Fr., Syst. Mycol. 1: 478. 1821.

= Clavaria fistulosa var. contorta (Holmsk.) Hohn., Ost. Bot. Z. 54: 425. 1904.

= Clavaria fistulosa f. contorta (Holmsk.) Bourdot & Galzin,

Hyménomyc. France: 121. 1928 [“1927”].

= Clavariadelphus fistulosus var. contortus (Holmsk.)

Corner, Ann. Bot. Mem. 1: 273. 1950.

= Macrotyphula fistulosa var. contorta (Holmsk.) Nannf. & L.

Holm, Publ. Herb. Univ. Uppsala 17: 8. 1985.

= Macrotyphula contorta (Holmsk. : Fr.) Rauschert, Feddes

Repert. Spec. Nov. Regni Veg. 98: 660. 1987.

LECTOTYPE (designated here): Holmsk., Beata Ruris 1: pl. 12. 1790.

The epithet contorta has traditionally been used for specimens with irregular,

wrinkled, or brain-like basidiomata. It has often been synonymized with

Typhula fistulosa (Julich 1984, Rauschert 1987, Berthier 1976) or considered a

variety of it (Knudsen 1997), due to high morphological plasticity (Holmskjold

1790, Bourdot & Galzin 1928). However, some authors have noted that

T. contorta-like basidiomata have larger basidiospores (Harper 1918, Corner

1950, Maas Geesteranus 1976, Breitenbach & Kranzlin 1986). Comparison of

the examined T! contorta material with typical T: fistulosa collections supports

this view. Accordingly, T. contorta is treated as an independent species in the

Iberian revision of the group (Olariaga 2009), although further focused studies

are desirable to confirm its identity. The reasons for accommodating this taxon

in Typhula (Pers. : Fr.) Fr. are given under T: fistulosa.

SPECIMENS EXAMINED—FRANCE. LANDES (40): Tartas, bois mort, 16/11/1987, G

110719, Gilles, [LY-Cl 349, as Macrotyphula fistulosa f. contorta]. SPAIN. ARABA:

Clavulina, Ramariopsis & Typhula combs. & noms. noy. (Iberia) ... 41

Gillerna, 30TQH1881, 750 m, Alnus glutinosa twigs, 7/XI/1985, Martinez-Irigoyen,

BIO-Fungi 12614, BIO-Fungi 12615, Alnus glutinosa twigs, 18/1/1986, BIO-Fungi 12616,

Alnus glutinosa twigs, 6/12/1986, BIO-Fungi 12617, Corylus avellana twigs, 30/XI/1985,

BIO-Fungi 12613; Oleta, Aramaio, 30T WN3189, 600 m, Corylus or Alnus glutinosa twigs,

15/X/2004, Salcedo, BIO-Fungi 10516. AviLa: Barquillo, El Loscar, 29TOF8776, 970 m,

Alnus glutinosa twig, 8/X1/2004, Sarrionandia & Olariaga, BIO-Fungi 10608. BizKata:

Getxo, Martiartu, 30TWN0298, 100 m, Alnus glutinosa twigs, 22/XI/2003, Olariaga,

BIO-Fungi 10107, BIO-Fungi 10105. CAcERgEs: Hervas, La Garganta, 30TTK5963,

790 m, Alnus glutinosa wood, 9/XI/2004, Sarrionandia & Olariaga, BIO-Fungi 10620.

ASTURIAS: Pigiieces, 30TQH1881, 750 m, Alnus stump, 6/X/2005, Olariaga, BIO-Fungi

11191. SALAMANCA: Candelario, 30TTK6571, 100 m, Corylus avellana twig, 8/X1/2004,

Sarrionandia & Olariaga, BIO-Fungi 10641. SWEDEN. BLEKINGE: Karlskrona, Vastra

Mark, strandsnaret, pa murkna algrenar, 10/XI/1946, Wikland 243, UPS F-124250 [as

Clavaria contorta]. VASTERGOTLAND: Hisingen, Goteborg, Rya skog, pa Alnus glutinosa,

26/XI & 9/XH/1999, Nordin, UPS F-124213 [as Macrotyphula fistulosa var. contorta].

Typhula fistulosa (Holmsk. : Fr.) Olariaga, comb. nov.

MycoBAnk 516731

= Clavaria fistulosa Holmsk., Beata Ruris 1: 15. 1790 : Fr.,

Syst. Mycol. 1: 479. 1821 [non Tode 1783].

= Eriocladus fistulosus (Holmsk. : Fr.) Lév., Ann. Sci. Nat., Bot., Sér. 3, 5: 159. 1846.

= Clavariella fistulosa (Holmsk. : Fr.) P. Karst., Rev. Mycol. (Toulouse) 3(9): 21. 1881.

= Clavariadelphus fistulosus (Holmsk. : Fr.) Corner, Ann. Bot. Mem. 1: 272. 1950.

= Macrotyphula fistulosa (Holmsk. : Fr.) R.H. Petersen, Mycologia 64: 140. 1972.

LEcTOYPE (designated here): Holmsk., Beata Ruris 1: pl. 6. 1790.

This species was proposed as the type of Macrotyphula R.H. Petersen (Petersen

1972), a currently used generic name. Morphological comparison between

Macrotyphula fistulosa and Typhula phacorrhiza [selected type species of

Typhula (Donk 1933)] revealed the following morphological similarities:

1. Basidiomata with similar appearance, ochre brown-coloured, pubescent at

the base.

2. Stipe surface formed by thin hyphae, slightly gelatinized, and with similar

caulotrichomes.

3. Basal tomentum formed by scarcely septate, thick-walled hyphae

4. A slight hyaline zebra-like striped encrustation on the medulla hyphae.

Based on these similarities, Macrotyphula fistulosa and T: phacorrhiza appear

to be closely allied species, as molecular phylogenetic inference also suggests

(Pine et al. 1999, Dentinger & McLaughlin 2006, Hibbett 2007). Therefore, we

reduce Macrotyphula to synonymy under Typhula to which we transfer several

Macrotyphula species.

SPECIMENS EXAMINED—ANDORRA. Pal, 31TCH7511, under Pinus, Populus, Betula

pendula, and Corylus avellana, 15/X/2002, Olariaga, BIO-Fungi 12772. FINLAND.

ETELA-HAME: Mustiala, X/1867, P.A. Karsten, UPS F-124239 [as Clavaria fistulosa].

LATVIA. BALDONE: Kekava, Birzuli, ad terram, in alnetum glutinosae, 30/X/1955, A.

Abolina, UPS F-552624 [as Clavaria fistulosa]. SPAIN. LEOn: Puerto del Pontén, Riafo,

30TUN3677, 1000 m, branches of Fagus sylvatica, 01/X/2003, Olariaga, BIO-Fungi

42 ... Olariaga & Salcedo

9914. MADRID: Dehesa de Somosierra, 30T VL5153, 1500 m, buried remnants of Corylus

avellana, 2/X1/2007, J.C. Zamora, BIO-Fungi 12611. asturrAs: Somiedo, 29TQH3373,

1150 m, among litter in acidophilous Fagus sylvatica forest, 27/X/2002, Olariaga, BIO-

Fungi 9711. SWEDEN. GASTRIKLAND: Gavle, Lovudden, on fallen decaying log of

Alnus (2), 10/X1/1957, R. Nannfeldt, UPS F-124234 [as Clavaria contorta]. UPPLAND:

Vange, Fiby urskog, S. Lundell n. 581, 09/IX/1932, UPS F-124218 [as Clavaria fistulosa].

VASTERGOTLAND: Toreboda, Gastorp, asplunden séder om Sommarhemmet, J.

Lundberg 27/X/1955, UPS F-124212 [as Clavaria fistulosa]. SWITZERLAND. vatals:

Mayoux, 30T VL5206, 1500 m, Corylus avellana branch, 31/VII/2007, Felipe & Olariaga,

BIO-Fungi 12610.

Typhula phacorrhiza (Reichard : Fr.) Fr., Observ. Mycol. 2: 298. 1818 : Fr., Syst.

Mycol. 1: 495. 1821, as “phacorrhiza’.

= Clavaria phacorrhiza Reichard : Fr., Schriften Berlin. Ges. Naturf. Freunde 1: 315. 1780.

= Phacorhiza filiformis Grev., Scott. Crypt. Fl. 2: 93. 1824.

LECTOTYPE (designated here): Sowerby, Col. Fig. Eng. Fung. 2: tab. 233. 1798, as

“phacorhiza”.

To our knowledge, the type species of Typhula has not previously been typified.

We select a lectotype according to Articles 9.2 and 9.10 in the forthcoming

“International Code of Nomenclature for algae, fungi, and plants (Melbourne

Code)” (ICN 2012, in prep.). Fries (1821: 495) referred to the lectotype as

“t. 253”, a typographic error for t. 233. The sanctioned epithet spelling (Fries

1821) has been adopted, in agreement with the proposal by Demoulin (2010;

subsequently ratified at the 18th International Botanical Congress, Melbourne,

2011).

SPECIMENS EXAMINED— Without locality, E. Coemans, [Rabenh. Fungi Eur. 418], UPS

F-000023. FRANCE. HAUTES PYRENEES ATLANTIQUES: near Candanchu, 30TYN0041,

1500 m, Chaerophyllum aureum leaves, 10/IX/2004, Olariaga, BIO-Fungi 10521.

SCOTLAND. Fire: Balmuto, 01/X/1822, Berkeley, E 218190 [as Phacorhiza filiformis].

SWEDEN. vASTERGOTLAND: Goteborg, Stora Anggarden, on decaying leaves, Sphagna

etc. in a bog, T. Nathorst-Windahl, 29/IX/1937, [Lundell & Nannfeldt, Fungi Exs. Suec.

550], UPS F-011279. SWITZERLAND. va.alts: Schwartzsee, bare soil under Petasites,

Chaerophyllum aureum and Cirsium, 5/1X/2007, Felipe & Olariaga, BIO-Fungi 12618.

UNITED STATES OF AMERICA. NEw york: Whetzel’s garden, Fores Home, on

buckwheat straw, 17/IV/1937, Remsberg, UPS F-552623.

Typhula tremula (Berthier) Olariaga, comb. nov.

MycoBAank 516732

= Macrotyphula tremula Berthier, Bull. Mens. Soc. Linn. Lyon 43: 187. 1974.

Typhula tremula shares with T: phacorrhiza the characteristics explained above,

as well as slender fruitbodies that make the two species appear very similar

macroscopically.

SPECIMENS EXAMINED—FRANCE. HAUTE-SAVOIE: Hte. Savoie, Samoéns, les Saix, sur

rachis d ‘Athyrium filix-femina, 08/1X/1965, G 110723, [Holotype, LY-Cl 14]. SPAIN.

LEON: Soto de Sajambre. Vegabafio, 30TUN8187, 1500 m, dead remants of Oreopteris

Clavulina, Ramariopsis & Typhula combs. & noms. nov. (Iberia) ... 43

limbosperma, Olariaga, 3/X/2004, BIO-Fungi 10727. sorta: Laguna Negra, 30T WM1649,

1190 m, dead remnants of Athyrium filix-femina, Olariaga, 06/X1/2004, BIO-Fungi

10609. SWEDEN. JAMTLAND: Tannas socken, Svansk6klappen, 31SDE9209, stems of

Athyrium distentifolium, Olariaga, 16/VII/2006, BIO-Fungi 11702. SWITZERLAND.

BERN: Grimselpass, 30TUN8187, 1500 m, on Athyrium distentifolium remnants,

Olariaga, 02/IX/2007, BIO-Fungi 12609, BIO-Fungi 12699.

Acknowledgments

We are grateful to S. Ryman and I. Melo for peer-reviewing this work. We thank

the curators of G (P. Clerc) and MA-Fungi (M. Duefas) for sending us the requested

material on loan. We wish to express our gratitude to Luis A. Parra for his nomenclatural

advice in the course of this work; to John McNeill for supplying the draft text of Articles

9.2 and 9.10 of the Melbourne Code, and to Shaun Pennycook for the thorough

nomenclatural revision that improved the manuscript. We thank S. Arauzo, B. Llamas,

J. Fernandez Vicente, P. Iglesias, J.L. Pérez-Butron, A. Terrén, and specially, P. Pérez

Daniéls and J.C. Zamora, for their assistance in the field trips and providing us with

interesting collections for this study. This work has been supported by a “grant for the

Training of Researchers from the Basque Government (2002/2003).

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MY COTAXON

http://dx.doi.org/10.5248/121.45

Volume 121, pp. 45-52 July-September 2012

A new species of Stigmidium (Mycosphaerellaceae)

on Aspicilia from North America

JANA KOCOURKOVA™ & KERRY KNUDSEN?”

‘Department of Ecology, Faculty of Environmental Sciences, Czech University of Life Sciences

Prague, Kamycka 129, Praha 6 - Suchdol, CZ-165 21, Czech Republic

*The Herbarium, Department of Botany and Plant Sciences, University of California,

Riverside, CA 92521-0124, U.S.A.

CORRESPONDENCE TO: kocourkovaj@fzp.czu.cz*, Knudsen@ucr.edu

ABSTRACT — Stigmidium lendemeri is described from North America on probably five species

of Aspicilia. The new species belongs to the Stigmidium placynthii group in Stigmidium s. lato,

distinguished from Stigmidium s. str. by long 3-to-5 celled periphysoids.

Key worps — lichenicolous fungi, pseudoparaphyses, taxonomy

Introduction

Stigmidium Trevis. 1860 is a genus of predominately lichenicolous taxa

comprising over ninety binominals (Mycobank 2012; Knudsen & Kocourkova

2010; Kocourkova & Knudsen 2009a; Pérez-Ortega et al. 2010) and currently

included in Mycosphaerellaceae (Lumbsch & Huhndorf 2010). The type species

is S. schaereri (A. Massal.) Trevis., a lichenicolous species occurring on sterile

thalli of Solorina species (Roux & Triebel 1994; Crous et al. 2007). Stigmidium

is heterogeneous (Calatayud & Triebel 2003; Kocourkova & Knudsen 2009b).

Stigmidium s. str. has perithecioid ascomata with external periphyses sensu

Roux & Triebel 1994, short 2-celled periphysoids (pseudoparaphyses “Type

a’ sensu Roux & Triebel 1994), and 1-septate ascospores and lacks interascal

filaments (Roux & Triebel 1994; Kocourkova & Knudsen 2009b; Pérez-Ortega

et al. 2010). Stigmidium s. str. comprises at least 24 species (Knudsen &

Kocourkova 2010; Etayo & Osorio 2004; Pérez-Ortega et al. 2010).

A distinct group in Stigmidium differs from Stigmidium s. str. in having

long periphysoids of 3-5 cells (pseudoparaphyses “Type b” sensu Roux

& Triebel 1994), no interascal filaments, and more or less well-developed

external periphyses (Roux & Triebel 1994; Kocourkova & Knudsen 2009b).

This group currently comprises five species: S. clauzadei Cl. Roux & Nav.-Ros.

46 ... Kocourkova & Knudsen

(Roux & Navarro-Rosinés 1994), S. epistigmellum (Nyl. ex Vouaux) Kocourk.

& K. Knudsen (Kocourkova & Knudsen 2009a), S. hesperium Kocourk. et al.

(Kocourkova & Knudsen 2009b), S. placynthii Cl. Roux & Nav.-Ros. (Roux &

Triebel 1994), and S. squamariae (B. de Lesd.) Cl. Roux & Triebel (Roux &

Triebel 1994, 2005). The long periphysoids are of diagnostic value but often hard

to observe, apparently breaking off and dissolving when asci fill the ascomatal

cavity in mature ascomata. Practically speaking, species with long periphysoids

can be keyed out by host and ascospore size after they have been described.

In this paper we describe a sixth species of Stigmidium s. lato with long

periphysoids from North America and found on several Aspicilia species.

Materials & methods

Specimens were supplied by the Cryptogamic Herbarium of the New York Botanical

Garden (NY) and the Lichen Herbarium of the University of California at Riverside

(UCR). Specimens have been examined using standard microscopical techniques.

Hand-made sections were studied in water and 10% KOH [K] and lactophenol cotton

blue [LPB]. Amyloid reactions were tested in Lugol's iodine 1% with and without pre-

treatment with 5% KOH [K/I]. Metachromatic reactions were tested with 1% solution of

brilliant cresyl blue [BCr]. Ascospore measurements were made in water with accuracy

of 0.5 um and given in the form “(minimum-) mean minus standard deviation-

mean-mean plus standard deviation (-maximum)” and followed by the number of

measurements (n); the length/breadth ratio of ascospore is indicated as 1/b and given

in the same form.

Macro and microphotographs were taken with a digital camera Olympus DP72

on Olympus SZX 7 Stereomicroscope and Olympus BX 51 fitted with a Nomarski

differential interference contrast.

The species

Stigmidium lendemeri Kocourk. & K. Knudsen, sp. nov. PLATE 1 & 2

MycoBank MB 518075

Differs from other members of the Stigmidium placynthii group by its larger ascospores

and its Aspicilia hosts.

Type: U.S.A. Pennsylvania. Lackawanna Co., Moosic Mountains, Montage Heath

Barrens, sandstone and conglomerate outcrops on west slope, 41°21'05"N 75°40'18" W,

396-426 m, on Aspicilia species, Sept. 30, 2005, Lendemer 5544 w/ Schuyler (NY,

holotype).

ErymMoLoecy: ‘The species is named for the North American lichenologist James

C. Lendemer (NY), who brought the taxon to our attention, in honor of his work

on the lichen biota of North America, on Lepraria, and on the journal OpUsCULA

PHILOLICHENUM. We value him as both a colleague and good friend.

INFECTION notvisible. MyCELIAL HYPHAE not observed. ASCOMATA perithecioid,

black, globose to subglobose, ostiolate, 60-100(-120) um high and 60-100(-

120) um wide, partially immersed in the host areoles, usually dispersed, one

Stigmidium lendemeri sp, nov. (USA) ... 47

Pia »

PiaTE 1. Fics A-D. Stigmidium lendemeri (Lendemer 5544, holotype). A, Infected thallus of

Aspicilia sp. B, Ostiole with fringe of external periphyses. C, D Ascomatal section with pendent

periphyses (pseudoparaphyses “Type b”). Scale bars: A = 1 mm; B-D = 20 um.

48 ... Kocourkova & Knudsen

or two per areole, causing no visible sign of infection but infected areoles not

usually producing apothecia. ExcrpLe dark brownish-black in upper 1/5-1/3,

lighter brown or reddish brown in lower 4/5-2/3, but still dark even in thin

sections, never hyaline, 10-15 um thick in optical section, often thickest (up to

15 um) in ostiolar area, of 2-4 compressed cell layers, textura angularis, cells

mostly 4-9 x 3.5-4 um in optical section, I-, BCr-. EXTERNAL PERIPHYSES

visible as a dense fringe when viewed from above in mature ascomata, derived

from ascomatal wall, 8-14 um long, brown at base to pale yellowish-brown

or almost hyaline at tip, mostly 1-1.5 um wide, septate, cells 3-5 um long,

comprised of 2 to 3 cells, the base cell usually widest. HaMATHECIUM of

pendant periphysoids (pseudoparaphyses “Type b”) originating from the upper

wall of ascomatal cavity, hyaline, rarely branching, 12-15(-20) um long, mostly

2 um wide, septate, formed of 3 or more cells, cells 3-5 um long, uneven in

length. Lacking interascal filaments. Hymenial gel I-, K/I-, BCr+ light violet.

Ascl originating from lower wall of ascomatal cavity, fissitunicate, narrowly

saccate, endoascus thickened in upper 1/3, sessile to stipitate, 8-spored, with

ascospores distichously arranged, 20-30 x 10-12(-15) um, I-, ascoplasma

I+ red, exoascus BCr-, endoascus BCr+ blue-violet. Ascospores 1-septate,

hyaline, brownish when over-mature and with 1 pseudoseptum (possibly

also pseudotetrablastic but not seen), non-halonate, not constricted at septa,

when 2-celled, cells usually equal in length, (14.0-)15.5-17.3-19.0(-21.0) x

(4.0-)5.0-5.8-6.5(-7.0) um (n = 40), I/b = (2.3-)2.6-3.0-3.4(-4.5); epispore

BCr+ dark blue; perispore BCr+ very pale violet. Contpiomata 30-45 um in

diam., conidiogenous cells ampuliform 4.5-5.0 high, 4.0-4.5 um wide, conidia

cylindrical, strait or slightly curved, 4.5-6.0 x 1.0-1.5 um.

SUBSTRATE AND ECOLOGY — On probably five different Aspicilia species

with stictic or norstictic acid or lacking extrolites that typically occur on

non-calcareous rocks in open habitats (talus slopes, glades, rock ledges) with

constant high humidity and on boulders in or near rivers or streams. We are

unsure of the exact Aspicilia species in many cases as the genus is in need of

revision. In a few cases the lack of conidia precluded accurate identification of

the host.

DISTRIBUTION — North America (California, Missouri, Ohio, Pennsylvania).

Because it is host specific on genus level on multiple species we expect more

reports from throughout North America. A taxon with similar ascospores

causing a black infection was seen from California (UCR) and Kansas (NY)

and needs further study.

PLATE 2. Fics E-I. Stigmidium lendemeri (Lendemer 5544, holotype). E. Pycnidium, conidiogenous

cells and conidia. F Young ascus with apical typically short beak. G. Ascus with ascospores.

H. Ascus releasing content. I. Nearly mature and mature ascospores. Scale bars: 20 um.

Stigmidium lendemeri sp, nov. (USA) ... 49

50 ... Kocourkova & Knudsen

SELECTED SPECIMENS EXAMINED — U.S.A. CALIFORNIA. MARIPOSA Co., Sierra Nevada

Mountains, Yosemite National Park, Merced River near Highway 140, 37°40'19"N

119°47'32"W, 653 m, on Aspicilia species (no extrolites) on boulders in river, Sept. 20,

2009, Knudsen 11680 (UCR); TUOLUMNE Co., Sierra Nevada Mountains, Yosemite

National Park, Inspiration Point along Hetch Hetchy Road, Poopanaut, 37°54'12"N

119°50'01" W, 1601 m, on Aspicilia species (no extrolites), Sept. 19, 2009, Oregon State

University Survey, Knudsen 11556 (UCR); Missouri. IRON Co., Ketcherside Mountain

Conservation Area, Royal Gorge Nature Area, Big Creek, 37°32'21"N 90°41'01" W, on

sterile Aspicilia species (stictic acid), March 26, 2006, Buck 49747 (UCR); SHANNON Co.,

Ozark National Scenic Riverways, Rocky Creek, mesic forest, 37°07'N 91°12'W, 200-215

m, on Aspicilia species (norstictic acid) on rhyolite, April 16, 1997, Buck 31872 (NY);

Wayne Co., Sam A. Baker State Park, Mudlick Mountains, 37°15'27"N 90°31'23”W, 300

m, on Aspicilia species (stictic acid), Oct. 15, 2003, Buck 45324 (NY); WASHINGTON Co.,

Mark Twain National Forest, Little Pilot Knob, 37°58'57"N 90°58'40"W, on Aspicilia

species (stictic acid), May 24, 2003, Harris 47935 (NY); Onto. ScroTo Co., along Pond

Lick Run, Shawnee State Forest, sandstone outcrops along stream in oak woodland,

38°42'20"N 83°08'25"W, 198-213m, on Aspicilia species, May 21, 2006, Lendemer 7175

(NY); PENNSYLVANIA. BEDFORD Co., Pleasant Valley, Buchanan State Forest, west-

facing slope with granite and sandstone boulders, 40°03'18"N 78°30'43"W, 518-548 m,

on sandstone, on Aspicilia species, May 18, 2006, Lendemer 7294 (NY); BERKS Co., west

slope of Mt. Pleasant, French Creek State Park, 40°11'33"N 75°47'10"W, 213-243 m,

on Aspicilia species, Nov. 19, 2006, Lendemer 8036 (NY); Bucks Co., Ringing Rock

County Park, diabase boulder field along stream, 40°33'43"N 75°07'42"W, 137-152 m,

on Aspicilia species, Sept. 15, 2005, Lendemer 4982 w/ Schulyer (NY); Carbon Canyon,

east shore of Leigh River, Leigh Gorge State Park, cliff along railroad track, 40°57'58"N

75°42'22"W, 304 m, on cliff, on Aspicilia cf. laevata, Nov. 9, 2003, Lendemer 1491 w/

Rhoads (NY).

Discussion

The presence of long periphysoids with 3 or more cells places Stigmidium

lendemeri in the S. placynthii group (Roux & Triebel 1994) in which we

currently recognize five other species. “The small differences in ascospores

size are assumed to be an important morphological expression of phylogenetic

distance.” (Kocourkova & Knudsen 2009b). These small differences (compared

with S. lendemeri) are summarized below, but in addition all differ in the host

genera and species infected.

(1) Stigmidium placynthii on Placynthium nigrum has smaller halonate

ascospores, (7.5-)9.5-11.1-12(-13.5) x 3.0-3.3-3.5(-4.0) um.

(2) Stigmidium epistigmellum on maritime Caloplaca species has narrower

ascospores, (14.5—)15.8-17.5-19.1(-21.5) x (3.5—-)3.9-4.2-4.7(-5.0) um.

(3) Stigmidium squamariae on several Lecanora and Rhizoplaca species has

shorter halonate and constantly hyaline ascospores, (8.5—)9-10.9-

12.5(-14) x (4-)5-5.3-5.5(-6) tum, and lacks reactions with BCr.

(4) Stigmidium clauzadei on Verrucaria nigrescens and V. viridula

has generally shorter and wider 1-septate hyaline and halonate

Stigmidium lendemeri sp, nov. (USA) ... 51

ascospores, (10)-12.5-15.1-19 x (4.5)5-5.5-6.5(-7.5) tum,

and all parts BCr- except perispore BCr+ pale blue.

(5) Stigmidium hesperium on Caloplaca species has shorter and narrower

ascospores, (12.5—)13.0-13.75-14.5(-15.5) x (3.5-)4.0-4.5-5.0(-5.5) um.

The only Stigmidium previously reported on Aspicilia is Stigmidium aggregatum

[= Pharcidia aspiciliae| on Circinaria calcarea [= Aspicilia calcarea], which has

much larger ascospores, 22-30 x 4.5-9 um and is apparently known only

from two lost types (Kocourkova & Knudsen 2010). Based on our study of the

problem, S. aggregatum may not even be a Stigmidium.

Acknowledgments

We thank our reviewers, David Hawksworth (UK), M. Gokhan Halici (Turkey), and

M. Zhurbenko (Russia) for their valuable comments. The work of Jana Kocourkova was

supported financially by the grant “Environmental aspects of sustainable development

of society” (42900/1312/3166) from the Faculty of Environmental Sciences, Czech

University of Life Sciences Prague.

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on Acarospora and Squamarina. Lichenologist 35: 103-116. http://dx.doi.org/10.1016/S0024-

2829(02)00097-X

Crous PW, Braun U, Groenewald JZ (2007). Mycosphaerella is polyphyletic. Studies in Mycology

58: 1-32. http://dx.doi.org/10.3114/sim.2007.58.01

Etayo J, Osorio HS. 2004. Algunos hongos liquenicolas de Sudamérica, especialmente del Uruguay.

[Some lichenicolous fungi from South America, especially from Uruguay]. Comunicaciones

Botanicas Museos Nacionales de Historia Natural y Antropologia 6 (129): 1-19.

Knudsen K, Kocourkova J. 2010. A new species of Stigmidium from corticolous Caloplaca in

southern California (U.S.A.) and Baja California (Mexico). Bibliotheca Lichenologica 105:

29-31,

Kocourkova J, Knudsen K. 2009a. Stigmidium epistigmellum (Mycosphaerellaceae), a lichenicolous

fungus from maritime Caloplaca in North America. Bryologist 112: 578-583.

Kocourkova J, Knudsen K. 2009b. A new species of Stigmidium (Mycosphaerellaceae) from western

North America. Czech Mycology 61: 73-80.

Kocourkova J, Knudsen K. 2010. Stigmidium eucline is not a synonym of Stigmidium aggregatum.

Opuscula Philolichenum 8: 101-105.

Lumbsch HT, Hundorf 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

MycoBank. 2012. Fungal databases: nomenclature and species banks: online taxonomic novelties

submission. Administered by the International Mycological Association. http://www.mycobank.

org/ (Accessed 25 March 2012).

Pérez-Ortega S, Halici MG, Knudsen K, Candan M. 2010. A new species of Stigmidium sensu

stricto on Thelenella muscorum. Lichenologist 42: 397-403. http://dx.doi.org/10.1017/

$0024282910000150

Roux C, Navarro-Rosinés P. 1994. Stigmidium clauzadei sp. nov., nelikeniginta fungo likenloga

(Ascomycetes). Bulletin de la Société linnéenne de Provence 44: 443-450.

52 ... Kocourkova & Knudsen

Roux C, Triebel D. 1994. Révision des espéces de Stigmidium et de Sphaerellothecium (champignons

lichénicoles non lichénisés, Ascomycetes) correspondant a Pharcidia epicymatia sensu Keissler

ou a Stigmidium schaereri auct. Bulletin de la Société linnéenne de Provence 45: 451-542.

Roux C, Triebel D. 2005. Ehamathécium de Stigmidium squamariae, ascomycete lichénicole non

lichénisé - conséquences systématiques. Mycotaxon 91: 133-136.

MY COTAXON

http://dx.doi.org/10.5248/121.53

Volume 121, pp. 53-62 July-September 2012

Notes on some Eurasian species of Anthracoidea and Entyloma

KyRYLO G. SAVCHENKO’”*, VASYL P. HELUTA?,

IVAN S. HIRYLOVICH?, SOLOMON P. WASSER?” & EVIATAR NEVO!

"Department of Evolutionary and Environmental Biology and the Institute of Evolution,

Faculty of Natural Sciences, University of Haifa, Mt. Carmel, Haifa 31905, Israel

°M.G. Kholodny Institute of Botany of the National Academy of Sciences of Ukraine,

2 Tereshchenkivska St., Kyiv 01601, Ukraine

*Belorusian State University, 4 Nezavisimosti Av., Minsk 220030, Belarus

* CORRESPONDENCE TO: savchenko.kyryll@gmail.com

ABSTRACT — Newrecords of smut fungi representing Anthracoidea and Entyloma from Israel,

Belarus, Kyrgyzstan, and Russia are presented. Each record is followed by a comprehensive

description and illustrations based on original material. Additionally, a revision of Entyloma/

Ranunculus records from Israel is given.

Key worps — Anthracoideaceae, biodiversity, Entylomatales, Middle East

Introduction

Five new records of Anthracoidea species are reported, three for Belarus, one

for Kyrgyzstan, and one for Russia; and three new records of Entyloma species,

one new for Belarus and two for Israel. In addition, records of Entyloma on

Ranunculus in Israel are revised.

Materials & methods

Sorus and spore characteristics were studied using dried herbarium material. The

specimens were examined by light microscopy (LM) and scanning electron microscopy

(SEM).

The sorus structure was studied under a Carl Zeiss Stemi Dv4 stereo microscope.

For light microscopy (LM), spores were mounted in lactic acid, gently heated to the

boiling point and cooled, and then examined at 1000x under a Carl Zeiss Axiostar light

microscope. LM photographs were taken with a Canon Power Shot G10 camera. At

least 50 spores were measured from each collection, and the variation is presented as a

range, with extreme values given in parentheses. In the descriptions, mean and standard

deviations (SD) calculated from spores measured in all specimens are listed after spore

size ranges. For scanning electron microscopy (SEM), spores were attached to metal stubs

by double-sided adhesive tape and coated with gold. The surface structure of spores was

54 ... Savchenko & al.

observed at 15 kV and photographed with a scanning electron microscope JEOL JSM-

6700F. Specimens used in this study are stored in the Herbarium of Haifa University

(HAI), Herbarium of the Hebrew University of Jerusalem (HUJ), National Herbarium

of Ukraine (KW), and HeKS, a personal, working collection of Kyrylo Savchenko.

Taxonomy

Anthracoidea angulata (Syd.) Boidol & Poelt, Ber. Bayer. Bot. Ges. 36: 23 (1963)

Fics. 1-2

Sort in the ovaries of the inflorescences, forming black, globose, subglobose

bodies around the achenes, 2-3 um in diameter, dusty on the surface. SPORES

medium-sized, flattened, medium reddish brown, angular, irregular, elongated,

(15-)17-20(-22) x 13-19(-20) um [av. + SD, 18 + 1.5 x 16.5 + 2 um]. SPORE

WALL evenly to unevenly thickened, ca. 1.2-2.5 um thick, thickest at the angles,

usually with several protuberances, light refractive spots and internal swellings;

surface finely verruculose with densely situated, rounded warts. Spore profile

wavy.

DISTRIBUTION: Europe, Asia.

SPECIMENS EXAMINED: BELARUS. Minsk region, Minsk district, near Volchkovichi,

oak-grove, 20.VII.2005, on Carex hirta L., leg. LS. Hirylovich (HAI 2864, HeKS 223);

Smolevichi district, forest near Zhazhelka, 25.VII.2005, on C. hirta, leg. I.S. Hirylovich

(HAI 2865). Mogilev region, Osipovichi district, near Daraganovo, on the railway

embankment, on C. hirta, 18.VII.2010, leg. 1.S. Hirylovich (HAI 2866).

Note: Anthracoidea angulata is a new species for the Belarusian mycobiota.

Anthracoidea echinospora (Lehtola) Kukkonen, Ann. bot. Soc. Zool.-Bot. fenn.

“‘Vanamo’ 34(3): 72 (1963) Fics. 3-4

Sor! in the ovaries of the inflorescences, forming carbonaceous, globose,

subglobose bodies around the achenes, 2-3 um in diameter, covered by a thin

silvery-greyish membrane; during maturation the spore mass cracks down.

SPpoRES small, flattened, olivaceous-brown to brown, subglobose, ovoid,

irregular, sometimes elongated, (12-)13-20(-22) x (11-)12-15(-17) um

[av. + SD, 16 + 2.5 x 13.4 + 2.1 um]. SPORE WALL evenly thickened, ca. 1 um,

without any light refractive spots or internal swellings; surface echinate with

irregularly dispersed, apically enlarged and flattened, often confluent spines

< 1.5 um high. The wall between the spines striate-rugulose; the base of the

warts longitudinally crumpled. On the tips of the warts the remnants of a thin

membrane cover some parts of the spore surface.

DISTRIBUTION: Europe, Asia.

SPECIMEN EXAMINED: BELARUS. Gomel region, Zhlobin district, near Zhlobin, on

Carex acuta L., 18.V1I.2009, leg. I.S. Hirylovich (HAI 2867, HeKS 224).

Note. Anthracoidea echinospora is a new species for the Belarusian

mycobiota.

Anthracoidea & Entyloma spp. new for Eurasia ... 55

IS.0kKV X2,300 104m WD 8.0:

Z barr iy

a i * J XS-666 Sem O616

os =

Fics 1-6. 1-2: spores of Anthracoidea angulata on Carex hirta. 3-4: spores of A. echinospora on

C. acuta. 5-6: spores of A. heterospora on C. nigra. 1,3,5 = LM; 2, 4,6 = SEM. Scale bars: 1-3, 5 = 10

um; 4 = 2 um; 6 = 5 um.

56 ... Savchenko & al.

Anthracoidea elynae (Syd.) Kukkonen, Ann. bot. Soc. Zool.-Bot. fenn. “Vanamo’

34(3): 65 (1963) Figs. 7-9

Sort in the ovaries of the inflorescences, forming black, globose, hard bodies

around the achenes, 1.5-2.5 mm in diameter, dusty on the surface, partly hidden

by the perigynium. Spores medium-sized, flattened, medium to dark reddish

brown, subglobose, disc-shaped, (15-)16.5-19.5(-20.5) x (13-)14-18(-19) um

[av. + SD, 18.2 + 1.6 x 15.9 + 2.2 um]. Hyaline sheaths on the flat sides of the

spores present. SPORE WALL evenly thickened, ca. 1-2 um thick, thickest at the

angles, no light-refractive areas, internal swellings were not observed; surface

almost smooth to finely verruculose on the flat side. Spore profile smooth.

DISTRIBUTION: Europe, Asia, North America.

SPECIMENS EXAMINED: KYRGYZSTAN. Issyk Kul Province, Ak-Suu district, near

Karakol, on Kobresia capillifolia (Decne.) C.B. Clarke, 5.VIII.1933, leg. M.V. Klokov

(KW 37520).

Note. Anthracoidea elynae is a new species for Kyrgyzstan.

Anthracoidea heterospora (B. Lindeb.) Kukkonen, Ann. bot. Soc. Zool.-Bot. fenn.

“‘Vanamo’ 34(3): 63 (1963) Fics. 5-6

Sort in the ovaries of the inflorescences, forming black, globose, subglobose

to ovoid bodies, 1-3 mm in diameter, powdery on the surface. SpoREs small,

flattened, medium reddish brown, globose, subglobose, angular, (10-)12-19

(-21) x (10-)11-17 um [av. + SD, 16 + 2.5 x 13.2 + 2.1 um]. SPORE WALL ca.

1-2 um thick, thickest at the angles, usually with several internal swellings;

surface finely and densely verruculose with densely situated, rounded, often

confluent low warts. Spore profile slightly wavy.

DISTRIBUTION: Europe, Asia.

SPECIMENS EXAMINED: BELARUS. Minsk region, Minsk district, near Zacep,

27.VI1I.2008, on Carex nigra (L.) Reichard, leg. I.S. Hirylovich (HAI 2871); Valozhyn

district, near Krazhino, on the forest edge, on C. nigra, 22.V1.2005, leg. I.S. Hirylovich

(HAI 2868, HeKS 225).

Note. Anthracoidea heterospora is a new species for the Belarusian mycobiota.

Anthracoidea tomentosae Vanky, Bot. Not. 132: 227 (1979) Fras. 10-12

Sort in the ovaries of the inflorescences, forming black, globose, subglobose,

hard bodies around the achenes, 1-2 mm in diameter, when young covered

by a white-silvery membrane; spore mass agglutinated to semi-agglutinated,

partly hidden by the perigynium. Sporgs large, flattened, medium to dark

reddish brown, subglobose, sub-angular, irregular, (19-)21-27(-28) x

(14-)15.5-22(-24) um [av. + SD, 23.5 + 2.3 x 18.1 + 2.6 um]. SPORE WALL

unevenly thickened, ca. 1.5-2.5(-3.5) um wide, thickest at the angles, usually

with several protuberances, light refractive spots and internal swellings

sometimes present; surface finely punctate-verruculose with densely situated,

Anthracoidea & Entyloma spp. new for Eurasia ... 57

13,66o

x37 BBB Shr &

Fics 7-12. 7-8: spores of Anthracoidea elynae on Kobresia capilliformis. 9: spore surface of

A. elynae on K. capilliformis. 10-11: spores of A. tomentosae on Carex tomentosa. 12: spore surface

of A. tomentosae on C. tomentosa. 7, 10 = LM; 8, 9, 11, 12 = SEM. Scale bars: 7, 8, 10 = 10 um;

9,12=1 um; 11 =5 um.

58 ... Savchenko & al.

often confluent, 0.2-0.5 um high, rounded warts. The spaces between the warts

minutely verruculose. Spore profile serrulate.

DISTRIBUTION: Europe, Asia.

SPECIMEN EXAMINED: RUSSIA. Krasnodar Krai, Gelendzhik district, near Kabardinka,

on Carex tomentosa L., 15.V1.1986, leg. N. Chernov (KW 37611).

Note. Anthracoidea tomentosae is a new species for the Russian mycobiota.

Entyloma ficariae A.A. Fisch. Waldh., Bull. Soc. nat. Moscou, Biol. 52: 309 (1877)

Fics. 13-15

Sor! (Fic. 13) in leaves as flat, round, or angular spots, 2-7 mm in diameter,

first white, later pale creamy-brown. SporEs (FIGs. 14-15) globose, subglobose,

ovoid, (11—)12-15(-16) x (9-)10-13(-15) um [av. + SD, 14+ 2.1 x 11+ 1.8 um],

subhyaline. SPORE WALL even, smooth, ca. 1-2.5 um thick. ANAMORPH present

on both sides of leaves.

DISTRIBUTION: Europe, Asia, Africa.

SPECIMEN EXAMINED: ISRAEL. Golan Heights, Hermon National Park, 400 m

northeast from Neve Ativ, 1180 m. alt., 32°26'45"N 35°74'83"E, on Ranunculus ficaria

L., 26.IV.2011, leg. K.G. Savchenko (HAI 2863, HeKS 228; GENBANK JQ586199).

Note. Entyloma ficariae is a new species for the Israeli mycobiota.

The closely related E. majewskii Vanky & M. Lutz is also distributed in the

Irano-Turanian floristic region, particularly in Iran, but differs from E. ficariae

by thicker spore walls (< 7 um), the absence of the anamorph, and bullate

rather than flattened sori (Vanky & Lutz 2010). Molecular analyses confirmed

the assignment of the specimen from Mount Hermon to E. ficariae.

Entyloma gaillardianum Vanky, Mycotaxon 16: 104 (1982) Figs. 19-21

Sori in leaves, as rounded, circular spots, 1-5 mm in diameter, or larger

by confluence, first pale yellowish-green, later brownish-green, with a thin

yellowish margin around the spots. Spores globose, subglobose, irregular,

subhyaline to lemon yellow, 10-14 x 9-13(-14) um [av. + SD, 12.5 + 1.6 x 11.4

+ 1.4 um]. SPoRE WALL 2-layered, ca. 1-3 um. Spore surface smooth.

DISTRIBUTION: worldwide.

SPECIMEN EXAMINED: ISRAEL. Upper Jordan Valley, Tiberias, 20.IV.2005, on cultivated

Gaillardia aristata Pursh, leg. S. Voytyuk (HAI 2869, HeKS 229).

Note. Entyloma gaillardianum is new for the Israeli mycobiota and probably

was imported into the country with Gaillardia plants.

Entyloma hieracii Syd. & P. Syd. ex Cif., Bull. Soc. Bot. Ital. 1924: 50 (1924)

Fics. 22-24

Sor! in leaves, as rounded, angular spots, 1-7 mm in diameter, first pale

yellowish-white, later yellowish-brown, with a thin pale green margin around

Anthracoidea & Entyloma spp. new for Eurasia ... 59

oo Sem @645)

Fics 13-18. 13: sori of Entyloma ficariae on Ranunculus ficaria. 14-15: spores of E. ficariae on

R. ficaria. 16: sori of E. microsporum on Ranunculus asiaticus. 17-18: spores of E. microsporum

on R. asiaticus. 14, 17 = LM; 15, 18 = SEM. Scale bars: 13, 16 = 5 mm; 2, 3, 5,6 =5 um.

60 ... Savchenko & al.

SEI 15.0kV X2,300 10pm WD 7.8mm

Fics 19-24. 19: sori of Entyloma gaillardianum on Gaillardia aristata. 20-21: spores of

E. gaillardianum on G. aristata. 22-23: sori of E. hieracii on Hieracium sylvularum, adaxial and

abaxial sides respectively. 24: spores of E. hieracii on H. sylvularum. 20, 24 = LM; 21 = SEM.

Scale bars: 19 = 10 um; 20, 21, 24 = 10 um; 22 = 4 mm; 23 = 3 mm.

Anthracoidea & Entyloma spp. new for Eurasia ... 61

the spots. Spores globose, subglobose, irregular, sub-hyaline to yellow, (9-)

10-15(-16) x 8-12(-14) um [av. + SD, 12.3 + 2.5 x 9.9 + 1.9 um]. SPORE WALL

even, ca. 1-3 um wide. Spore surface smooth.

DISTRIBUTION: Europe, Asia, North America.

SPECIMEN EXAMINED: BELARUS. Minsk region, Minsk district, near Volchkovichi, oak-

grove, on Hieracium sylvularum Boreau, 20.VII.2005, leg. I.S. Hirylovich (HAI 2870,

HeKS 230).

Note. Entyloma hieracii is a new smut fungus for the Belarusian mycobiota,

and H. sylvularum is a new host for this species.

Entyloma microsporum (Unger) J. Schrét., in Rabenhorst, Fungi Europ. Exsicc.

no. 1872 (1874) Figs. 16-18

Sort in leaves, as wart-like swellings, different in shape, 1-5 mm in diameter,

first yellowish-white, when dry adaxially brownish, abaxially ocher-brown,

swollen, with cracked surface. Spores densely situated, globose, subglobose,

irregular, subhyaline to light yellowish, (12—) 14-19(-21) x (10-)12-17(-18)

um [av. + SD, 16 + 2.1 x15 + 1.8 um], with granular matrix. SPORE WALL 2-

layered, with yellow, even, ca. 0.5 um thick inner layer, and hyaline, uneven,

ca. 1-6 um thick outer layer. Spore surface smooth, often covered by separated

remnants of spore walls.

DISTRIBUTION: worldwide.

SPECIMENS EXAMINED: ISRAEL. Samaria, Wadi Fedjaz, 24.11.1957, on Ranunculus

asiaticus L., leg. A. Grizi (HUJ); Upper Galilee, Mount Meron (= Mount Jermak), on R.

asiaticus, 27.11.1951, leg. C. Rayss (HUJ, as E. ranunculorum).

Note. This taxon has previously been recorded from Israel under the names

Entyloma microsporum and “E. ranunculorum” (Rayss 1952, 1959; Savchenko

et al. 2010). Although the invalid name “E. ranunculorum” Liro is usually

equated with E. ranunculi-repentis Sternon (Vanky 2011: 201-202), accurate

microscopical examination of soral structure and spore morphology indicated

that the Mount Meron collection should be assigned to E. microsporum. The

Mount Meron collection has smaller spores [(12-)12.5-17(-18) x (10.5-)

11-16(-17) um] with thinner walls [1.5-5 um], compared with the Samaria

collection [(13-)13.5-19.5(-21) x (11-)13-17.5(-18) um, with 2.5-6 um

walls].

Acknowledgments

We thank Matthias Lutz and Marcin Piatek for peer-reviewing the manuscript,

Matthias Lutz for confirming our identification of Entyloma ficariae with molecular

methods, Shaun Pennycook for a number of useful corrections, curators of the

herbaria HUJ and KW for loaning specimens, and Vitalii Sapsai for help with the SEM

microscopy.

62 ... Savchenko & al.

Literature cited

Rayss T. 1952. Etudes de quelques Ustilaginées récoltées en Palestine. Palestine Journal of Botany

Jerusalem Series 5: 229-236.

Rayss T. 1959. Quelques additions a la mycoflore d’Israel. Bulletin Research Council of Israel 8D :

1-14.

Savchenko KG, Heluta VP, Wasser SP, Nevo E. 2010. Smut fungi of Israel: a preliminary check-list.

Mycologia Balcanica 7: 111-116.

Vanky K. 2011 (“2012”). Smut fungi of the world. American Phytopathology Society, St. Paul MN.

1458 p.

Vanky K, Lutz M. 2010. Entyloma majewskii sp. nov. (Entylomataceae) on Ranunculus ficaria from

Iran. Polish Botanical Journal 55(2): 271-279.

MY COTAXON

http://dx.doi.org/10.5248/121.63

Volume 121, pp. 63-67 July-September 2012

Lactarius annulocystidiatus sp. nov. from India

SAMIDHA SHARMA, MUNRUCHI KAUR & N. S. ATRI*

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

*CORRESPONDENCE TO: narinderatri04@yahoo.com

AsstRAcTt — Lactarius annulocystidiatus, a new species of subgenus Tristes, is found

associated with Quercus leucotrichophora. It is characterized by its granular pleurocystidia,

with granulations clustered in annular rings around the cystidial tips. Macro- and microscopic

features and data on ecological distribution are presented.

KEY worps — ectomycorrhizal, macrofungi, Russulaceae, taxonomy

Introduction

Lactarius Pers. is an agaricoid member of family Russulaceae, represented by

450 species worldwide (Kirk et al. 2008) and approximately 71 taxa from India

(Atri et al. 1994, Natarajan et al. 2005, Das & Sharma 2005). The macroscopic

and microscopic characters of a collection from Himachal Pradesh were

compared with previously described species (Pearson 1950, Hesler & Smith

1979, Eberhardt & Verbeken 2004, Das et al. 2004, Das & Sharma 2005, Buyck

et al. 2007, Le et al. 2007, Wang 2007, Stubbe et al. 2008, Montoya & Bandala

2008, Stubbe et al. 2010, Das & Verbeken 2011, 2012) and found to be sufficiently

different to warrant description as a new species.

Material & methods

The macromorphology of the examined collection recorded on gross external

morphology and micromorphology was studied in accordance to the methodology

given by Atri et al. (2005). The terminology used for describing the color tone of the

carpophore parts and spore print is after Kornerup and Wanscher (1978). Microscopic

line drawings were made with the aid of a camera lucida at 1000x. Basidiospore

measurement excludes the height of ornamentation and length of apiculus. Basidium

length excludes the length of sterigmata. The spore shape quotient (Q=L/W) was

calculated considering the mean value of length and width of 20 basidiospores. The

holotype specimen has been deposited in the Herbarium of Botany Department (PUN),

Punjabi University.

64 ... Sharma, Kaur & Atri

Taxonomy

Lactarius annulocystidiatus S. Sharma, M. Kaur & Atri sp. nov. Fics 1-6

MycoBANkK MB 562761

Differs from Lactarius argillaceifolius by its smaller basidiospores and smaller

pleurocystidia with granulations in annular rings towards the tips.

Type: India, Himachal Pradesh: Andretta, solitary carpophore on humicolous soil

in association with Quercus leucotrichophora A. Camus (Fagaceae), 31 August 2009,

Samidha 4314 (PUN, holotype)

Erymo oey: The species epithet is based on the presence of granulations in an annular

pattern around the cystidial tips.

Fructification < 6.5 cm in height. Pileus < 6 cm broad, convex, feebly zoned

concentrically around the periphery, shiny viscid; margin regular, unbearded;

surface light yellow (4A,), olivaceous to spotted pinkish to reddish brown;

viscid; apex depressed; surface browning on bruising. Latex grayish milky,

unchanging, mild; cuticle not peeling; flesh < 0.2 cm broad, light yellow (4A,),

brownish on exposure, mild. Lamellae adnexed, unequal, crowded, light yellow

(4A), brownish on bruising, narrow (0.3 cm broad), edges smooth, fragile.

Taste mild, odour fruity. Stipe central, 4 cm long, 2 cm broad above and 1.2 cm

broad below, tapering downwards, concolorous with the pileus, slimy viscid,

brownish on bruising, hollow, smooth.

Basidiospores 5.7-8.6 x 5.7-7.2 um, globose to broadly ellipsoidal (Q =

1.0-1.2), warty, warts < 1.4 um high, strongly amyloid, ornamentation in the

form of coarse banded ridges forming incomplete reticulation and some warts

isolated, at places 2-3 warts connected, ornamentation type I, IIb, IV, VI (sensu

Singer, 1986), plage amyloid and distinct, apiculate; apiculus < 1.4 um long.

Basidia 40-50 x 5.7-8.6 um, 4-spored, clavate, hyaline to densely granular,

6 basidia/100 um, abundant, sterigmata 2.9-7.2 um long. Pleurocystidia:

macrocystidia 43-71.5 x 5.7- 11.4 um, granular, thick walled, clavate, fusoid

to ventricose in shape with moniliform, flame-shaped, capitate to mucronate

or tubular pointed tips, granulated with granular contents aggregated in rings

mostly towards the tips, which are prominently present around and near the

gill edges; pseudocystidia 57.2-65.8 x 5.7-7.2 um, vermiform, deeply seated,

not projecting beyond basidia. Cheilocystidia 32.9-38.6 x 5.7-7.2 um, similar

in details to pleurocystidia. Hymenophoral trama hyphal, irregularly arranged,

subhymenium indistinct. Pileus cuticle 150-200 um thick, partially gelatinized,

made up of horizontally running, branched closely septate 4.3- 5.7 um broad

hyphae, some ampulliform, some H-shaped with rectangular cells; pilear trama

made up of rosettes of sphaerocysts intermingled with < 5.7 um broad hyphae

and laticifers. Stipe cuticle gelatinized, made up of longitudinally running

<4.3-7.2 um broad hyphae; stipe trama made up of rosettes of sphaerocysts

intermingled with < 5.7 um broad hyphae. Laticifers present throughout the

context of carpophores. Clamp connection absent.

Lactarius annulocystidiatus sp. nov. (India) ... 65

4 5

Figs 1-5. Lactarius annulocystidiata: microscopic structures.

1. Carpophore; 2. basidiospores; 3. basidia; 4. pleurocystidia; 5. cheilocystidia

REMARKS— Lactarius annulocystidiatus is characterized by its viscid, light

yellow pileus surface with pinkish to reddish brown tinge, unbearded margins,

grayish milky mild unchanging latex in young fruit body and all parts of the

66 ... Sharma, Kaur & Atri

Fic. 6. Lactarius annulocystidiata in situ.

fungus becoming brownish on bruising. These features are typical of subgenus

Tristes, section Pseudomyxacium, stirps Argillaceifolius (sensu Hesler & Smith,

1979). It seems to be a close relative of L. argillaceifolius Hesler & A.H. Sm.

var. argillaceifolius, which can be separated in the field by its pubescent margin

and decurrent closely placed lamellae. Microscopically, the basidiospores of

L. argillaceifolius var. argillaceifolius are larger [(7—)8-10(-11) x 7-8 um], the

pleurocystidia and cheilocystidia, which are larger and clavate, lack granulations

around the tips, and the pileus surface lacks ampulliform, H-shaped branched

hyphae.

Acknowledgements

The authors thank Dr. Kanad Das, Botanical Survey of India, Sikkim Himalayan

Regional Centre, Gangtok, India and Prof. B.M. Sharma, Department of Plant Pathology,

COA, CSKHPAU, Palampur, H.P., India for peer review and Dr. Shaun R. Pennycook,

Manaaki Whenua Landcare Research, Auckland, New Zealand, for nomenclature

review. Head, Department of Botany, Punjabi University, Patiala, for providing research

facilities. We are indebted to UGC & DST, New Delhi for financial assistance.

Literature cited

Atri NS, Saini MK, Saini SS. 1994. Indian Russulaceae Roze —a check list. In: Current Researches in

Plant Science. Sarma TA, Saini SS, Trivedi ML, Sharma M (eds). Bishan Singh Mahendra Pal

Singh Dehradun. Pp. 81-93.

Lactarius annulocystidiatus sp. nov. (India) ... 67

Atri NS, Kaur A, Kaur H. 2005. Wild mushrooms - collection and identification. 9-26. in:RD Rai

et al. (eds). Frontier in mushroom biotechnology. National Research Centre for Mushroom,

Chambaghat Solan.

Buyck B, Verbeken A, Eberhardt U. 2007. The genus Lactarius in Madagascar. Mycol. Res. 111:

787-798. http://dx.doi.org/10.1016/j.mycres.2007.04.006

Das K, Sharma JR. 2005. Russulaceae of Kumaon Himalaya. Botanical Survey of India. Ministry of

Environment and Forests, Kolkata.

Das K, Verbeken A. 2011. Three new species of Lactarius (Russulaceae) from Sikkim, India.

Cryptogamie, Mycologie 32: 365-381.

Das K, Verbeken A. 2012. New species of Lactarius subg. Plinthogalus and new records of Lactifluus

subg. Gerardii (Russulaceae) from Sikkim, India. Taiwania 57: 37—48.

Das K, Sharma JR, Montoya L. 2004. Lactarius (Russulaceae) in Kumaon Himalaya.1.New species

of subgenus Russularia. Fungal Diversity 16: 23-33.

Eberhardt U, Verbeken A. 2004. Sequestrate Lactarius species from tropical Africa: L. angiocarpus

sp. nov. and L. dolichocaulis comb. nov. Mycol. Res. 108: 1042-1052.

http://dx.doi.org/10.1017/S0953756204000784

Hesler LR, Smith AH. 1979. North American species of Lactarius. The University of Michigan

Press, USA.

Kirk PM, Cannon PF, Minter DW, Stalpers JA (eds). 2008. Dictionary of the fungi, 10" edition.

CABI, UK.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour, 3 ed. Eyre Methuen, London,

252 p.

Le HT, Stubbe D, Verbeken A, Nuytinck J, Lumyong S, Desjardin DE. 2007. Lactarius in northern

Thailand: 2 Lactarius subgenus Plinthogali. Fungal Diversity 27: 61—94.

Montoya L, Bandala VM. 2008. A new species and new records of Lactarius (subgenus Russularia)

in a subtropical cloud forest from eastern Mexico. Fungal Diversity 29: 61—72.

Natarajan K, Kumaresan V, Narayanan K. 2005. A checklist of Indian agarics and boletes

(1984-2002). Kavaka 33:61—128.

Pearson AA. 1950. The genus Lactarius. The Naturalist: 81-99.

Singer R. 1986. The Agaricales in modern taxonomy, 4th ed. Bishan Singh Mahendra Pal Singh

Dehradun. 981 p., 88 plates.

Stubbe D, Nuytinck J, Verbeken A. 2008. Lactarius subgenus Plinthogalus of Malaysia. Fungal

Diversity 32: 125-156.

Stubbe D, Nuytinck J, Verbeken A. 2010. Critical assessment of Lactarius gerardii species complex

(Russulales). Fungal Biology 114: 271-283. http://dx.doi.org/10.1016/j.funbio.2010.008

Wang XH. 2007. Type studies of Lactarius species published from China. Mycologia 99: 253-268.

http://dx.doi.org/10.3852/mycologia.99.2.253

MYCOTAXON

http://dx.doi.org/10.5248/121.69

Volume 121, pp. 69-74 July-September 2012

A new species of Lentinus from India

GUNASEKARAN SENTHILARASU’ & SANJAY K SINGH?

National Facility for Culture Collection of Fungi, MACS’ Agharkar Research Institute,

G. G. Agarkar road, Pune-411 004, India

CORRESPONDENCE TO: ’ senthilarasug@rediffmail.com, * singhsksingh@rediffmail.com

AxBstTRAcT—Lentinus alpacus sp. nov. is described, illustrated, and discussed based on

collections made in Maharashtra State, India. Lentinus alpacus is tentatively placed in

subg. Panus sect. Panus, where it is distinguished by its small, reddish brown, squarrose

basidiomata.

Key worps—Basidiomycota, Polyporaceae, Polyporales, taxonomy

Introduction

The genus Lentinus sensu Pegler (1983b; Polyporaceae) is widespread in the

tropics. A review of literature (Manjula 1983; Natarajan et al. 2005; Kumaresan

& Senthilarasu unpubl. data) revealed that the genus is well represented in

India. Several species have been described and reported from Kerala State

alone (Manimohan & Leelavathy 1995; Joseph et al. 1995; Manimohan et al.

2004; Kumar & Manimohan 2005), but only one species, L. cochleatus, has been

reported from Maharashtra State (Trivedi 1972). In this paper, a new species of

Lentinus collected from Pune, Maharashtra State, is described, illustrated, and

discussed.

Materials & methods

Thin, handmade sections were made from dried specimens, revived in 10% KOH,

and stained in 3% Phloxine. Fifty basidiospores were measured for evaluation of the

range of spore-size and extreme values are given in parentheses followed by mean spore

measurements in parentheses. Camera lucida diagrams were made using a Nikon Y-IDT

prism attached to a Nikon E200 microscope. Microphotographs were made using Zeiss

Axio Imager A2 microscope. The colour names and notations used are from Kornerup &

Wanscher (1978). The examined collections cited are deposited at Ajrekar Mycological

Herbarium (AMH), MACS’ Agharkar Research Institute, Pune, India.

70 ... Senthilarasu & Singh

Taxonomy

Lentinus alpacus Senthil. & S.K. Singh, sp. nov. PLATES 1, 2

MycoBAank MB563805

Differs from L. courtetianus in a squarrose red pileus and thin, hispid stipe.

Type: India, Maharashtra State, Pune, Pune University Campus (18°31'18.4"N

73°49'53.6"E), on decaying twigs, solitary, 21.07.2011, G. Senthilarasu (holotype, AMH

9442).

ErymMo.oey: alpacus (Sanskrit), small.

PILEus 6-25 mm diam., orbicular, plano-concave or infundibuliform; surface

Venetian red (8D7) to Persian red (8E8), squarrose: scales reddish brown (9F8),

more towards the disc, at first often with purplish tints; margin incurved, thin,

entire, ciliate, not striate. LAMELLAE subdecurrent, white to orange white (5A2)

to pale orange (5A3) to grayish orange (6B3), becoming dark brown (8D4-

8D5) on drying, crowded, with numerous lamellulae, thin, less than 1 mm

broad; edge entire, concolorous with the sides. ST1PE central to excentric, rarely

lateral, 14-25 x 1-1.5 mm, equal, cylindric; surface reddish brown (8E8-9F8),

hispid, solid, arising from concolorous basal tomentum. PILEUS CONTEXT <1

mm thick, orange white (5A2) to pale orange (5A3).

BASIDIOSPORES (5-)5.5-6.5(-7) x (2—)2.5-3(-3.5), (6.140.31 x 2.9+0.14)

um, Q = 2.1, oblong-ellipsoid to subcylindric, hyaline, smooth, with a few

guttules. Basip1A 18-25 x 5-6.5 um, cylindrico-clavate to clavate, tetrasporic;

sterigmata up to 5 um long. LAMELLA EDGE sterile, with scattered cheilocystidia

together with metuloids. CHEILocysTiIp1a 20-30 x 8-12 um, clavate to

subclavate with broadly rounded apex, hyaline, thin-walled. PLEUROCYSTIDIA

abundant, gloeocystidioid, 18-45 x 5-11 um, cylindrico-clavate with obtusely

rounded apex, often constricted, initially embedded within the hymenium and

projecting 20 um beyond the basidia. METULOIDs scattered, on sides and edge

of lamellae, 19-25 x 7-11.5 um, thick-walled, 1-3.5 um thick, subventricose

with an obtusely rounded apex, projecting 15 um beyond the basidia. LAMELLA

TRAMA irregular, consisting of thick walled, hyaline, skeletal hyphae 2-2.5 um

diam. SUBHYMENIUM indistinct. PILEAL SURFACE a regular cutis consisting of

radially repent hyphae, often disrupting to form pileal squamules at the margin,

distinctly trichoderm at the disc due to more squamules; squamules up to 840

x 80 um, composed of fascicles of monomitic hyphae, up to 5 um diam., often

containing brown vacuolar sap, thick-walled, septate with clamp connections.

PILEAL CONTEXT consisting of dimitic hyphal system; generative hyphae up

to 3 um diam., not inflated, hyaline, thin-walled, with clamp connections,

often collapsing and difficult to observe; skeletal hyphae up to 2.5 um, hyaline,

unbranched, thick-walled, with a narrow lumen. HyPHAL PEGs absent.

ADDITIONAL SPECIMENS EXAMINED: INDIA, MAHARASHTRA STATE, Pune, Pune

University Campus (18°31'18.4"N 73°49'53.6"E), 03.08.2009, (AMH 9457), 26.07.2012,

(AMH 9526), 01.08.2012 (9527) G. Senthilarasu.

Lentinus alpaca sp. nov. (India) ... 71

— — ‘ ,

p e

7 N 7

' 7 >

10 pm

10 pm ,

PLATE 1. Lentinus alpacus: Under natural conditions on Pune University Campus. A. Surface view.

B. Gill view. c. Metuloids. p. Basidiospores. E. Gloeocystidia. F. Pileipellis generative hypha with

clamp connection. Photo: Senthilarasu G.

Lentinus alpacus is a fairly common and widespread species in the Pune

University Campus but usually found solitary on decaying twigs every year

during monsoon. The habit is also (rarely) connate, with a single basidiome

arising from the stipe apex of partly decayed basidiome.

72... Senthilarasu & Singh

PLATE 2. Lentinus alpacus:

A. Basidiospores. B. Basidia. c. Cheilocystidia. p. Pleurocystidia. £. Skeletal hyphae.

F. Generative hyphae with clamp connections. Scale bar = 10 um.

The generic demarcation and phylogenetic relationships of Panus Fr. and

Lentinus Fr. are controversial (Corner 1981; Pegler 1983b; Singer 1986; Hibbett

& Vilgalys 1991, 1993, Grand et al. 2011). These have been accepted as separate

genera by many modern authors (Corner 1981; Moser 1978; Singer 1975,

1986), but were reduced by Pegler (1983b) to subgenera within a broader

Lentinus alpaca sp. nov. (India) ... 73

concept of Lentinus. Redhead & Ginns (1985) created the genus Neolentinus,

characterised by bipolar mating systems and the ability to cause brown rot,

to accommodate species from Lentinus sections Pulverulenti, Cirrhosi, and

Squamosi sensu Pegler (1983b). Lentinus sensu stricto is characterized by

species having radiate, descending, or intermediate tramas with ligative hyphae

and hyphal pegs in the hymenium, whereas Panus sensu stricto is distinguished

by strongly radiate hymenophoral trama with dimitic hyphae and lacking

hyphal pegs. Additionally, in Panus young basidiomata are brightly pigmented

in purple and fade with age (Hibbett et al 1993). At present it is not clear to

which segregate genus L. alpacus belongs. However, it could be accommodated

in subgenus Panus sensu Pegler (1983b) as it lacks both skeleto-ligative hyphae

in the context and hyphal pegs in the hymenium, and in section Panus based on

its hymenial cystidia, hymenophoral trama of radiate construction and entire

lamella edge. Therefore, we have followed the broad Lentinus concept of Pegler

(1983b).

In sect. Panus, one African species, L. courtetianus Har. & Pat. (Pegler

1983b), resembles L. alpacus in having more or less similar sized basidiomata

and cheilo- and pleurocystidia. However, L. courtetianus has a smooth, pure

white or lemon yellow tinted pileus and thicker (3-10 mm diam.), smooth,

glabrous, subbulbous stipe.

Lentinus alpacus closely resembles the neotropical species L. tephroleucus

Mont. (Pegler 1983b) in having small, infundibuliform pileus with short erect

fasciculate hairs that become squamulose towards margin and similar sized

basidiospores and basidia. However, L. tephroleucus has a pale yellowish to

grayish brown or cinnamon brown pileus with glabrescent disc and cream

coloured lamellae. Microscopically, L. tephroleucus is clearly unrelated from

L. alpacus because of its sclerocystidia.

The southeastern Asian species Lentinus ciliatus Lév. (Pegler 1983b) also

shares a squamulose pileus and similar-sized spores and basidia but differs in

its larger basidiomata, cinnamon to fuscous brown pileus with densely ciliate

margin, and thick-walled sclerocystidia.

Lentinus leprieurii Mont. (Pegler 1983a), which resembles L. alpacus in its

brown pileus with fasciculate hairs, short stipe, and similar-sized basidiospores

and basidia, differs in its larger pileus, brown lamellae, thicker stipe (< 6 mm),

cylindric cheilocystidia, and absence of pleurocystidia. Moreover, L. leprieurii

grows in tufts, not solitary.

Acknowledgments

We thank Prof. R.H. Petersen and Prof. P. Manimohan for critically reviewing the

manuscript and suggesting appropriate modifications. GS personally thanks Prof. P.

Manimohan for providing literature on Lentinus species. We thank Dr. $.R. Pennycook

for nomenclatural review and Dr. L.L. Norvell for final review of the manuscript. Sincere

74 ... Senthilarasu & Singh

thanks are extended to the Director, Agharkar Research Institute for providing all

laboratory facilities. We extend our sincere thanks to the Department of Science and

Technology (DST), Government of India, New Delhi, for providing financial support

under the IRPHA Programme for setting up a National Facility for Culture Collection

of Fungi at Agharkar Research Institute, Pune, India.

Literature cited

Corner E. 1981. The agaric genera Lentinus, Panus, and Pleurotus with particular reference to

Malaysian species. Beih. Nova Hedwigia 69: 169 p.

Grand EA, Hughes KW, Petersen RH. 2011. Relationships within Lentinus subg. Lentinus

(Polyporales, Agaricomycetes) with emphasis on sects. Lentinus and Tigrini. Mycological

Progress 10: 399-413. http://dx.doi.org/10.1007/s11557-010-0711-4

Hibbett DS, Vilgalys R. 1991. Evolutionary relationships of Lentinus to the Polyporaceae: evidence

from restriction analysis of enzymatically amplified ribosomal DNA. Mycologia 83: 425-439.

http://dx.doi.org/10.2307/3760353

Hibbett DS, Vilgalys R. 1993. Phylogenetic relationships of Lentinus (Basidiomycotina)

inferred from molecular and morphological characters. Systematic Botany 18(3): 409-433.

http://dx.doi.org/10.2307/2419417

Hibbett DS, Murakami S, Tsuneda A. 1993. Sporocarp ontogeny in Panus (Basidiomycotina):

Evolution and Classification. American Journal of Botany 80(11): 1336-1348.

http://dx.doi.org/10.2307/2445719

Joseph AV, Abraham TK, Vrinda KB, Pradeep CK. 1995. New agarics from southern India.

Mushroom Research 4: 1-6.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3rd edn. Methuen and Co., Ltd.,

London. 243 p.

Kumar TKA, Manimohan P. 2005. A new species of Lentinus from India. Mycotaxon 92: 119-123.

Manimohan P, Leelavathy KM. 1995. A new variety of Lentinus caespiticola from southern India.

Mycological Research 99(4): 451-452. http://dx.doi.org/10.1016/S0953-7562(09)80644-8

Manimohan P, Divya N, Kumar TKA, Vrinda KB, Pradeep CK. 2004. The genus Lentinus in Kerala

State. Mycotaxon 90: 311-318.

Manjula B. 1983. A revised list of the agaricoid and boletoid basidiomycetes from India and Nepal.

Proceedings of Indian Academy of Sciences (Plant Science) 92: 81-213.

Moser M. 1978. Keys to agarics and boleti. R. Philips, London, England. 535 p.

Natarajan K, Kumaresan V, Narayanan K. 2005. A checklist of Indian agarics and boletes

(1984-2002). Kavaka 33: 61-128.

Pegler DN. 1983a. Agaric flora of the Lesser Antilles. Kew Bulletin Additional Series 9: 668 p.

Pegler DN. 1983b. The genus Lentinus: a world monograph. Kew Bulletin Additional Series 10:

281 p.

Redhead SA, Ginns JH. 1985. A reappraisal of agaric genera associated with brown rots of wood.

Transactions of Mycological Society of Japan 26: 349-381.

Singer R. 1975. The Agaricales in modern taxonomy (3rd ed.). Vaduz: J. Cramer. 912 p.

Singer R. 1986. The Agaricales in modern taxonomy (4th ed.). Sven Koeltz Scientific Books.

Koenigstein, Germany. 981 p.

Trivedi TK. 1972. Agaricales of Nagpur-I. The Botanique (Nagpur) 3: 53-59.

MY COTAXON

http://dx.doi.org/10.5248/121.75

Volume 121, pp. 75-79 July-September 2012

Two new species in the Graphidaceae

(Ostropales, Ascomycota) from China

ZE-FENG JIA’, RUI-FANG WANG! & JIANG-CHUN WEI”

‘College of Life Sciences, Shandong Agricultural University,

Taian 271018, China

*Key Laboratory of Systematic Mycology and Lichenology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing 100101, China

“CORRESPONDENCE TO: weijc2004@126.com

AxsstTRAcT— During an examination of subtropical and tropical lichen collections from

China, two species, Fissurina isidiata collected from Hainan Island and Graphis wangii

collected from Yunnan province, were discovered and are reported as new to science.

Key worps— lichenized fungi, taxonomy

Introduction

During the study of the lichen family Graphidaceae from China, two

interesting corticolous species of Fissurina and Graphis were found that are

reported here as new to science following the new generic concept of the

Graphidaceae proposed by Staiger (Staiger 2002). The types of the new species

are deposited in HMAS-L, LHS and KUN-L in China.

Material & methods

The examined material was collected by the author from Hainan Island and preserved

in HMAS-L LHS and KUN-L. Morphological characters were studied on dry specimens

using dissecting microscope (Tech XTS-20) and anatomical characters were examined

in hand-cut sections of lirellae mounted in water using a research microscope (Olympus

CHB-213). The lichen compound was detected by thin-layer chromatography (TLC)

(Culberson & Kristensson 1970; Culberson 1972).

Taxonomy

Fissurina isidiata Z.F. Jia, sp. nov. Piet

FUNGAL NAME FN570009

Sicut Fissurina astroisidiata, sed sporis longioribus et acido stictico continente differt.

76 ... Jia, Wang & Wei

Cc . D E

PLaTE 1 Fissurina isidiata. A. Thallus with isidia (bar = 10 mm); B. Cross section of an apothecium

(bar = 50 mm); C-E. Ascospores in asci (bar = 50 um).

Type: China. Hainan Island, Mountain Wuzhi, 18°46’N 109°31’E, alt. 950 m, 30.XI.2010,

Ze-Feng Jia 10-612 (Holotype, HMAS-L 117919; isotype, LHS).

Erymo oey: from the Latin isidium, referring to the isidiate thallus.

THALLUS corticolous, crustose, pale green to olive-green, uneven, isidiate, isidia

simple, concolorous with the thallus, 0.2-0.3 mm in diameter and 0.3-1.2 mm

high, distributed over the thallus.

AscomMarta lirelline, lirellae greyish white, 1-10 mm long, usually simple,

sometimes branched, more branched at the end of lirellae, fissured, immersed

to slightly raised. Disc slit-like, narrow to moderately broad, epruinose,

sometimes open when aged. ExcIpLE complete, present below, non-striate,

non-carbonized, yellow to dark orange-brown, sometimes becoming darkened

laterally or apically, convergent, covered by a thalline margin. HyYMENIUM

hyaline, not inspersed, 60-80 um high, I-, KI-. HypoTHEcium distinct, thin,

pale, 6-8 um high. ParapuysEs simple, 1.5-2.0 um thick. PERIPHysorDs short

to moderately elongate, 3-6 um long, with warty tips. Asci 8-spored, 50-75

x 5-12.5 um. Ascospores hyaline, oval or broadly ellipsoidal, transversely

Fissurina & Graphis spp. nov. (China) ... 77

septate when younger, submuriform to muriform, becoming 2-6 x 1-2-locular

with age, mature spores 15-20 x 5.5-7.5 um, sometimes with a halo, I+ reddish

violet.

CHEMISTRY: stictic acid present (TLC).

SUBSTRATE & DISTRIBUTION: The type specimen was collected in the central

region of the Nature Reserve of Mountain Wuzhi with a tropical forest located

in the central part of Hainan Province, Southern China. Fissurina isidiata is so

far known only from the type material. Associated species are Graphis japonica

(Mull. Arg.) A.W. Archer & Liicking and other graphidean lichens.

Notes: Fissurina isidiata is characterised by the presence of an isidiate thallus,

fissurine apothecia, a non-carbonized exciple, a clear hymenium, small

submuriform ascospores, and the presence of stictic acid. It is the second isidiate

species found in the genus Fissurina, the first being F. astroisidiata Herrera-

Camp. & Liicking described from Mexico (Lumbsch & al. 2011). The Mexican

species is readily distinguished by shorter, stellately branched lirellae, smaller

ascospores (12-15 um long), and the absence of lichen compounds. Fissurina

abdita (A.W. Archer) A.W. Archer also possesses small ascospores and stictic

acid but is differentiated by a non-isidiate thallus and larger ascospores (18-35

x 8-10 um and 8-10/2-3 locular as reported by Archer; 2006).

Graphis wangii Z.F. Jia, sp. nov. Pi)

FUNGAL NAME FN570011

Species nova Graphis biferae similis sed ascosporis submuriformibus differt.

Type: China, Yunnan province, Xinping county, Mont. Mopanshan, 23°55'N 101°58’E,

alt. 2350 m, on bark of Quercus sp.. 3.1.2009. coll. Li-Song Wang 09-30091 (Holotype,

KUN-L).

ErymMo ocy: The new species is named in honour of the esteemed Prof. Li S. Wang, a

famous collector and conservator of lichens and mosses from China.

THALLUS corticolous, crustose, pale white to whitish grey, surface dull, unevenly

thickened, tightly attached to the substratum, without isidia and soralia.

APOTHECIA lirelliform, elongate to sinuous, simple or rarely branched, 0.5-5

mm long, 0.4—0.5 mm wide, sessile, lacking a thalline margin, black, curved and

straight, often rounded at the ends, not striate, scattered over the thallus, disc

concealed. PROPER EXCIPLE conspicuous, completely carbonised. EPITHECIUM

3-5 um thick, brownish. HyMENtvuM colorless, not inspersed, 180-230 um high,

I-. HyPOTHECIUM colourless to brownish, 30-40 um high. PARAPHYSES simple,

up to 1-1.5 um wide, apices unbranched. Asci cylindrical to clavate, 110-160 x

15-25 um, 2-8-spored. Ascospores hyaline, oblong to ellipsoid, transversely

septate or with a few central longitudinal septa, 14—20/1-2-locular, 70-103 x

13-20.5 um, I+ blue-violet.

CHEMISTRY: C-, K-, P-; no lichen compounds detected by TLC.

78 ... Jia, Wang & Wei

—

Si =

af ;

. , : bifse &

"ha iii ~ Se

. | s i) tan a

| {SS Att

at . ] ¢ ‘= = ih eee

‘i WES} FG

mast \e at vi sy 7 ET

eh - Wes a ee

: ‘ ’ ’ : ~ ee =—_

\ : ee \ ts) cus /

\ ala Tar :

SUS OO EGE Qk ey ‘.

AS ‘ \ oi] ha EF | Pal +.

PLATE 2 Graphis wangii. A. Habit (bar = 2 mm); B. Lirellae showing completely carbonized exciple

(bar = 2 mm); C. Cross section of an apothecium (bar = 100 um); D-F. Asci with ascospores (bars

= 50 um); G. Ascospore (bar = 50 um).

Notes: Graphis wangii is characterized by simple or seldom branched,

sessile, lirellae, lacking a thalline margin, nuda morph ascomata, a completely

carbonized exciple, and a clear hymenium, with submuriform ascospores and

lacking lichen compounds.

The new species has similar thallus and lirella morphology and similar

chemistry to G. bifera Zahlbr., which differs in having ascospores with 15-21

transverse septa (Liicking et al. 2009).

Graphis wangii also resembles G. lourdesina Aptroot, described from Brazil

(Licking et al. 2009). Both species have short, simple sessile lirellae, muriform

ascospores, and lack thalline margins and lichen compounds, but G. lourdesina

differs in larger ascospores, 100-120 x 30-35 um.

At present, the new species is known only from the Mopanshan National

Forest Park. It grows on the trunks of Quercus sp.

Acknowledgements

The authors are deeply grateful to Dr. Alan W. Archer and Prof. Klaus Kalb for

pre-submission reviews and their valuable comments. The authors thank the National

Natural Science Foundation of China (No. 39899400) and the Ministry of Science and

Technology of China (No. 2006FY110500-5), for their financial support.

Fissurina & Graphis spp. nov. (China) ... 79

Literature cited

Archer AW. 2006. The lichen family Graphidaceae in Australia. Bibliotheca Lichenologica 94:

1-191.

Culberson CE. 1972. Improved conditions and new data for the identification of lichen products by

a standardized thin-layer chromatographic method. Journal of Chromatography 72: 113-125.

http://dx.doi.org/10.1016/0021-9673(72)80013-X

Culberson CF, Kristensson H. 1970. A standardized method for the identification of lichen products.

Journal of Chromatography 46: 85-93. http://dx.doi.org/10.1016/S0021-9673(00)83967-9

Licking R, Archer AW, Aptroot A. 2009. A world-wide key to the genus Graphis (Ostropales:

Graphidaceae). Lichenologist 41(4): 363-452. http://dx.doi.org/10.1017/S0024282909008305

Lumbsch HT, et al. 2011. One hundred new species of lichenised fungi: a signature of undiscovered

global diversity. Phytotaxa 18: 1-127.

Staiger B. 2002. Die Flechtenfamilie Graphidaceae. Bibliotheca Lichenologica 85: 1-526.

MY COTAXON

http://dx.doi.org/10.5248/121.81

Volume 121, pp. 81-92 July-September 2012

Tricholomopsis in Europe — phylogeny, key, and

notes on variability

JAN HOLEc & MIROSLAV KOLARIK?

"National Museum, Mycological Department, Cirkusova 1740, CZ-193 00 Praha 9, Czech Republic

*Department of Botany, Faculty of Science, Charles University,

Bendtska 2, CZ-128 01 Praha 2, Czech Republic

* CORRESPONDENCE TO: jan_holec@nm.cz

AsstrRact— Tricholomopsis (Basidiomycota, Agaricales) species in Europe were studied

by classical and molecular methods (morphology, anatomy; sequences of nuclear rDNA

containing ITS1, ITS2, 5.8S and part of 28S regions). The identity of four species was

confirmed: T! rutilans, T. flammula, T. decora, T. osiliensis. Within T. flammula, two well-

supported groups were recovered. An identification key based on a better understanding

of the species limits was prepared. All species are briefly characterized and their variability

is discussed. Tricholomopsis ornata should be rejected; it is considered a doubtful, poorly

documented taxon, and the name has been variously interpreted.

KEY worps— Fungi, macromycetes, taxonomy, nomenclature

Introduction

The genus Tricholomopsis Singer is represented in Europe by a rather

small number of species. Most authors recognise only the well-known species

T. rutilans and T: decora (e.g., Boekhout & Noordeloos 1999, Gréger 2006,

Vesterholt 2008). Some identification keys give two more species, namely

T. flammula and T. ornata (Moser 1983, Bon 1995, Horak 2005). The identity

of T: flammula was documented recently using both classical and molecular

methods (Holec 2009, Holec & Kolarik 2011). In 2009, the new species

T. osiliensis was described by Vauras (2009) from the Estonian island Saaremaa

(Osilia in Latin). Four species (T: rutilans, T: ornata, T: flammula, T. osiliensis)

are reported from Estonia (Kalamees 2011). Altogether, five species have been

reported from Europe. However, T: ornata is poorly documented as there are

no well-described records agreeing with the original description by Fries (1838)

and a detailed DNA study comparing the species is absent.

In the molecular phylogenetical study by Moncalvo & al. (2002)

Tricholomopsis, represented by the nuclear LSU rDNA sequence of T. rutilans,

82 ... Holec & Kolatik

was placed in one phylogenetic clade together with Clavaria fusiformis and

Marasmius rhyssophyllus. In a multigene dataset (RPB1, RPB2, ncRNA)

another representative species, T: decora, fell into a pluteoid clade, outside of

any conventional families, close to Amanitaceae (Matheny & al. 2006). To date,

a detailed DNA study has been published only on T: flammula (Holec & Kolarik

2011).

The aim of this study is to verify the identity of Tricholomopsis species in

Europe by classical and molecular methods and to prepare an identification key

based on a better understanding of species limits.

Material & methods

Morphological study

Fresh and dried collections of Tricholomopsis species from Europe were studied.

Voucher specimens are kept in herbaria PRM, WU, TU, and TUR-A (formerly TURA).

For herbarium acronyms see Thiers (2012). All collections were studied by traditional

taxonomic methods (generally: Singer 1986, Bas & al. 1988; specifications: Holec 2009).

Spore sizes are presented as the main data range (c. 10-90 percentile values), flanked by

extreme values in parentheses, of all spores measured (20 measurements per collection).

The spores were measured directly in the microscope using the eyepiece micrometer

with the basic scale division of 0.8 um and distinguishable difference of 0.4 um. Thus,

the scale reading was made in the following steps (in um): 1.6, 2.0, 2.4, etc.

DNA study

Genomic DNA from nine herbarium specimens (Tas. 1) was isolated using

ArchivePure DNA Yeast and Gram-+ Kit (5 PRIME, Inc. Gaithersburg, MD) with

modified time of the incubation with Lytic Enzyme Solution (2h, 37 °C) and Cell Lysis

Solution (4h, 64 °C). Nuclear rDNA containing internal transcribed spacers (ITS1 and

ITS2), 5.8S and part of 28S regions was amplified with primer set ITS1/LR6 (Gardes &

Bruns 1993; Vilgalys & Hester 1990). The reaction mixtures and amplification protocols

for rDNA regions were made according to Hulcr & al. (2007) except of polymerase,

PerfectTaq Plus DNA Polymerase (5 PRIME, Inc. Gaithersburg, MD) which was used

here to overcome amplifications problems. Custom purification of PCR products and

sequencing was done at Macrogen Inc. (Seoul, South Korea) using the primers ITS1,

ITS4S, LR6, NL1 and NL4 (Gardes & Bruns 1993; O'Donnell 1993). Sequences were

combined with the most related published Tricholomopsis rDNA (Tas. 1) sequences

from well documented specimens and aligned in MAFFT v6.861b using Q-INS-i

strategy considering secondary structure of rRNA (Katoh & Toh 2008). To filter both

gaps and hyper variable regions, we used Gblocks version 0.91b (Talavera & Castresana

2007) with less stringent selection allowing smaller final blocks and gap positions within

the final blocks. There were a total of 19 sequences and 1119 positions in the final dataset

(unpruned dataset contained 1786 positions) from which 843 were conservative and

274 variable. Maximum likelihood (ML) and Minimum Evolution (ME) analyses were

conducted with 1000 bootstrap replicates and default parameters in MEGA 5.0 (Tamura

& al. 2011). In both analyses, all positions with less than 5% site coverage were eliminated.

Tricholomopsis in Europe ... 83

TABLE 1. Tricholomopsis collections used for DNA analysis.

SPECIES COUNTRY

T. flammula

Czech Rep.

Slovakia

Austria

Pakistan

T. decora

Czech Rep.

Slovakia

USA

T. osiliensis Estonia

Estonia

Slovakia

T. rutilans

Czech Rep.

Canada

Czech Rep.

LOCALITY

Boubinsky

prales

Cernava

Na Stiibrné

Badinsky prales

Dobroésky

prales

Dobroésky

prales

Eichberg

Rote Au

Diirradmer

Diamir, Fairy

Meadows

Olginka

Mala Niva

Badinsky prales

Massachusetts,

Harvard

Forest

Saaremaa

Vormsi

Dobroésky

prales

Mt. Kamenna

Kamenny vrch

hill

BC, Capilano

River

REFERENCE

Holec (2009), Holec

& Kolarik (2011)

Holec & Kolatik

(2011)

Holec & Kolatik

(2011)

This study

Holec & Kolatik

(2011)

Holec (2009)

This study

Razaq & Khalid

(unpublished)

Holec & Kolatik

(2011)

Holec & Kolatik

(2011)

This study

Petersen & Hughes

(2012)

Vauras (2009), this

study

This study

Holec (2009)

This study

Johnston (2006)

GENBANK

FN554893

FN554894

FN554896

HE649939

HE649940

HE649941

FN554897

FN554892

HE652866

FR822742

FN554890

FN554891

HE649942

DQ404384

HE649943

HE649944

HE649945

FN554895

HE649946

EF530929

VOUCHER

PRM 899108

(JH 149/2008)

PRM 899459

(JH 227/2008)

PRM 909608

PRM 899162

(JH 140/2009)

PRM 899180

(JH 162/2009)

PRM 899190

(JH 172/2009)

WU 12087

WU 25091

WU 13075

SR 161

PRM 882317

PRM 898238

(JH 218/1997)

PRM 899160

(JH 145/2009)

TENN 62342

(PBM 2482)

PRM 899461

(isotype)

TU 101571

PRM 899184

(JH 166/2009)

PRM 889120

(JH 362/1996)

PRM 899460

(JH 91/2009)

UBC F16251

Bayesian searches (MB) were conducted with MrBayes 3.0 (Ronquist & Huelsenbeck

2003) and 1.5 million replicates estimated together with burn-in value in Tracer v1.5

(Rambaut & Drummond 2007). Pluteus romellii, known as related to the Tricholomopsis

group (Matheny & al. 2006), was chosen as outgroup. Sequences obtained in this study

were submitted to GenBank (Tas. 1).

Abbreviations

bp: base pairs, JH: Jan Holec, Q: quotient of length/width of the spores, MB: Bayesian

analyses using MrBayes, ME: Minimum Evolution analysis, ML: Maximum Likelihood

analysis, Qav: mean value of Q in each of the collections studied.

84 ... Holec & Kolatik

Results and discussion

DNA study

The phylogenetic analysis computed by all three methods consistently

recovered five major supported clades (Fic. 1). ME showed the highest

statistical supports from all methods. The first clade consists of T: decora

sequences, where specimen PRM 899160 (1521 bp) showed one base pair

difference from two other European specimens and nine base pairs difference

from the USA specimen. The next two clades consist of T: flammula specimens.

The major clade is divided further into two moderately supported subgroups

differing in four positions from 1521 bp. Specimen PRM 899190 differs in seven

positions in the ITS regions and eleven in the LSU region from the specimen

PRM 899108, which represents the second clade. Sequences from T. rutilans

specimens formed a clade with material from Canada exhibiting ten different

positions in the ITS region and six positions in the LSU region. ‘The last clade

consists of the little variable group of T- osiliensis specimens.

Phylogenetic analysis recovered lineages corresponding with our morpho-

logical observations and showing T. decora, T: flammula, T. osiliensis and

T: rutilans as well defined taxa. Two of the studied species (T. decora and T. rutilans),

known also in North America, showed the presence of vicariant populations

resulting evidently from the running allopatric speciation. Our DNA analysis

also revealed two well-supported groups (“species”) within T. flammula.

Delimitation of species based solely on molecular data of a single locus,

especially in closely related taxa, is tricky. Additional distinguishing characters

obtained from more collections could elucidate whether the specimen PRM

899108, previously shown as morphologically different from other T: flammula

collections (Holec & Kolarik 2011), really represents a separate species.

Key to critically revised Tricholomopsis species in Europe

Fa: 'Fibrils‘or-scales on pileus surface réd:toiviolet-+.i0;,n22-05.42¢-03 bre op begins Vertis wage 2

1b. Fibrils or scales on pileus surface without red to violet colour .................. 3

2a. Spores broadly ellipsoid to subglobose, Q = 1.14-1.55, Qav = 1.30-1.35, width

4.0-6.5 um; mature basidiocarps large and fleshy, stipe always covered with

rece vao letdibtils to'sealegs. Sse Poy a haribet Gite ht Fed T. rutilans

2b. Spores ellipsoid, Q = 1.33-2.11(-2.44), Qav = 1.50-1.90, width 3.2-4.8 um;

mature basidiocarps slender, mostly (not usually) small to medium-sized,

stipe mostly yellow, rarely covered with red-violet fibrils........... T. flammula

3a. Pileus covered with yellow-green, bronze, olive-brown to olive-black,

distinctly upraised scales on brightly yellow to orange ground......... T. decora

3b. Pileus covered with yellow-ochre to orange-brown, fine to coarse, adpressed

fibrils or fine scales on less bright, yellow to ochre ground .......... T. osiliensis

1.00/100/100

0.96/94/99

0.91/83/98

0.85/62/84

Tricholomopsis in Europe...

T. decora PRM899160 Slovakia

T. decora PRM898238 Czech R.

T. decora PRM882317 Czech R.

T. decora TENN62342 USA

T. flammula PRM909608 Czech R.

T. flammula WU25091 Austria

T. flammula WU12087Austria

T. flammula SR161 Pakistan

T. flammula \WU13075 Austria

T. flammula PRM899459 Czech R.

T. flammula PRM899180 Slovakia

T. flammula PRM899190 Slovakia

T. flammula PRM899162 Slovakia

T. flammula PRM899108 Czech R.

T. rutilans UBCF16251 Canada

1.00/100/100 T. rutilans PRM899460 Czech R.

0.91/91/91

1.00/100/100

T. rutilans PRM889120 Czech R.

T. osiliensis PRM899461 Estonia

T. osiliensis PRM899184 Slovakia

T. osiliensis TU101571 Estonia

Pluteus romellii AY854065 AY634279

Figure 1. ITS-LSU rDNA based phylogram obtained from Bayesian inference depicting

relationships among European Tricholomopsis species. The numbers from left to right refer to

posterior probabilities of the Bayesian Markov chain Monte Carlo analysis, percentage bootstrap

values of Maximum Likelihood and Minimum Evolution analysis. Values smaller than 50% are not

shown. Branch lengths are scaled in terms of expected numbers of nucleotide substitutions per site.

Sequences printed in bold were obtained during this study.

86 ... Holec & Kolatik

Survey of critically revised Tricholomopsis species in Europe

Tricholomopsis rutilans (Schaeff. : Fr.) Singer 1939

SELECTED DESCRIPTIONS AND ICONES. Breitenbach & Kranzlin (1991: no. 445), Ryman

& Holmasen (1992: 281), Boekhout & Noordeloos (1999: 151-152), Ludwig (2000: no.

85.3.B, C; 2001: 680), Roux (2006: 380), Vesterholt (2008: 334-335).

DIAGNOSTIC CHARACTERS — Basidiocarps large and fleshy when mature,

pileus and stipe densely covered with red-violet fibrils to scales. Spores broadly

ellipsoid to subglobose, rarely broadly obovoid or ellipsoid, 5.0-8.5 x 4.0-6.5

um, Q = 1.14-1.55, Qav = 1.30-1.35; pleurocystidia absent or rare.

DISTRIBUTION AND ECOLOGY — Common throughout Europe; living on

dead wood of conifers, often buried in soil; in all forest types, in clearings, along

forest roads, in parks and gardens.

Comments — Tricholomopsis rutilans is easily recognizable by the large and

fleshy basidiocarps (when mature), red-violet fibrils or scales on pileus and

stipe surface, and above all by the broadly ellipsoid to subglobose spores. A

typical T. flammula mostly differs by slender basidiocarps with yellow stipes

and more elongated (ellipsoid), more slender spores. However, in some cases its

basidiocarps reach almost the same size as those of T’ rutilans and the stipe is

covered by violet fibrils, especially when young. In such cases, the spore shape

and size (see the key above) and abundance of pleurocystidia are decisive for

correct identification of T. flammula.

COLLECTIONS STUDIED — CZECH REPUBLIC. Krkonofe Mts., Rokytnice n. Jizerou:

Dvoratky, decayed trunk of Picea abies, 13.X.1946 leg. J. Kubicka (PRM 520341).

Central Bohemia, Dobrichovice, on bark of Picea abies, 15.X.1996 leg. M. Svréek (PRM

889808). Sumava Mts., Mt. Kamenné, on soil under Picea, 28.VIII.1996 leg. J. Holec

(PRM 889120). Nova Bystrice, stream valley S of Kamenny vrch hill, on stump of Picea

abies, 17.1X.2009 leg. J. Holec (PRM 899460). SLOVAKIA. Zapadné Tatry Mts., Mt.

Osobita, on decayed trunk of Picea abies, 7.1X.1979 leg. R. Leontovyé (PRM 821774).

UKRAINE. Eastern Carpathians, Dilove (“Trebusany”), Mt. Menchul (“Menc¢ul”), Abies

alba, V11I.1934 leg. A. Pilat (PRM 497334).

Tricholomopsis flammula Métrod ex Holec 2009

SELECTED DESCRIPTIONS AND ICONES. Métrod (1946: p. 77, pl. 1: fig. 5), Cetto

(1995: fig. 1010), Krisai-Greilhuber & Voglmayr (2000), Holec (2009), Holec &

Kolafik (2011).

DIAGNOSTIC CHARACTERS — Basidiocarps slender, not fleshy, small to medium-

sized, rarely large (pileus < 11 cm), young pileus densely covered with fine red-

violet to purplish brown fibrillose scales which become scarce on mature pileus,

stipe mostly pale yellow, lemon-yellow to bright yellow, rarely (especially when

young) covered with fine, red-violet to violet-brown fibrils. Spores variable

in length, rather elongated, ellipsoid, (5.2-)5.6-8.0(-8.8) x 3.2-4.8 um,

Q = 1.33-2.11(-2.44), Qav = 1.50-1.90, pleurocystidia abundant, with pale

yellow refractive content.

Tricholomopsis in Europe ... 87

DISTRIBUTION AND ECOLOGY — Widespread but rare throughout Europe;

on dead conifer wood, mostly fallen decaying trunks of Abies and Picea,

rarely on wood of deciduous trees (Fagus), in forests, shrubs and abandoned

orchards; from the hilly country to the mountains. In the Czech Republic and

Slovakia, the species clearly prefers natural to virgin forests; however, records

from habitats strongly influenced by man are known, too.

ComMENTS — In most cases, T’ flammula is a small to medium-sized fungus

with a pileus < 5(-8) cm broad. Rarely the pileus can reach a diameter of

11 cm (PRM 899191) but even then the basidiocarp remains slender (less

fleshy) than similarly sized T’ rutilans basidiocarps. In previous publications

(Krisai-Greilhuber & Voglmayr 2000, Holec 2009, Holec & Kolarik 2011) a

yellow stipe lacking red-violet fibrils or scales is emphasized as one diagnostic

character of T’ flammula. Ample material collected in Slovakia in 2009 (see

below) showed that in some collections the young stipe is almost completely

covered with a fine violet felt which is present even in older basidiocarps as

sparse violet-brown fibrils. Simultaneously, spores were unusually elongated in

the Slovak collections, reaching a Qav of 1.72-1.90 in comparison with Qav of

1.51-1.63 in the previously studied collections (Holec & Kolarik 2011). These

collections (including PRM 899459 from the Moravian Carpathians), grouped

together also in our phylogram (Fic. 1), differ from sister “western and central

European clade” in four positions in the entire ITS-LSU region. This group

seems to represent a slightly deviating “Carpathian” T. flammula subpopulation

characterized by larger basidiocarps and more elongated spores (with Q often

reaching 2.00-2.44 and Qav = 1.72-1.90). The “western and central European”

subpopulation has smaller basidiocarps and less elongated spores (with Q =

1.33-2.00; Qav = 1.51-1.63). In any case, the differences in DNA sequences are

negligible and show at most the beginnings of a vicariance.

Consequently, basidiocarp size and stipe surface color are not stable

characters and have a limited value for distinguishing T. flammula and

T. rutilans. The reliable distinguishing characters of T: flammula are the spore

shape and width and the abundance of pleurocystidia (see the key above for

details).

Tricholomopsis flammula is a variable species where three infraspecific

subdivisions were revealed by molecular methods (i.e. two well-supported

groups, one with two moderately supported subgroups, representing the

subpopulations discussed above). This fact is phenotypically expressed by

variable macrocharacters (see above), a variability previously discussed in

Holec & Kolarik (2011) based on a smaller number (5) of molecularly studied

collections than here (10). However, the problem remains unresolved. The

first clade revealed here is represented by a morphologically variable group

of specimens comprising the two subpopulations discussed above and having

88 ... Holec & Kolatik

both small and medium to large basidiocarps and larger (especially longer)

spores [5.6-8.0(-8.8) x 3.2-4.8 um]. The second clade is represented by

only one collection (PRM 899108: Boubinsky prales) with rather small, pale

basidiocarps (Holec & Kolarik 2011: fig. 4) and small spores [(5.2-)5.5-6.4 x

3.2-4.0 um]. More collections are needed to understand the limit between the

two clades.

COLLECTIONS STUDIED — See Holec (2009) and Holec & Kolarik (2011) for collections

from CZECH REPUBLIC, AUSTRIA and FRANCE.

ADDITIONAL COLLECTIONS. SLOVAKIA. Kremnické vrchy Hills, virgin forest Badinsky

prales, fallen decaying trunk of Abies alba, 28.1X.2009 leg. J. Lederer (PRM 899162,

899164). Veporské vrchy Mts., virgin forest Dobro¢sky prales, fallen trunk (Picea?

Abies?), 29.1X.2009 leg. S. Kominkova (PRM 899180); fallen decaying trunk of Abies

alba, 30.1X.2009 leg. J. Holec (PRM 899190, 899191).

Tricholomopsis decora (Fr. : Fr.) Singer 1939

SELECTED DESCRIPTIONS AND ICONES. Breitenbach & Kranzlin (1991: no. 444), Ryman

& Holmasen (1992: 281), Boekhout & Noordeloos (1999: 151-152), Ludwig (2000: no.

85.1; 2001: 678), Roux (2006: 381), Vesterholt (2008: 335).

DIAGNOSTIC CHARACTERS — Basidiocarps medium-sized, ground color yellow

to orange, pileus covered with densely arranged, upraised, yellow-green,

bronze, olive-brown to olive-black scales. Spores variable in size and shape,

broadly ellipsoid to ellipsoid, rarely obovoid or slightly phaseoliform in side

view, (6.0-)6.5-9.0(-10) x (4.0-)4.5-6.5(-7.0) um, Q = 1.20-1.75, Qav = 1.40-1.54,

pleurocystidia absent to rare.

DISTRIBUTION AND ECOLOGY — Scattered to common throughout Europe;

living on dead conifer wood, especially on old stumps and decaying fallen

trunks; in moist and shady mixed and coniferous forests, above all in the

mountains.

Comments — Tricholomopsis decora is a species easily recognized especially

from the bright colours and densely arranged upraised scales on pileus surface.

For differences from T. osiliensis see below.

COLLECTIONS STUDIED. CZECH REPUBLIC. Ceské Svycarsko national park, site called

Babylon, fallen decaying trunk of Picea abies, 16.VIII.2011, leg. J. Holec (PRM 899309).

Sumava Mts., nature reserve Mala Niva, decaying trunk of Picea abies, 6.VIII.1997 leg.

J. Holec (PRM 898238). Novohradské hory Mts., virgin forest Zofinsky prales, decayed

trunk of Abies alba, 30.1X.2008 leg. J. Borovicka et J. Kubrova (PRM 915260). Hruby

Jesenik Mts., nature reserve Pradéd, fallen decaying trunk of Picea abies, 25.VUI.2011

leg. J. Holec (PRM 899339). Nizky Jesenik Mts., nature monument ReSsovské vodopady,

fallen trunk of Picea abies, 23.VIII.2011 leg. J. Holec (PRM 899318). SLOVAKIA.

Kremnické vrchy hills, virgin forest Badinsky prales, fallen decaying trunk of Abies alba,

28.1X.2009 leg. J. Holec (PRM 899160). Klenovec, Mt. Klenovsky Vepor, decaying trunk

of Abies alba, 31.V1II.1982 leg. F. Kotlaba (PRM 828487).

Tricholomopsis in Europe ... 89

Tricholomopsis osiliensis Vauras 2009

SELECTED DESCRIPTIONS AND ICONES. Vauras (2009).

DIAGNOSTIC CHARACTERS — Basidiocarps medium-sized, yellow, yellow-

ochre to brownish yellow, pileus 3.5-9 cm in diam., without distinct upraised

scales but with concolorous minute squamules at margin (Vauras 2009) or

with coarse, adpressed, ochre-brown fibrils or cords of fibrils (PRM 899184),

becoming orange-brown when old. Spores variable in size and shape, broadly

ellipsoid, ellipsoid, obovoid, rarely subglobose, 6.0-9.5 x 4.5-6.0 um, Q = 1.14-

1.85, Qav = 1.25-1.54; pleurocystidia present, scarce to frequent.

DISTRIBUTION AND ECOLOGY — Apparently a rare species, known from two

Estonian islands in the Baltic Sea (Saaremaa: Vauras 2009, Vormsi: TU 101571)

and Slovakia (virgin forest Dobro¢sky prales, PRM 899184). The collections

originate from fallen decaying trunks of Picea abies (Saaremaa, Vormsi) and

Abies alba (Dobroésky prales). It is possible that T: osiliensis also occurs in

France, as a collection fitting its description was mentioned by Bon (1995) in

his remark on T. decora.

Comments — ‘The three T. osiliensis collections mentioned here form a well-

supported clade with little molecular variability. Macro- and microscopically,

the Slovak collection clearly differs from the Estonian ones in a pileus covered

with coarse fibrils darker (orange-brown) than the ground; the Estonian

basidiocarps exhibit almost smooth to finely fibrillose pilei with minute

squamules that are present only at the margin. The striking nature of the Slovak

fibrils was probably increased by their age and weather conditions (old and

partly dried basidiocarps influenced by first autumn frosts). Additionally,

spores in the Slovak basidiocarps were distinctly larger (8.0-9.5 x 5.2-6.0 um)

than in the Vormsi (6.0-7.2 x 4.8-5.6 um) and Saaremaa (6.0-8.2 x 4.5-6.0

um) collections (see Vauras 2009). However, a rather large spore size variability

is typical for all European Tricholomopsis species (see above). The spores of

T. osiliensis have a similarly broad range of spore length values.

Tricholomopsis osiliensis is a recently described, little known species. It

is necessary to observe basidiocarp variability in future collections. Based

on current data, T. osiliensis differs from T. rutilans and T. flammula by the

absence of red-violet fibrils or scales and from T’ decora by its almost smooth to

fibrillose pileus without distinctly upraised scales and less bright pileus cuticle

color. It is possible that some collections published as T: ornata represent in

fact T. osiliensis (e.g. Courtecuisse & Duhem 2000: no. 419; see the discussion

below).

COLLECTIONS STUDIED — ESTONIA. Island Saaremaa, Salme, Kaugatoma-Loéu, on

fallen mossy stem of Picea abies, 18.[X.2008 leg. J. Vauras (isotype: PRM 899461). Laane,

island Vormsi, Saxby, rotten trunk of Picea abies, 19.1X.2010 leg. I. Saar (TU 101571).

90 ... Holec & Kolatik

SLOVAKIA. Veporské vrchy Mts., virgin forest Dobroésky prales, fallen trunk of Abies

alba, 29.1X.2009, leg. S. Malec et P. Zitiian (PRM 899184).

Notes on Tricholomopsis ornata

Tricholomopsis ornata (Fr.) Singer is mentioned by some European authors

(e.g. Moser 1983, Riva 1988, Bon 1995, Horak 2005). According to the original

description by Fries (1838: 130), it is a medium-sized to large, fleshy fungus

(pileus 5-12.5 cm broad, stipe about 1.2 cm thick) similar to T! rutilans but

with a pileus covered with brown flocculose scales (“squamulis flocculosis

fulvofuscescentibus”). No red-violet color is mentioned. We do not know

any collection corresponding with this description. In the recent book on

Scandinavian fungi (Vesterholt 2008), T: ornata is not mentioned, although it

was originally described from the Uppsala vicinity growing on Pinus trunks.

In our opinion, T: ornata is a doubtful, poorly documented taxon. It is possible

that Fries (1838) described abnormal or old T. rutilans basidiocarps, which is

supported by his note that T’ ornata is similar to T: rutilans.

Bon (1995) is the only recent author describing T’ ornata in detail and

mentioning the material studied. However, his description differs from the

original. Bon’s fungus is smaller [pileus 3-6 cm, stipe 4-6(-8) x 0.3-0.5 cm]

with adpressed, rusty brown to reddish scales on pileus surface and pale

lemon-yellow stipe. These characters, including the rather large ellipsoid spores

[(6-)7-9(-10) x 4.6-6.0 um] and the presence of pleurocystidia, suggest that

his T: ornata could represent T! osiliensis. A revision and molecular study of

Bon’s collections would be desirable to confirm this hypothesis.

Similarly, the illustrations of T: ornata by Courtecuisse & Duhem (2000:

fig. 419, rather small yellow fungus with adpressed, fibrillose, vividly fulvous

brown scales) do not correspond to the original T. ornata of Fries but do

correspond well with our Slovakian collection of T: osiliensis.

Judging from our field experience with Tricholomopsis species, the

Rebaudengos illustration of T: ornata published in Riva (1988: 61) most

probably represents aberrant T: decora basidiocarps. The scales agree with the

Friesian description (small, floccose, fulvous brown), but the slender stature

and bright yellow ground color are more typical of T: decora.

There are some recent photographs labeled as T. ornata. Those by Dominguez

(2008) are closest to the original description by Fries; however, as they are not

accompanied by an analysis of microcharacters, their identity remains unclear.

On the other hand, the microcharacters presented by Musumeci (2004) clearly

show that his T’ ornata is in fact T. flammula.

The discussion shows that different interpretations of the name T: ornata

exist. It seems that the original T: ornata represents aberrant basidiocarps of

T. rutilans, and since them the name has been erroneously used for collections

of T: osiliensis, T: flammula and T. decora. For these reasons we recommend

rejecting the name T. ornata.

Tricholomopsis in Europe... 91

Acknowledgements

We thank J. Vauras (Finland) and I. Saar (Estonia) for loans of the voucher specimens

of T: osiliensis and the curators of the herbarium WU for loans of T: flammula material.

I. Krisai-Greilhuber (Austria), V. Antonin (Czech Republic), and S. Pennycook (New

Zealand) are acknowledged for careful reviews of the manuscript. J. Holec was financially

supported by the Ministry of Culture of the Czech Republic (DKRVO MK-S 760/2012

OVV). M. Kolafik was sponsored by the grant MSM 6007665801.

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MY COTAXON

http://dx.doi.org/10.5248/121.93

Volume 121, pp. 93-132 July-September 2012

Ganoderma in Brazil: known species and new records

MABEL GISELA TORRES-TORRES?”, LAURA GUZMAN-DAVALOS?*

& ADRIANA DE MELLO GUGLIOTTA3

"Universidad Tecnologica del Chocé, Ciudadela Medrano, Quibd6, Chocé, Colombia,

*Departamento de Botanica y Zoologia, Universidad de Guadalajara,

Apdo. postal 1-139, Zapopan, Jal., 45101, Mexico

*Secdo de Micologia e Liquenologia, Instituto de Botanica,

Caixa Postal 3005, CEP 01061-970, Sado Paulo, SP, Brazil

*CORRESPONDENCE TO: lguzman.cucba@gmail.com

ABSTRACT — Eighteen species of Ganoderma (Basidiomycota, Polyporales, Ganodermataceae)

are recorded from Brazil based on specimens deposited at EMBRAPA and SP. Twenty type

specimens from Brazil, Belize, Colombia, Cuba, Ecuador, Finland, France, Grenada, Guinea,

Mexico, Nicaragua, Panama, USA, and Venezuela were studied in order to establish the

proper identity of specimens. Three species (G. mexicanum, G. perzonatum, G. pulverulentum)

are new reports for Brazil, G. weberianum is recorded for the first time for America, and

G. mexicanum, G. perturbatum, and G. perzonatum, are recorded for the first time since their

original description. All species with laccate pileus (except G. vivianimercedianum) and the

non-laccate G. amazonense and G. brownii are described in detail and illustrated.

Key worps — basidiospores, pileipellis cells, taxonomy

Introduction

Ganoderma P. Karst. is a genus with a cosmopolitan distribution, with

many tropical species and some restricted to temperate regions. According to

Ryvarden (2004) the genus is represented in the neotropics by 20 species, a

large number of which are known only from their type locality. Although many

Ganoderma species have been recorded from Brazil, a tropical country with a

great mycological diversity, some have been incorrectly determined and the

occurrence of others has not been confirmed. Except for papers by Torrend

(1920), Loguercio-Leite et al. (2005) and Gomes-Silva et al. (2011), there have

been no detailed studies of Ganoderma in Brazil, and taxa not described by them

are only recorded in lists, so they have been neither described nor illustrated.

Previous records from Brazil have been listed under the following names:

G. amazonense, G. annulare, G. applanatum, G. australe, G. bibadiostriatum,

94 ... Torres-Torres, Guzman-Davalos & Gugliotta

G. brownii, G. citriporum, G. chalceum, G. colossus, G. concinnum, G. curtisii,

G. dorsale, G. elegantum, G. lobatoideum, G. lobatum, G. lucidum, G. multicornum,

G. multiplicatum, G. multiplicatum var. vitalii, G. neurosporum, G. nitens,

G. nitidum, G. oerstedii, G. opacum, G. orbiforme, G. parvulum, G. perturbatum,

G. pygmoideum, “G. reniformis” [Elfvingia reniformis], G. resinaceum, G. sessile,

G. sessiliforme, G. stipitatum, G. subamboinense, G. subfornicatum, G. testaceum,

G. tornatum, G. tropicum, G. vivianimercedianum, and G. zonatum (Baltazar &

Gibertoni 2009; Bononi et al. 1981, 2008; Da Silva & Minter 1995; de Jesus

1993, 1996; de Meijer 2001, 2006; Drechsler-Santos et al. 2008, 2009; Fidalgo

1968; Furtado 1967; Gerber 1996; Gerber & Loguercio-Leite 1997; Gibertoni

& Cavalcanti 2003; Gibertoni & Drechsler-Santos 2010; Gdes-Neto 1999;

Goées-Neto et al. 2003; Gomes-Silva & Gibertoni 2009; Gomes-Silva et al. 2011;

Gottlieb et al. 1998; Gottlieb & Wright 1999a,b; Groposo & Loguercio-Leite

2005; Loguercio-Leite & Wright 1991; Loguercio-Leite et al. 2005; Moncalvo

& Buchanan 2008; Patouillard 1889; Rajchenberg & de Meijer 1990; Rick 1960;

Ryvarden 2004; Ryvarden & de Meijer 2002; Sotao et al. 1991, 2002; Steyaert

1962, 1980; Torrend 1920; Torres-Torres et al. 2008; Westphalen et al. 2010). Of

these, the most commonly reported species are G. applanatum and G. lucidum

sensu lato. As a continuation of the work by Torres-Torres & Guzman- Davalos

(2005, 2007, 2008) and by Torres-Torres et al. (2008), the goal of the present

study is to contribute to the knowledge of Ganoderma through reviewing

species occurring in Brazil by examining the specimens deposited at SP and

EMBRABA herbaria.

Materials & methods

Specimens studied

Materials studied were from EMBRAPA (personal collection of A.A.R. de Meijer)

and SP herbaria. Type specimens were requested on loan to herbaria BPI, ENCB, FH,

NY, O, PC, and UPS. Herbarium abbreviations follow Holmgren et al. (1990).

Macro and micromorphological observations

Key colors are from Kornerup & Wanscher (1963). Micromorphological observations

were made from material mounted in 10% KOH and Melzer’s reagent using a 100x oil-

immersion objective in Zeiss K7 or Zeiss Axioskop 40 microscope. Basidiospore shape

is based on the Q ratio (length/broad ratio; Bas 1969: 320-321) of 20 randomly selected

mature basidiospores. Although basidiospores were examined using Axio Vision 4

software with the Zeiss Axioskop 40 microscope with 1600x magnification, the details

can also be observed with 1250x magnification).

Main macromorphological features for delimiting species included spongy versus

woody basidiomata; pileus crust thickness and hardness; width, color stratification

(homogeneous, not fully homogeneous, or duplex), and resinous deposits of the context

(Gottlieb & Wright 1999a,b; Steyaert 1980). In duplex context there is an abrupt change,

Ganoderma (Brazil) ... 95

with two separate, contrasting colors; ‘not fully homogeneous’ is used when there is

an evident but gradual color difference between the upper and lower parts without

abrupt color changes. Resinous deposits are hard and brittle substances, which may be

dull or shiny and continuous (lines) or discontinuous (incrustations) (Torres-Torres &

Guzman-Davalos 2012). Main micromorphological features taken into consideration

were basidiospore size and apex, disposition and size of basidiospore pillars, and

size, shape, and incrustations of the pileipellis cells (Bazzalo & Wright 1982; Gottlieb

& Wright 1999a,b; Steyaert 1967, 1980). Basidiospore pillars can be free, subfree,

partially anastomosed, or anastomosed. Free pillars appear as independent dots on the

basidiospore surface; subfree is when free dots are mixed with two fused pillars or with

shortly elongated structures. The term partially anastomosed is used when 2 two pillars

grow together to form an irregular surface. Terms and concepts used in the descriptions

are defined in Torres-Torres & Guzman-Davalos (2012).

Results

We reviewed Brazilian specimens associated with almost all vegetation types

present in Brazil and deposited in EMBRAPA and SP and further checked

specimens (including 20 types) deposited in other herbaria. Specifically, four

Brazilian type specimens were compared with 16 types from Belize, Colombia,

Cuba, Ecuador, Finland, France, Grenada, Guinea, Mexico, Nicaragua, Panama,

USA, and Venezuela.

We report 18 Ganoderma species from Brazil, including three (G. mexicanum,

G. perzonatum, G. pulverulentum) that are new records for Brazil and one

(G. weberianum) recorded for the first time in America. Ganoderma mexicanum,

G. perturbatum and G. perzonatum are recorded for the first time since their

original description. An identification key to the Ganoderma species studied for

this paper is presented. Complete descriptions and illustrations for species with

laccate pileus surface (excluding G. vivianimercedianum, which was recently

described elsewhere) are given; only micromorphological data are provided

for all species with dull surfaces, except that G. amazonense and G. brownii

are completely described because both species lack modern descriptions.

Ganoderma applanatum and G. australe are not described because both are

quite common and widely covered in the literature (Baltazar & Gibertoni 2009,

Gibertoni & Cavalcanti 2003, Loguercio-Leite et al. 2005, Pegler & Young 1973,

Steyaert 1975, Telleria 1980).

Key to Ganoderma spp. from Brazil studied in this work

OPTUS ULE ee, Fe ee Sen ee ey are, CEC been, Cee Petey, Seen Oe ae eran Sete Fr, 2

ba Piletisrobecay ster eM nore yh te Tee cea Rela Api eae ee Cee tn Meade 3 eee Ds te ts nel 5

Pt CCONLERT DEO WIth err delat ante dah Wain dak Wain ded Sveti cb deedons dad hoa had hog ted cho Ded chy tt phe Pei ted 3

2. Context pale, not fully homogeneous; basidiospores 7-9 x 5-7 um,

mentite gaptl lars xc Meee, eur ene, ay Me, IG hac hae eM La eet ty AMR a G. amazonense

96 ... Torres-Torres, Guzman-Davalos & Gugliotta

Ra an vB zB

13:

13,

14.

15:

15.

16.

Pileus crust very hard, generally = 0.5 mm thick; context with resinous bands;

basidiosporessS— 13:4 67s A re oem ean ees etapa rea being aed dahoe 4

. Pileus crust hard, generally < 0.5 mm thick; context without resinous bands;

basidiospores! 7-9; % Da: Uia vests, Soret Lowa she Mere see Mera wee tora he G. applanatum

. Basidiospores with free, 0.3-0.4 um thick inter-walled pillars........... G. australe

. Basidiospores with anastomosed, 0.5-0.8 um thick inter-walled pillars... G. brownii

. Basidiomata stipitate; basidiospores with subacute apex.............. 20. eee ee ee 6

. Basidiomata sessile to stipitate; basidiospores with truncate apex................. 8

. Context not fully homogeneous, brown; pileipellis cells without granulations ...... 7

. Context duplex, light orange or brownish orange above and light brown to brown

below; pileipellis cells with or without granulations ................. G. dorsale

. Pileipellis cells entire; basidiospores 10-13 x 8-10 um............. G. perturbatum

. Pileipellis cells with up to four protuberances;

basidiospores 0=13 XS =7 tas ee se siesta teet Fh suet Pane e aaeed tks G. elegantum

. Context pale in general, sometimes with a narrow darker zone towards the tubes .... 9

ON TEME TSE DRO WIC: DROW eis ge wsste ge sessed paaeicle raise ratings sea ape oie Be ees 14

. Context homogeneous to not fully homogeneous but not duplex;

basidiospores 8-10 x 6-7 um, with free or subfree pillars................... 10

. Context duplex; basidiospores 11-14 x 7-9 um, with free pillars.......... G. sessile

. Context without resinous deposits; pileipellis cells clavate,

WAthOUite ran Ul At ONS. sy sic cei Poets «2 oily Touatc ute fastest 4 G. sessiliforme

. Context with resinous deposits; pileipellis cells cylindrical to

narrowlyclavate, witheoratiul ations: 1.8.2 praslee sracene caiate arate atts oe Bisa eons 11

. Pileus surface soft, easy to penetrate with the fingernail....................... 12

. Pileus surface hard, difficult to penetrate with the fingernail................... 13

. Pileipellis cells with concentric elongate granulations in the apex;

basidiospores 8-10 x 6-7 um, with free pillars ................. G. perzonatum

Pileipellis cells without concentric elongate granulations in the apex;

basidiospores 9-12 x 6-8 um, with subfree pillars ...... G. vivianimercedianum

Context changing to yellow when cut, not fully homogeneous;

basidiospores. witlksubiree pillars... 8.5 een tat es ae ana, a G. weberianum

Context unchanging, homogeneous; basidiospores with free pillars. ...G. parvulum

Context homogeneous; pileipellis cells entire, almost cylindrical to cylindrical;

basidiospores 11-13 x 6-7 um, with free pillars ................ G. resinaceum

Context not till whomoceneOus sy 4 o25 nko ds eit d welt edd Fhe be foe baat 9 a 15

Pileipellisccell Men tines a. '. vei ore shears in suse e atin aan auee an eee are ameengee ett 16

Pileipellis-cells-with:protuberances*or: branches. «42.5 eating ea klne ee anes eee 17

Basidiospores 9-11 x 6-7 um, with subfree pillars ................ G. mexicanum

Basidiospores 9-13 x 6-8 um, with partially anastomosed pillars..... G. pulverulentum

Ganoderma (Brazil) ... 97

17. Pileipellis cells with up to 14 lateral or apical protuberances;

basidiospores 8-10 x 6-7 um, with free pillars ............... G. multiplicatum

17. Pileipellis cells with up to 10 lateral or apical protuberances or branches;

basidiospores 2h3 i680 4.4% isan ge eng eh eI ely Oe heh ae Raders 18

18. Pileipellis cells usually with a constriction, generally with <5 protuberances

and 1-2 branches, apex with ferruginous granulations;

Basidiospores withitree pillars. nl. gle ewgelnnogeles cane nana G. subfornicatum

18. Pileipellis cells irregular, with <10 lateral or apical protuberances or branches,

without granulations in the apex; basidiospores with subfree pillars ..... G. orbiforme

Brazilian species

Ganoderma amazonense Weir, U.S. Dep. Agr. Bull. 1380: 12, 84 (1926).

BASIDIOMATA 7.5-9.5 x 5-9.5 x 2-3 cm, perennial, sessile to substipitate,

usually with a narrow base, occasionally imbricate, woody. PiLEus round-

flabelliform to circular, slightly convex to generally applanate; surface glabrous,

bumpy, dull; with a hard crust, difficult to penetrate with the fingernail; surface

reddish-brown (8E6); margin white, lobulate, thin. CoNTEXT 1.5-2 cm thick,

fibrous, not fully homogeneous, pale in general, pale orange to light orange

(5A3, 5A4) above, gradually changing to light brown (6D7) toward tubes,

zonate; without resinous deposits. PoREs 3-5 per mm, angular to round, woody;

pore surface yellow (3A2) to chrome-yellow (2A8) when fresh, darkening to

ochraceous when aging and drying; tubes 0.4-0.5 cm thick, unstratified to

stratified, concolorous with the lower part of the context. HyPHAL SYSTEM

dimitic. CONTEXTUAL TRAMA: no generative hyphae were observed; skeletal

hyphae 3-5 um diam., generally solid to thick-walled, non-septate, unbranched,

hyaline to light yellow. PILEIPELLIs an irregular crustotrichoderm; hyphal ends

impregnated inaresinous substance, entire, solid, golden-yellow. BASIDIOSPORES

8-9 x (5-)6-7 um, Q = 1.23-1.38, widely ellipsoid to ellipsoid, apex truncate,

light yellow, inamyloid; perisporium wrinkled, hyaline; exosporium with

inter-walled pillars up to 0.3 um thick, free. BAsip1A not observed. CysTIDIA

absent.

Hasirat: Tropical secondary vegetation, tropical rainforest, plantation of

Hevea brasiliensis.

MATERIAL EXAMINED: BRAZIL, ParA, Cocal Grande, on Spondias lutea in Hevea

brasiliensis plantation, 20 August 1923, J.R. Weir s.n. (BPI 62043, lectotype).

COLOMBIA, Cuoco, Municipality of Atrato, Yuto, 27 September 2002, M.G. Torres-

Torres 131 (XAL, CHOCO), COSTA RICA, CARTAGO, Turrialba, La Central, without

date, A. Jiménez s.n. (USJ 66156); PUNTARENAS, Osa, Sierpe, Corcovado National Park,

24 February 2000, E. Fletes 1296 (INB).

REMARKS: The remarkable features of G. amazonense are its dull pileus, pale

context, and small light yellow basidiospores. Although we did not check other

materials from Brazil except for the type specimen, we examined collections

98 ... Torres-Torres, Guzman-Davalos & Gugliotta

Fics. 1-15. Basidiomata of Ganoderma. 1. G. brownii (SP214649). 2-3. G. dorsale (SP45190,

SP45819). 4. G. elegantum (SP211483). 5. G. mexicanum (SP109514). 6. G. multiplicatum

(SP211484). 7. G. orbiforme (SP211911). 8. G. parvulum (NY type). 9. G. perturbatum (SP60278).

10. G. perzonatum (SP50687). 11. G. pulverulentum (SP103257). 12. G. resinaceum (SP61535).

13. G. sessiliforme (SP61432). 14. G. subfornicatum (SP375899). 15. G. weberianum (SP61099).

Bar = lcm.

Ganoderma (Brazil) ... 99

from Colombia and Costa Rica. Slightly smaller basidiospores were described

by Gottlieb & Wright (1999b; 7-8 x 5-6 um), Ryvarden (2004; 7-8 x 5-6

(-7) um), and Gomes-Silva et al. (2011; (6-)7-8 x 5-6(-7) um), while Welti

& Courtecuisse (2010) cited measurements (8-9 x 6-7 um) that agree with

ours. Ganoderma amazonense has been recorded from other Brazilian localities

as well as from several neotropical countries and Africa (e.g., Furtado 1967;

Gomes-Silva & Gibertoni 2009; Gomes-Silva et al. 2011; Ryvarden 2004;

Steyaert 1980; Welti & Courtecuisse 2010). The Asian records (e.g., Corner

1983) are doubtful (see Welti & Courtecuisse 2010).

Ganoderma applanatum (Pers.) Pat., Hyménomyc. Eur.: 143 (1887) [nom. cons.].

= Ganoderma lipsiense (Batsch) G.E. Atk., Ann. Mycol. 6: 189 (1908).

BASIDIOSPORES 7-9.6 x 5.6-6.4 um, Q = 1.4-1.57, ellipsoid, apex

truncate, yellowish-brown, inamyloid; perisporium wrinkled, reddish-brown;

exosporium with inter-walled pillars 0.3-0.4 um thick, free.

Hasirat: Rain forest, secondary forest.

MATERIAL EXAMINED: BRAZIL, PARANA, Curitiba, SAo José dos Pinhais, ADEA Reserva

Biolégica Cambui, on dead palm trunk, 4 March 1980, A.A.R. de Meijer 386 (EMBRAPA);

Antonina, Marumbi, Parque Marumbi, Rio do Nune, on dead dicotyledonous trunk, 12

December 1987, A.A.R. de Meijer 962A (EMBRAPA).

REMARKS: A very common species with a wide distribution, G. applanatum

is characterized by a pileus crust <0.5 mm thick, a context without resinous

deposits, and small basidiospores. For a complete description, refer to Kotlaba

& Pouzar (1971), and Telleria (1980).

Ganoderma australe (Fr. : Fr.) Pat., Bull. Soc. mycol. Fr. 5: 71 (1889).

= Polyporus australis Fr. : Fr., Elench. Fung. 1: 108 (1828).

= Ganoderma tornatum (Pers.) Bres., Hedwigia 53: 55 (1912).

BASIDIOSPORES 8.4—12 x 6-7.2 um, Q = 1.4-1.57, ellipsoid, apex truncate,

yellowish-brown, inamyloid; perisporium wrinkled, reddish-brown;

exosporium with inter-walled pillars 0.3-0.4 um thick, free.

Hasirat: Dense ombrophilous forest.

MATERIAL EXAMINED: BRAZIL, PARANA, Curitiba, SAo José dos Pinhais, ADEA Reserva

Biolégica Cambui, 3 February 1979, A.A.R. de Meijer 23 (EMBRAPA); on stump, 15 July

1979, A.A.R. de Meijer 98 (EMBRAPA).

REMARKS: The main characteristics of G. australe are a very hard pilear crust

thicker than 0.5 mm (although young specimens may have a thinner but hard

crust) and a context with resinous deposits. Ganoderma applanatumdiffers

in smaller basidiospores and a thinner pilear crust. Although a cosmopolitan

species, occurrences of G. australe are often underestimated because of wrong

determinations. Studied specimens agree with the descriptions [some cited

as G. tornatum] of Gottlieb & Wright (1999b), Kotlaba & Pouzar (1971),

100 ... Torres-Torres, Guzman-Davalos & Gugliotta

Melo (1986), Nufiez & Ryvarden (2000), Pegler & Young (1973), Ryvarden &

Johansen (1980), Steyaert (1967, 1975), Telleria (1980); refer to these authors

for a complete description.

Ganoderma brownii (Murrill) Gilb., Mycologia 53: 505 (1962 [“1961”]). Fig. 1

BASIDIOMATA 6.5-16 x 6.5-15 x 1.4-2 cm, perennial, sessile, occasionally

imbricate, woody. PiLEus round-flabelliform to circular, generally applanate;

surface glabrous, bumpy, dull, concentrically sulcate; with a 0.6-0.8 mm

thick crust, not cracking, very hard, difficult to penetrate with the fingernail;

surface brown (7F7); margin concolorous, entire, thin to thick, obtuse, sulcate.

CONTEXT 0.3-0.9 cm thick, fibrous, homogeneous, reddish-brown (9F8),

azonate; generally with resinous deposits close to the pileus’ base. PorEs

4-5 per mm, angular to round, woody; pore surface yellow (3A2) to chrome-

yellow (2A8); tubes 0.4-1.5 cm thick, unstratified to stratified, concolorous

with the lower part of the context. HyPHAL sySTEM trimitic. CONTEXTUAL

TRAMA: no generative hyphae were observed; skeletal hyphae 3.7-9.3 um

diam., generally solid to thick-walled, non-septate, arboriform, yellowish-

brown; no binding hyphae were observed. PILEIPELLIS a crustotrichoderm;

terminal elements 6.8-9.3 um wide, with apex 7-10 um wide, entire, solid,

golden-yellow. BAsip1osPpoREs 9.3-11.2 x 6.6-7.4 um, Q = 1.25-1.5, widely

ellipsoid to ellipsoid, apex truncate, yellowish-brown, inamyloid; perisporium

wrinkled, reddish-brown; exosporium with inter-walled pillars 0.5-0.8 um

thick, anastomosed. Basrp1A4 not observed. CysTip1A absent.

Hasirat: Atlantic dense ombrophilous rain forest.

MATERIAL EXAMINED: USA, CALIFORNIA, Strawberry Canyon, on dead Umbellularia,

November 1910, V.S. Brown 307 (NY, lectotype). BRAZIL, SAo PauLo, Reserva

Bioldgica de Paranapiacaba, 25 August 1987, M. Capelari 1723 (SP214649).

REMARKS: This species is characterized by its very hard, >0.5 mm thick pileus

crust, reddish-brown context, yellow pore surface, and large broadly ellipsoid

to ellipsoid basidiospores. Although Gilbertson & Ryvarden (1986) suggest

that G. brownii is restricted to California, the Brazilian specimen agrees with

the lectotype, which has basidiospores 9-12 x 6.5-8 um. As Bononi et al.

(2008) recorded the species from Mato Grosso do Sul, this represents a second

record for Brazil. Gottlieb & Wright (1999b) cite G. brownii as a synonym of

G. lipsiense, but we consider G. applanatum/G. lipsiense as separate (cf. the two

descriptions).

Ganoderma dorsale (Lloyd) Torrend, Brotéria, sér. Bot. 18: 32 (1920). Figs 2-3, 16-17

= Ganoderma concinnum Ryvarden, Mycologia 92: 183 (2000).

BASIDIOMATA 2-5.5 x 3-10 x 0.5-1.5 cm, annual, stipitate, woody-corky.

Piteus round-flabelliform to reniform, convex to generally applanate; surface

glabrous, bumpy, semiglossy to glossy, concentrically sulcate; with a laccate

Ganoderma (Brazil) ... 101

Fic. 16. Ganoderma dorsale (SP45190). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores. c. Skeletal hyphae of crustohymeniderm.

crust, not cracking, difficult to remove, easy to penetrate with the fingernail;

surface garnet-red (11E8) or violet-brown (11F8), almost homogeneous in

the adult; margin yellowish to concolorous, entire, acute to obtuse, smooth to

sulcate. STIPE 3.5-10 x 0.3-1.5 cm, lateral, cylindrical, solid, context duplex as

basidiomata context; surface smooth to tuberculate, generally very shiny, red-

wine almost black, generally darker than the pileus. CONTEXT 0.2—-0.6 cm thick,

up to 1.2 cm in pileus’ base, fibrous, duplex, pale-orange to light-orange (5A3,

5A4) to brownish orange or caramel (6C6) above, brown (lighter than 6F8) to

light brown (6D7) toward tubes, azonate; generally with resinous bands very

thin and inconspicuous, only one specimen with thick lines; stipe context also

duplex. Pores 4-8 per mm, angular to round, woody; pore surface yellow (3A2);

tubes 0.1-0.7 cm thick, unstratified, concolorous with the lower part of the

context. HYPHAL SYSTEM trimitic. CONTEXTUAL TRAMA: no generative hyphae

were observed; skeletal hyphae 3.1-12 um diam., solid to generally thick-walled,

apex septate, arboriform, yellow to yellowish-brown; binding hyphae 1.9-3.1

um diam., thick to solid, non-septate, yellowish, scarce. HyYMENOPHORAL

TRAMA as the contextual trama. PILEIPELLIS a crustohymeniderm, cells

46-72 x 7.2-13.6 um, narrowly clavate to clavate, entire or with one lateral

protuberance; thick-walled, golden-brown, with or without granulations in

the apex; dextrinoid. BAsip1osporgs 11.8-14 x 8-9.3(-10) um, Q = 1.4-1.65,

ellipsoid, apex subacute, yellowish-brown, inamyloid; perisporium wrinkled,

102 ... Torres-Torres, Guzman-Davalos & Gugliotta

Fic. 17. Ganoderma dorsale (SP45819). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c-d. Crustohymeniderm: c. Skeletal hyphae. d. Binding hyphae.

Ganoderma (Brazil) ... 103

reddish-brown; exosporium with inter-walled pillars 0.5-0.6 um thick, partially

anastomosed; with apical germ pore difficult to observe. BAsip14 not observed.

CysTIDIA absent.

HasitaT: Seasonal semi-deciduous forest, seasonal semi-deciduous

submontane forest, seasonal semi-deciduous alluvial forest, tropical rain

forest.

MATERIAL EXAMINED: BRAZIL, R10 GRANDE DO SUL, on buried wood, without date,

RJ. Rick s.n. (BPI 303197, holotype); PARANA, Curitiba, Centro Politécnico, Jardim

das Americas, on buried roots of living dicotyledonous tree, 18 January 1989, A.A.R.

de Meijer 1194 (EMBRAPA); Parque Boriqui, on soil, 5 March 1993, A.A.R. de Meijer

2533 (EMBRAPA)); Reserva Biolégica Cambui, on forest soil, 24 November 1994, A.A.R.

de Meijer 2961 (EMBRAPA); Reserva Biolodgica Cambui, on buried dicotyledonous, 20

May 1995, A.A.R. de Meijer 2278 (EMBRAPA); Jundiai do Sul, Fazenda Monte Verde, on

decayed dicotyledonous, on buried wood, 4 February 1996, A.A.R. de Meijer 3253 (O,

EMBRAPA); R10 GRANDE DO SUL, Pelotas, Horto Florestal, buried, 9 June 1959, E.C.

Santos 29 (SP45190); SAo PAULO, lepe, about 5 km of Porto Alvarado, along of the Rio

Paranapanema, Fazenda CAPI, 9 February 1965, G. Eiten, L.T. Eiten & H. Kimura 6009

(SP102607). COLOMBIA, Cuoco, Municipality of Riosucio, Sautata, Parque Nacional

Katios, 28-30 June 1978, L. Ryvarden 18640 (O, holotype of G. concinnum).

REMARKS: Ganoderma dorsale is characterized by its stipitate basidiome,

duplex context, and basidiospores with subacute apex. Examined specimens

agree with the holotype and with the description by Gottlieb & Wright (1999a,

as “Ganoderma lucidum var. dorsale”). Although Moncalvo & Ryvarden

(1997) reported this species as common in Brazil and recorded it also from

Malaysia and Philippines, it was not included in the most recent treatment of

neotropical polypores (Ryvarden 2004). Baltazar & Gibertoni (2009) reported

it from the Brazilian Atlantic forest. The species was considered a synonym of

G. perturbatum by Steyaert (1967) and of G. lucidum s.l. by Ryvarden (1990).

The two specimens in SP (SP45190, SP102607) were labeled G. lucidum and

possibly published under the same name. Ganoderma concinnum is considered

a synonym of G. dorsale because it also has a stipitate basidiome, duplex

context, and basidiospores with subacute apex. The only differences found are

granulations of the pileipellis cells and absence of resinous bands in the context

in G. dorsale, not enough to support two separate species. Ryvarden (2000)

suggested that the long stipe distinguished G. dorsale from G. concinnum;

however, there are specimens with very short stipes that agree well with the

G. dorsale concept (e.g., SP45190 and SP102607). Moreover, Ryvarden (2000)

described larger (12-14 x 7-8 um) basidiospores than we measured (8-10 um

wide) in the G. concinnum holotype. Ganoderma concinnum has been recorded

from Brazil by Baltazar & Gibertoni (2009), de Meijer (2006), and Ryvarden

& de Meijer (2002). Ganoderma perturbatum, which is also stipitate and

produces basidiospores with a subacute apex and partially anastomosed pillars,

is otherwise easily distinguished (cf. G. perturbatum).

104 ... Torres-Torres, Guzman-Davalos & Gugliotta

Ganoderma elegantum Ryvarden, Syn. Fung. 19: 81 (2004). Fics 4, 18

BASIDIOMATA 1.7 x 2 x 1 cm, annual, stipitate, woody-corky. Pileus round-

flabelliform, applanate; surface glabrous, glossy, concentrically sulcate; with

a laccate crust, not cracking, easy to penetrate with the fingernail; surface

Fic. 18. Ganoderma elegantum (SP211483). a. Pileipellis cells. b. Basidiospores.

c. Crustohymeniderm: skeletal hyphae.

Ganoderma (Brazil) ... 105

homogeneous reddish-brown (11F8); margin whitish, entire, obtuse to acute,

smooth. STrpE 8.7 x 1.3 cm, lateral, cylindrical, solid; surface smooth, very

shiny, red-wine almost black, darker than pileus. CONTEXT 0.9 cm thick,

spongy-fibrous, not fully homogeneous, yellowish-brown (5E8) to eye-brown

(7F7), azonate; with resinous bands that do not extend up to the margin. PoRES

6-7 per mm, angular to round, woody; pore surface yellowish (2A3); tubes 0.1

cm thick, unstratified, concolorous with the lower part of the context. HYPHAL

SYSTEM dimitic. CONTEXTUAL TRAMA: no generative hyphae were observed;

skeletal hyphae 2.5-8.7 um diam., solid to generally thick-walled, non-septate,

arboriform, yellow to yellowish-brown. HYMENOPHORAL TRAMA: No generative

hyphae were observed; skeletal hyphae 2.5-6.2 um diam., solid to generally

thick-walled, non-septate, arboriform, yellow to yellowish-brown. PILEIPELLIS

a crustohymeniderm, cells 29.8-52.7 x 6.2-10.5 um, clavate, entire or with up

to six lateral or apical branches or protuberances; subthick-walled, golden-

yellow, without granulations in the apex; negative or apex slightly amyloid.

BASIDIOSPORES 10-12 x 5-7 um, Q = 1.4-1.65, oblong to ellipsoid, apex

subacute, yellowish-brown, inamyloid; perisporium wrinkled, reddish-brown;

exosporium with inter-walled pillars up to 0.3 um thick, subfree; endosporium

wrinkled; with apical germ pore difficult to observe. BAsip1a not observed.

CysTIpIA absent.

Hasirat: Amazonian rain forest.

MATERIAL EXAMINED: ECUADOR, Yasuni National Park, Yasuni Scientific Reserve,

on dead hardwood log, 12 March 2002, Ryvarden 44573 (O, holotype). BRAZIL,

RONDONIA, Jart, near the airport, on wood, 10 October 1986, M. Capelari & R. Maziero

962 (SP211483).

REMARKS: Ganoderma elegantum was recently described from Ecuador

(Ryvarden 2004) and recorded in a checklist from Brazilian Amazonia by

Gomes-Silva & Gibertoni (2009; based on a specimen determined by Torres-

Torres in 2005). Although the Brazilian specimen cited above has many

immature basidiomata with few basidiospores, the features agree in general

with the holotype. Ryvarden (2004) described apically widened pileipellis cells

without protuberances, but we observed up to six lateral or apical branches or

protuberances per cell in both the holotype and Brazilian specimens.

Ganoderma mexicanum Pat., Bull. Soc. mycol. Fr. 14: 54 (1898). Fics 5, 19

BASIDIOMATA 6.2-11.2 x 9-19 x 1.6-2.5 cm, annual, sessile, woody, but

light in weight when dry. Piteus round-flabelliform to semicircular, convex

to applanate; surface glabrous, rugose, semiglossy; with a laccate crust, thin,

not easily cracked or removed, but easy to penetrate with the fingernail,

concentrically sulcate; surface reddish-black in a 80% of the surface to brown-

violet (11F5) towards the margin, almost homogeneous; margin henna (7E8)

to concolorous with the pileus, slightly lobulate, thin to thick, obtuse, smooth.

106 ... Torres-Torres, Guzman-Davalos & Gugliotta

Fic. 19. Ganoderma mexicanum (SP109514). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c-d. Crustohymeniderm: c. Generative hyphae. d. Skeletal hyphae.

CONTEXT 0.4-0.8 cm thick, fibrous, not fully homogeneous, pale-orange to

light brown (5A3, 5B6) in the upper portion, brown or cognac (6E7) toward

the tubes, azonate; with resinous bands up to half of the context. Pores 3-5

Ganoderma (Brazil) ... 107

per mm, round, woody; pore surface pale yellow (2A3); tubes 0.8-1.1 cm thick,

unstratified, concolorous with the lower part of the context. HyYPHAL SYSTEM

dimitic. CONTEXTUAL TRAMA: generative hyphae 1.8-5 um diam., thin-walled,

hyaline, non-branched; skeletal hyphae 1.9-9.3 um diam., thick-walled to

generally solid, non-septate, arboriform, yellowish. HYMENOPHORAL TRAMA

as the contextual trama. PILEIPELLIS a crustohymeniderm, cells 35.2-72.4 x

6.8-10.5 um, clavate to narrowly clavate, some apically widened, entire or rarely

with one lateral protuberance; very thick-walled, golden-brown to yellowish-

brown, without granulations in the apex; dextrinoid. BAstp1osPpoREs 9.3-10.6

x 6.2-7.4 um, Q = 1.42-1.54, ellipsoid, apex truncate, yellowish-brown,

inamyloid; perisporium wrinkled, reddish-brown; exosporium with inter-

walled pillars 0.3 um thick, subfree. BAsip1a not observed. Cystrip1A absent.

Hapsirat: Not specified.

MATERIAL EXAMINED: MEXICO, EstTapo DE MExiIco, D. de Jonacatepec, Tepalcingo,

22 October 1890, s. coll. (FH, lectotype). BRAZIL, SAo PauLo, Tremembé, Horto

Florestal de Cantareira, on trunk of Araucaria angustifolia, 3 March 1953, C.D.F

Pacheco s.n. (SP109514).

REMARKS: The most important features of G. mexicanum are a not fully

homogeneous context, apically widened pileipellis cells, and small basidiospores

with subfree pillars. The studied specimen agrees with the lectotype, which,

however, “seems” to have a homogenous context without resinous bands.

A possible explanation is that the type is poorly conserved with most of the

context destroyed. This species was previously known only from the type locality

(Patouillard 1898). This is the second record of the species for the world.

Ganoderma multiplicatum (Mont.) Pat., Bull. Soc. mycol. Fr. 5: 74 (1889). Fras 6, 20

BASIDIOMATA 5-7 x 4-12 x 1-3 cm, perennial, sessile to substipitate, woody.

PitEus round-flabelliform with a contracted base, slightly convex to applanate;

surface glabrous, smooth to bumpy, glossy; with a laccate crust, thin, not easily

cracked or removed, but easy to penetrate with the fingernail, concentrically

sulcate; surface very dark reddish-black to dark violet-brown (11F5) in

80-90% of the pileus, the remaining area rust-brown (6E8) to dark orange

(5B8); margin whitish, entire to slightly lobulate, obtuse, smooth. CONTEXT

0.6-0.9 cm thick, fibrous, not fully homogeneous, light yellowish-brown

changing to light brown (6D7) toward the tubes, zonate; with resinous bands.

Pores 4-6 per mm, angular to round, woody; pore surface pale yellow (2A3);

tubes 0.4-0.8 cm thick, unstratified, concolorous with the lower part of the

context. HYPHAL SYSTEM dimitic. CONTEXTUAL TRAMA: no generative hyphae

were observed; skeletal hyphae 3.8-7.5 um diam., subthick-walled to generally

solid, non-septate, arboriform, yellow. HYMENOPHORAL TRAMA: generative

hyphae 2-3.8 um diam., thin-walled, hyaline, abundant; skeletal hyphae

108 ... Torres-Torres, Guzman-Davalos & Gugliotta

Fic. 20. Ganoderma multiplicatum (SP211484). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c. Crustohymeniderm: skeletal hyphae.

Ganoderma (Brazil) ... 109

2.2-3.8 um diam., subthick-walled to generally solid, non-septate, arboriform,

yellow to yellowish-brown. PILEIPELLIS a crustohymeniderm, cells 38.5-62 x

5.6-10 um, clavate, narrowly clavate or irregular, with up to 14 lateral or apical

protuberances; very thick-walled, golden-yellow, without granulations in the

apex; amyloid. BAsIpIosPoREs 8-9.9 x 6.2-6.8 um, Q = 1.3-1.47, ellipsoid,

apex truncate, yellowish-brown, inamyloid; perisporium wrinkled, reddish-

brown; exosporium with inter-walled pillars up to 0.3 um thick, free. BASIDIA

not observed. Cystip1 absent.

Hasirat: Atlantic rain forest, Amazonian rain forest.

MATERIAL EXAMINED: BRAZIL, ALAGOAS, Maceid, Reserva do Ibama, 28 August 1992,

R. Maziero s.n. (SP250605); R10 DE JANEIRO, Mata de Tijuca, 4 May 1966, J.S. Furtado

s.n. (SP91404); RONDONIA, Jaru, right margin of Rio Jaru, 3 October 1986, M. Capelari

& R. Maziero 693 (SP211484); SAo PAULO, Parque do Estado, Instituto de Botanica, on

wood, 23 January 1970, B. Skvortzov s.n. (SP10746); SERGIPE, Estacao Ecoldgica Santa

Isabel, 9 December 2003, R.H. Marino s.n. (SP375895).

REMARKS: Ganoderma multiplicatum is characterized by a reddish-black

pileus, a not fully homogenous context, small basidiospores, and pileipellis cells

with many protuberances. The cited specimens agree with Gottlieb & Wright

(1999a) and Ryvarden (2000, 2004), except that the last author described

pileipellis cells lacking or with only a few small apical protuberances. However,

Gottlieb & Wright (1999a) described pileipellis cells in the type as “elongated

acanthophysis-like elements,’ and the cells we found in the Brazilian specimens

are very similar to their illustrations. This species was described from French

Guiana and recorded from Angola, Argentina, Brazil, India, Indonesia, Ivory

Coast, Seychelles, Sierra Leone, Zaire and Zambia (Baltazar & Gibertoni 2009;

Gomes-Silva & Gibertoni 2009; Gomes-Silva et al. 2011; Gottlieb & Wright

1999a; Ryvarden 2000; Steyaert 1980). According to Ryvarden (2000), it has

a pantropical distribution. Some EMBRAPA and SP specimens were labeled

G, lucidum.

Ganoderma orbiforme (Fr.) Ryvarden, Mycologia 92: 187 (2000). Fics 7, 21

BASIDIOMATA 4-6.5 x 5-8 x 1.5-2 cm, perennial, sessile, occasionally

imbricate, woody. PrtEus round-flabelliform, convex to applanate; surface

glabrous, bumpy, glossy; with a laccate crust, thin, not easily cracked or removed,

but easy to penetrate with the fingernail, concentrically sulcate; surface very

dark reddish-black in 80-90% of the surface to light-brown (6D8) toward the

margin; margin light-brown (6D8), entire, obtuse, generally sulcate. CONTEXT

0.8-1 cm thick, fibrous, not fully homogeneous, light golden-brown (5D) to

rust-brown (6E8) toward the tubes, zonate; with discontinuous resinous bands.

Pores 4-8 per mm, round, woody; pore surface yellow (3A6); tubes 0.7-0.9

cm thick, unstratified to stratified, concolorous with the lower part of the

context. HYPHAL SYSTEM trimitic. CONTEXTUAL TRAMA: no generative hyphae

110 ... Torres-Torres, Guzman-Davalos & Gugliotta

Fic. 21. Ganoderma orbiforme (SP211911). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c-d. Crustohymeniderm: c. Generative hyphae. d. Skeletal hyphae.

were observed; skeletal hyphae 2.5-8.8 um diam., generally solid, non-septate,

arboriform, yellow to yellowish-brown. HyMENOPHORAL TRAMA: generative

hyphae 2.5-5 um diam., thin-walled, hyaline, branched; skeletal hyphae 2.5-8

um diam., generally solid, non-septate, arboriform, golden-yellow to yellowish-

brown; binding hyphae 1.9-2.5 um, thick-walled, hyaline to yellowish, scarce.

PILEIPELLIS a crustohymeniderm, cells 33.5-62 x 4.4-9.3 um, irregular, with

up to 10 lateral or apical protuberances or branches; thick-walled, golden-

yellow to golden-brown, without granulations in the apex; strongly amyloid.

BASIDIOSPORES 9-11.2(-13) x 6.9-8(-8.6) um, Q = 1.38-1.5, ellipsoid, apex

truncate, yellowish-brown, inamyloid; perisporium wrinkled, reddish-brown;

Ganoderma (Brazil) ... 111

exosporium with inter-walled pillars 0.4-0.5 um thick, subfree. BAsIp1A not

observed. Cystip1a absent.

Hasirat: Atlantic rain forest, artificial Pinus forest.

MATERIAL EXAMINED: GUINEA, without data, A. Afzelius s.n. (UPS, holotype).

BRAZIL, Rio DE JANEIRO, Angra de Reis, Ilha da Gilpoia, 4 March 1956, O. Fidalgo

F-365 (SP95346); PARANA, Paranagua, Ilha do Mel, on dead standing trunk of Ocotea

pulchella, 6 May 1989, A.A.R. de Meijer 1234 (EMBRAPA); SAo Pauto, Agua Funda,

Secretaria de Agricultura, on trunk of Pinus sp., 31 March 1986, M.A. de Jesus s.n.

(SP211911); Trilha do Jardim Botanico de Sao Paulo, 12 February 2004, A.M. Gugliotta

& G.R. Leal 1205 (SP382045).

REMARKS: Ganoderma orbiforme is characterized by a very dark (almost black)

pileus, not fully homogenous context, pileipellis cells with many protuberances

and branches, and basidiospores with subfree pillars. The Brazilian specimens

agree with the holotype, although it comprises only one poorly conserved

basidiome with only rust-brown (6E8) context remnants, from which it is

impossible to elucidate structure or presence of resinous bands. Ryvarden (2000,

2004) described broadly ellipsoid basidiospores, but this shape agrees with

neither his measurements (10-11 x 6-7 um) nor the ellipsoid basidiospores of

the type specimen (9-11 x 7-8 um). Ganoderma orbiforme is morphologically

similar to G. multiplicatum, but the latter has smaller basidiospores (8-9.9 x

6.2-6.8 um) with free pillars and pileipellis cells with only short protuberances.

Although recorded from the states of Amazonas and Parana, Brazil by de Meijer

in 2006 (Baltazar & Gibertoni 2009), Gomes-Silva et al. (2011), and Ryvarden

& de Meijer (2002), G. orbiforme probably has a wider tropical distribution. We

record this species from the states of Rio de Janeiro and Sao Paulo here for the

first time.

Ganoderma parvulum Murrill, Bull. Torrey bot. Club. 29: 605 (1902). —— Fias 8, 22

= Ganoderma stipitatum (Murrill) Murrill, N. Amer. Fl. 9: 122 (1908).

BASIDIOMATA 1.7-5.5 x 2.2-4 x 0.7-1.3 cm, annual, substipitate to sessile,

with a contracted base, occasionally imbricate, woody. PILEUs semicircular,

round-flabelliform to flabelliform, convex to applanate; surface glabrous,

rivulose to slightly radially rugose, hard, glossy to dull; with a laccate crust,

not cracked, hard (difficult to penetrate with the fingernail), but easily lost

and then leaving the surface relatively homogeneously dull, zonate; surface

reddish-black to violet-brown (11F4) in 80-90% of the pileus, reddish-brown

(9E8) towards the margin; margin pure white or yellowish, entire to slightly

lobulate, thin, smooth. SuBsTIPE when present 0.9-1.5 x 1-1.5 cm, short and

thick, thinner toward the base, horizontal, slightly darker than pileus, solid.

CONTEXT 0.4-0.8 cm thick, fibrous, homogeneous, cream (4A3), azonate;

with resinous dark bands up to half of the context. PoREs 5-6 per mm, round,

woody; pore surface yellowish-white (3A2) to light yellow (444) when fresh,

112 ... Torres-Torres, Guzman-Davalos & Gugliotta

darkening to ochraceous or yellowish-brown (6E8) when aging and drying;

tubes 0.2-0.4 cm long, unstratified, dark brown contrasting with the context.

HYPHAL SYSTEM dimitic. CONTEXTUAL TRAMA: generative hyphae 2.4-5.6

um diam., thin-walled, with conspicuous clamps, non-branched, hyaline to

yellowish; skeletal hyphae 5.6-12 um diam., thick-walled to generally solid,

septate to non-septate, arboriform with few branches, moderately branched,

golden-yellow. HYMENOPHORAL TRAMA as the contextual trama. PILEIPELLIS a

crustohymeniderm, cells 48-60 x 7.2-9.6 um, cylindrical to narrowly clavate,

entire or rarely with one lateral protuberance, thick-walled to solid, apex with

granulations, golden-yellow, amyloid. BAsipIosporREs 8-9 x 6-6.8 um, Q =

1.18-1.38, broadly ellipsoid, apex truncate, with apical germ pore, yellowish-

brown, inamyloid; perisporium wrinkled, reddish-brown; exosporium with

inter-walled pillars up to 0.3 um thick, free; endosporium wrinkled. BAsIDIA

not observed. CysTip1A absent. CHLAMYDOSPORES 9.6-10.5 um, globose,

thick-walled, with inter-walled very thick pillars, yellow.

Hasirat: Frondose forest, mixed ombrophilous alluvial forest.

MATERIAL EXAMINED: NICARAGUA, without data, C.L. Smith s.n. (NY, lectotype

of G. parvulum; BR, isolectotype); without data, C.L. Smith s.n. (NY, lectotype of G.

stipitatum; BR, isolectotype). BRAZIL, PARANA, Curitiba, Sao José dos Pinhais, ADEA-

reserve, Reserva Biolégica Cambui, on decayed tree, 7 July 1979 A.A.R. de Meijer 91

(EMBRAPA); 10 June 1987, A.A.R. de Meijer 850 (EMBRAPA); on dead dicotyledonous

trunk, 14 December 1989, A.A.R. de Meijer 1418 (EMBRAPA).

REMARKS: Ganoderma parvulum may be recognized by its generally small

basidiomata, hard pileus crust, resinous dark bands contrasting with the

pale context, cylindrical pileipellis cells with apical granulations, and small

basidiospores with free pillars. The species is morphologically similar to

G. perzonatum and G. weberianum, but G. perzonatum has larger basidiomata,

a soft pilear laccate crust (easy to penetrate with the fingernail), a not fully

homogeneous context, and pileipellis cells with concentric elongate granulations

while G. weberianum has a not fully homogeneous context that changes to

yellow when cut and basidiospores with subfree pillars. In his description of

G. parvulum, Ryvarden (2000, 2004) reported “the lower part of context dark

brown, darker than the intermediate and upper paler brown parts,’ which, in

our terminology, is the equivalent to a not fully homogenous context. He also

noted pileipellis cells without incrustations. He synonymized G. parvulum

with G. stipitatum, recorded by him for Brazil (Ryvarden 2000). We agree

with Ryvarden (2004) that they are synonyms, but the name G. parvulum has

priority over G. stipitatum. Steyaert (1980) cited a wrong reference for the

G. parvulum protologue (“North. Am. Fl. 9: 13, 1908”), and this was apparently

followed by Ryvarden (2004). Ryvarden (2004) cited G. stipitatum from Bolivia,

Brazil, Costa Rica, Nicaragua, Peru, Suriname, and Venezuela, but some of the

cited specimens have a dark brown context and larger basidiospores and may

Ganoderma (Brazil) ... 113

Fic. 22. Ganoderma parvulum (NY type). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c. Crustohymeniderm: skeletal hyphae. d. Chlamydospores.

114 ... Torres-Torres, Guzman-Davalos & Gugliotta

represent another species (unpublished observations by M.G. Torres-Torres).

Gomes-Silva et al. (2011) and Welti & Courtecuisse (2010) cited this species

(as G. stipitatum) from Brazil, Belize, and French Guiana. The specimens

from USA (NY) noted by Welti & Courtecuisse (2010) and revised by Torres-

Torres in 2008 correspond to G. parvulum [= G. stipitatum]. Rajchenberg &

de Meijer (1990) cited G. parvulum from Parana, Brazil (specimen de Meijer

850); but Ryvarden & de Meijer (2002) reported de Meijer 850 and 1418 as

G. resinaceum. Our examinations show both collections to be very different

from G. resinaceum and with the cream context and smaller basidiospores that

corresponds to G. parvulum.

Ganoderma perturbatum (Lloyd) Torrend, Brotéria, sér. Bot. 18: 34 (1920). Fries 9, 23

BASIDIOMATA 4 X 5 x 1.2 cm, perennial, stipitate, woody-corky, light in

weight. PILEus reniform, convex; surface glabrous, bumpy, semiglossy to glossy,

concentrically sulcate; with a laccate crust, not cracking, difficult to remove,

easy to penetrate with the fingernail; surface almost homogeneous violet-

brown (darker than 11F7 or 11F8); margin whitish to yellow, entire, obtuse,

sulcate. Stipe 4.7 x 1 cm, lateral, cylindrical, solid, very dark violet-brown

(11F8) to black, concolorous to generally darker than pileus; surface smooth

to tuberculate, shiny, red-wine almost black, darker than pileus. CONTEXT 0.4

cm thick, fibrous, not fully homogenous, raw-sienna or light brown (6D7),

to slightly darker next to the tubes, azonate; resinous incrustations not very

visible. Pores 3-4 per mm, angular to round, woody; pore surface yellow

(3A2); tubes up to 0.8 cm thick, stratified, concolorous with the lower part of

the context. HYPHAL SYSTEM trimitic. CONTEXTUAL TRAMA: generative hyphae

3.2 um diam., thin-walled, hyaline, scarce; skeletal hyphae 2.5-7.5 um diam.,

generally solid to thick-walled, non-septate to septate in the apex near to the

pileipellis, arboriform, golden-yellow; binding hyphae 1.9-2.2 um diam.,

thick-walled, yellowish, scarce. HYMENOPHORAL TRAMA as the contextual

trama. PILEIPELLIS a crustohymeniderm, cells 43.5-83.8 x 6.2-14.9 um, clavate

to narrowly clavate, entire or rarely with one lateral protuberance or branch;

thick-walled, multistratified, golden-brown, without granulations in the apex;

slightly amyloid. BAsip1osPporEs (9.9—)10.8-12.4(-13) x 8-9.3(-9.9) um, Q =

1.14-1.43, broadly ellipsoid to ellipsoid, few subglobose, apex subacute, with

apical germ pore difficult to observe, yellowish-brown, inamyloid; perisporium

wrinkled, reddish-brown; exosporium with inter-walled pillars 0.5-0.6 um

thick, partially anastomosed. Basip1a not observed. Cystip1a absent.

Hasirat: Not specified.

MATERIAL EXAMINED: BRAZIL, R10 GRANDE DO SUL, Lageado, without date, R. Rick

s.n. (BPI, holotype); Santa Maria, Distrito de Boca do Monte, on dead buried wood, 22

April 1961, J.P. da Costa Neto s.n. (SP60278).

Ganoderma (Brazil) ... 115

Fic. 23. Ganoderma perturbatum (SP60278). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c-d. Hyphal system of crustohymenoderm: c. Generative hyphae. d. Skeletal hyphae.

116 ... Torres-Torres, Guzman-Davalos & Gugliotta

REMARKS: Distinctive features of G. perturbatum are the stipitate basidiome,

a not fully homogeneous context, entire pileipellis cells with multi-stratified

walls, and basidiospores with a subacute apex and partially anastomosed

pillars. Steyaert (1967) synonymized it with G. dorsale and Ryvarden (1990)

considered it in the G. lucidum complex. Ganoderma dorsale is similar, but

differs by a duplex context and generally incrusted pileipellis cells. Although

only a small fragment of the G. Jucidum neotype remains [F. Fennici Exsiccati

239, Finland, Aboa (Turku), Runsala, n. hosp, P.A. Karsten 1858 (H, neotype)],

it demonstrates a pale and homogeneous (maybe not fully homogeneous)

context, and basidiospores with sub-free pillars that distinguish it from

G. perturbatum. The specimen SP60278 was labeled as G. lucidum in SP. This is

the second world record for G. perturbatum. Although known only from Brazil,

this species likely has a wider tropical distribution.

Ganoderma perzonatum Murrill, N. Amer. Fl. 9: 121 (1908). Fries 10, 24

BASIDIOMATA 3.5-10 x 3-12 x 0.6-1.8 cm, annual, substipitate, occasionally

imbricate, woody-corky, light in weight. Pireus flabelliform to round-

flabelliform, convex to applanate; surface glabrous, bumpy, glossy to semiglossy,

concentrically sulcate; with a laccate crust, not cracking, difficult to remove,

easy to penetrate with the fingernail; surface almost homogeneous violet-brown

(11F8), to garnet-red (11E8) towards the margin; margin whitish to generally

concolorous, entire, thin, smooth. CONTEXT 0.3-0.9 cm thick, 0.7 cm average,

fibrous, not fully homogeneous, pale in general, pale-orange to light orange

(5A3, 5A4) upwards, gradually changing to light brown (6D7) toward the tubes

but in a narrow zone, zonate; with resinous bands. PorEs 4-6 per mm, angular

to round, woody; pore surface pale-yellow (2A3) to sun-yellow (2A5); tubes

0.5-0.7 cm thick, unstratified to stratified, concolorous with the lower part

of the context. HyPHAL SYSTEM dimitic. CONTEXTUAL TRAMA: no generative

hyphae were observed; skeletal hyphae 5-13 um diam., generally solid to thick-

walled, non-septate, arboriform, yellowish to golden-yellow. HYMENOPHORAL

TRAMA as the contextual trama. PILEIPELLIS a crustohymeniderm, cells 50-86

x 6.2-13.6 um, cylindrical, apex obtuse to rarely subcapitate, entire or with one

lateral protuberance; thick-walled, generally multi-stratified only in the apex,

golden-yellow, with concentric elongate granulations in the apex; dextrinoid.

BASIDIOSPORES 8.3-10.2 x 5.9-7.4 um, Q = (1.25-)1.33-1.53, widely ellipsoid

to ellipsoid, apex truncate, with apical germ pore, light yellowish-brown,

inamyloid; perisporium wrinkled, hyaline; exosporium with inter-walled pillars

up to 0.3 um thick, free; endosporium wrinkled. Basrp1a 15.5-18.6 x 8-11 um,

utriform, hyaline. CystTrp1A absent.

Hasitat: Mangrove, Amazonian forest.

Ganoderma (Brazil) ... 117

Fic. 24. Ganoderma perzonatum (a, c—e: SP50687, b: SP233313). Bar = 8 um.

a-b. Pileipellis cells c. Basidiospores. d. Crustohymeniderm: skeletal hyphae. e. Basidiola.

118 ... Torres-Torres, Guzman-Davalos & Gugliotta

MATERIAL EXAMINED: CUBA, La HABANA, Santiago de Las Vegas, on mango log, ES.

Earle 309 (UPS, lectotype). BRAZIL, AmaPpA, Grande do Curua, Bailique, Igarapé, on

dead wood, 22 September 1988, H. Sotao & coll. 88.18.13 (SP233313); mangrove, Ilha

de Maraca, on dead wood, January 1989, H. Sotao & coll. 88.21.06 (SP233306); PARA,

Santarém, 21 November 1965, B. Lowy 228B (SP92143); Rio GRANDE DO SUL, Porto

Alegre, Parque Farroupilha, on Salix babylonica trunk, 4 Jun 1960, J.C. Paim Costa s.n.

(SP50687).

REMARKS: Ganoderma perzonatum can be identified by its pale context, small

basidiospores, and cylindrical pileipellis cells with apical elongate concentric

granulations. The studied specimens agree with the lectotype, which, however,

has inconspicuous resinous bands in the context. Ryvarden (2000) described

“oblong ellipsoid” basidiospores but with the same dimensions as described

here. This is the second world report for G. perzonatum, previously known

only from Cuba. Although Mycobank (http://www.mycobank.org/) cites

G. perzonatum as a synonym of G. parvulum, we regard them as independent

(see REMARKS under G. parvulum). For other related species see Torres-Torres

et al. (2008). Three of the studied specimens were labeled as G. lucidum in SP.

Ganoderma pulverulentum Murrill, N. Amer. Fl. 9: 121 (1908). Figs 11, 25

BASIDIOMATA 10 x 15 x 1.5 cm, annual, sessile, imbricate, woody-corky,

light in weight. PiLeus round-flabelliform, applanate; surface glabrous,

bumpy, soft-corky, semiglossy, concentrically sulcate; with a laccate crust, not

cracked, generally difficult to remove, easy to penetrate with the fingernail;

surface very dark reddish-black, almost black close to the base of the pileus,

to photo-brown (9F8), henna (7E8) to oxide-red (8E8) towards the margin;

margin whitish, lobulate, thin, smooth. CoNTEXT 0.7-0.9 cm, fibrous-corky,

not fully homogenous, ochre-yellow (5B6) to dark brown (7F7) close to the

tubes, zonate; with discontinuous resinous bands. PorEs 4-5 per mm, angular

to round, woody; pore surface cream to pale-yellowish (3A3), darkening when

bruising or aging; tubes 0.7-1 cm thick, stratified, concolorous with the lower

part of the context. HYPHAL SYSTEM trimitic. CONTEXTUAL TRAMA: generative

hyphae 4-4.8 um, thin-walled, with conspicuous clamps, hyaline; skeletal

hyphae 2.8-12.8 um diam., thick-walled to solid, non-septate, arboriform or

not, very branched, yellowish to golden-yellow, predominant; binding hyphae

1.6-4 um diam., subthick-walled to solid, non-septate, hyaline to yellowish,

scarce. HYMENOPHORAL TRAMA as the contextual trama. PILEIPELLIS a

crustohymeniderm, cells 37.2-59 x 8-16 um, narrowly clavate to clavate,

generally entire; thick-walled, apex with scarce granulations, golden-yellow to

yellowish-brown, amyloid. BAsIp1osPoREs 9.6-12.8 x 6.2-8 um, Q = 1.41-1.64,

ellipsoid, apex truncate, with apical germ pore, yellowish-brown, inamyloid;

perisporium semi-wrinkled, reddish-yellow; exosporium with inter-walled

pillars 0.4-0.5 um thick, partially anastomosed; endosporium smooth. BAsIDIA

not observed. CysTip1A absent.

Ganoderma (Brazil) ... 119

Fic. 25. Ganoderma pulverulentum (SP103257). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c-d. Crustohymeniderm: c. Skeletal hyphae. d. Binding hyphae.

120 ... Torres-Torres, Guzman-Davalos & Gugliotta

Hasirat: Tropical secondary vegetation, near the city.

MATERIAL EXAMINED: GRENADA, on dry manchinell, W.E. Broadway s.n. (NY,

lectotype). BRAZIL, Banta, Correntina, on trunk, 28 January 1967, D.M. Vital s.n. (SP

103257).

REMARKS: Distinctive features of G. pulverulentum are the lightweight

basidiomata, entire pileipellis cells with granulations, and basidiospores

with partially anastomosed pillars. The Brazilian specimen agrees with the

lectotype, except that the latter has narrower pileipellis cells (8-11.2 um diam.)

with conspicuous granulations. This species is morphologically similar to

G. resinaceum and G. praelongum Murrill, and Ryvarden (2004) synonymized it

with G. resinaceum. However, both have a context without resinous deposits and

basidiospores with free pillars. Furthermore, G. resinaceum has a homogenous

light reddish-brown (7E7) context, while G. praelongum has a pale duplex

context, ochre-yellow (6D7) above and rust-brown (6E8) close to the tubes (ES.

Earle & L. Murrill 536, lectotype, NY). Ganoderma pulverulentum was known

only from the type locality, until recently recorded from French West Indies

(Martinique) by Welti & Courtecuisse (2010). This is the first record of the

species for Brazil. This fungus may have a wider neotropical distribution.

Ganoderma resinaceum Boud. ex Pat., Bull. Soc. mycol. Fr. 5: 72 (1889). Fras 12, 26

= Ganoderma chaffangeonii Pat., Bull. Soc. mycol. Fr. 5: 74 (1889).

BASIDIOMATA 2.4-5 x 3-6.5 x 0.9-1.5 cm, annual, sessile, woody-spongy.

Piteus round-flabelliform, convex to applanate; surface glabrous, uniform to

slightly bumpy, slightly soft, glossy, broadly concentrically sulcate; with a laccate

crust, cracked, easy to remove and to penetrate with the fingernail; surface

violet-brown (darker than 11F7) to dark red in almost all the pileus, gradually

changing to orange (5A7) toward the margin or fully violet-brown (11F7),

darker in adult basidiomata; margin whitish, brownish-orange to concolorous,

entire, thin to obtuse, smooth. ConTEXT 0.4-1.3 cm thick, fibrous-spongy,

homogeneous, light reddish-brown (7E7), with an apricot (5B6) thin fringe

below the laccate crust, azonate; without resinous bands. Pores 4-5 per mm,

angular to circular, woody; pore surface paste yellow (2A4) to yellow (3A2)

when fresh, darkening to ochraceous or yellowish-brown (6C5) when aging

or drying; tubes 0.1-0.5 cm long, stratified, concolorous with the context.

HYPHAL SYSTEM trimitic. CONTEXTUAL TRAMA: no generative hyphae were

observed; skeletal hyphae 1.9-6.8 um diam., thick-walled, generally thick-

walled to solid, non-septate, arboriform or not, moderately branched, golden-

brown; no binding hyphae were observed. HYMENOPHORAL TRAMA: generative

hyphae 1.9-3.1 um diam., thin-walled, hyaline, scarce; skeletal hyphae 2.8-7.5

um diam., thick-walled to solid, non-septate, arboriform or not, moderately

branched, golden-brown; binding hyphae 2.8-4.4 um diam., generally thick-

Ganoderma (Brazil) ... 121

¥, b

Fic. 26. Ganoderma resinaceum (SP61535). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c-e. Crustohymeniderm: c. Generative hyphae. d. Skeletal hyphae. e. Binding hyphae.

walled to solid, non-septate, golden-yellow to yellowish-brown. PILEIPELLIS

a crustohymeniderm, cells 34-59 x 6.2-9.3 um, clavate, narrowly clavate, or

almost cylindrical, generally without protuberances or branches, few with one

lateral branch, thick-walled to solid, apex with granulations, brownish-yellow,

amyloid. BAsip1ospoREs 11.2-12.5 x 6.5-7.4 um, Q = 1.5-1.72, ellipsoid to

oblong, apex truncate, with apical germ pore, yellowish-brown, inamyloid;

perisporium smooth, hyaline; exosporium with inter-walled pillars up to 0.4

um thick, free; endosporium wrinkled, reddish-brown. Basrp1a not observed.

CysTipIA absent.

Hasirat: Not specified.

MATERIAL EXAMINED: FRANCE, Lorr-ET-CHER, Blois, on Quercus trunks in mature

forest, J.L. Boudier s.n. (PC, holotype). VENEZUELA, Orinoco region, on trunks of

trees, 1885, J. Chaffangeon s.n. (PC, holotype of G. chaffangeonii; FH, isotype). BRAZIL,

MarTo Grosso, Rio Alto Juruena, August 1962, M. Mee s.n. (SP61535).

REMARKS: This species presents a generally very dark pileus, homogenous

brown context, smooth cylindrical pileipellis cells, and relatively large

basidiospores with free pillars. Although G. resinaceum is a rare species, many

authors have used this name for organisms with very different features. For

122 ... Torres-Torres, Guzman-Davalos & Gugliotta

example, Ryvarden & de Meijer (2002) listed as G. resinaceum de Meijer 1418

(EMBRAPA), which properly represents G. parvulum, and de Meijer 2776 and

3238 (EMBRAPA), which correspond to a different, unidentified species. For

this reason, based on the literature, its distribution is uncertain. Steyaert (1980)

proposed synonymizing G. chaffangeonii with G. resinaceum; we examined

type specimens of both taxa and agree with this opinion.

Ganoderma sessiliforme Murrill, Bull. New York Bot. Gard. 8: 149 (1912).

Fics 13, 27

BASIDIOMATA 4-7 x 5.7-10 x 1.1-1.2 cm, annual, sessile to substipitate,

occasionally imbricate, woody, but light in weight. Prteus flabelliform to

semicircular, somewhat conchate to convex; surface glabrous, rugose to radially

rugose, hard, semi-glossy, slightly concentrically sulcate; with a laccate crust,

not cracking, easy to remove and to penetrate with the fingernail; surface darker

than violet-brown (9F8) in the 80% of the pileus, wine-red (11D8) towards the

margin; margin whitish to mandarin-orange (6B8), entire to slightly lobulate,

acute to obtuse, smooth. SusBsTIPE 3 x 1-1.5 cm, horizontal, flattened, solid,

surface shiny, red-wine to almost black, darker than the pileus. CONTEXT

0.5-0.7 cm, fibrous, homogeneous to not fully homogeneous, pale in general,

orange-white (5A2), cocoa (6E6) toward the tubes, with a deep yellow (4A8)

fringe below the laccate crust, azonate; without resinous bands. PorEs 3-4 per

mm, angular, woody; pore surface pale-yellow to pastel-yellow (3A3, 3A4),

darkening to brown (6D8) when bruising or aging; tubes 0.4—0.6 cm thick, darker

than context. HyPHAL sySTEM dimitic. CONTEXTUAL TRAMA: no generative

hyphae were observed; skeletal hyphae 2.5-11.2 um diam., mainly solid to

some thick-walled, septate to non-septate, arboriform, with few branches to

branched, yellowish to golden-yellow, predominant. HYMENOPHORAL TRAMA

as the contextual trama. PILEIPELLIS a crustohymeniderm, cells 52.7-68.2 x

6.8-14.8 um, narrowly clavate to clavate, generally smooth or with one lateral

protuberance, thick-walled, golden-yellow, without granulations, slightly

amyloid. BAsIDIOSPORES (8.7-)9.9-10.3 x 6.3-8.8 um, Q = 1.31-1.57, ellipsoid,

apex truncate, with apical germ pore, yellowish-brown, inamyloid; perisporium

wrinkled, reddish-brown; exosporium with inter-walled pillars 0.3-0.4 um

thick, subfree, in some free; endosporium wrinkled. BAsip1A not observed.

CysTIDIA absent. CHLAMYDOSPORES 11-16.7 x 7.4-13 um, Q = 1.35-1.6(-2),

ellipsoid, with one or both apexes mucronate, thick-walled, yellowish.

Hapsirat: Not specified.

MATERIAL EXAMINED: MEXICO, More.os, Municipality of Cuernavaca, on dead

wood, 24-27 December 1909, W.A. & E.L. Murrill 392 (NY, holotype). BRAZIL, Rio

GRANDE DO SUL, Porto Alegre, Parque Saint Hillaire, on wood, 30 April 1960, M.H.

Homrich s.n. (SP61432).

Ganoderma (Brazil) ... 123

Fic. 27. Ganoderma sessiliforme (SP61432). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores. c. Crustohymeniderm: skeletal hyphae. d. Chlamydospores.

124 ... Torres-Torres, Guzman-Davalos & Gugliotta

REMARKS: ‘This species has generally a conchate basidiome, pale context

without resinous deposits, and relatively small basidiospores. Ryvarden (2004)

did not include G. sessiliforme in his treatment of neotropical polypores. ‘This

is the second record of the species for Brazil, where it was previously recorded

by Gottlieb & Wright (1999a). It is a rare fungus, known only from Argentina,

Brazil, and Mexico.

Ganoderma subfornicatum Murrill, N. Amer. Fl. 9: 121 (1908). Figs 14, 28

BASIDIOMATA 3.4-10.5 x 4-8 x 1.3-1.8 cm, perennial, substipitate, rarely

with a long stipe, with a contracted base, woody. PiLeus round-flabelliform,

generally applanate; surface glabrous, bumpy, slightly to radially rugose,

very shiny, remarkably concentrically sulcate; with a laccate crust, difficult to

remove, hard but easily to penetrate with the fingernail; surface totally reddish-

brown (darker than 9F8) almost black, homogenous; margin concolorous

with the pileus, entire, thick, obtuse to truncate, sulcate. SUBSTIPE 1.4-2(-5.3)

x 1.2-2.3 cm, generally short and thick, cylindrical, horizontal or lateral,

concolorous with the pileus, solid. CONTEXT 0.4-0.8 cm thick, fibrous, not

fully homogeneous, pale in general, yellowish-orange (4B7) upwards, gradually

changing to rust-brown (6E8) next to the tubes; with resinous bands. PorEs

4-6 per mm, round, woody, very small; pore surface brown (6D8); tubes

0.6-1 cm long, stratified, concolorous with the base of the context. HyPHAL

SYSTEM trimitic. CONTEXTUAL TRAMA: generative hyphae 2.4-4.3 um diam.,

thin-walled, with large and conspicuous clamps, hyaline to yellowish; skeletal

hyphae 5.6-11.2 um diam., generally solid, not-branched or with few branches,

golden-yellow to yellowish-brown, predominant; binding hyphae 1.2-3.7 um

diam., solid, non-septate, hyaline to yellowish, scarce, notably thinner and

paler than skeletal hyphae. HYMENOPHORAL TRAMA as the contextual trama.

PILEIPELLIS a crustohymeniderm, cells 37.2-62 x 5-9.3 um, cylindrical to

irregular, not clavate, commonly with a constriction near to the apex, generally

with one to two branches (occasionally three) and up to five protuberances,

mainly solid to thick-walled, not multi-stratified-wall, apex with ferruginous

granulations, yellowish, slightly amyloid. Basip1ospoREs 9.6-11.8 x 6.4-8

um, Q = 1.41-1.69, ellipsoid to oblong, apex truncate, slightly visible apical

germ pore, yellowish, inamyloid; perisporium semi-wrinkled, reddish-brown;

exosporium with inter-walled pillars up to 0.4 um thick, free; endosporium

semi-wrinkled. Basip14 not observed. CystTip1A absent.

Hasitat: Seasonal forest, Amazonian ombrophilous rain forest, Atlantic

rain forest.

MATERIAL EXAMINED: BELIZE, s.loc., on dead wood, 1906, M.E. Pecks.n. (NY, lectotype).

BRAZIL, PARANA, Paranagua, Pontal do Sul, on wood, 23 October 1968, G. Hatschbach

20106 (SP8633); Antonina, Parque Marumbi, Rio do Nunes, on dead dicotyledonous

trunk, 12 December 1987, A.A.R. de Meijer 962 (EMBRAPA); PERNAMBUCO, Recife,

Ganoderma (Brazil) ... 125

Fic. 28. Ganoderma subfornicatum (SP375899). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores.

c-e. Crustohymeniderm: c. Generative hyphae. d. Skeletal hyphae. e. Binding hyphae.

Dois Irmaos, 1 May 1956, s.coll. (SP39359); RONDONIA, Rio Ji-Parana, Acampamento

JP-15, on wood, 8 December 1987, M. Capelari 1770 (SP214815); SAo PAULO, Cananéia,

Ilha Comprida, 2 km from the beach, on wood, 25 February 1983, O. Yano & J.R. Pirani

5977 (SP214844); SERGIPE, SE Estacao Ecoldgica Santa Isabel, 11 November 2003, U. da

Silva-Aragao s.n. (SP375899).

REMARKS: This species has a shiny and dark pileus that contrasts with a generally

pale context with resinous bands. Micromorphologically, it is distinguished by

its pileipellis cells with many protuberances. Ryvarden (2004) did not include

126 ... Torres-Torres, Guzman-Davalos & Gugliotta

G. subfornicatum in his treatment of neotropical polypores. According to

Moncalvo & Ryvarden (1997), G. subfornicatum has been reported throughout

the tropics. Rajchenberg & de Meijer (1990) cited this species from Brazil

(specimens de Meijer 962 and 1234); however, later Ryvarden & de Meijer

(2002) mentioned that those specimens corresponded to G. orbiforme. We

studied them and conclude that de Meijer 1234 is G. orbiforme while de Meijer

962 comprises two basidiomata, one representing G. subfornicatum and the

other (marked as de Meijer 962A) G. applanatum (see above).

Ganoderma vivianimercedianum M.G. Torres, Mycotaxon 105: 448 (2008).

For a complete description see Torres-Torres et al. (2008).

Hasirat: Secondary tropical forest.

MATERIAL EXAMINED: MEXICO, Estapo DE MExiIco, Valle del Tepeite, 10 km NE of

Santa Maria, 10 August 1986, E. Bastidas-Varela s.n. (ENCB, holotype). BRAZIL, R1o

GRANDE DO SUL, Guaiba, Fazenda da Faculdade de Agronomia e Veterinaria, em lenho

morto, 29 March 1963, J.P. de Costa-Neto s.n. (SP70533); SAo PAULO, Tremembé da

Cantareira, Villa Amalia, Horto Forestal SPSF, podridao da madeira, 20 May 1952, C.D.F.

Pacheco s.n. (SP109563); AMAPA, Ilha de Maraca, tronco de madeira en descomposic¢ao,

24 October 1988, H. Sotao et al. 88.21.26 (SP233297).

REMARKS: Ganoderma vivianimercedianum is recognized by the flabelliform

basidiomata with a contracted base, not fully homogenous light brown context,

narrowly clavate to clavate pileipellis cells, entire or with up to two lateral

protuberances, unbranched or rarely so, with scattered granulations in the

apex, and 8.8-11.2(-12) x 6.4-8 um basidiospores (Torres-Torres et al. 2008).

This species is morphologically close to G. argillaceum Murrill, G. perzonatum,

and G. resinaceum. Ganoderma argillaceum has larger basidiospores (10.4—13.6

x 7.2-9.6 um) and pileipellis cells without granulations; G. perzonatum has

slightly smaller basidiospores (8.4-10.4 x 6.4-7.2 um) and remarkable large

cylindrica pileipellis cells with concentric elongate granulations only in the

apex; G. resinaceum has 11.2-13.6 x 6.5-7.4(-8.1) um basidiospores, narrowly

clavate to almost cylindrical pileipellis cells, and a fibrous-spongy homogeneous

light reddish-brown context without resinous bands.

Ganoderma weberianum (Bres. & Henn. ex Sacc.) Steyaert, Persoonia 7: 79 (1972).

Fics 15, 29

BASIDIOMATA 4-5.5 x 4-5 x 0.9-1.3 cm, annual, substipitate, with a

contracted base, occasionally imbricate, woody. PILEUs semicircular, round-

flabelliform to flabelliform, convex to applanate; surface glabrous, rivulose to

slightly radially rugose, hard, glossy to dull; with a laccate crust, not cracking,

difficult to penetrate with the fingernail, easily lost leaving the surface relatively

homogeneously dull, zonate; surface reddish-black to violet-brown (11F7) or

darker in almost all the pileus, except towards the margin where gradually

Ganoderma (Brazil) ... 127

changes to orange (5A7) or fully violet-brown (11F7); margin pure white or

yellowish, entire to slightly lobulate, thin, smooth. SuBstT1pe when present

0.9-1.5 x 1-1.5 cm, short and thick, thinner toward the base, horizontal,

slightly darker than pileus, solid. CONTEXT 0.6-1 cm thick, fibrous, not fully

homogeneous, light-yellow (4A4), darkening to reddish-brown (8F7) close to

the tubes, changing to yellow when cut, zonate; with resinous incrustations

throughout the context. Pores 4-5 per mm, angular to round, woody; pore

surface yellowish-white (3A2) to sun-yellow (2A5) when fresh, darkening to

ochraceous or yellowish-brown (6E8) when aging and drying; tubes 0.2-0.4 cm

long, unstratified, concolorous with lower part of the context. HYPHAL SYSTEM

dimitic. CONTEXTUAL TRAMA: generative hyphae 1.9-3.7 um diam., thin-walled,

with conspicuous clamps, non-branched, hyaline to yellowish; skeletal hyphae

1.6-8 um diam., thick-walled to generally solid, septate to non septate, non-

branched to arboriform with few branches, moderately branched, apex rounded

and slightly wider near to the pileipellis, golden-yellow. HyMENOPHORAL

TRAMA as the contextual trama. PILEIPELLIS a crustohymeniderm, cells 46.6-

65.7 x 6.2-10.6 um, cylindrical to narrowly clavate, entire, thick-walled to

solid, apex with granulations, golden-yellow, slightly amyloid (light bluish).

BASIDIOSPORES 8.4-9.3 x 6.8-7.2 um, Q = 1.29-1.5, ellipsoid, few broadly

ellipsoid, apex truncate, with apical germ pore, yellowish-brown, inamyloid;

perisporium wrinkled, reddish-brown; exosporium with inter-walled pillars up

to 0.5 um thick, subfree; endosporium wrinkled. BAstp1a 13.6-17.3 x 8-9.3

um, widely clavate, hyaline. Cystip1a absent. CHLAMYDOSPORES 9.3-10.5 um,

globose, thick-walled, with inter-walled, very thick pillars, yellow.

Hasirat: “Cerrado”.

MATERIAL EXAMINED: BRAZIL, SAo Pauto, S. Paulo Brotas, km 216-217 of Sao

Paulo-Brotas Road, Regiao de Cerrado, 11 January 1962, A. Milanez & D. Altimari s.n.

(SP61099).

REMARKS: The most important features of G. weberianum are a hard pileus

crust, pale context that changes to yellow when cut with resinous incrustations,

cylindrical pileipellis cells with granulations, and small basidiospores. The single

Brazilian specimen studied agrees with the specimen recorded from China

by Wang et al. (2005), except that they describe clavate pileipellis cells which

nonetheless appear cylindrical in their illustration. The examined specimen

matches Steyaert (1972) and Corner’s (1983) descriptions except for their

observations of the inter-pillars in the basidiospores as barely visible (“minutely

echinulate to apparently smooth;” Corner 1983). Our examination of the

basidiospore ornamentation with a higher magnification lens (1600x) enabled

us to detect how the pillars (“echinula” sensu Corner and Steyaert) relate to

one another, appearing in this case as free pillars mixed with some fused ones.

Steyaert (1972) described two forms of G. weberianum: one with long narrow

128 ... Torres-Torres, Guzman-Davalos & Gugliotta

Fic. 29. Ganoderma weberianum (SP61099). Bar = 8 um.

a. Pileipellis cells. b. Basidiospores. c-d. Crustohymeniderm:

c. Generative hyphae. d. Skeletal hyphae. e. Chlamydospores. f. Basidium and basidiola.

Ganoderma (Brazil) ... 129

pileipellis cells (30 x 7-8 um) and without or with few chlamydospores, and

the other with short broad pileipellis cells (20 x 10-12 um) and with abundant

chlamydospores. Nevertheless, Steyaert’s pictures show pileipellis cells that are

longer than he described. On the other hand, Corner (1983) described long

narrow pileipellis cells and abundant chlamydospores.

Ganoderma weberianum has been reported from Africa, Asia and Samoa

Island (Steyaert 1972), Australia (Smith & Sivasithamparam 2000), China

(Wang et al. 2005), and Singapore (Corner 1983), but Nufiez & Ryvarden (2000)

did not include it in their treatment of East Asian Ganodermataceae. This is the

first record of G. weberianum in America.

Discussion

Although G. annulare (Lloyd) Boedijn, G. bibadiostriatum Steyaert, G. citriporum

Ryvarden & Iturr., G. chalceum (Cooke) Steyaert, G. colossus (Fr.) C.F. Baker,

G. curtisii (Berk.) Murrill, G. lobatoideum Steyaert, G. lobatum (Schwein.) G.F.

Atk., G. lucidum (Curtis) P. Karst., G. multicornum Ryvarden, G. multiplicatum

var. vitalii Steyaert, G. nitens (Fr.) Pat., G. nitidum Murrill, G. oerstedii (Fr.)

Murrill, G. pygmoideum Steyaert, “G. reniformis” [Elfvingia reniformis (Morgan)

Murrill], G. subamboinense (Henn.) Bazzalo & J.E. Wright ex Moncalvo &

Ryvarden, G. testaceum (Lév.) Pat., G. tropicum (Jungh.) Bres., and G. zonatum

Murrill have been cited from Brazil in the literature (e.g., Drechsler-Santos et

al. 2008; Gerber & Loguercio-Leite 1997; Gomes-Silva et al. 2011; Gottlieb &

Wright 1999a; Loguercio-Leite et al. 2005; Patouillard 1889; Steyaert 1962, 1980;

Westphalen et al. 2010), they are not included in either key or descriptions

because we were unable to examine any material representing them.

As a specimen of G. sessile Murrill was found in SP, the species is included

in the key but not described; it lacked data and thus we are uncertain whether

it was collected in Brazil. Gottlieb & Wright (1999a) tentatively determined

BAFC 34352 from Brazil as G. sessile.

We exclude G. neurosporum J.S. Furtado [= Haddowia neurospora (J.S.

Furtado) Teixeira] because basidiospores of the type specimen [Panama, Buenos

Aires, on decaying root, 8 August 1945, S.L. Mayer s.n. (BPI, lectotype)] have

crested ornamentations and do not correspond with Ganoderma.

Likewise, the type specimen of G. opacum (Berk. & Mont.) Pat. [Brazil, Bahia,

B & M 32 (K, lectotype)] has Humphreya-like basidiospores. According to

phylogenetic analysis with morphological and molecular data, the monophyletic

Humphreya differs from Ganoderma (Torres-Torres et al. unpublished). Of the

30 specimens kept at EMBRAPA and SP labeled as G. lucidum, none of them

belong to this species. Furthermore, it is probable that many Brazilian records

under G. lucidum in the literature represent a minimum of nine different

species, but this should be carefully verified.

130 ... Torres-Torres, Guzman-Davalos & Gugliotta

Acknowledgements

The authors are grateful to Lorelei Norvell and Shaun Pennycook for their valuable

comments and corrections, especially Dr. Pennycook for his nomenclatural guidance.

Tatiana B. Gibertoni and Mario Rajchenberg are thanked for the critical review of this

paper. We would like to express our gratitude to curators from BPI, EMBRAPA, ENCB,

FH, INB, NY, O, PC, SP, and UPS herbaria, and to R.R.A. de Meijer for provide us his

personal collection. Adriano Spielmann is thanked for taking the pictures of Brazilian

specimens (SP) and Miguel de Santiago for inking the drawings. The senior author

thanks to Red Latinoamericana de Botanica for the internship scholarship (RLB-05-

P5) at Instituto de Botanica da Secretaria de Estado do Meio Ambiente, Sao Paulo. She

also thanks Universidad Tecnoldgica del Chocé for grants for Mexicos internships.

Funds were obtained from CONACYT (project CONACYT-SEP-2003-C02-42957)

and Universidad de Guadalajara (projects 62935, 72640, 88682). Roberto Quintero and

Alfonso Langle are thanked for the manuscript’s English improvement.

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MY COTAXON

http://dx.doi.org/10.5248/121.133

Volume 121, pp. 133-138 July-September 2012

New records of pyrenocarpous lichenized fungi

from Bulgaria

VESELIN V. SHIVAROV. & DIMITAR Y. STOYKOV

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

2 Gagarin St., 1113 Sofia, Bulgaria

* CORRESPONDENCE TO: v.shivarov@abv.bg

ABSTRACT — Five pyrenocarpous species of lichenized fungi, Acrocordia salweyi, Staurothele

hymenogonia, Verrucaria praetermissa, V. viridula and Wahlenbergiella striatula, are reported

for the first time from Bulgaria. Detailed descriptions, illustrations, and comments are

provided.

Key worps — Pyrenulales, lichen taxonomy, Verrucariales

Introduction

Pyrenocarpous lichens comprise lichenized fungi with perithecioid ascomata.

The taxa discussed here belong to the orders Pyrenulales and Verrucariales,

characterized by having usually crustose, immersed or superficial thallus,

variously colored and inhabit rocks, soil and bark.

Several lichenologists have taxonomically studied Acrocordia species in

Bulgaria (e.g. Kazandzhiev 1906; Szatala 1929; Zhelezova 1963; Pisut 1969,

2001), while many specialists (see Mayrhofer et al. 2005) have surveyed

Verrucaria and Staurothele, with more recent records provided by Vondrak

(2006) and Krzewicka et al. (2007). Recently five additional species were found

for the first time in Bulgaria.

Material & methods

This study is based on specimens collected by the authors during 2006, 2007, and

2011 and now housed in the Mycological Collection at the Institute of Biodiversity and

Ecosystem Research, Bulgarian Academy of Sciences, Sofia (SOMF). Determination

of species follows Smith et al. (2009). Measurements of ascospores, conidia, and

hymenial algal cells are given as follows: (min—) mean + standard deviation (—max).

Macrophotographs were made by means of a Windaus Labortechnik D-38678 dissecting

microscope with Canon PowerShot A630 digital camera.

134 ... Shivarov & Stoykov

Taxonomic descriptions

Acrocordia salweyi (Leight. ex Nyl.) A.L. Sm., Monogr. Brit. Lich. 2: 315, 1911. Pt. 1A

THALLUS immersed or superficial, greenish-grey to pale brown, thin.

PERITHECIA 620-800 um diam., one-quarter to halfimmersed. INVOLUCRELLUM

tightly incurved around the exciple and continuous below. AscosPorgs (19.7-)

22.4 + 1.3(-25.8) x (8.3-)10.0 + 0.7(-11.3) um, |/w ratio usually c. 2.2 (n = 25),

colorless, ellipsoid, 1-septate. PycNip1A 0.2-0.3 mm diam. CONIDIA (3.0-)3.5 +

0.4(-4.6) x (1.2-)1.5 + 0.1(-2.0) um, I/w ratio usually >2.2 (n = 25).

SPECIMEN EXAMINED — BULGARIA, Shoumen Plateau, Bukaka Nature Reserve, on

sedimentary rocks in a shady habitat, 25 October 2007, D.Y. Stoykov (SOME 27 992).

ECOLOGY & DISTRIBUTION — In shaded habitats on soft calcareous rocks.

Acrocordia salweyi occurs in the sub-Atlantic region of the temperate zone over

the sub-Mediterranean region to the Mediterranean area (Wirth 1995). Known

from Europe (Santesson et al. 2004, Liska et al. 2008, Smith et al. 2009).

COMMENTS — Similar to A. conoidea (Fr.) Korb., but differing in the

outwardly spreading base of the involucrellum and smaller ascospores (12-19

x 6-9 um). Another species, A. macrospora A. Massal., has a laterally spreading

involucrellum, but it never continues below the exciple.

Staurothele hymenogonia (Nyl.) Th. Fr., Bot. Not. 1865: 40, 1865. PrlB

THALLUS endolithic, grey. PERITHECIA 270-420 um diam., superficial,

leaving small shallow pits in the rock, dusted with rock fragments. OSTIOLE

plane or in a small depression. INVOLUCRELLUM present. HYMENIAL ALGAL

CELLS (3.0-)4.5 + 0.9(-6.6) x (2.3-)2.8 + 0.5(-3.6) um, I/w ratio usually >1.6

(n = 50). Asci 8-spored. Ascospores (19.1-)22.4 + 1.6(-24.5) x (10.5-)12.1 +

0.9(-13.8) um, I/w ratio usually >1.8 (n = 25), colorless, ellipsoid, muriform.

SPECIMENS EXAMINED — BULGARIA, Forebalkan, Vratsa distr., near Stoyanovo village,

on limestone rocks, 8 June 2006, D.Y. Stoykov (SOMF 27 986); Liutajik village, on

calcareous rock, 7 July 2006, D.Y. Stoykov (SOMF 28 005); Stara Planina Mts (western),

Lakatnik, along the path to the Temnata Doupka Cave, on pebbles, 6 September 2006,

D.Y. Stoykov (SOME 27 980); Strandzha Mts, Kovach locality near Zvezdets village, on

rocks, 24 May 2007, D.Y. Stoykov (SOMF 27 995).

ECOLOGY & DISTRIBUTION — On limestone as well as porous granular substrata

(calcareous sandstone, sandy dolomite, chalk pebbles) in lightly shaded to

well-lit habitats. From the boreal pine belt southwards to the Mediterranean

area (Wirth 1995). Reported from Asia and Europe (Grillo & Caniglia 2004,

Santesson et al. 2004, Seaward et al. 2004, Spribille et al. 2006, Liska et al. 2008,

Smith at al. 2009).

ComMENnts — Staurothele hymenogonia differs from other members of this

genus by its smaller colorless spores.

Acrocordia, Staurothele, Verrucaria & Wahlenbergiella spp. new to Bulgaria... 135

PiaTE 1. A. Acrocordia salweyi (SOMF 27 992). B. Staurothele hymenogonia (SOMF 27 986).

C. Verrucaria praetermissa (SOMF 27 977). D. Verrucaria viridula (SOME 27 991).

E. Wahlenbergiella striatula (SOMF 28 001). Scale bars = 1 mm.

136 ... Shivarov & Stoykov

Verrucaria praetermissa (Trevis.) Anzi, Comment. Soc. Crittog. Ital. 2(1): 24, 1864.

PL. 1C

THALLUS 40-260 um thick, superficial, non-gelatinous, pale green or

grey-brown; surface smooth, with numerous cracks when well developed.

PROTHALLUS white. PERITHECIA immersed in thallus, at most forming very

low projections, which are too ill-defined to measure; apex visible as a pink

or brown dot. INVOLUCRELLUM conical, extending laterally and fusing with

dark basal layer below. Asc1 60-85 x 13-25 um, 8-spored, clavate. ASCOSPORES

(15.5-)18.8 + 1.8(-21.3) x (6.2-)8.0 + 0.9(-10.0) um, I/w ratio usually >2

(n = 25), colorless, ellipsoid, simple. CONIDIA (4.0-)5.0 + 0.6(-6.2) x (0.9-)1.2 +

0.1(-1.5) um, |/w ratio usually >4 (n = 50), rod-shaped, some slightly curved.

SPECIMEN EXAMINED — BULGARIA, Shoumen Plateau, near the entrance of the Zandana

Cave, on frequently inundated sedimentary rock in a shady habitat, 23 October 2007,

D.Y. Stoykov (SOME 27 977).

EcoLocy & DISTRIBUTION — Amphibious in the splash water zone, rarely

submerged for long periods. On hard and stable slightly acidic to basic siliceous

or calcareous substrata near freshwater sources. Tolerating a wide range of

illumination. From sea level to high mountain areas, very rarely reaching alpine

sites (Thtis & Schultz 2009). Known from North and Central America, Asia,

Australia, Europe, New Zealand (Aptroot & Seaward 1999, Smith et al. 2009).

ComMMENTS — Similar to Verrucaria hydrela Ach., which usually develops

thin subgelatinous crust with the basal involucrellum and exciple generally

separated. There is no free space between the involucrellum and the exciple

in V. praetermissa. Another close species, V. elaeina Borrer, has a thin epilithic

thallus and prominent perithecia but lacks a black basal layer.

Verrucaria viridula (Schrad.) Ach., Method. Lich., Suppl.: 16, 1803. Pu. 1D

THALLUS immersed or superficial, pale brown to whitish, areolate, divided

by cracks. PERITHECIA 200-500 um diam., half to almost completely immersed

in thallus, appearing as convex to conical-hemispherical projections, with basal

part immersed in the substratum. INVOLUCRELLUM poorly developed, seen

only in the upper half of the exciple. AscosPpoREs (27.9—)32.6 + 2.9(-38.5) x

(15.3-)19.1 + 1.5(-22.1) um, |/w ratio usually >1.7 (n = 25), ellipsoid to broadly

ellipsoid, sometimes with perispore. PYCNIDIA appearing as dark dots. CONIDIA

(8.5-)10.0 + 1.2(-13.2) x (0.9-)1.3 + 0.2(-1.8) tm, I/w ratio usually >7.5 (n = 25),

straight to slightly curved.

SPECIMEN EXAMINED — BULGARIA, Mt Vitosha, above Douhlata Cave, on calcareous

rock, 22 May 2011, V.V. Shivarov (SOME 27 991).

ECOLOGY & DISTRIBUTION — In foothill and submontane sites. On calcareous

rocks (including limestone). Widely distributed in Europe (from temperate

regions to the Mediterranean area), it is also known from Africa, North and

Acrocordia, Staurothele, Verrucaria & Wahlenbergiella spp. new to Bulgaria ... 137

Central America, Asia, Australia, and Macaronesia (Wirth 1995, Santesson et

al. 2004, Spribille et al. 2006, Lika et al. 2008, Smith et al. 2009).

ComMENts — Verrucaria viridula is variable but distinguished by its large

ascospores and the apex of the perithecium, which often forms a beak. The

thallus is typically superficial, cracked, and occasionally scarcely apparent.

Verrucaria hochstetteri Fr. has similar-sized spores but differs by its endolithic

thallus and absence of involucrellum.

Wahlenbergiella striatula (Wahlenb.) Gueidan & Thiis, Taxon 58: 199, 2009. PL. 1E

THALLUS superficial, subgelatinous, green to dark dull green, translucent

when wet, very thin or up to 250 um, cortex absent. PERITHECIA 180-350

um diam., forming prominent conical projections, with often irregular shape,

usually divided by large irregular ostiole. ExcrpLe dark brown, but paler at base.

INVOLUCRELLUM thick, appressed to exciple and broadened at base. HYMENIAL

GEL I+ blue, then light purple. AscosporEs (6.2-)8.7 + 1.1(-11.0) x (4.2-)6.0 +

0.8(-7.1) um, |/w ratio usually c. 1.4 (n = 25), ellipsoid. Pycnip1A not seen.

SPECIMEN EXAMINED — BULGARIA, Black Sea coast, Sinemorec village, near the North

Beach, on rocks periodically flooded by water, 13 September 2011, V.V. Shivarov (SOMF

28 001).

ECOLOGY & DISTRIBUTION — Maritime species in mid-littoral zone on rocky

seashores. Reported from America (North), Australia, Europe, and New

Zealand (Santesson et al. 2004, Orange 2008).

Comments — Wahlenbergiella has recently been separated from Verrucaria

to accommodate the two marine species, V. mucosa Wahlenb. and V. striatula

Wahlenb. (Gueidan et al. 2009). Verrucaria halizoa Leight., the only close

species recorded from the Bulgarian Black Sea coast (Krzewicka et al. 2007),

differs in its thin olive-green thallus, different perithecial shape, and oblong-

ellipsoid ascospores with I/w ratio up to 3.5.

Acknowledgments

We are grateful to Dr Volker John (Pfalzmuseum fiir Naturkunde, Germany) and Dr

Laszl6 Lékés (Department of Botany, Hungarian Natural History Museum, Hungary)

for critically reading the manuscript and serving as pre-submission reviewers, and to

Prof. Mark R.D. Seaward (University of Bradford, Bradford, UK) for checking a part of

the manuscript and for useful suggestions. We would like to thank Dr Beata Krzewicka

(W. Szafer Institute of Botany, Krakow, Poland) for her help with identification of

Verrucaria praetermissa.

Literature cited

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57-101.

Grillo M, Caniglia GM. 2004. A check-list of Iblean lichens (Sicily). Flora Mediterranea 14: 219-251.

138 ... Shivarov & Stoykov

Gueidan C, Savi¢ S, Thiis H, Roux C, Keller C, Tibell L, Prieto M, Heidmarsson S, Breuss O,

Orange A, Fréberg L, Wynns AA, Navarro-Rosinés P, Krzewicka B, Pykala J, Grube M, Lutzoni

FE 2009. Generic classification of the Verrucariaceae (Ascomycota) based on molecular and

morphological evidence: recent progress and remaining challenges. Taxon 58: 184-208.

Kazandzhiev S. 1906. Additions to the lichen flora of Bulgaria. Godishnik na Sofiiskiya Universitet

2: 125-137. (In Bulgarian).

Krzewicka B, Stoykov DY, Nowak J. 2007. New and noteworthy species of Verrucaria from Bulgaria.

Mycologia Balcanica 4: 131-134.

Liska J, Palice Z, Slavikova $. 2008. Checklist and red list of lichens of the Czech Republic. Preslia

80: 151-182.

Mayrhofer H, Denchev CM, Stoykov DY, Nikolova SO. 2005. Catalogue of the lichenized and

lichenicolous fungi in Bulgaria. Mycologia Balcanica 2: 3-61.

Orange A. 2008. British pyrenocarpous lichens. Department of Biodiversity and Systematic Biology,

National Museum of Wales. Cardiff, Wales. accessed on 10 Oct. 2011.

[online at www.museumwales.ac.uk/media/13849/British-Pyrenocarpous-Lichens.pdf]

Pisut I. 1969. Beitrag zur Kenntnis der Flechten Bulgariens. II. Acta Rerum Naturalium Musei

Nationalis Slovenici 15(1): 27-37.

Pisut I. 2001. Beitrag zur Kenntnis der Flechten Bulgariens. III. Acta Rerum Naturalium Musei

Nationalis Slovenici 47: 21-26.

Santesson R, Moberg R, Nordin A, Tonsberg T, Vitikainen O. 2004. Lichen-forming and

lichenicolous fungi of Fennoscandia. Museum of Evolution, Uppsala.

Seaward MRD, Sipman HJM, Schultz M, Maassoumi AA, Haji Moniri AM, Sohrabi M. 2004.

A preliminary lichen checklist for Iran. Willdenowia 34: 543-576.

http://dx.doi.org/10.3372/wi.34.34218

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA (eds). 2009. The

lichens of Great Britain and Ireland. 2nd ed. The British Lichen Society, London.

Spribille T, Schultz M, Breuss O, Bergmeier E. 2006. Notes on the lichens and lichenicolous fungi

of western Crete (Greece). Herzogia 19: 125-148.

Szatala O. 1929. Beitrage zur Flechtenflora von Bulgarien. I. Magyar Botanikai Lapok 28: 82-99.

This H, Schultz M. 2009. Freshwater flora of Central Europe. Vol. 21/1, Fungi, Part 1, Lichenes.

Spektrum, Heidelberg.

Vondrak J. 2006. Contribution to the lichenized and lichenicolous fungi in Bulgaria. I. Mycologia

Balcanica 3: 7-11.

Wirth V. 1995. Die Flechten Baden-Wirttembergs. Teil. 1 & 2. E. Ulmer, Stuttgart.

Zhelezova B. 1963. Materials on the lichen flora of Bulgaria. Izvestiya na Botanicheskiya Institut

(Sofia) 12: 245-265. (In Bulgarian)

MY COTAXON

http://dx.doi.org/10.5248/121.139

Volume 121, pp. 139-145 July-September 2012

Lichenological notes 5: Neotypification of

Sarcogyne magnussonii (Acarosporacedae)

KERRY KNUDSEN” & JANA KOCOURKOVA’

"The Herbarium, Department of Botany and Plant Sciences, University of California,

Riverside, CA 92521-0124, U.S.A.

*Department of Ecology, Faculty of Environmental Sciences, Czech University of Life Sciences,

Prague, Kamycka 129, Praha 6 - Suchdol, CZ-165 21, Czech Republic

CORRESPONDENCE TO: Knudsen@ucr.edu '*, kocourkovaj@fzp.czu.cz *

ABSTRACT — Sarcogyne magnussonii is neotypified and its description revised based on new

specimens from Canada.

Key worps — biodiversity, calciphiles, North America, typification

Introduction

The holotypes for four species of Sarcogyne described from single specimens

from North America (Magnusson 1935) were missing when the first author

began studying the genus as part of the Sonoran lichen flora project in 2004.

Three of these were in the herbarium of Maurice Bouly de Lesdain and were

presumed destroyed in the bombing of Dunkirk in France during WW2:

Sarcogyne novomexicana H. Magn., Sarcogyne integra B. de Lesd. ex H. Magn.,

and Sarcogyne magnussonii. The holotype of the fourth species, Sarcogyne

athroocarpa H. Magn., was in the Farlow Herbarium (FH), but the Sarcogyne is

missing from the rock of the type on which there only remains a Lecidea taxon

that was misidentified as Sarcogyne “privigna” auct. non (Ach.) A. Massal. and

unambiguously fails to match the protologue (Magnusson 1935). No Sarcogyne

athroocarpa was found on a duplicate of this collection in NY, only another

specimen of the misidentified Lecidea species.

Knudsen & Lendemer (2005) designated a neotype for Sarcogyne

novomexicana that was deposited in the Uppsala Herbarium (UPS) with

a duplicate deposited in the New York Botanical Garden (NY). Sarcogyne

novomexicana has proved to be a rare calcicolous species known currently only

from California from Joshua Tree National Park in Riverside County, the San

Bernardino Mountains in San Bernardino County, and Yosemite National Park

140 ... Knudsen & Kocourkova

in Mariposa County (Knudsen & Kocourkova, unpublished data; Knudsen &

Standley 2007).

The types of S. integra and S. magnussonii were from the mountains of New

Mexico and S. athroocarpa was from the mountains of Colorado. Despite our

attention to these taxa, we found no material from western North America

that could be attributed to these three species. We hypothesized they belonged

to a distinctive North American biogeographic unit of lichen distribution we

have noted during our taxonomic studies. It encompasses northeastern North

America from the Rockies to the Atlantic with the ranges of some species

extending south into non-Sonoran Mexico and west into Sonoran Arizona.

For instance, Acarospora chrysops (Tuck.) H. Magn., A. janae K. Knudsen, and

A. tuckerae K. Knudsen share this distributional pattern (Knudsen 2007;

Knudsen et al. 2011; Lendemer 2010). In 2009 and 2010, the lichenologist Colin

Freebury (CANL) sent us specimens of a Sarcogyne from the northern border of

this biogeographical unit on a calcareous conglomerate boulder in Grasslands

National Park, in Saskatchewan, Canada (PLATE 1). We were surprised

when these specimens matched the description of Sarcogyne magnussonii

(Magnusson 1935). We revise Magnusson’s description and designate a neotype

to be deposited at CANL and isoneotype at UCR.

Materials & methods

Specimens were examined from CANL, COLO, FH, NY, and UCR. Specimens have

been examined using standard microscopical techniques. Structures were studied in

water and 10% KOH [K]. Amyloid reactions were tested in Lugol's iodine [I] with and

without pretreatment with K. Ascospore measurements were made in water with an

accuracy of 0.5 um. Macrophotographs were taken with a digital camera Olympus DP72

mounted with Quick Photo Camera 2.3 on an Olympus SZX 7 Stereomicroscope. The

illustrations were prepared using Adobe Photoshop. The pictures of neotype locality

were taken by Colin Freebury.

The species

Sarcogyne magnussonii B. de Lesd., Ann. Crypt. Exot. 5: 107 (1932). PLATES 1, 2

Type: U.S.A. NEw MExIco. environs of Las Vegas, Brouard s.n. (Hb. B. de. Lesd, n.v.,

presumed destroyed). CANADA. SASKATCHEWAN. Grasslands National Park, sloped

grassland with scattered boulders, 49°10.903'N 107°41.346'W, 795 m, common on a

conglomerate boulder of glacial erratic, composed of mostly rounded quartz pebbles

held together by calcite May 2, 2010. C. Freebury 1260 (CANL, neotype designated

here; UCR, isoneotype).

DESCRIPTION — ‘Thallus epilithic, white, about 0.3 mm thick, covering

several cm, cracked not areolate, farinose, coating the substrate, unstratified,

ecorticate of gelatinized hyphae or polysaccharide secretions mixed with

abundant large crystals not dissolving in K, with some distinct hyphae 4-5 um

Sarcogyne magnussonii, neotypified ... 141

PLATE 1. Sarcogyne magnussonii. A, Locality of S. magnussonii in the Grasslands National Park in

Saskatchewan, Canada. B, Solitary boulder with S. magnussonii.

wide, and with scattered algal cells, sometimes forming a thin continuous to

broken layer (some non-fertile parts of the thallus are necrotic lacking distinct

hyphae or lichenized algae or pycnidia, formed only of gelatinized hyphae

or polysaccharide secretions and crystals). Apothecia dispersed, emergent,

0.3-1.5 mm in diam., broadly attached with thin algal layer beneath the

hypothecium; disc usually smooth, reddish-black and epruinose, becoming

only slightly redder when wetted, with thin, smooth, black margins, slightly

elevated above the disc, not becoming convex, stipitate, or immarginate and

142 ... Knudsen & Kocourkova

PLATE 2. Sarcogyne magnussonii (C. Freebury 1260, isoneotype, UCR), thallus with apothecia.

Scale bar = 1 mm.

with no signs of vegetative division. Exciple 100-150 um thick, of radiating

hyphae, outer layer blackish, internally reddish-brown to hyaline, apices

expanded up to 6 um. Hymenium 80-100(-120) um tall, I+ blue turning

yellow-green or red, K/I+ blue, epihymenium 10-15 um tall, yellowish-brown,

paraphyses mostly 2 um thick, septate, rarely constricted at septa, cells up to 10

um long, the upper three or four becoming shorter, 3-5 um long, apices firmly

conglutinate, expanded to 3.0-3.5 um or barely expanded, clavate or not. Asci

clavate, 60-70 x 18-20 um, 100-200 ascospores per ascus. Ascospores various,

4-6 x 2-3 um, mostly 2 um wide, often with 1 or 2 oil drops. Subhymenium

poorly differentiated from hymenium 30-50 um thick, I+ blue. Hypothecium

narrow, prosoplectenchymatous, c. 10-15 um thick, poorly differentiated,

merging into the exciple. Algal layer beneath the apothecium, usually thin,

20-30 um thick, algal cells mostly 10 um diam. Medulla prosoplectenchymatous

and gelatinized, continuous with attaching hyphae and thallus, obscure with

crystals not dissolving K, and with scattered algal cells, up to 200 um high.

Pycnidia visible as black dots, globose, 50-100 um in diam. Conidia hyaline,

simple, either globose (1 x 1 um) or minutely bacilliform (1.0 x 1.5 um). Spot

tests negative, containing no apparent exolites.

Sarcogyne magnussonii, neotypified ... 143

OTHER SPECIMEN EXAMINED — CANADA. SASKATCHEWAN: Grasslands National

Park, sloped grassland with scattered boulders, 49° 10.903’ N 107°41.346' W, 795 m,

common on a calcareous conglomerate boulder, June 18, 2009, C. Freebury 829B w/ M.

Freebury (CANL).

Discussion

Sarcogyne magnussonii is abundant on a single calcareous conglomerate

boulder of glacial erratic in the grasslands of Saskatchewan. The specimens

from Canada match well both the scant protologue (Bouly de Lesdain 1932)

and the more detailed description of Magnusson (1935).

Ecologically, like Sarcogyne novomexicana, S. magnussonii was originally

collected on silicate rock but probably is predominately a calciphile. Like

S. “privigna, S. magnussonii probably occurs on silicate rock in drainages or

seeps where calcium is deposited during evaporation (Knudsen & Kocourkova

2011).

Magnusson (1935) compared Sarcogyne magnussonii only with S. similis

H. Magn. He noted that the difference between S. magnussonii and the

widespread S. similis was broader ascospores (5-6.5 x 2.5-3 vs. 4-6 x 1.5 um).

We feel this is a poor character as the ascospore sizes of both species actually

overlap and are variable (4-6 x 2-3 vs. 4-6 x 1.5-2.5 um). The apothecia

resemble some apothecia of S. similis, but do not become convex, stipitate,

or immarginate and show no signs of vegetative division, all secondary

characteristics of S. similis. The real difference between the two species is

the well-developed farinose epilithic thallus of S$. magnussonii with frequent

pycnidia and emergent apothecia.

The only other North America species that sometimes produces a white

or dun-colored epilithic thallus is Sarcogyne arenosa (Herre) K. Knudsen &

Standl. (Knudsen & Standley 2007; Lendemer et al. 2009). That species usually

has a chasmolithic thallus that is ecorticate and whitish when visible, often as

a ring subtending the exciple. It rarely becomes epilithic, sub-corticate, and

sub-areolate on crumbling rock, and can even turn a light brown in full sun. It

has a firm texture, while the thallus of S. magnussonii is farinose. The thallus of

S. arenosa is also thin, usually < 0.1 mm thick, while the thallus of S. magnussonii

is thicker, usually 0.3 mm. Sarcogyne arenosa also has mostly narrowly ellipsoid

ascospores (3.5-5 x 1-1.5 um), while the ascospores of S. magnussonii are

usually wider (2-3 um). Sarcogyne arenosa occurs on both silicate rock and

soil as well as calcareous rock and soil, while S. magnussonii probably prefers

calcareous rock and silicate rock drainages with calcium deposits. The two

species may be sympatric in the Midwest or in New Mexico or Texas, but

S. arenosa is only apparently common in California, from where it was originally

described. The two species can be easily separated based on the differences in

ascospore sizes and texture and thickness of the thallus.

144 ... Knudsen & Kocourkova

Six Sarcogyne species from Europe and Asia have thinner white ecorticate

epilithic or chasmolithic thalli than S. magnussonii and occur on calcareous

rocks (Knudsen et al. 2009). They all differ especially in having larger or globose

ascospores (see the “nivea group” table in Knudsen et al. 2009).

Sarcogyne cretacea Poelt grows on limestone in the Alps in Europe and

produces a thicker epilithic thallus than S. magnussonii, 5.0 vs. 0.3 mm thick

(Knudsen et al. 2009; Poelt 1964). The S. cretacea thallus is chalky and firm

with distinct rounded contours, confluent individuals, and a dun-colored edge.

The S. magnussonii thallus is loosely organized without any stratification and

without a distinct edge. It is soft and easily abraded. The farinose ecorticate

structure of S. magnussonii thallus would not support a thallus 5 mm thick as

in S. cretacea. Otherwise their apothecial size, hymenium height, and ascospore

size overlap and do not significantly differ. But S. cretacea has longer conidia

than S. magnussonii (2.0-2.5 x 1.0 vs. 1.0 x 1.0-1.5 um). The two species can

be distinguished by the difference in the thallus structure and thickness and

conidial size. Other Sarcogyne species are mainly identified by their thallus

structure and conidia, for instance S. crustacea K. Knudsen & Kocourk. from

California and S. brunnea K. Knudsen & Flakus from South America (Knudsen

& Kocourkova 2010; Knudsen et al. 2012).

The neotypification of Sarcogyne magnussonii is important for the further

study of the North American lichen biota. Its revision will hopefully lead to

the discovery of new populations. We are happy to designate the neotype of

S. magnussonii and now have some hope of discovering populations of S. integra

and S. athroocarpa.

Acknowledgements

We thank our reviewers, Erin Tripp (RSBG) and James C. Lendemer (NY). We thank

Colin Freebury (CANL) who re-discovered Sarcogyne magnussonii. The work of Jana

Kocourkova was supported financially by the grant “Environmental aspects of sustainable

development of society” 42900/1312/3166 from the Faculty of Environmental Sciences,

Czech University of Life Sciences Prague.

Literature cited

Bouly de Lesdain, M. 1932. Lichens de letat de New-Mexico (USA) recueillis par le frere G. Arsene

Brouard. Annales de Cryptogamie Exotique 5: 89-139.

Knudsen K. 2007 [“2008”]. Acarospora. 1-38, in: TH Nash II/ et al. (eds.) Lichen flora of the Greater

Sonoran Desert Region. Volume 3. Lichens Unlimited, Arizona State University, Tempe.

Knudsen K, Flakus A, Kukwa M. 2012. A contribution to the study of Acarosporaceae in South

America. Lichenologist 44: 253-262. http://dx.doi:10.1017/S00242829 11000703.

Knudsen K, Halici MG, Kocakaya M. 2009. Sarcogyne magnispora (Acarosporaceae), a new species

in the Nivea group from Turkey. Mycotaxon 107: 413-417. http://dx.doi.org/10.5248/107.413

Knudsen K, Kocourkova J. 2010. Lichenological notes 1: Acarosporaceae. Mycotaxon 112: 361-366.

http://dx.doi.org/10.5248/112.361

Sarcogyne magnussonii, neotypified ... 145

Knudsen K, Kocourkova J. 2011. Lichenological notes 3: Sarcogyne plicata in California. Mycotaxon.

118: 423-431. http://dx.doi.org/10.5248/118.423

Knudsen K, Lendemer JC. 2005. Changes and additions to the checklist of North American lichens

— IL. Mycotaxon 93: 277-281.

Knudsen K, Lendemer JC, Harris RC. 2011. Studies in lichens and lichenicolous fungi - no. 15:

miscellaneous notes on species eastern North America. Opuscula Philolichenum 9: 45-75.

Knudsen K, Standley SM. 2007. Sarcogyne. 289-296, in: TH Nash III et al. (eds.): Lichen flora of

the Greater Sonoran Desert Region. Volume 3. Lichens Unlimited, Arizona State University,

Tempe.

Lendemer JC. 2010. Contributions to the lichen flora of Pennsylvania: further new and interesting

reports of lichens and lichenicolous fungi. Evansia 27: 47-64.

Lendemer JC, Kocourkova J, Knudsen K. 2009. Studies in lichens and lichenicolous fungi: more notes

on taxa from North America. Mycotaxon 110: 373-378. http://dx.doi.org/10.5248/110.373

Magnusson AH. 1935. On the species of Biatorella and Sarcogyne in America. Annales de

Cryptogamie Exotique 7: 115-145.

Poelt J. 1964. Mitteleuropaische Flechten VIII. Mitteil. der Bot. Staatssammlung Miinchen 5:

247-265.

MY COTAXON

http://dx.doi.org/10.5248/121.151

Volume 121, pp. 147-151 July-September 2012

New species of Humicola and Endophragmiella

from China

YUE-MING Wu & TIAN-YU ZHANG

Department of Plant Pathology, Shandong Agricultural University,

Taian, 271018, China

Key Laboratory of Agricultural Microbiology,

Shandong Province, Taian, 271018, China

*CORRESPONDENCE To: tyzhang1937@yahoo.com.cn

Axsstract —Three new species, Humicola jilongensis, Humicola shannanensis and

Endophragmiella zhangmuensis, are described and illustrated from soil in China. The type

specimens (dried cultures) and living cultures are deposited in the Herbarium of Shandong

Agricultural University, Plant Pathology (HSAUP). Isotypes are kept in the Herbarium of

Institute of Microbiology, Academia Sinica (HMAS).

KEY worps — dematiaceous hyphomycetes, soil fungi, taxonomy

Introduction

During the course of a survey of soil dematiaceous hyphomycetes in China,

several unusual species of Humicola and Endophragmiella were collected. Two

Humicola and one Endophragmiella species are described and illustrated as new.

The genus Humicola was established by Traaen (1914) and is characterized by

possession of micronematous or semi-macronematous conidiophores, which

are cylindric or slightly inflated. The conidiogenous cells swell apically to

form globose to ovoid aleuriospores. Of the 66 taxa listed by Index Fungorum

(2012), many are infraspecific, and Seifert et al. (2011) estimate that the genus

may contain only 20 valid species. Endophragmiella was erected by Sutton

(1973) with E. pallescens as the type species. The genus, which was emended

by Hughes (1979), is characterized by solitary, acrogenous, septate conidia

seceding rhexolytically from monoblastic, integrated, terminal, determinate

or percurrently proliferating conidiogenous cells. More than 80 species are

recorded in this genus (Index Fungorum 2012).

148 ... Wu & Zhang

Fic. 1. Humicola jilongensis (ex holotype).

Conidia, conidiophores and conidiogenous cells. (Bars = 25 um).

Humicola jilongensis Y.M. Wu & T.Y. Zhang, sp. nov. FIG. 1

MycoBank MB 563887

Differs from H. globosa and H. indica by its larger conidia with numerous melanin

granules giving a rough appearance to the conidia.

Type: China, Tibet: Jilong, from a mountain soil, altitude 4550 m, 20 Sept. 2007, Y.M.

Wu (Holotype HSAUP II_,1485; isotype HMAS 196260).

ETyMoLoey: in reference to the type locality.

Colonies on PDA at 26°C for 7 days 3-5 cm diam., effuse, velvety, brown to

dark brown. Mycelium superficial or immersed: hyphae branched, septate,

smooth, subhyaline to light brown, 2-3 um wide. Conidiophores subhyaline

to light brown, mononematous, septate, smooth, 3-7 um wide. Conidia

globose, solitary or forming short chains, golden-yellow, thick-walled, 15-24

(commonly 19) um in diameter, covered by numerous melanin granules giving

the conidial surface a rough appearance. Chlamydospores intercalary, globose.

Phialospores not seen.

ComMeEnts -Morphologically, H. jilongensis resembles H. globosa De Bert. (De

Bertoldi 1976) and H. indica S.C. Agarwal (Agarwal 1983; nom. illegit., non

Haware & Pavgi 1971) but its conidia are larger than those of H. globosa (12-13

um) or H. indica (8-10 um). In addition, H. jilongensis conidia are covered by

Humicola & Endophragmiella spp. nov. (China) ... 149

numerous melanin granules giving the conidial surface a rough appearance,

while the conidia of H. indica are slightly roughened and those of H. globosa

are smooth.

Fic. 2. Humicola shannanensis (ex holotype).

Conidia, conidiophores and conidiogenous cells. (Bars = 25 um).

Humicola shannanensis Y.M. Wu & T.Y. Zhang, sp. nov. FIG. 2

MycoBank MB 563889

Differs from Humicola lutea by its verrucose conidia.

Type: China, Tibet: Shannan, from a grassland soil, altitude 3850 m, 28 Jun. 2007, Y.M.

Wu (Holotype HSAUP II ,.0597; isotype HMAS 196262).

EryMoLoey: in reference to the type locality.

Colonies on PDA at 26°C for 7 days 2-3 cm diam.., effuse, velvety, greyish brown

to dark brown. Mycelium superficial and immersed: hyphae branched, septate,

smooth, subhyaline to light brown, 1-3 um wide. Conidiophores subhyaline,

mononematous, septate, verrucose above, 3-5 um wide. Conidia solitary,

apical and lateral, globose, verrucose, golden-yellow, 8-10 (commonly 9) um

in diameter, thin-walled, surrounded by many melanin granules, forming a pile

or coat (3-5 um) thick. Phialospores not seen.

COMMENTS —Humicola shannanensis is similar to H. lutea De Bert. (De Bertoldi

1976) in conidium size, but H. lutea has smooth conidia.

Endophragmiella zhangmuensis Y.M. Wu & T.Y. Zhang, sp. nov. FIG. 3

MycoBank MB 563891

Differs from Endophragmiella theobromae by its smaller, 1-septate conidia, and from E.

ovoidea by its narrower conidia.

150 ... Wu & Zhang

Via

pa aes

[p>

rags

Fic. 3. Endophragmiella zhangmuensis (ex holotype).

Conidia, conidiophores and conidiogenous cells. (Bars = 25 tm).

TyPE: China, Tibet: Zhangmu, from a grassland soil, altitude 2250 m, 14 Sept. 2007, Y.M.

Wu (Holotype HSAUP II ,1249; isotype HMAS 196263).

EryMoLoey: in reference to the type locality.

Colonies on PDA at 25°C for 7 days 3-5 cm diam., effuse, hairy, dark blackish

brown to black. Mycelium superficial and immersed: hyphae branched,

septate, pale brown, smooth, 2-4 um wide. Conidiophores macronematous,

synnematous or mononematous, erect, straight or flexuous, smooth, septate,

light brown, paler towards the apex, 40-200 x 3-5 um. Conidiogenous cells

monoblastic, integrated, terminal, percurrent, cylindrical, tapered to a truncate

apex. Conidia mostly clavate, smooth, 1-septate, pale brown, with a small and

protuberant thin-walled peg at the base, 15-20 x 3-5 um.

Comments - In conidial shape, E. zhangmuensis resembles E. theobromae

M.B. Ellis (Ellis 1976) and E. ovoidea P.M. Kirk (Kirk 1981), which differ in

conidial dimensions (16-30 x 8-11 um in E. theobromae and 14-16 x 5.5-6.5

uum in E. ovoidea). In addition, the conidia of E. theobromae are 2-3-septate

while those of E. ovoidea and E. zhangmuensis are 1-septate.

Acknowledgments

The authors are grateful for pre-submission comments and suggestions provided

by Dr. Eric McKenzie, Prof. Y. L. Guo, and Dr. Shaun Pennycook. This project was

supported by the National Science Foundation of China (no. 30970011 & 30499340).

Humicola & Endophragmiella spp. nov. (China) ... 151

Literature cited

Agarwal SC. 1983 (“1982”). A new species of Humicola from Indian alkaline soils. Indian J. Mycol.

Plant Pathol. 12(2): 222-223.

De Bertoldi M. 1976. New species of Humicola: an approach to genetic and biochemical

classification. Can. J. Bot. 54: 2755-2768.

Ellis MB. 1976. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey,

England. 507 p.

Haware MP, Pavgi MS. 1971. New species of Humicola from Varanasi India soil. Sydowia 24:

129-130.

Hughes SJ. 1979. Relocation of species of Endophragmia auct. with notes on relevant generic names.

New Zealand J. Bot. 17: 139-188.

Index Fungorum. 2012. http://www.indexfungorum.org/Names/Names.asp; accessed 22 Mar.

2012.

Kirk PM. 1981. New or interesting microfungi I. Dematiaceous hyphomycetes from Devon. Trans.

Br. Mycol. Soc. 76: 71-87. http://dx.doi.org/10.1016/S0007-1536(81)80010-1

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

Biodiversity Series. CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands. 997 p.

Sutton BC. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycol. Pap. 132. 143 p.

Traaen EA. 1914. Untersuchungen tiber die Bodenpilze aus Norwegen. Nyt. Mag. Naturvid. 32:

20-121. http://dx.doi.org/10.1016/S0007-1536(61)80031-4

MY COTAXON

http://dx.doi.org/10.5248/121.153

Volume 121, pp. 153-157 July-September 2012

Pholiota virescens, a new species from China

EN-JING TIAN & TOLGOR BAu*

Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,

Jilin Agricultural University, Changchun 130118, China

*CORRESPONDENCE TO: junwusuo@126.com

ABSTRACT — Pholiota virescens, a new species of Pholiota is described from China. It is

characterized by its pileus with the green tone at maturity, and its prominent and conspicuously

projecting pleurocystidia with thin wall.

KEY worps — Agaricales, Strophariaceae, taxonomy

Introduction

Pholiota (Fr.) P. Kumm. (Agaricales, Strophariaceae), established by Kummer

in 1871, is a widespread genus (especially in the North Temperate Zone) with

about 150 species worldwide (Kirk et al. 2008; Smith & Hesler 1968; Holec

2001; Noordeloos 2011).

There have been 55 taxa of Pholiota recorded in China (Tian & Bau 2004,

2005, 2011a,b; Bau et al. 2005), among which 20 species are edible (Dai et al.

2010) and 9 species are medicinal (Dai & Yang 2008). Recently, an additional

new species was discovered during the research on the genus based on

morphological examinations of collections.

Materials & methods

Specimens were examined by traditional taxonomic methods (Singer 1975). 5%

KOH solution and Melzer’s reagent were used as the mountants when examining

the microstructure. Morphological characteristics of the species were described and

illustrated according to the observations on the materials. Colour description for the new

species refer to Kornerup & Wanscher (1978). The specimen studied is now deposited in

the Herbarium of Mycology of Jilin Agricultural University (HMJAU).

Taxonomy

Pholiota virescens T. Bau & E.J. Tian, sp. nov. PLATE 1, FIGURE 1

MycoBank MB563114

Differs from Pholiota velaglutinosa by the green tone of its mature pileus and by the

thin-walled pleurocystidia.

154 ... Tian & Bau

PLATE 1 Pholiota virescens.

Habit.

Type: China, Jilin Province, Hunchun City, Shengli Village, 5 km to the east, on decayed

hardwood, 10 July 2010, Tolgor Bau & En-Jing Tian (Holotype, HMJAU 22498).

Erymo ocy: The species epithet virescens refers to the pileus color becoming green with

maturity.

PILEus 2.8-6.4 cm broad, convex, then becoming plane, even at the margin,

viscid, dull vinaceous-brown over the disc at first, but becoming dull green

towards the margin, glabrous except for brown floccose veil-remnants

adhering along the margin. LAMELLAE adnate, near avellaneous, becoming

dull cinnamon-brown to dull greenish-brown at maturity, dense, medium

broad, edges even. STIPE 2-4.5 cm long, 0.4-0.5 cm thick, equal, solid, white

above, becoming pale vinaceous to brownish below, dingy brown at base, with

scattered patches of fibrils to fibrillose-squamulose below; Veil brown, fibrillose,

evanescent, leaving an evanescent ring. CONTEXT thin, brownish, odor and

taste mild. SPoRE PRINT dull cinnamon-brown.

SPORES 6.2-7.2(-7.7) x 3.8-4.3(-5.3) um, ellipsoid, oblong to ovate in face

view, phaseoliform in side view, smooth, pale rusty to cinnamon-brown in

KOH, slightly paler in Melzer’s reagent, germ pore minute. BAsip1a 17-20 x

5-6 um, (2-)4-spored, narrowly clavate, hyaline in KOH. CHRYSOCYSTIDIA

absent. PLEUROCYSTIDIA 50-80 x 11-15 um, fusoid-ventricose with obtuse

to slightly acute apex, upper part elongated (up to 44 um), wall thin, smooth,

content homogeneous to collapsed, hyaline to tawny in KOH. CHEILOCYSTIDIA

24-44 x 8.5-10 um, clavate, irregularly clavate to fusoid-ventricose, thin-

walled, smooth, content homogeneous, hyaline to pale yellow in KOH. GILi

TRAMA Of parallel to subparallel hyphae hyaline in KOH and with smooth

Pholiota virescens sp. nov. (China) ... 155

Figure | Pholiota virescens.

a, Pleurocystidia; b, Basidia; c, Basidiospores; d, Cheilocystidia; e, Pileipellis.

Bars = 10 um.

walls, the cells inflated, up to 25 um in diam.; subhymenium of gelatinized,

interwoven hyphae. PILEUS CUTIS an ixocutis of hyphae 2.5-5 um in diam.,

thin-walled, smooth to slightly encrusted, yellowish to pale tawny in KOH.

156 ... Tian & Bau

ConTEXT hyphae thin-walled, smooth, hyaline to yellowish in KOH. Clamp

CONNECTIONS present.

Hasit-Scattered on dead wood of hardwoods in summer.

ComMENTS-This species belongs to the subgenus Flammuloides (Smith &

Hesler 1968) based on its viscid pileus, prominent and conspicuously projecting

cheilo- and pleurocystidia, and typically gelatinous subhymenium.

Pholiota virescens is similar to P. velaglutinosa A.H. Sm. & Hesler in young

pileus color and basidiospore size and shape (Smith & Hesler 1968), but differs

in the green tone in the mature pileus and the thin-walled pleurocystidia. It is

also close to P. vinaceobrunnea A.H. Sm. & Hesler (Smith & Hesler 1968; Tian

& Bau 201 1a), which differs by its thick-walled pleurocystidia. Pholiota lubrica

(Pers.) Singer is also similar, but its pileus is not dull vinaceous-brown but

bright orange-brown (Noordeloos 1999; Holec 2001; Jacobsson 1990). Pholiota

velaglutinosa, P. vinaceobrunnea, and P. lubrica are all known from China (Tian

& Bau 2004, 201 1a).

Acknowledgments

This study was supported by the National Natural Science Foundation of China

(No. 31070013). We thank Dr. Takahito Kobayashi of Hokkaido University Museum,

Japan, Dr. Jan Holec of National Museum, Mycological Department, Czech Republic and

Dr. T.H. Li of Guangdong Institute of Microbiology, China, for revising the manuscript

patiently and meticulously.

Literature cited

Bau T, Tian EJ, Wang H. 2005. Strophariaceae of China (I) Pholiota. Journal of Fungal Research

3(3): 1-50 (in Chinese).

Dai YC, Yang ZL. 2010. A revised checklist of medicinal fungi in China. Mycosystema 27: 801-824

(in Chinese).

Dai YC, Zhou LW, Yang ZL. 2010. A revised checklist of edible fungi in China. Mycosystema 29:

1-21 (in Chinese).

Holec J. 2001. The genus Pholiota in central and western Europe. Libri botanici 20: 1-220.

Jacobsson S. 1990. Pholiota in northern Europe. Windahlia 19: 1-86.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10th edition. CAB International, Wallingford (UK). 771 p.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour, third edition. Eyre Methuen Ltd.,

London. 252 p.

Noordeloos ME. 1999. Pholiota (Fr.) Kumm., Fihr. Pilzk.: 84. 1871. 80-107, in: C Bas et al. (eds).

Flora Agaricina Neerlandica, vol. 4. Balkema, Rotterdam & Brookfield.

Noordeloos ME. 2011. Strophariaceae s.1. Fungi Europaei, vol. 13. Edizioni Candusso, Alassio.

648 p.

Singer R. 1975. The Agaricales in modern taxonomy (3rd ed.). J.Cramer, Vaduz. 912 p.

Smith AH, Hesler LR. 1968. The North American species of Pholiota. Hafner Publishing Company,

New York. 402 p.

Pholiota virescens sp. nov. (China) ... 157

Tian EJ, Bau T. 2004. Known species of Pholiota (s. 1.) from China and their distribution. Journal of

Fungal Research 2(1): 25-34 (in Chinese).

Tian EJ, Bau T. 2005. A new record of Pholiota from China. Mycosystema 24(2): 310-311 (in

Chinese).

Tian EJ, Bau T. 2011a. New records of Pholiota collected from Changbai Mt. in China. Mycosystema

30(3) : 408-413 (in Chinese).

Tian EJ, Bau T. 2011b. New Chinese records of Pholiota collected from Inner Mongolia and

Heilongjiang. Mycosystema 30(4) : 649-652 (in Chinese).

MY COTAXON

http://dx.doi.org/10.5248/121.159

Volume 121, pp. 159-163 July-September 2012

Conocybe hausknechtii, a new species of sect. Pilosellae

from the Western Caucasus, Russia

EKATERINA FE MALYSHEVA

Komarov Botanical Institute,2 Prof. Popov Str., Saint Petersburg RUS-197376 Russia

CORRESPONDENCE TO: ef.malysheva@gmail.com

ABSTRACT — A new species, Conocybe hausknechtii, from the Western Caucasus, Russia is

proposed here with detailed descriptions and illustrations.

Key worps — Bolbitiaceae, taxonomy

Introduction

Members of the genus Conocybe Fayod are characterized by mycenoid habit,

usually orange or brownish basidiocarp color, adnexed lamellae, lecythiform

cheilocystidia, a hymeniderm pileipellis, and variously shaped spores usually

with a prominent germ-pore (Arnolds 2005; Hausknecht 2009).

In an earlier study (Malysheva 2011) twenty-five species belonging to

Conocybe were reported for the Western Caucasus. Detailed macroscopic and

microscopic examination of two collections from this territory found some

remarkable differences from known species; these collections are proposed as

a new species.

Materials & methods

Material collected by the author from the Teberda State Biosphere Reserve (Western

Caucasus, Russia) in August, 2009, was documented and dried using standard techniques.

Macroscopic characters were recorded from fresh material in the field. Microscopic

structures were determined from dried material observed under a light microscope

Micmed 2-2 in squash preparations of small parts of the basidiocarp in 5% KOH. Size

ranges for microstructures, except for basidiospores, are given based on measurement

of at least 10 structures from each collection. In the descriptions of basidiospores, Q =

mean length/width ratio of an individual spore and Q* = average Q from the total of 20

spores per collection. For observation of spore surface in Scanning Electron Microscopy

(SEM), the spore material was prepared following Pegler & Young (1972). The images

were captured using a “JEOL’ JSM-6390LA Analytical Scanning Electron Microscope.

The specimens examined are deposited in the Mycological Herbarium of the

Komarov Botanical Institute (LE). The nrITS sequences were submitted to GenBank.

160 ... Malysheva

PLATE 1. Conocybe hausknechtii: mature basidiocarps.

Taxonomy

Conocybe hausknechtii E.F. Malysheva, sp. nov. PL. 1-3

MycoBank MB 564193

Differs from Conocybe rostellata by its much wider pileus, stouter stipe, and slightly

shorter spores.

Type: Russia, Karachaevo-Cherkesia, Teberda State Biosphere Reserve, vicinity of

Teberda town, broad-leaved forest with Fagus, on strongly decayed wood of deciduous

tree (probably Fagus), 6.VIII.2009, E. Malysheva (Holotype, LE 253789; GenBank

JQ247194).

Erymotoey: Named after Dr. Anton Hausknecht in honor of his exceptional contribution

to knowledge of the Bolbitiaceae.

PiLEus 25-40 mm, at first obtusely conical with broad umbo, then conico-

convex to applanate, hygrophanous, translucently striate to center when moist,

in fresh condition pale to dark reddish brown, sometimes with ochraceous tint,

slightly paler towards margin, in dry condition dull brown, clay-color to ochre,

with center remaining darker, surface smooth or almost invisibly pruinose,

especially at center; LAMELLAE moderately crowded, narrowly adnate,

ventricose, pale yellow-brown to rusty brown, with concolorous edge and

lamellulae; Stipe 40-50 x 3-4 mm, cylindrical or slightly thickened towards

base, without distinct bulb, pale cream-yellowish or ochraceous at apex to

yellow-brown or red-brown in lower part, longitudinally striate and entirely

pubescent.

Conocybe hausknechtii, sp. nov. (Russia) ... 161

PLATE 2. Conocybe hausknechtii (holotype): A - elements of pileipellis with pileocystidia;

B - basidium; C - spores; D - cheilocystidia; E - caulocystidia. Scale bar = 10 um.

162 ... Malysheva

15kV X16,000 11m 15kV X15,000 1m

PLATE 3. Conocybe hausknechtii: SEM photographs of spores.

BASIDIOSPORES 7.6-8.0 x 4.6-5.4 um, Q = 1.6-1.7, Q* = 1.6, narrowly

ellipsoid, some proportion limoniform to amygdaliform, not lentiform,

sometimes papillate, with distinct germ-pore, pale yellow-brown in KOH,

slightly thick-walled, smooth (even in scanning electron microscope); BASIDIA

4-spored, 27-30 x 8-12.5 um, broadly clavate; CHEILocystip1A lecythiform,

18-30 x 8-12 um, with short neck and small head, 4-5.5 um in diameter;

PILEIPELLIS a hymeniderm, consisting of clavate or spheropedunculate

elements, 25-40 x 15-35 um, often pigmented and thick-walled at base;

PILEOCYSTIDIA numerous, mostly lageniform with long flexuous neck, 37-55

x 5.5-8 um, more rarely lecythiform with long neck and small head, 27-40 x

7-11 um, thin-walled, hyaline or with yellow-brown content; CAULOCYSTIDIA

numerous, forming a continuous layer, variable in shape and size, mostly

represented by long hyaline hairs <100 um in length and <4 um wide as well

as lageniform, cylindrical, utriform or clavate elements, 12-60 x 8-14 um;

CLAMP CONNECTIONS present.

ECOLOGY & DISTRIBUTION — On strongly decayed wood of deciduous tree

or on soil in forest. Only known from two places in the Western Caucasus.

ADDITIONAL SPECIMEN EXAMINED — RUSSIA, KARACHAEVO-CHERKESIA, Teberda

State Biosphere Reserve, vicinity of Teberda town, mixed forest (with Fagus, Carpinus,

Pinus), on soil, 7. VIII.2009, T. Svetasheva (LE 253998; GenBank JQ247195).

Discussion

Conocybe hausknechtii is characterized by the relatively dark basidiocarp

color, strongly striate pileus, small limoniform or amygdaliform spores,

presence of lageniform and lecythiform pileocystidia, and a stipitipellis

lacking lecythiform caulocystidia. Based on this combination of characters,

C. hausknechtii belongs to sect. Pilosellae Singer, series Siennophylla Hauskn. &

Krisai [as “Sienophylla”| (Hausknecht, Krisai-Greilhuber 2006). Microscopically

it is rather close to some species of the C. siennophylla group: C. siennophylla

Conocybe hausknechtii, sp. nov. (Russia) ... 163

(Berk. & Broome) Singer, C. rostellata (Velen.) Hauskn. & Svréek., and C. ochro-

striata var. favrei Hauskn.

Conocybe siennophylla differs from C. hausknechtii mainly in brighter

basidiocarp color, larger ellipsoid-ovoid (never limoniform) spores, and usually

inconspicuous pileocystidia.

Conocybe rostellata is the species closest to C. hausknechtii, having similar

shaped but slightly longer spores. It also differs from the new species by more

slender habit and smaller basidiocarp size with a 5-27 mm broad pileus and

stipe measuring 25-70 x 0.5-2 mm (Hausknecht 2009, Arnolds 2005).

Conocybe ochrostriata var. favrei differs from C. hausknechtii in its smaller

and brighter colored basidiocarps, larger spores, and absence of pileocystidia.

The spore surface as seen in SEM shows that C. hausknechtii indeed is a

member of the series Siennophylla, characterized by smooth spores.

Acknowledgments

I am grateful to Prof. A. Hausknecht for detailed examination of the holotype and

valuable comments. I am also grateful to my reviewers, Prof. E. Arnolds, Prof. R.H.

Petersen, and Prof. M. Prydiuk, for improving the text, to Prof. A.E. Kovalenko and

staff of the Teberda State Biosphere Reserve for organization of the expedition and to

L.A. Kartseva for help with making the SEM- photographs of spores. This research was

supported by the Ministry of Education and Science of the Russian Federation (contract

N 16.518.11.7071) and a grant of Russian Foundation for Basic Research (N 10-04-01189-a).

Literature cited

Arnolds E. 2005. Genus Conocybe. 120-179, in: ME Noordeloos et al. (eds.), Flora Agaricina

Neerlandica, vol. 6. Taylor & Francis: Boca Raton, London, New York, Singapore.

Hausknecht A. 2009. Bolbitiaceae. A monograph of the genera Conocybe Fayod and Pholiotina

Fayod in Europe. Fungi Europaei, vol. 11. Edizioni Candusso: Alassio. 968 p.

Hausknecht A, Krisai-Greilhuber I. 2006. Infrageneric division of the genus Conocybe - a classical

approach. Osterr. Z. Pilzk. 15: 187-212.

Malysheva EF. 2011. Studies on Conocybe (Bolbitiaceae, Agaricomycetes) in the Western Caucasus,

Russia. Nova Hedwigia 93(1-2): 249-273.

http://dx.doi.org/10.1127/0029-5035/2011/0093-0249

Pegler DN, Young TWK. 1972. Basidiospore form in the British species of Inocybe. Kew Bull. 26(3):

499-537. http://dx.doi.org/10.2307/4120316

MY COTAXON

http://dx.doi.org/10.5248/121.165

Volume 121, pp. 165-170 July-September 2012

Additions to the smut fungi of Pakistan. 1.

CvETOMIR M. DENCHEV’, MUHAMMAD FIAZ?, TEODOR T. DENCHEV,'

HABIB AHMAD? & ABDUL NASIR KHALID*

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

2 Gagarin St., 1113 Sofia, Bulgaria

*Department of Botany, Hazara University Mansehra, Pakistan

*Department of Genetics, Hazara University Mansehra, Pakistan

‘Department of Botany, University of the Punjab, Lahore, Pakistan

* CORRESPONDENCE TO: cmdenchev@yahoo.co.uk

ABSTRACT — A new smut fungus, Sporisorium pakistanense on Pennisetum lanatum, is

described and illustrated from Pakistan. Two species, Bauhinus tenuisporus on Persicaria

maculosa and Sporisorium dinteri on Bothriochloa bladhii, are reported for the first time from

Pakistan.

Key worps — Microbotryales, taxonomy, Ustilaginomycetes

Introduction

During collecting trips for parasitic fungi in Mansehra District (Khyber

Pakhtunkhwa Province, Pakistan), carried out by M. Fiaz in 2009-10, 20

specimens of smut fungi were collected. Based on the study of these specimens,

we propose here a new smut fungus, Sporisorium pakistanense on Pennisetum

lanatum, and report two records new for Pakistan: Bauhinus tenuisporus on

Persicaria maculosa and Sporisorium dinteri on Bothriochloa bladhii.

Material & methods

Dried specimens from the Hazara University Herbarium (HUP), Pakistan were

examined under light (LM) and scanning electron (SEM) microscopes. For LM

observations, spores were mounted in lactophenol solution on glass slides, gently heated

to boiling point to rehydrate the spores, and then cooled. Spore measurements are given

in the form: min-max (mean + 1 standard deviation). For SEM, spores were attached

to specimen holders by double-sided adhesive tape and coated with gold with an ion

sputter. The surface structure of spores was observed at 10 kV and photographed with a

JEOL SM-6390 scanning electron microscope.

166 ... Denchev & al.

Taxonomy

Bauhinus tenuisporus (Cif.) Denchev & R.T. Moore, in Denchev et al., Mycologia

Balcanica 3: 74, 2006. FIGs 1-2

= Microbotryum tenuisporum (Cif.) Vanky, Mycotaxon 67: 50, 1998.

Sorr in swollen flowers; systemic infection, all flowers attacked. SPORE

MASS powdery, purplish chestnut (based on the Anonymous (1969) Colour

identification chart). SporEs globose, subglobose, broadly ellipsoidal or ovoid,

8-12.5 x 7.5-9.5 (9.7 + 0.9 x 8.6 + 0.6) um (n = 50); wall 1.3-1.6 um thick (incl.

reticulum), reticulate, meshes irregularly polygonal, 4-5(-6) meshes per spore

diameter, muri 0.8-1.3 um high; in SEM the interspaces with a hemispherical

tuberculum.

SPECIMEN EXAMINED — On Persicaria maculosa Gray: PAKISTAN, KHYBER

PAKHTUNKHWA PROVINCE, Mansehra District, Dhodial Mansehra, October 2009, leg.

M. Fiaz, no. FS-07 (HUP 301).

DISTRIBUTION — On Persicaria spp. (Polygonaceae), Asia, North & South

America, Australia, and New Zealand (Vanky 2011). Persicaria maculosa is a

new host record for this species.

Sporisorium dinteri (Syd. & P. Syd.) Vanky, Fungal Diversity 15: 232, 2004. Fics 3-4

Sort destroying the whole inflorescence, ca. 2.5 cm long, partly hidden by the

leaf sheath; initially covered by a yellowish brown peridium which later ruptures

irregularly, exposing semi-agglutinated, dark reddish brown mass of spores and

sterile cells surrounding a central, stout, simple or branching columella with

longitudinal furrows. STERILE CELLS in chains or irregular groups, subglobose,

broadly ellipsoidal or slightly irregular, sometimes collapsed, 6.5-12(-14) um

long, hyaline, wall 0.5-0.7 um thick, smooth. Spores subglobose, broadly

ellipsoidal, ovoid or slightly irregular, 9-14 x 8.5-12.5 (11.9 + 0.9 x 10.4 + 0.9)

um (n = 50), yellowish brown; wall 0.5-0.7 um thick, in LM densely echinulate,

spore profile not or slightly affected, in SEM echinulate.

SPECIMEN EXAMINED — On Bothriochloa bladhii (Retz.) S.T. Blake: PAKISTAN, KHYBER

PAKHTUNKHWA PROVINCE, Mansehra District, Damgala Mansehra, 3 September 2009,

leg. M. Fiaz, no. FS-04 (HUP 302).

DISTRIBUTION — On Bothriochloa spp., Dichanthium spp. (Poaceae), South

Asia, Africa, and Australia (Vanky 2011, Vanky et al. 2011).

Sporisorium pakistanense Denchey, T. Denchev & Fiaz, sp. nov. FIGs 5-6

MycoBank MB 564488

Differs from Sporisorium cenchri and S. sphacelatum by a central stout columella with

longitudinal furrows and large spores, from S. pennisetinum by a single columella, and

from S. penniseticola by a single columella and large spores.

Sporisorium pakistanense sp. nov. (Pakistan) ... 167

Fics 1-2. Spores of Bauhinus tenuisporus on Persicaria maculosa (HUP 301) in LM and SEM.

Fics 3-4. Spores and sterile cells of Sporisorium dinteri on Bothriochloa bladhii

(HUP 302) in LM and SEM. Scale bars: 1, 3 = 10 um, 2, 4=5 um.

168 ... Denchev & al.

Type on Pennisetum lanatum Klotzsch (Poaceae): Pakistan, Khyber Pakhtunkhwa

Province, Mansehra District, Naran Jalkhud, August 2009, leg. M. Fiaz, no. FS-15

(HOLOTYPE, SOME 28 008; IsoTYPE, HUP 303).

EryMo_Loey: named after Pakistan, the country from which it was collected.

Sort destroying the distal part of sterile shoots, 1-3 cm long, 1.5-3 mm wide,

partly hidden by the uppermost leaf sheath; initially covered by a greyish

brown peridium which later ruptures irregularly, exposing semi-agglutinated,

dark reddish brown spore mass surrounding a central, stout columella with

longitudinal furrows. SPORE BALLS and STERILE CELLS not seen. SPORES

subglobose, broadly ellipsoidal, ellipsoidal, ovoid or slightly irregular, 11-15 x

9-12.5 (12.5 + 0.8 x 10.5 + 0.9) um (n = 50), not dimorphic, light (to middle)

reddish brown; wall 0.5-0.8 um thick, in LM punctate to echinulate, spore

profile slightly or not affected, in SEM echinulate.

COMMENTS — Apparently, the spore balls of this S. pakistanense are early

disintegrating.

Nine Sporisorium species have been described on Pennisetum (Vanky 2009,

2011). The sori of two of them, S. ehrenbergii (Kithner) Vanky and S. tothii

Vanky, are restricted to the ovaries, while the sori of three species, S. divisum

Vanky, S. penniseti (Rabenh.) Ershad, and S. penniseti-japonici (Henn.) Vanky,

are restricted to the spikelets. The remaining species possess sori that destroy

the whole inflorescence or are located on the top of sterile shoots: (i) S. cenchri

(Lagerh.) Vanky on Cenchrus and Pennisetum spp., with type on Cenchrus

sp. from Ecuador (sori destroying the whole inflorescence, with numerous,

thread-like columellae; spore balls persistent, opaque; spores 8-13 um long);

(ii) S. pennisetinum (S. Ahmad) Vanky on Pennisetum flaccidum Griseb.,

known only from the type collection from Pakistan (sori destroying the whole

inflorescence, with numerous, filiform columellae; spores densely verruculose-

echinulate); (iii) S. penniseticola Vanky on Pennisetum sphacelatum (Nees) T.

Durand & Schinz, known only from Ethiopia (sori on the top of sterile shoots,

with several, slender columellae; spores (7.5-)8-11(-13) um long); and (iv) S.

sphacelatum Vanky, with type on Pennisetum sphacelatum from South Africa

(sori on the top of sterile shoots, with several, long, filiform columellae; spores

5.5-7.5 um long, darker on one side, where the wall is thicker).

Sporisorium pakistanense differs from S. cenchri and S. sphacelatum by

having a central, stout columella with longitudinal furrows, and large spores.

We have not seen the type of S. pennisetinum, but based on its illustration in

Vanky (2011: 708), the spore ornamentation of S. pennisetinum, as seen in SEM,

consisted of more densely situated ornaments than those of S. pakistanense;

moreover, the sori of S. pennisetinum have numerous, filiform columellae while

these of S. pakistanense possess a central columella.

Sporisorium pakistanense sp. nov. (Pakistan) ... 169

Fics 5-8. Spores of Sporisorium pakistanense on Pennisetum lanatum

(holotype, SOMF 28 008) in LM and SEM. Scale bars: 5-6 = 10 um, 7-8 = 5 um.

170 ... Denchev & al.

The sori of S. penniseticola differ from these of S. pakistanense in being

situated on the top of sterile shoots comprising the whole inflorescence and

its stem, forming long, cylindrical, greyish brown bodies, 5-18 cm long, 1-3

mm wide, partly enclosed by the uppermost leaf sheath, at maturity, the distal

part of the sori splits longitudinally in several places, after the dispersion of the

spore balls with exposed long, slender columellae, whereas the proximal part of

the sori is still entire containing sporogenous hyphae with immature spore balls

(Vanky 2005: 93). Additionally, the spores of S. penniseticola are smaller.

Acknowledgements

We gratefully acknowledge Dr Kalman Vanky (Herbarium Ustilaginales Vanky,

Tiibingen, Germany) and Dr Roger G. Shivas (Biosecurity Queensland, Australia) for

critically reading the manuscript and serving as pre-submission reviewers; and Assist.

Prof. Jan Alam (Department of Botany, Hazara University Mansehra, Pakistan) for

identification of a host plant.

Literature cited

Anonymous. 1969. Flora of British fungi. Colour identification chart. Her Majesty’s Stationery

Office, Edinburgh.

Vanky K. 2005. Two new smut fungi (Ustilaginomycetes) on Pennisetum (Poaceae) from Ethiopia.

Mycologia Balcanica 2: 91-94.

Vanky K. 2009. Keys to smut fungi of selected host plant families and genera. Mycologia Balcanica

6:.1=36;

Vanky K. 2011[“2012”]. Smut fungi of the world. APS Press, St. Paul, Minnesota, USA, xvii + 1458 pp.

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

3.177,

MYCOTAXON

http://dx.doi.org/10.5248/121.171

Volume 121, pp. 171-179 July-September 2012

Two new microfungi from Portugal:

Magnohelicospora iberica gen. & sp. nov. and

Phaeodactylium stadleri sp. nov.

RAFAEL FE. CASTANEDA-RuIz', MARGARITA HERNANDEZ-RESTREPO’,

JOSEPA GENE’, JOSEP GUARRO’, DAvip W. MINTER’,

& MASATOSHI SAIKAWA‘4

"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

*Unitat de Micologia, Facultat de Medicina i Ciéncies de la Salut, Universitat Rovira i Virgili,

43201 Reus, Spain

°CABI, Bakeham Lane, Egham, Surrey, TW20 9TY, United Kingdom

‘Department of Biology, Tokyo Gakugei University,

Nukuikita-machi, Koganei-shi, Tokyo 184-8501, Japan

CORRESPONDENCE TO *: josepa.gene@urv.cat

ABSTRACT — Two new microfungi are described and illustrated from a forest in Portugal.

Magnohelicospora iberica gen. & sp. nov. is distinguished by polyblastic, integrated, sympodial

conidiogenous cells and solitary, doliiform or conical, multi-euseptate, brown conidia tightly

coiled in three planes. Phaeodactylium stadleri sp. nov. is characterized by obovoid to clavate,

0- to 2-septate, coarsely verruculose, subhyaline to very pale brown conidia. A key to accepted

Phaeodactylium species is provided.

Key worps — Ammophila arenaria, anamorphic fungi, leaf litter, systematics

During the twenty-fifth mycological foray for study of the Iberian mycobiota

held in the Minho and Braga provinces of Portugal, two undescribed anamorphic

fungi were collected, one in a forest, the other near a beach. Individual samples

of plant material were placed in paper and plastic bags, taken to the laboratory,

and treated using the methods of Castafieda (2005). Mounts were prepared

in polyvinyl alcohol-glycerol (8 g in 100 ml of water, plus 5 ml of glycerol)

and measurements made at a magnification of x1000. Photomicrographs were

obtained with a Zeiss AXIO imager M1 microscope (G6ttingen, Germany).

The pure culture was obtained by transferring single conidia observed under a

stereo-microscope on to oatmeal agar in Petri dishes, then incubated at 25°C

172 ... Castafieda-Ruiz & al.

under near-UV, 12 h alternating cycles of light/dark. One new genus and two

new species are described below.

Taxonomy

Magnohelicospora R.F. Castafieda, Hern.-Rest., Gené & Guarro, anam. gen. nov.

MycoBank MB563847

Differs from Helicoon and Inesiosporium by differentiated conidiophores; from

Rogergoosiella by sympodial proliferations of the conidiogenous cells; and from

Troposporium and Laocoon by differentiated conidiophores, polyblastic conidiogenous

cells and brown pigmented conidiophores and conidia.

TYPE SPECIES: Magnohelicospora iberica R.F. Castafeda et al.

ErymMoLoecy: Latin , magno-, meaning great, Latin, -helicospora, referring to the

helicospore, a coiled conidium that may turn in one or more complete rotations in two

or three dimensions.

Anamorphic fungi. CoLoNnres on the natural substratum effuse, hairy,

brown to dark brown. Mycelium superficial and immersed. CONIDIOPHORES

macronematous, mononematous, erect, smooth or verruculose, brown to dark

brown. CONIDIOGENOUS CELLS polyblastic, integrated, sympodial, flattened,

indeterminate. Conidial secession schizolytic. Conrp1a solitary, tightly coiled

in three dimensions, doliiform to conical, multi-euseptate, acrogenous to

acropleurogenous, brown to dark brown or olivaceous, smooth or verrucose.

Teleomorph unknown.

Magnohelicospora iberica R.F. Castafieda, Hern.-Rest., Gené & Guarro,

anam. sp. nov. BiGse Li 2

MycoBank MB563848

Differs from Helicoon spp., Inesiosporium spp., Laocoon paradoxus Rogergoosiella

roystoneicola, and Troposporium album by its brown, differentiated conidiophores and

sympodially proliferating conidiogenous cells.

Type: Portugal, Minho province, “Lagoas de Bertiandos’, 41°46'N 8°38’ W, C11/57, on

rotten leaf of unidentified plant, 9 November 2011, R.E Castafieda, M. Hernandez-

Restrepo, J. Gené & J. Mariné-Gené (Holotype, HAL 2447 F; Isotype, FMR 12184).

EtTyMoLoGy: Latin, iberica, in reference to the Iberian Peninsula.

CoLonizs on the natural substratum, effuse, hairy, hypophyllous, brown.

Mycelium mostly immersed. Hyphae septate, branched, 2 um diam.,

smooth, brown. CONIDIOPHORES macronematous, mononematous, erect,

straight, simple, 3- to 5-septate, smooth, up to 150 um tall, 5-8 um wide at

the base, brown below, pale brown towards the apex. CONIDIOGENOUS CELLS

polyblastic, terminal, indeterminate, sympodially proliferating, 15-25 x 4-5

um, integrated, pale brown. Conidial secession schizolytic. Conrp1a solitary,

acrogenous or acropleurogenous, compactly circinate in three dimensions,

doliiform to somewhat conical, with a truncate basal cell, 2-3 um wide and

Magnohelicospora gen. & sp. nov. and Phaeodactylium sp. nov. (Portugal) ... 173

Fic. 1. Magnohelicospora iberica (ex holotype HAL 2447F)

a. Conidia. b. Conidiogenous cells and conidia. c. Conidiogenous cells. Scale bars = 10 um.

rounded to obtuse apical cell, multiseptate, 35-50 x 23-30 um, brown, smooth,

dry, composed of filaments tightly coiled 7-9 times in 3-dimensions, brown or

olivaceous-brown, euseptate, smooth, 4-5 um wide. Teleomorph unknown.

NoTE: According to the key given by Kendrick in Seifert et al. (2011), the genera

Helicoon Morgan, Inesiosporium R.F. Castaheda & W. Gams, Rogergoosiella

174 ... Castafieda-Ruiz & al.

Fic. 2. Magnohelicospora iberica (ex holotype HAL 2447F).

Conidiophores, conidiogenous cells and conidia. Scale bars = 10 um.

A. Hern. Gut. & J. Mena, and Troposporium Harkn. are superficially similar to

Magnohelicospora. Helicoon does not have differentiated conidiophores, but the

conidiogenous cells are polyblastic or monoblastic and denticulate, giving rise

to euseptate, hyaline or dark pigmented conidia coiled in three planes (Zhao

et al. 2007, Seifert et al. 2011). Inesiosporium has conidia strongly coiled in 3-

dimensions and olivaceous to brown, but conidiophores are not differentiated

and are mostly reduced to intercalary monoblastic conidiogenous cells each

with a short subulate, lateral neck (Castafeda & Gams 1997, Zhao et al. 2007,

Magnohelicospora gen. & sp. nov. and Phaeodactylium sp. nov. (Portugal) ... 175

Seifert et al. 2011). Rogergoosiella is characterized by macronematous, simple

or dichotomously branched dilute brown to hyaline conidiophores, with

monoblastic, percurrently proliferating conidiogenous cells (Hernandez-

Gutiérrez & Mena 1996, Seifert et al. 2011). In Troposporium the conidiophores

are undifferentiated, hyaline and bear monoblastic conidiogenous cells

which produce hyaline, aseptate conidia coiled in 3-dimensions (Goos 1978,

Seifert et al. 2011). The genus Laocoon J.C. David is also somewhat similar to

Magnohelicospora but, Laocoon has conidia formed by filaments loosely coiled

1-5 times and the conidiogenous cells are polyblastic, each with a convex,

cicatrized, melanized scar (David 1997), making them easy to differentiate

from Magnohelicospora.

Phaeodactylium stadleri R.F. Castafieda, Hern.-Rest., Gené & Guarro,

anam. sp. nov. Figs 3-5

MycoBank MB563850

Differs from Phaeodactylium alpiniae by its smaller, mostly 1-septate, obovoid, truncate,

verruculose conidia.

Type: Portugal. Braga: “Playa de Ofir’, Esposende, C11/66, on decaying stem of

Ammophila arenaria (L.) Link (Poaceae), 10 November 2011, R.E Castafieda, M.

Hernandez-Restrepo, J. Gené & J. Mariné-Gené (Holotype, HAL 2448 F; ex-holotype

culture, IMI 501220, CBS 132715, FMR 12185).

ETyMo.Loay: Latin, stadleri, in honor of Dr Marc Stadler (Helmholtz-Center for infection

Research, Germany) who has collaborated with us in the study of fungal diversity.

CoLonigs on the natural substratum caespitose to pustule-like or pilose, gray.

Mycelium superficial and immersed. Hyphae septate, branched, smooth, very

pale brown to subhyaline, 1-3 um diam. CONIDIOPHORES macronematous,

congested fasciculate, tree-like, more or less dichotomously branched, with

branches 120-150 um long, erect, straight or flexuous, slightly geniculate to

sinuate, multiseptate, very pale brown at the base, subhyaline towards the

apex, up to 200-280 x 4-8 um, smooth. CONIDIOGENOUS CELLS polyblastic,

denticulate, intercalary and terminal, subhyaline or hyaline, 10-30 x 2-3

um, discrete, slightly geniculate, branched, indeterminate, with numerous

holoblastic sympodial proliferations. Conidial secession schizolytic. CONIDIA

solitary, acropleurogenous, mostly obovoid, truncate and delicately cicatrized

at the base, (0—)1(-2)-septate, strongly verruculose, 13-18 x 5-6 um, very pale

brown to subhyaline, dry. Teleomorph unknown.

Culture from the holotype: Colonies on oat meal agar, attaining 20-28 mm

after two weeks at 25°C, white to gray. Reverse white. Mycelium immersed

or aerial, arachnoid. Hyphae thin-walled, septate, hyaline, 1.5-2 um diam.

Sporulation obtained after 2 weeks, conidia similar to those observed in nature,

1-2-septate, verruculose, 14-23 x 4-6 um, subhyaline. Cultures deposited:

IMI 501220, CBS 132715, and FMR 12185.

176 ... Castafieda-Ruiz & al.

Fic. 3. Phaeodactylium stadleri (ex holotype HAL 2448F).

a. Conidia. b. Conidia and conidiogenous cells.

c. Conidiophore, conidiogenous cells and conidia. d. Conidiogenous cells.

Scale bars: a, b, d = 10 um; c = 40 um.

Note: The genus Phaeodactylium Agnihothr., as discussed by Castaneda et

al. (2009), included three accepted species: Phaeodactylium alpiniae (Sawada)

M.B. Ellis [= P venkatesanum Agnihothr., the generic type], P biseptatum R.F.

Castafieda et al., and P. curvularioides Matsush. Another taxon P. cephalotaxi

Magnohelicospora gen. & sp. nov. and Phaeodactylium sp. nov. (Portugal) ... 177

Fic. 4. Phaeodactylium stadleri (ex holotype HAL 2448F).

Conidiophores, conidiogenous cells and conidia. Scale bar = 40 um.

Fic. 5. Phaeodactylium stadleri (ex holotype HAL 2448F).

Conidiogenous cells and conidia. Scale bar = 10 um.

178 ... Castafieda-Ruiz & al.

Y.D. Zhang & X.G. Zhang has subsequently been added (Zhang et al. 2011).

Only P. alpiniae is at all similar to P stadleri, particularly in terms of the

pigmentation of conidia, but P alpiniae has conidia that are clavate, 16-25 x

6-9 um, 3-septate and smooth, and the two taxa can be easily separated.

Key to accepted Phaeodactylium species

1 Gonidia smiGgothewallede. Awe igs legs heer ys Raed ak tee eae A «ad oe le ee oe 2

Conidia closely verruculose, (0-)1-(-2)-septate, 13-18 x 5-6 um,

very pale brown to-subhyaline: 20 cscs. eke qece te mone melee Mies» P. stadleri

DIG ALORA SEPLALES, 15 4 Nociag dh scteg dit osteg tin actadinschadingeh-adinad-adin ein ting wenger 3

Conidia sesepiatess © fis 7, -,Gi.F. ASPs eh eae Gia seine eos «ley oly SMe Sie 4

3(2) Conidia clavate, fusiform or narrowly ellipsoid, but slightly ventricose at center,

10-14 x 2-3 um, pale brown to subhyaline ..................... P. biseptatum

Conidia narrowly ellipsoidal to clavate, pale brown to brown,

T5320 5% SHVO WM 4 os «ties x ona 2 ne ttn Sse seat Ae Og P. cephalotaxi

4(2) Conidia ellipsoidal or clavate, tapered to the base, 16-25 x 6-9 um,

colorless or Sublyalitie rs: bag ikke a da dial hata Made OM P. alpiniae

Conidia obovoid, 13-22 x 5-10 um, with brown median cells

and subhyalineends, smooth t.ho ee nee eens P. curvularioides

Acknowledgments

The authors express their sincere gratitude to Dr. Lori Carris and Dr. De-Wei Li for

their critical review of the manuscript. This study was supported by the Ministry of

Science and Innovation of Spain, grant CGL 2011-27185. We thank the Cuban Ministry

of Agriculture for facilities. We thank Drs. Uwe Braun, De-Wei Li, Vadim Melnik, Pedro

Crous, Kazuaki Tanaka and Raghvendra Singh, Shaun Pennycook Cony Decock and

Martina Reblova. We also acknowledge the facilities provided by Dr P.M. Kirk and

Drs. V. Robert and G. Stegehuis through the IndexFungorum and Mycobank websites.

Dr. Lorelei L. Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclature review

are greatly appreciated.

Literature cited

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

do V Congresso Latino Americano de Micologia. Brasilia.

Castafieda-Ruiz RF, Gams W. 1997. Inesiosporium, a new genus of helicosporous hyphomycetes.

Nova Hedwigia 64: 485-490.

Castafieda-Ruiz RF, Iturriaga T, Minter DW, Heredia-Abarca G, Stadler M, Saikawa M, Fernandez

R. 2009. Two new anamorphic fungi and some microfungi recorded from “El Avila’, Venezuela.

Mycotaxon 107: 225-237.

David JC. 1997. A contribution to the systematics of Cladosporium: revision of the fungi previously

referred to Heterosporium. Mycol. Pap. 172: 1-157.

Goos RD. 1987. Fungi with a twist: the helicosporous hyphomycetes. Mycologia 79: 1-22.

http://dx.doi.org/10.2307/3807740

Magnohelicospora gen. & sp. nov. and Phaeodactylium sp. nov. (Portugal) ... 179

Hernandez Gutiérrez A, Mena Portales J. 1996. A new helicosporous hyphomycete collected on

Roystonea regia in Cuba. Mycol. Res. 100: 1483-1484.

http://dx.doi.org/10.1016/S0953-7562(96)80082-7

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

Biodiversity Series 9: 997 p.

Zhang YD, Ma J, Castafieda-Ruiz RF, Zhang XG. 2011. New species of Phaeodactylium and

Neosporidesmium from China. Sydowia 63: 125-130.

Zhao G, Liu X, Wu W. 2007. Helicosporous hyphomycetes from China. Fungal Diversity 26:

313-524.

MY COTAXON

http://dx.doi.org/10.5248/121.181

Volume 121, pp. 181-186 July-September 2012

Ellisembia karadkensis sp. nov.

from southern Western Ghats, India

KUNHIRAMAN C. RAJESHKUMAR’, SWAPNIL C. KAJALE,

SOMNATH A. SUTAR & SANJAY K. SINGH?

National Facility for Culture Collection of Fungi,

Agharkar Research Institute, G.G. Agarkar Road, Pune, India

CORRESPONDENCE TO: ' rajeshfungi@gmail.com & ? singhsksingh@gmail.com

ABSTRACT— A new species, Ellisembia karadkensis is described from Bambusa bambos

culms as a saprobe. It differs from other Ellisembia species in possessing simple or (often)

Y-shaped branched conidia with long flagelliform apical appendages. A morphological

comparison is made with similar Ellisembia species, and with those restricted to bamboo.

A new combination, E. magnibrachypus, is proposed for Sporidesmium magnibrachypus.

Key worps— anamorphic fungi, dematiaceous hyphomycete, Kerala

Introduction

Subramanian (1992) erected the genus Ellisembia during his reappraisal of

Sporidesmium Link and morphologically similar taxa. Subramanian considered

the type of septation of conidia, the nature, regularity and other features of

the percurrent proliferations of conidiophores, and the presence and absence

of conidiophores as important diagnostic criteria. Thus, Ellisembia with

distoseptate conidia was separated from Sporidesmium with euseptate conidia,

with E. coronata (Fuckel) Subram. (= Sporidesmium coronatum Fuckel)

designated as type species (Subramanian 1992). After Ellis (1971, 1976) made

significant studies on Sporidesmium, many distoseptate species were later

transferred to Ellisembia. Wu & Zhuang (2005), who expanded the generic

concept of Ellisembia to include species with typically lageniform, ovoid or

doliiform percurrently extending conidiogenous cells, also synonymized Imicles

Shoemaker & Hambl. (Shoemaker & Hambleton 2001) with Ellisembia. Shenoy

et al. (2006) recognized that species of the Sporidesmium complex (including

Ellisembia) were polyphyletic. Since Subramanian (1992) established the genus

Ellisembia, 47 species epithets have been added (MycoBank 2012).

182 ... Rajeshkumar & al.

The pristine natural forests, microhabitats, and tropical warm humid climate

that prevail in the Western Ghats make it rich and diverse in fungal diversity

(Bhat & Kendrick 1993, Rajeshkumar 2007). During 2010-11, expeditions were

conducted to explore the microfungal diversity in natural forests and plantations

of southern and northern Western Ghats (Rajeshkumar et al. 2011a,b, Singh et

al. 2010). During one survey, an unusual saprobic Ellisembia species (sensu Wu

& Zhuang 2005) was discovered on bamboo culms.

Materials & methods

ISOLATES AND MORPHOLOGY— Conidia of the fungus were directly isolated from the

surface of a dead culm and observed under a Nikon binocular stereomicroscope (Model

SMZ-1500 with Digi-CAM, Japan). Cultures from single conidia were established on

2% potato dextrose agar plates (PDA; Crous et al. 2009). For morphotaxonomic studies

and photomicrographs, Carl Zeiss (AXIO Imager 2, Germany) and Olympus (Model

CX-41, Japan) microscopes were used. Conidia and conidiophores were mounted in

lactic acid cotton blue and measured using an ocular micrometer, with 30 observations

per structure. The measurements were also confirmed with the software available with

the Carl Zeiss (AXIO Imager 2, Germany) microscope. Colony characteristics in culture

were studied on two different media: 2% malt extract agar (MEA) and PDA (Crous et al.

2009). Colony colours were determined using Methuen Hand book of Colour (Kornerup

& Wanscher 1981). A herbarium specimen was deposited in the Ajrekar Mycological

Herbarium (AMH); a culture was accessioned and preserved in the National Fungal

Culture Collection of India (NFCCI [WDCM-932]), Agharkar Research Institute, Pune,

India.

Taxonomy

Ellisembia karadkensis Rajeshkumar & S.K. Singh, sp. nov. Fis 1-2

MycoBank MB 564202

Differs from all other Ellisembia species by its simple or Y-shaped branched conidia with

long flagelliform apical appendages.

Type: India, Kerala, Kasaragod, Karadka. On decaying culms of Bambusa bambos

(L.) Voss, November 2011, K.C. Rajeshkumar (Holotype, AMH 9446; ex-type culture

NFCCI 2664.)

Erymo.oey: karadkensis; referring to the collection locality Karadka, a village in the

Kasaragod District.

Colonies brownish black, erumpent, mycelium consisting of pale to dark

brown, septate, thin-walled, smooth hyphae, 2-3.5 um wide. Conidiophores

macronematous, mononematous, stout, straight or flexuous, erect, dark to

blackish brown, arising from creeping hyphae, 1-6-septate, 17.5-81 x 3.5-8

um. Conidiogenous cells monoblastic, integrated, terminal, lageniform or

cylindrical, pale or dark brown. Conidial secession schizolytic. Conidia solitary,

simple or branched, obclavate, mostly curved, tapering towards narrow whip-

Ellisembia karadkensis sp. nov. (India) ... 183

Fic. 1. Ellisembia karadkensis (holotype): a-b. Distoseptate conidial bases. c. Conidium with

rudimentary branch initiation. d—e. Conidia with branches. f, i. Simple conidia with flagelliform

apical appendages. g—h. Branched Y-shaped conidia.

like apical tip, 10-35-distoseptate, basal cells (up to 25th cell from base) are

pale or olivaceous brown with dark septa, but apical cells are pale or hyaline

with thin septa, walls thin and smooth; simple conidia, 150-345 x 9.5-12.5 um,

184 ... Rajeshkumar & al.

Bow

oT!

Fic. 2. Ellisembia karadkensis (holotype): a-b. Conidia attached to conidiophores. c-d. Conidio-

phores and conidial development. e-h. Variation in conidia. i. Conidium with curved base.

j. Conidiophore with conidiogenous cell. Bars= 20um.

Ellisembia karadkensis sp. nov. (India) ... 185

conidial tip 1.5-2.7 um wide, conidial base truncate, 3.2-5.5 um wide; branched

conidia Y-shaped, 190-250 x 9-10.7 um, conidial tip 2-2.7 um wide, conidial

base 4-5.5 um wide.

TELEOMORPH: not observed.

Colonies on PDA and MEA slow growing, 9-20 mm diam. at 25+2°C in 12

hours dark and 12 hours light conditions after 15 days, pale gray to dark gray

(2D1), floccose, centrally umbonate; margin even; reverse dark grayish (1F1).

Sporulation was not observed even after 30 days of growth.

CoMMENTS— Bamboo supports many Ellisembia species (McKenzie 1995,

Mena et al. 2000, Wu & Zhuang 2005), and some species are known exclusively

from Bambusa spp. These species are compared morphologically in TaBLe 1.

TABLE 1. Comparison of Ellisembia species recorded on bamboos

Connoionnonss um) | Como im)

E. bambusae 30-70 x 4-5 65-100 x 14-16, Wu & Zhuang (2005)

base 3-4.5

E. bambusicola 55-105 x 4-7 65-125 x 11-14, Mena-Portales et al.

base 4—4.5 (2000)

E. bambusina 33-120 x 4.5-5.5 33-48 x 9-12, McKenzie (1995)

base 3-4

E. karadkensis 17.5-29.5 x 5.3-6.4 150-345 x 9.5—-12.5, This paper

base 3.2—5.5

The conidia of Ellisembia karadkensis are morphologically similar to those

of E. podocarpi Jian Ma & X.G. Zhang, E. photiniae Jian Ma & X.G. Zhang,

E. flagelliformis (Matsush.) W.P. Wu, and E. magnibrachypus [ = Sporidesmium

magnibrachypus| (Matsushima 1975, Ma et al. 2010) in shape —especially

in possessing simple hyaline flagelliform apices— and coloration. However,

E. karadkensis conidia are longer and the Y-shaped branches have not been

observed in these other species. The conidia of E. magnibrachypus may branch,

but only in the hyaline, non-septate apical part (Matsushima 1975), whereas,

in E. karadkensis, branched conidia are Y-shaped with a hyaline distoseptate

apical tip. Ellisembia magnibrachypus also has shorter conidia (48-62 um,

excluding apical appendages that are < 65 um long; Matsushima 1975); in that

species Matsushima (1975) considered the apical hyaline non-septate region as

an apical appendage.

Neither Subramanian (1992) nor Wu & Zhuang (2005) considered S. magni-

brachypus in their reassessments of Sporidesmium, even though it is a disto-

septate species. Therefore, we propose a new combination here.

Ellisembia magnibrachypus (Matsush.) Rajeshkumar & S.K. Singh, comb. nov.

MycoBank MB 564216

= Sporidesmium magnibrachypus Matsush., Icon. Microfung. Matsush. Lect.: 138. 1975.

186 ... Rajeshkumar & al.

Acknowledgements

We are indebted to Drew Minnis (Systematic Mycology & Microbiology Laboratory,

Beltsville, Maryland, USA) and Eric McKenzie (Landcare Research, Auckland, New

Zealand) for commenting on this manuscript. RKC thanks Prof. Uwe Braun for his

valuable advice and suggestions. Thanks are also due to the Department of Science and

Technology (DST), Government of India, New Delhi for providing financial support for

setting up the National Facility for Culture Collection of Fungi (No. SP/SO/PS-55/2005)

at Agharkar Research Institute, Pune, India and the Director, ARI for providing facility.

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.

Crous PW, Verkley GJM, Groenewald JZ, Samson RA (eds). 2009. Fungal Biodiversity. CBS

Laboratory Manual Series. Centraalbureau voor Schimmelcultures, Utrecht, Netherlands.

Ellis MB. 1971. Dematiaceous hyphomycetes. CMI, Kew, UK. 608 p.

Ellis MB. 1976. More dematiaceous hyphomycetes. CMI, Kew, UK. 507 p.

Ma J, Zhang YD, Ma LG, Ren SC, Zhang XG. 2010. Taxonomic studies of Ellisembia from Hainan,

China. Mycotaxon 114: 417-421. http://dx.doi.org/10.5248/114.417

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Kobe, Japan. 209 p.

McKenzie EHC. 1995. Dematiaceous hyphomycetes on Pandanaceae. 5. Sporidesmium sensu lato.

Mycotaxon 56: 9-29.

Mena-Portales J, Delgado-Rodriguez G, Heredia-Abarca G. 2000. Nuevas combinaciones para

especies de Sporidesmium sens. lat. Boletin de la Sociedad Micolégica de Madrid 25: 265-269.

MycoBank. 2012. Fungal databases: nomenclature and species banks: online taxonomic novelties

submission. Administered by the International Mycological Association.

http://www.mycobank.org/MycoTaxo.aspx (Accessed: 03/01/2012).

Kornerup A, Wanscher JH. 1981. Methuen handbook of colour. 3rd ed. London: Methuen. 282p.

Rajeshkumar KC. 2007. Diversity of plant pathogenic fungi in natural forests in the Western Ghats,

India. Ph.D thesis, FRI University, Dehradun, India. 201 p.

Rajeshkumar KC, Hepat RP, Gaikwad SB, Singh SK. 201 1a. Pilidiella crousii sp. nov. from northern

Western Ghats, India. Mycotaxon 115: 155-162. http://dx.doi.org/10.5248/115.155

Rajeshkumar KC, Sharma R, Hepat RP, Swami SV, Singh PN, Singh SK. 2011b. Morphology and

molecular studies on Pseudocercospora kamalii sp. nov. a foliar pathogen on Terminalia from

India. Mycotaxon 117: 227-237. http://dx.doi.org/10.5248/117.227

Shenoy BD, Jeewon R, Wu WY, Bhat DJ, Hyde KD. 2006. Ribosomal and RB2 DNA sequence

analyses suggest that Sporidesmium and morphologically similar genera are polyphyletic.

Mycological Research 110: 917-929. http://dx.doi.org/10.1016/j.mycres.2006.06.004

Shoemaker RA, Hambleton S. 2001. “Helminthosporium” asterinum, Polydesmus elegans, Imimyces

and allies. Canadian Journal of Botany 79: 592-599. http://dx.doi.org/10.1139/cjb-79-5-592

Singh SK, Yadav LS, Singh PN, Sharma R, Rajeshkumar KC. 2010. Anewrecord of Gliocephalotrichum

(Hypocreales) from India. Mycotaxon 114: 163-169. http://dx.doi.org/10.5248/114.161

Subramanian CV. 1992. A reassessment of Sporidesmium (Hyphomycetes) and some related taxa.

Proceedings of the Indian Academy of Science B 58: 179-190.

Wu WP, Zhuang WY. 2005. Sporidesmium, Endophragmiella and related genera from China. Fungal

Diversity Research Series 15. 531 p.

MY COTAXON

http://dx.doi.org/10.5248/121.187

Volume 121, pp. 187-191 July-September 2012

Mycoacia angustata sp. nov. (Basidiomycota, Meruliaceae),

the first Chinese hydnoid species

HatI-SHENG YUAN™ & XIAN-ZHEN WAN’?

"State Key Laboratory of Forest and Soil Ecology, Institute of Applied Ecology,

Chinese Academy of Sciences, Shenyang 110164, P. R. China

*Graduate University of the Chinese Academy of Sciences, Beijing 100049, China

* CORRESPONDENCE TO: yuanhs911@yahoo.com.cn

AsBstTRAcT — A new hydnoid basidiomycete, Mycoacia angustata sp. nov., is described

and illustrated from the tropical and subtropical forest of Hainan and Hubei Province,

southern China. It is the first new Mycoacia species described from China. The new species

is characterized by ceraceous basidiocarps, an odontioid to hydnoid hymenophore with

buff to cinnamon-buff surface, a monomitic hyphal system, and allantoid narrow hyaline

basidiospores. Relationships between the new and closely related species are discussed.

Key worps — Polyporales, wood-decaying fungi, taxonomy

Introduction

Mycoacia Donk was described by Donk based on the type species Hydnum

fuscoatrum Fr. The genus is characterized by ceraceous fruitbodies with a

hydnoid hymenophore with conical to cylindrical spines, monomitic hyphal

system, and narrowly ellipsoid, cylindrical, or allantoid basidiospores (Eriksson

& Ryvarden 1976). Mycoacia comprises species with a distinctly hydnoid

hymenophore but is closely related to Phlebia Fr., which mostly comprises

phlebioid species. Of the approximately 20 species encompassed in this genus

at present, three have been reported from China (Dai 2011).

China is very rich in wood-decaying fungi. Species diversity, taxonomy,

phylogeny, ecology and economically important wood-decaying species have

been recently studied extensively (Dai 2010, 2011, 2012, Dai et al. 2009, Zhou

& Dai 2012), and a number of new basidiomycetes in subtropical and tropical

forests have been described (Cui & Dai 2008, Cui et al. 2011, Dai et al. 2010, Dai

& Li 2010, Li et al. 2008, Wei & Dai 2008, Zhou & Dai 2008).

During the most recent surveys of the wood-decaying fungi in southern

China, three specimens of Mycoacia were collected from fallen angiosperm

188 ... Yuan & Wan

trunks. Here we describe and illustrate them as a new species. The relationships

between the new species and its closely related species are discussed.

Materials & methods

The specimens are deposited at the biological herbarium of Institute of Applied

Ecology, Chinese Academy of Sciences (IFP). The microscopic examinations followed

Dai (2010) and were performed on sections mounted in Cotton Blue (CB): 0.1 mg aniline

blue dissolved in 60 g pure lactic acid; CB+ = cyanophilic, CB- = acyanophilic. Amyloid

and dextrinoid reactions were tested in Melzer’s reagent (IKI): 1.5 g KI (potassium

iodide), 0.5 g I (crystalline iodine), 22 g chloral hydrate, aq. dest. 20 ml; IKI- = neither

amyloid nor dextrinoid reaction. 5% KOH was used as reagent. Sections were studied at

magnifications up to x1000 using a Nikon Eclipse E600 microscope and phase contrast

illumination, and dimensions were estimated to an accuracy of 0.1 um. The apiculus

was excluded from the spore measurements, and 5% of the spore dimensions at each

end of the range are shown in parentheses. Abbreviations include: L = mean spore

length (arithmetical mean of all spores), W = mean spore width (arithmetical mean of

all spores), Q = extreme values of the length/width ratios among the studied specimens,

and n = the number of spores measured from a given number of specimens. Special

color terms are from Petersen (1996).

Taxonomy

Mycoacia angustata H.S. Yuan, sp. nov. FIGURE 1

MycoBank MB 800172

Differs from Mycoacia subconspersa by its darker hymenophore color, its longer, more

widely spaced spines, and its longer basidiospores.

Type — China. Hainan Province, Changjiang County, Bawangling Nature Reserve, on

fallen angiosperm trunk, 12.XII.2007, Yuan 3963 (holotype, IFP).

ETYMOLOGY — angustata (Lat.): referring to the narrow basidiospores.

FruiITBopy — Basidiocarps annual, resupinate, adnate, ceraceous, without

special odor or taste when fresh, brittle when dry, up to 18 cm long, 5 cm wide

and 2 mm thick; sterile margin white, narrow or almost lacking. Hymenophore

odontioid to hydnoid, buff when fresh, cinnamon-buff to yellowish brown

upon drying; spines crowded, evenly distributed, 3-4 per mm, ceraceous when

young, corneous when old, cylindrical, with obtuse or acute apices, mostly

individual, occasionally confluent, up to 1 mm long. Subiculum homogenous,

azonate, cream to buff, 0.5-1 mm thick.

HYPHAL STRUCTURE — Hyphal system monomitic; generative hyphae

bearing clamp connections or simple septa, IKI-, CB-; tissue unchanged in

KOH.

SUBICULUM — Generative hyphae hyaline, bearing clamp connections or

simple septa, thin- to thick-walled, occasionally with short side-branches,

frequently branched at right angles, loosely interwoven, mostly 3-6 um diam,

occasionally inflated up to 12 um diam.

Mycoacia angustata sp. nov. (China) ... 189

== qeypdyed

FiGuRE 1. Microscopic structures of Mycoacia angustata (holotype).

a: Basidiospores. b: Basidia and basidioles.

c: Hyphae from spine trama. d: Hyphae from subiculum.

TuBEs — Generative hyphae hyaline, bearing clamp connections or simple

septa, thin- to slightly thick-walled, occasionally branched, subparallel to

interwoven, 2-5 um diam. Cystidia absent; basidia narrowly clavate, bearing

four sterigmata and a clamp connection at the base, 16-23 x 2.7-3.1 um;

basidioles in shape similar to basidia, but slightly smaller.

190 ... Yuan & Wan

Spores — Basidiospores, narrowly allantoid, hyaline, thin-walled, smooth,

IKI-, CB-, (4.7-)4.8-6.5(-7.2) x (0.9-)1-1.2 um, L = 5.52 um, W = 1.07 um,

Q = 4.83-5.51 (n = 60/2).

ADDITIONAL SPECIMENS EXAMINED — CHINA. HAINAN PROVINCE, CHANGJIANG

County, Bawangling Nature Resever, on fallen angiosperm trunk, 13.XIJ.2007, Yuan

4001 (IFP); HUBEI PROVINCE, WUFENG CouNTY, Houhe Nature Reserve, on fallen

angiosperm trunk, 28.[X.2004, Wei 2273 (IFP).

REMARKS — Mycoacia angustata is characterized by annual resupinate

ceraceous basidiocarps, odontioid to hydnoid, buff, cinnamon-buff to yellowish

brown hymenophore with white margin, presence of both clamp connections

and simple septa, narrowly allantoid basidiospores, and absence of cystidia. It is

closely related to Mycoacia aurea (Fr.) J. Erikss. & Ryvarden, M. uda (Fr.) Donk,

and M. subconspersa (Rick) Hjortstam & Ryvarden.

Mycoacia aurea also is characterized by a yellow-ochraceous hymenophore

surface, absence of cystidia, and allantoid to suballantoid basidiospores.

However, it is distinguished from M. angustata by its longer spines (2-3 mm)

and wider basidiospores (3.5-5.5 x 1.5-2 um) (Eriksson & Ryvarden 1976).

Mycoacia uda resembles M. angustata in having sordidly yellow to ochraceous

basidiocarps when old and conical to subcylindrical spines that are 1-2 mm

long. However, M. uda can be differentiated by the presence of subfusiform

cystidioles that are apically capped with non-crystalline matter, rod-like crystals

in the spine trama, and narrowly ellipsoid basidiospores. Moreover, parts of the

young basidiocarp turn deep red in KOH (Eriksson & Ryvarden 1976).

Mycoacia subconspersa also has similarly shaped basidiospores but differs

from M. angustata by having more crowded (5-8 per mm) and shorter (0.25-0.4

mm long) spines, paler hymenophore color, and shorter basidiospores (4—-4.5 x

1-1.25 um) (Hjortstam & Ryvarden 1982).

OTHER SPECIMENS EXAMINED: Mycoacia aurea — SPAIN. NAVARRA PROVINCE,

Lekunberri, about 30 km NW of Pamplona, on Quercus, 11.X1.1977, Ryvarden 15230 (O);

USA. TENNESSEE, Great Smoky Mts. National Park, Sugarland primitive campground,

substrate unknown, 6.[X.1977, Ryvarden 14096 (O).

M. subconspersa — CHINA. YUNNAN PROVINCE, MENGLA County, Xishuangbanna

Botanic Garden, on fallen angiosperm trunk, 30.1X.1993, 48251 (HKAS).

M. uda — ESTONIA. PArnu, Nigula Nature Reserve, on Pinus, 26.VIII.1989,

Ryvarden 27103 (O).

Acknowledgments

We appreciate Drs. Xiao-Yong Liu (IM, CAS, China) and Jun-Feng Liang (RITF,

China) who reviewed the manuscript. The author also thanks Drs. Yu-Lian Wei, Wen-

Min Qin (IFP, China) for the company in the field trips. The research was financed by the

National Natural Science Foundation of China (Project Nos. 31170022 & 31070023).

Mycoacia angustata sp. nov. (China) ... 191

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(Basidiomycota) from Wanmulin Nature Reserve, Fujian Province. Mycotaxon 105: 343-348.

Cui BK, Du P, Dai YC. 2011. Three new species of Inonotus (Basidiomycota, Hymenochaetaceae)

from China. Mycol. Prog. 10: 107-114. http://dx.doi.org/10.1007/s11557-010-0681-6

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Divers. 45: 131-343.

http://dx.doi.org/10.1007/s13225-010-0066-9

Dai YC. 2011. A revised checklist of corticioid and hydnoid fungi in China for 2010. Mycoscience

52: 69-79. http://dx.doi.org/10.1007/s10267-010-0068-1

Dai YC. 2012. Polypore diversity in China with an annotated checklist of Chinese polypores.

Mycoscience 53: 49-80. http://dx.doi.org/10.1007/s10267-011-0134-3

Dai YC, Li HJ. 2010. Notes on Hydnochaete (Hymenochaetales) with a seta-less new species

discovered in China . Mycotaxon 111: 481-487. http://dx.doi.org/10.5248/111.481

Dai YC, Yang ZL, Cui BK, Yu CJ, Zhou LW. 2009. Species diversity and utilization of medicinal

mushrooms and fungi in China (Review). Int. J Med. Mushrooms 11: 287-302.

Dai YC, Cui BK, Liu XY. 2010. Bondarzewia podocarpi, a new and remarkable polypore from

tropical China. Mycologia 102: 881-886. http://dx.doi.org/10.3852/09-050

Eriksson J, Ryvarden L. 1976. The Corticiaceae of North Europe 4. Hyphodermella-Mycoacia.

Fungiflora, Oslo. pp 549-886.

Hjortstam K, Ryvarden L. 1982. Studies in tropical Corticiaceae (Basidiomycetes) IV. Type studies

of taxa described by J. Rick. Mycotaxon 15: 261-276.

Li J, Xiong HX, Dai YC. 2008. Polypores from Shennongjia Nature Reserve in Hubei Province,

Central China. Cryptogamie Mycol. 29: 267-277.

Petersen JH. 1996. Farvekort. The Danish Mycological Society’s colour-chart. Foreningen til

Svampekundskabens Fremme, Greve. 6 p.

Wei YL, Dai YC. 2008. Notes on Elmerina and Protomerulius (Basidiomycota). Mycotaxon 105:

349-354.

Zhou XS, Dai YC. 2008. A new species of Megasporoporia (Polyporales, Basidiomycota) from China.

Mycol. Prog. 7: 253-255.

Zhou LW, Dai YC. 2012. Recognizing ecological patterns of wood-decaying polypores

on gymnosperm and angiosperm trees in northeast China. Fungal Ecol. 5: 230-235.

http://dx.doi.org/10.1016/j.funeco.2011.09.005

MYCOTAXON

http://dx.doi.org/10.5248/121.193

Volume 121, pp. 193-197 July-September 2012

A new species of Entoloma

from Liaoning Province, Northeast China

LIANG-LIANG QT’, XIAO-LAN HE”? & Yu L1'*

‘Jilin Agricultural University, Engineering Research Center of Edible and Medicinal Fungi,

Ministry of Education, Changchun 130118, China

South China Agricultural University, Guangzhou 510642, China

>Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application,

Guangdong Institute of Microbiology, Guangzhou 510070, China

CORRESPONDENCE TO: *yuli966@126.com, ** xl_he@qq.com

AsstRact —Entoloma liaoningense, a new species from Northeast China, is described,

illustrated, and discussed. It is characterized by mycenoid to marasmioid basidiomata,

conspicuously sulcate pileus, 2-spored basidia, isodiametric to subisodiametric basidiospores,

and absence of both cystidia and clamp connections.

Key worps — Agaricales, Basidiomycota, Entolomataceae, taxonomy

Introduction

Entoloma (Fr.) P. Kumm. (Entolomataceae, Agaricales, Basidiomycota)

is a large genus that is widely distributed from tropic to temperate regions.

Representatives of this genus are distinguished by their basidiospores that

appear angular in all views. The genus has been extensively studied in Europe,

North America, Africa, Indomalaya, and Australasia (Hesler 1967; Horak

1980, 2008; Gates & Noordeloos 2007; Largent 1994; Noordeloos 1992, 2004;

Noordeloos & Gates 2009; Romagnesi & Gilles 1979).

During a preliminary study on the entolomataceous mycota in Liaoning

Province, China, five specimens of an Entoloma species were collected from

Benxi Laotudingzi Nature Reserve, which proved to represent a new species. It

is formally described and discussed here.

Materials & methods

Specimens were collected from Laotudingzi Nature Reserve located in east Liaoning

Province of Northeast China, at 41°16'38"-41°21'10"N 124°49'06"-124°57'08"E at an

altitude of 600-1367 m. Fresh collections were photographed. Macro-morphological

194 ... Qi, He & Li

descriptions are based on field notes and photos. Color notations are according to

Kornerup & Wanscher (1978). Micro-morphological data were obtained from dried

material examined under a light microscope. Basidiospores, basidia, pileipellis, and

stipitipellis were observed in 5% KOH or 1% Congo Red, and KOH-soluble pigments

were determined in distilled water mountants. All measurements were made in 5%

KOH. For basidiospore dimensions, Q = “length/width ratio” of a basidiospore in side

view, Q. = average Q + standard deviation. Basidiospore dimensions exclude the hilar

appendix. The studied specimens are deposited in the Herbarium of Jilin Agricultural

University (HMJAU).

Taxonomy

Entoloma liaoningense Yu Li, L.L. Qi & Xiao Lan He, sp. nov. Fics 1-2

MycoBank MB 564420

Differs from Entoloma sericeum by its smaller basidiomata, its bisporic basidia, and its

habitat.

Type: China. Liaoning Province, Benxi, Laotudingzi National Nature Reserve, 12

September 2010, Qi Liang-Liang 5589 (HMJAU 23636, Holotype).

Erymo oey: liaoningense refers to Liaoning province, the collection location of the

holotype.

BASIDIOMATA mycenoid to marasmioid. PrLEus 10-15 mm broad, plano-convex

to hemispherical, slightly depressed at center, usually with a small papilla in the

depression, more applanate to umbilicate in older ones, smooth, radially sulcate

almost to the center, yellowish grey to pale brownish grey (3A2-3B2, 5B3),

paler towards margin, slightly darker in the grooves, somewhat pale brown at

the disc, dry, hygrophanous, sometimes zonate with one or two zones when

mature. LAMELLAE sinuate to adnate, with short decurrent tooth, ventricose,

moderately distant, thin, 1-3 mm broad, whitish grey, becoming somewhat pale

pinkish with age, with concolorous and somewhat eroded edge; with lamellulae

in two tiers. STIPE central, 35-45 x 1-2.5 mm, cylindrical, equal, pale brown,

concolorous with the pileus center, hollow, glabrous, smooth, polished, dry,

often with white tomentum at base. CONTEXT greyish white, very thin. ODOR

and TASTE not distinctive.

BASIDIOSPORES 8.0-10.5 x 7.0-8.5 um, 5-6-angled in side-view, Q=1.0-

1.25 (Q_=1.15+0.03), isodiametric to subisodiametric, thick-walled. Basmp1a

clavate, 25-37 x 8-12 um, 2-spored, sometimes 1-spored; base clampless.

LAMELLA EDGE fertile. CHEILOCYSTIDIA and PLEUROCYSTIDIA absent. LAMELLA

TRAMA subparallel, composed of cylindrical, 8-18 um wide, slightly encrusted

elements. PILEIPELLIS a cutis of repent hyphae; terminal elements cylindrical

to narrowly clavate, 5-11 um in diam, with pale brownish internally encrusting

and intracellular pigment; subpellis composed of yellow brown, cylindrical

elements, 6-13 um in diam. STIPITIPELLIS a cutis of cylindrical hyphae, 5-15

Entoloma liaoningense sp. nov. (China) ... 195

Fic. 1. Entoloma liaoningense (holotype HMJAU 23636): basidiomata.

um wide, with pale brown, intracellular and encrusting pigment. BRILLIANT

GRANULES absent. OLEIFEROUS HYPHAE rare. CLAMP CONNECTIONS absent in

all tissues.

Hasitat: Scattered or solitary on soil among pine needles in mixed forests

with Pinus koraiensis and Quercus mongolica or in coniferous forests with Pinus

koraiensis.

ADDITIONAL SPECIMENS EXAMINED: CHINA. LIAONING PROVINCE, BENXI, Laotudingzi

National Nature Reserve, 13 September 2010, Qi Liang-Liang 5659 (HMJAU 23637); 14

September 2010, Qi Liang-Liang 6076 (HMJAU 23638); Qi Liang-Liang 6144 (HMJAU

23639); Qi Liang-Liang 6176 (HMJAU 23640).

ComMMENTARY: The combination of characters such as the mycenoid to

marasmioid habit, radially sulcate and pale brownish pileus surface, bisporic

basidia, and isodiametric to subisodiametric basidiospores is diagnostic

for E. liaoningense. Its mycenoid habit, absence of clamp-connections, and

basidiospores that are never cuboid or cruciform, place it in subg. Nolanea,

where it may be placed in sect. Papillata based on the absence of cystidia and

presence of both incrusting and intracellular pigments.

196 ... Qi, He & Li

a 10m

Cc 10um

Fic. 2. Entoloma liaoningense (holotype): a. Basidia; b. Basidiospores; c. Pileipellis.

The basidioma stature, the pale brownish grey pileus, and the 2-spored

basidia reminds one of the Japanese species Entoloma bisporum (Hongo) Hongo

(Hongo 1957, as Rhodophyllus bisporus). However, E. bisporum can be easily

distinguished by the more heterodiametric (8.5-9.5(-10) x (5.5-)6-6.5(-7)

uum) basidiospores. In addition, the ecology of the two species also differs, with

E. bisporum always occurring among mosses, while E. liaoningense is found on

soil among pine needles.

Two other bisporic species described recently from China — E. mastoideum

T.H. Li & Xiao Lan He and E. praegracile Xiao Lan He & T-H. Li (He et al.

2011) — also differ from E. liaoningense. Entoloma mastoideum can be easily

distinguished by the pinkish to flesh-colored pileus, cylindrical to narrowly

clavate cheilocystidia, and larger basidiospores (9.5-12(-13) x 7-8(-8.8)

um). Entoloma praegracile can be differentiated by the wax-yellowish to pale

orange-yellow pileus and stipe and the heterodiametric basidiospores ((8-)9-

10.5 x 6.5-8(-8.5) um). The European E. bisporigerum (P.D. Orton) Noordel.

Entoloma liaoningense sp. nov. (China) ... 197

also has a brownish pileus and bisporic basidia but is separated by the larger

basidiospores (10-12(-13) x (7.0-)8.0-9.5(-10.5) um) and the presence of

abundant clamp connections in all tissues (Noordeloos 1992).

The macro- and microscopic characters of E. liaoningense are somewhat

similar to those of E. sericeum Quél.; nevertheless, E. sericeum can be separated

by larger basidiomata (pileus 2-7 cm broad), tetrasporic basidia, and occurrence

in grasslands or meadows (Noordeloos 1992).

Acknowledgements

The authors express sincere gratitude to Dr. PD. Manimohan (University of Calicut)

and Dr. Cui Bao-Kai (Beijing Forestry University) who reviewed this manuscript, and

also to Mr. Zhu Ye-Ping (Benxi Laotudingzi National Nature Reserve) for his help in field

collection. The research was financed by the National Natural Science Foundation of

China (No. 31170012) and the Earmarked Fund of Modern Agro-industry Technology

Research System.

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MY COTAXON

http://dx.doi.org/10.5248/121.199

Volume 121, pp. 199-206 July-September 2012

Beauveria lii sp. nov. isolated from

Henosepilachna vigintioctopunctata

SHENG-LI ZHANG, LING-MING HE, XUE CHEN & Bo HUANG*

Anhui Provincial Key Laboratory of Microbial Control, Anhui Agricultural University,

West Changjiang Road 130, Hefei, Anhui 230036, China

* CORRESPONDENCE TO: bhuang@ahau.edu.cn

ABSTRACT — This paper describes a previously unreported entomopathogenic Beauveria

species isolated from a lady beetle larva collected in Xunyi County, Shaanxi Province, China, in

September 2010. Its main distinguishing morphological feature is its ellipsoidal to cylindrical

conidia that are larger than those of other Beauveria species with cylindrical conidia.

Phylogenetic analysis of four nuclear loci uniquely differentiate it from known Beauveria

species that also produce cylindrical conidia, but place it with B. varroae and B. kipukae,

two cryptic species that constitute the sister lineage to B. bassiana sensu stricto, additionally

supporting its proposed species status. Integration of morphological and molecular evidence

support classification of this unknown organism as a new species, here named Beauveria lii.

KEY worDs — entomogenous fungi, taxonomy, multilocus phylogeny

Introduction

Beauveria Vuill. is one of the most ubiquitous anamorphic genera of

entomopathogenic fungi. The genus is characterized morphologically

by sympodial conidiogenous cells and sporulation on an indeterminate,

denticulate rachis. Because the broad overlap in conidial dimensions and shape

made it difficult to differentiate species, MacLeod’s (1954) broad-sense species,

B. bassiana (Bals.-Criv.) Vuill. and B. tenella (Sacc.) Siemaszko (= B. brongniartii

(Sacc.) Petch), were used for more than 50 years until molecular evidence

demonstrated that these were complex species consisting of morphologically

similar but cryptic lineages (Rehner & Buckley 2005, Rehner et al. 2011).

Evidence for cryptic diversification was first inferred from the nuclear

ribosomal internal transcribed spacer (ITS) and elongation factor 1-alpha

(EF1-a) (Rehner & Buckley 2005). Rehner et al. (2011) proposed a multilocus

phylogeny of Beauveria species based on partial sequences of RNA polymerase

II largest subunit (RPB1), RNA polymerase II second largest subunit (RPB2),

translation elongation factor-1 alpha (TEF), and the Bloc nuclear intergenic

200 ... Zhang & al.

region. They established a new classification system that distinguished 12

species, six of which produce ellipsoidal or cylindrical conidia (i.e., B. amorpha

(H6éhn.) Samson & H.C. Evans, B. malawiensis S.A. Rehner & Aquino de Muro,

B. caledonica Bissett & Widden, B. brongniartii, B. asiatica S.A. Rehner &

Humber, B. sungii S.A. Rehner & Humber; Samson & Evans 1982, Bissett &

Widden 1988, Rehner et al. 2006, Shimazu et al. 1988, Rehner et al. 2011).

Five other species producing globose to subglobose conidia are B. bassiana,

B. australis S.A. Rehner & Humber, B. kipukae S.A. Rehner & Humber, B.

pseudobassiana S.A. Rehner & Humber, and B. varroae S.A. Rehner & Humber

(Rehner et al. 2011). In addition, B. vermiconia de Hoog & V. Rao (de Hoog &

Rao 1975) produces comma-shaped conidia. The species position of B. sobolifera

Zuo Y. Liu et al. (Liu et al. 2001), anamorph of Cordyceps sobolifera (Hill ex

Watson) Berk. & Broome, has yet to be verified by the multilocus method.

A fungal epizootic was observed in a population of the lady beetle

Henosepilachna vigintioctopunctata in Shaanxi, China, caused by a Beauveria

species. Although this organism formed colonies that were similar to those

of B. bassiana, it had a unique microscopic morphology that differs from

B. bassiana and other species. In this study, we investigate the morphological

characteristics and molecular evidence to determine whether this organism is

a novel species.

Materials & methods

Fungal culture

20 Beauveria strains were isolated from specimens of the lady beetle species

Henosepilachna vigintioctopunctata (Coleoptera, Coccinellidae), collected in Shaanxi,

China. RCEF5500 was selected as a test subject from these isolates. Routine growth was

tested on full strength Sabouraud's dextrose agar. This isolate was incubated at 25°C

under 12:12 hour light-dark fluorescent illumination.

Colony growth and morphology

Colony description and measurement were determined on the 10th day of culturing.

Terms and notations used to describe colony coloration followed those of Kornerup &

Wanscher (1961). Microscopic measurements of conidiogenous cells and conidia were

taken from slide cultures on the 4-6th day. Images were acquired with a DP70 digital

camera mounted on an Olympus BX51 microscope, with DP-BSW software (Olympus,

Tokyo, Japan). Mean values for length, width, and length to width ratio (Q) are indicated

by L™, W”, and Q™.

DNA extraction, PCR, cloning, sequencing, and analysis

Conidia were inoculated on potato dextrose agar (PDA) Petri plates-overlaid with

a disc of sterilized cellophane, and incubated at 25°C for 5 days. Fungal mycelia were

obtained by scraping the cellophane. DNA extraction was performed using benzyl

chloride to disintegrate the cell wall chemically as in previously described methods (Zhu

et al. 1994, Wang et al. 2005). The concentration and quality of the purified DNA was

evaluated by 0.8% agarose gel electrophoresis and spectrophotometry. The extracted

Beauveria lii sp. nov. (China) ... 201

DNA was added to sterile distilled water at a final concentration of 100 ng per uL and

stored at -20°C for the following PCR reactions.

Nuclear ribosomal internal transcribed spacer region (ITS) was amplified by PCR

as described by White et al. (1990). For phylogenetic analysis of the isolated Beauveria

strains (Rehner et al. 2011), the partial sequences of four nuclear loci (TEF, Bloc,

RPB1, and RPB2) were amplified as described by Rehner & Buckley (2005), Rehner

et al. (2011). PCR products were purified with the AxyPrep DNA gel purification kit

(Axygene Biotechnology Hangzhou, Hangzhou, China) and were cloned using the

PMD18-T plasmid vector (TaKaRa Dalian Corporation, Dalian, China).

The DNA samples were sequenced by Invitrogen Company (Shanghai, China). The

derived ITS, TEE, Bloc, RBP1, and RBP2 sequences of RCEF5500 were submitted to

GenBank.

Sequence alignment and phylogenetic analyses

TEE, Bloc, RBP1, and RBP2 sequences from 68 taxa determined by Rehner et al.

(2011) (including 67 Beauveria isolates and one Isaria tenuipes Peck strain, which was

used as the outgroup) were downloaded from GenBank. Multiple sequence alignments

of these loci for the 68 isolates and from the isolate RCEF5500 was carried out by

the software Bioedit (Hall 1999). Editing of sequences was performed with Bioedit,

according to Rehner et al. (2011). The sequence files of the above four genes were

concatenated manually into a single file and submitted to European Molecular Biology

Laboratory-European Bioinformatics Institute for multiple alignment using fast Fourier

transform (http://www.ebi.ac.uk/Tools/msa/mafft/). The resulting output was in Nexus

file format to permit phylogenetic analysis by Molecular Evolutionary Genetics Analysis

4.0 (Tamura et al. 2007). The dataset was analyzed using both maximum parsimony and

Bayesian approaches.

The combined data set of the four genes was analyzed phylogenetically using PAUP*

4.0b10 (Swofford 2003) under maximum parsimony, employing a heuristic search with

100 random addition replicates, tree bisection-reconnection (TBR) branch swapping,

and saving multiple trees. Branch support was estimated by performing 1000 bootstrap

replicates, with a full heuristic search (Felsenstein 1985). Clades with bootstrap values

=70% were considered robustly supported by the data and are listed above branches in

the Beauveria phylogenetic tree (Fic. 1).

A Bayesian analysis was carried out in MrBayes 3.1.2 (Huelsenbeck et al. 2001)

applying a general time reversible model and gamma-distributed rate variation to

account for rates of variation across sites, with a proportion of them being invariant sites.

The remaining parameters were default values. One tree was saved to a file every 1000

generations for a total of 1,000,000 Markov Chain Monte Carlo (MCMC) generations;

the first 25% of trees were discarded as burn-in. Values of the posterior probabilities for

the branches supported by 295% of pseudo-replicates were listed below branches in the

phylogenetic tree.

Results

The derived ITS, TEF, Bloc, RBP1, and RBP2 sequences of RCEF5500

were accepted by GenBank with accession numbers of JN689372, JN689371,

JN689373, JN689374, and JN689370, respectively.

202 ... Zhang & al.

ARSEF 6213

ARSEF 7058

ARSEF 6215

ARSEF 6214

ARSEF 10277

ARSEF 10280

ARSEF 2271

| ARSEF 7376

1 ARSEF 2831

ARSEF 7268 B. brongniartii

ARSEF 10278

ARSEF 4362

ARSEF 4363

100/100 ||| ARSEF 617

ARSEF JE276

ARSEF 979

99/100 ARSEF 985

ARSEF 7517

100/100 A RSARSEF 4580

son ARSEF 4622

ARSEF 4598

100/100 ARSEF 4384 | : a

ARSEF 4850 |2-. asiatica

ARSEF 1564

100/100 ARSEF 7518

ARSEF 751

ARSEF 1478 ,

ARSEF 1040 B. bassiana

a ARSEF 300

ARSEF 1811

ARSEF 1848

100/100 | 199400| 100/100 ARSEF 8257

i ARSEF 8259 | B. varroae

ARSEF 2694 ;

Gara ARSEF 7032 — _ B.kipukae

RCEF 5500 — B. lii

ARSEF 4149

B518a

100/100 ARSEF 7542 |B. amorpha

70/100 ARSEF 1969

ARSEF 2641

ARSEF 3405

ARSEF 7242

ARSEF 3220

IL ARSEF 3529 B. pseudobassiana

| ARSEF 3216

ARSEF 1855

100/100 ARSEF 2997

100/100 ARSEF 6229

ARSEF 4933

ARSEF 7280

ARSEF 7043

1 ARSEF 7281 ch

ARSEF 7044 = ‘|B. sungii

100/100 {| ARSEF 5689

ARSEF 7279

ARSEF 1685

ARSEF 1567

/100 100/100 | ARSER S567

Fit F ARSEF 0254 B.caledonica

ARSEF 7117

ARSEF 4302

ARSEF 2922 — B. vermiconia

too/100 | ARSEF 4755

ARSEF 17613 |B.malawiensis

ARSEF 7760 ARSEF 7260

B. australis

100 changes Isaria tenuipes

Fic. 1. Phylogenetic tree of Beauveria based on maximum parsimony and Bayesian analysis of the

combined Bloc, TEF, RPB1, and RPB2 dataset. Bootstrap values (270%) and Bayesian posterior

probabilities (295%) are labeled above and below branches, respectively. Sequence data from all

strains except RCEF5500 are from Rehner et al. (2011). RCEF5500 is indicated in boldface.

Beauveria lii sp. nov. (China) ... 203

The ITS sequence of RCEF5500 differed from other Beauveria species.

Online BLAST searches of the National Center for Biotechnology Information

databases showed that the RCEF5500 isolates are most similar to B. varroae

(ARSEF 8257, HQ880800, 98%), B. kipukae (ARSEF 7032, HQ880734, 97%),

and B. bassiana (ARSEF 1564, HQ880761, 97%), making it hard to determine

the species position. Further comparative multilocus analysis was undertaken

to determine its taxonomic and phylogenetic status.

The combined four-gene, 69 taxon data set published by Rehner et al.

(2011) included 7750 bp of sequence data (Bloc: 1645; TEF: 1010; RPB1: 2928;

RPB2: 2167). After excluding ambiguously aligned regions, the final alignment

comprised 7569 bp (Bloc: 1558; TEF: 986; RPB1: 2861; RPB2: 2164), of which

1261 were parsimony informative. The sequence alignments are available from

TreeBASE as submission 11144. The bootstrap values from maximum parsimony

analyses and the posterior probability values from one of the Bayesian analyses

are shown in FiG.1.

Phylogenetic analysis with four nuclear loci supports RCEF5500 as closely

related to two species, B. varroae and B. kipukae but in an independent lineage

(Fic. 1). Morphologically, RCEF5500 is easily distinguishable from these

two species by its large, ellipsoidal to cylindrical conidia. Additionally, the

phylogenetic analysis showed that RCEF5500 and three morphologically similar

species (B. amorpha, B. caledonica, B. malawiensis) all clustered separately into

different clades and are clearly distinguishable from each other. Therefore, the

phylogenetic data support the recognition of RCEF5500 as a distinct species.

Taxonomy

Beauveria lii Sheng L. Zhang & B. Huang, sp. nov. PLATES 2-11

MycoBank MB563695

Differs from all other Beauveria species by its larger ellispoidal to cylindrical conidia,

with a larger mean length/width ratio.

Type: China, Shaanxi Province, Xunyi County, isolated from a larva of Henosepilachna

vigintioctopunctata, 23 Sep 2010, coll. Ling-ming He (Holotype, RCEF5500; GenBank

JN689370-689374). A living ex-type culture ARSEF 11741 is deposited in USDA-ARS

Collection of Entomopathogenic Fungal Cultures (Ithaca, NY).

Erymo oey: /ii is named in honor of Zengzhi Li, in recognition of his contributions to

the study of entomopathogenic fungi, especially B. bassiana.

Colony growth and appearance similar on full strength Sabouraud's dextrose

and potato dextrose agars, 28-29 mm in diam. after 10 d at 25°C, non-odorous,

cottony, powdery while sporulating; white to cream, with a white margin.

Reverse colorless, sometimes pale yellow in older portions. On Czapek-Dox

agar, colonies slow growing, 19 mm in diameter, thin, sparse, with reverse

colorless; sporulating poorly. Vegetative hyphae septate, branched, hyaline,

smooth walled, 1.3-3.4 um wide. Conidiogenous cells solitary but occasionally

204 ... Zhang & al.

occurring in tight clusters of two to three, the base ellipsoidal to cylindrical

4.8-9.0 x 1.7-2.6 um (FIGs 2, 4, 7), or sometimes globose to subglobose (Fics 5,

6) and produced laterally on aerial hyphae or from subtending cells mostly 4.8-

9.0 x 1.7-2.6 um (Fics 3, 10) Conidiogenous cell apex with an indeterminate,

geniculate, denticulate rachis less than 1m wide. Conidia (3.1—)4.3-6.5(-10.1)

x (1.4-)2.1-2.6(-3.6) um, Q = 2.1-2.7 (L™ = 5.2 um, W™ = 2.3 um, Q™ = 2.3),

mostly ellipsoidal to cylindrical (Fic. 8), occasionally obovoid (Fics 6, 10)

and very occasionally slightly flattened on one side, usually produced from

conidiogenous cells but occasionally directly from hyphal tips or laterally from

hyphae (Fics 9, 11). Teleomorph not observed.

ADDITIONAL MATERIAL EXAMINED: RCEF5183—5195, RCEF5887—5862.

Microscopically, B. lii is distinct from other Beauveria species mainly by its

large ellipsoidal to cylindrical conidia. The six other Beauveria spp. with

similarly shaped conidia all produce smaller conidia: B. amorpha (mean = 3.3

x 2.0 um), B. asiatica (mean =3.2 x 2.3 um), B. brongniartii (mean = 3.0 x 1.6

um), B. caledonica (mean = 3.6 x 1.8 um), B. malawiensis (mean = 3.9 x 1.9

um), B. sungii (mean = 3.0 x 2.1 um); the mean conidial length/width ratios

(Q™) of these six species are also smaller (ranging from 1.4 to 2.1; Rehner et al.

2011, Humber & Rehner 2007) than that of B. lii, which has the largest Q™ in

the genus.

Discussion

In addition to the cryptic species within the B. bassiana sensu lato and

B. brongniartii sensu lato, additional new species (e.g., B. asiatica, B. australis,

B. kipukae, B. pseudobassiana, B. sungii, B. varroae) have been differentiated

chiefly on the strength of molecular data. However, morphological identification

of these species remains difficult in the absence of molecular data. For example,

morphological overlap makes difficult the physical differentiation among

B. bassiana, B. pseudobassiana, B. australis, B. kipukae, and B. varroae. The

multilocus phylogeny approach should be applied to identification of both

cryptic and unknown species of Beauveria. This study has demonstrated that

the four-locus phylogenetic method of Rehner et al. (2011) can be used to

provide a practical solution for distinguishing complex species and establishing

a taxonomic and systematic position for an unknown species, such as B. Jii.

China is a country rich in fungal species diversity; various Beauveria species

have been recorded in different geographical locations, including the dry

northwest region. Beauveria lii was discovered in such an area, suggesting that

this vast zone should be further investigated for entomopathogenic fungi.

Acknowledgments

We are grateful to Drs. Stephen A. Rehner and Long Wang for reviewing the

manuscript. We also thank Dr. Stephen A. Rehner for his kind help with molecular

Beauveria lii sp. nov. (China) ... 205

Fics 2-11. Beauveria lii (holotype, RCEF5500). 2, 4, 7. Ellipsoidal or cylindrical conidiogenous

cells. 3, 10. Globose conidiogenous cell originating from subtending cells. 5-6. Globose

conidiogenous cell. 8. Conidia. 9. Conidia originating from mycelium directly. 11. Conidia

forming from a hyphal tip. Bar = 5 um.

206 ... Zhang & al.

identification. This work was supported by a grant from the Natural Science Foundation

of China (Nos. 30972368 and 31070009).

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Mycosystema 13: 34-40.

MY COTAXON

http://dx.doi.org/10.5248/121.207

Volume 121, pp. 207-213 July-September 2012

Kuehneola warburgiana comb. nov. (Phragmidiaceae, Pucciniales),

causing witches’ brooms on Rosa bracteata

YOSHITAKA ONO

Faculty of Education, Ibaraki University, 2-1-1 Bunkyo, Mito, Ibaraki 310-8512 Japan

CORRESPONDENCE TO: herb-iba@m«x.ibaraki.ac.jp

ABSTRACT—Caeomatoid rust infection has been observed on Rosa bracteata plants at a

single site in Ishigaki Island, Okinawa, Japan, since 1995. The fungus (not previously known

in Japan) was identified as Caeoma warburgiana by its characteristic systemic infection

causing witches’ brooms and its spore morphology. Uredinial and telial sori were found on

the leaves of the witches’ brooms of the infected rose plants at the same site in 2009. The

urediniospores were pedicellate and echinulate. The teliospores were composed of two to

four linearly arranged, thin-walled cells on a short pedicel. Caeoma-type aecia, Uredo-type

uredinia and pedicellate teliospores with two to four linearly arranged cells are characteristic

of the genus Kuehneola. Identical telia and teliospores were found in the lectotype of

C. warburgiana. Caeoma warburgiana is recombined as Kuehneola warburgiana.

Key worps —Asia, life cycle, nomenclature, taxonomy

Introduction

Rosa bracteata is an evergreen perennial shrub, native of southern regions

of China, growing in mixed forests, scrub, and sandy hills at low altitudes and

seashores (Wu & Raven 2003). The plants also occur at coastal areas of Taiwan

and adjacent islands of south Japan (Satake et al. 1989, Liu et al. 2000, Wu &

Raven 2003). Although this rose, called “McCartney rose,” has been introduced

to the United States for breeding new cultivars like “Mermaid” (Beales et al.

1998), it has become a noxious weed in southern States (Texas Invasive Plant

and Pest Council 2011).

In the native regions of this rose species, Caeoma “rosae-bracteatae’,

C. warburgiana, and Kuehneola japonica have been reported to occur

(Anonymous 1979; Hiratsuka & Chen 1991; Sawada 1943; Zhuang 1983);

however, only the last fungus has previously been known in Japan (Hiratsuka

et al. 1992, Kobayashi 2007). In December 1995, a caeomatoid rust infection

was found on R. bracteata plants at a single site at Hirakubo Peninsula, Ishigaki

Island, Okinawa. The caeomatoid fungus causing witches’ brooms on roses was

208 ... Ono

previously not known in Japan. Through continued field observations, uredinial

and telial sori were found on the leaves of the witches’ brooms of the infected

rose at the same site in February 2009.

This paper describes the morphology of the whole life cycle of the R. bracteata

rust fungus and discusses its taxonomic relationships to C. “rosae-bracteatae.’

Materials & methods

Small sorus-bearing pieces were cut out from the herbarium specimens and thin-

sectioned with a razor blade under a binocular dissecting microscope. Spores and

paraphyses were scraped from sori. Thin-sections, scraped spores, and paraphyses

were mounted on microscopic slides and treated by the method described by Ono

(2000). The slide preparations were then examined both by bright-field and differential

interference contrast microscopy (DIC) with an Olympus BH2 microscope (Olympus,

Tokyo, Japan), and measurements were made with an ocular micrometer. Fifty or twenty

randomly selected spores and paraphyses were measured for each specimen and five or

ten spermogonia were measured. For observation of urediniospore germ pores, spores

were mounted in lactic acid on glass slides and heated to boiling point for a few seconds,

after which a drop of lactophenol solution with aniline blue was added onto the boiled

spores.

For scanning electron microscopy (SEM), spores scraped from herbarium specimens

were placed on double-sided adhesive tape on a specimen holder and coated with

platinum-palladium at 25 nm thicknesses under a Hitachi E-1030 ion sputter (Hitachi,

Tokyo, Japan). Samples were observed with a Hitachi $-4200 SEM operating at 15 kV.

Taxonomy

Kuehneola warburgiana (Henn.) Y. Ono comb. nov. Fics. 1-3

MyYCOBANK 563065

= Caeoma warburgiana Henn., in Warburg, Monsunia 1: 4, 1899 [“1900”].

LECTOTYPE (designated here): on Rosa sp., China, Zhejian, Ningbo, 0 + I + II, Dec

1887, O. Warburg (B).

Caeoma “rosae-bracteatae” Sawada, Rep Dept Agr Gov Res

Inst Formosa 86:131, 1943, nom. inval.

AECIAL INFECTION systemic, causing witches’ brooms on shoots spreading

a large area. SPERMOGONIA on juvenile shoot tips, leaf stipules and petioles

(but not on leaf body), subepidermal, of determinate growth, dome-shaped

or broadly convex with flat hymenium, 139-206 um wide, and 58-104 um

high. Axcra strictly on shoots, subepidermal in origin becoming erumpent, of

indeterminate growth, neither peridiate nor paraphysate, bright orange-colored,

and powdery. AECIOsPoRES formed in basipetal succession, but separated soon

after maturation, thus not forming chains; ellipsoid to broadly ellipsoid, often

angular, and 20-29 x 12-18 um in size; wall 1-2 um thick at sides, apically

thickened, colorless, and verrucose with no apparent germ pore.

Kuehneola warburgiana sp. nov. (Japan) ... 209

“ \

: ie

: ’ ‘

. . > ,

“ \

FiGuRE 1 Kuehneola warburgiana (IBAR-7744). A. Witches’ brooms exhibited by Rosa bracteata

shoots with spermogonial/aecial infection. B. Early stage of witches’ broom development. Leaf

petioles and stipules are densely covered with orange-colored spermogonia. C. Mature caeomatoid

aecia on shoots with powdery orange-colored spores. D. Vertical section of a subepidermal

spermogonium with a flat hymenium. E. Vertical section of caeomatoid aecium. Aeciospores are

formed in basipetal succession. F. Aeciospores. G. Surface structure of an aeciospore. Bars: D = 50

um; E, F= 10 um; G= 5um

UrepInIA on abaxial leaf surface, scattered or in loose groups, bright orange-

colored and powdery, subepidermal in origin and erumpent, peripherally

paraphysate, and often mixed with teliospores; paraphyses irregularly

cylindrical, weakly to strongly incurved, apically and dorsally thick-walled,

colorless, 25-30 um high, and 6-9 um wide. UREDINIOSPORES formed singly

on a short pedicel, ellipsoid, broadly ellipsoid, obovoid or pyriform, and 18-25

x 11-16 um in size; wall thin, colorless, and echinulate with no apparent germ

pore.

2O.. Oe

FIGURE 2 Kuehneola warburgiana (IBAR-10130). A. Orange-colored, powdery uredinia and

white cottony telia formed on the abaxial leaf surface of Rosa bracteata. B. Incurved paraphyses.

C. Urediniospores. D. Vertical section of a telium. The sorus is surrounded by incurved paraphyses.

The wall is dorsally and apically thickened. E. Two- or three-celled teliospores with a short

pedicel (fragment remained at the base). F. Metabasidium development from a distal end cell.

G. Basidiospores formed from a four-celled metabasidium (one spore is missing). Bars = 20 um

TELIA on abaxial leaf surface, white, cottony, peripherally paraphysate,

and often intermixed with uredinia. TELIOspPORES composed of 2-4 linearly

arranged cells formed on a short pedicel; each telial cell ovoid, ellipsoid

or broadly ellipsoid, and 21-34 x 10-17 um in size; wall thin and colorless;

germinating in situ to form four basidiospores on a four-celled metabasidium.

ADDITIONAL SPECIMENS EXAMINED: On Rosa sp.: CHINA, FUJIAN, Futschau, Yuenfuthal,

0 + I, Dec. 1887, O. Warburg (B).

On Rosa bracteata J.C. Wendl.: JAPAN, OKINAWA, Ishigaki, Hirakubo Peninsula, 0

+ I, 10 Dec 1995, Y. Ono (IBAR-7744); 0 + I, 7 Nov 1997, Y. Ono (IBAR-7961); 0 + I, 10

Nov 2002, Y. Ono (IBAR-8888); 0 + I + II + III, 17 Feb 2009, Y. Ono (IBAR-10130)

EcoLtoGcy—tThe rust infected R. bracteata plants spread in the steep slope at

the peninsula in Ishigaki. The habitat is heavily utilized for grazing and the

Kuehneola warburgiana sp. nov. (Japan) ... 211

FIGURE 3 Caeoma warburgiana (Lectotype, deposited in B). A. Witches’ brooms exhibited by

unidentified rose shoots with spermogonial/aecial infection. B. Mature caeomatoid aecia with

powdery spores. C. Aeciospores. D. White cottony telia formed on the abaxial leaf surface.

E. Vertical section of a telium. Bars = 20 um

prickly bushes are the property by which the rose is selected for in the habitat,

otherwise it is not commonly observed in the island covered with subtropical

evergreen forests.

ComMMENTS—Kuehneola japonica (Dietel) Dietel is the only other Kuehneola

species to occur on Rosa spp. This fungus is microcyclic and forms only telia on

15 species and 5 varieties of Rosa, being widely distributed in east and southeast

Asia (Anonymous 1979; Arthur & Cummins 1936; Hiratsuka & Chen 1991;

Hiratsuka et al. 1992; Kobayashi 2007; Sawada 1918, 1944; Teodoro 1937).

This species forms pulvinate bright orange-colored telia, not associated with

spermogonia, on both surfaces of leaves, petioles and shoots (Hiratsuka et al.

1992). The sori often become confluent, particularly along the leaf veins and on

shoots; however, the telial infection is not systemic. Teliospores are 2-4-celled

with a short, persistent pedicel. Each teliospore cell is 22-38 x 15-22 um in size

and thin-walled.

Phylogenetic relationships between K. warburgiana and K. japonica are not

apparent. The non-systemic nature of telial infection of K. japonica suggests

that the “correlated species” concept (Arthur 1934) and its evolutionary

interpretation, Tranzschel’s Law (Shattock & Preece 2000; Cummins &

Hiratsuka 2003), may not apply to a possible relationship between the two

fungi, i.e., a direct evolutionary derivation of microcyclic K. japonica from

macrocyclic K. warburgiana.

212) Ono

In his protologue of C. warburgiana, Hennings (1899) cited two collections

— “China, Fitschau und Ningpo: auf Zweigen von Rosa sp. Dez. 1887.

(O. Warburg).” These collections are syntypes, because the protologue did not

designate either as the holotype. In B, the collection from Ningpo (Zhejiang) is

labeled as “holotypus” and the collection from Fitschau (Fujian) as “isotypus.”

Yet Hein (1989) listed the Fitschau collection as the “holotypus” and did not

mention the other collection. In the Ningpo collection, the telia and teliospores

are present (Fics. 3 D,E) and morphologically the same as those found on

the Ishigaki collection (IBAR-10130). Therefore, the Ningpo collection is

designated here as the lectotype (although it is possible that this collection may

already have been designated as lectotype, by a corrective interpretation of the

misapplied term “holotypus’” on the label of the B herbarium specimen).

Caeoma warburgiana has been reported on R. bracteata in Fujian (Zhuang

1983), R. banksiae W. T. Aiton var. banksiae in Yunnan (Tai 1979), R. banksiae

var. normalis Regel in Yunnan (Tai 1979), R. hugonis Hemstl. in Fujian (Zhuang

1983), and R. longicuspis Bertol. in Yunnan (Tai 1979).

Caeoma “rosae-bracteatae” Sawada was published (without Latin diagnosis)

for a fungus on R. bracteata var. bracteata (as R. bracteata “var. typica” Lindl.)

(Sawada 1943). Four specimens were cited, all from Hsinchu, Taiwan. The sori

were described as elongated up to 70 mm and becoming naked with orange-

yellow spores; the spores were in chains of 3-4, ellipsoid, globoid or obovoid-

ellipsoid, and 22-36 x 11-16 um in size; the wall was densely verrucose,

2 um thick and up to 7-8 um apically. Although none of Sawada’s collections

were available for the study, all the details described for the fungus support its

biological identity with C. warburgiana.

Spermogonia of Kuehneola species are characterized by their subcuticular

origin with a flat hymenium and indeterminate growth (Type 11 of Cummins

& Hiratsuka 2003). However, intraepidermally formed spermogonia of

indeterminate growth was reported for K. loeseneriana (Henn.) H.S. Jacks.

& Holw. (Hernandez & Hennen 2003). Kuehneola warburgiana forms Type

6 spermogonia (subepidermal with a flat hymenium of determinate growth),

a type previously known only in Gymnoconia (Hiratsuka & Hiratsuka 1980;

Cummins & Hiratsuka 2003).

Acknowledgments

I would like to express my gratitude to Dr. Robert Vogt, curator, the Mycological

Herbarium, Botanischer Garten und Botanisches Museum Berlin-Dahlem for the loan

of the type material examined in this study and Dr. Burghard Hein at the same institution

for providing me with the data of the Hennings’ fungal collections at B. I also thank

peer reviewers, Dr. Stephan Helfer, Royal Botanic Garden, Edinburgh and Dr. Anibal A.

de Carvalho, Jr., Instituto de Pesquisas Jardim Botanico do Rio de Janeiro and editors,

Dr. Shaun Pennycook and Dr. Lorelei Norvell, for their technical assistance.

Kuehneola warburgiana sp. nov. (Japan) ... 213

Literature cited

Anonymous 1979. List of plant diseases in Taiwan. Plant Protection Society, Republic of China.

Arthur JC. 1934. Manual of the rusts in United States and Canada. Purdue Research Foundation.

Arthur JC, Cummins GB. 1936. Philippine rusts in the Clemens collection 1923-1926, II. Philipp

J Sci 61: 463-488.

Beales P, Cairns T, Duncan W, Fagan G, Grant W, Grapes K, Harkness P, Hughes K, Mattock J,

Ruston D, Sutherland P, Williams T. 1998. Botanica’s roses: The encyclopedia of roses. Milsons

Point, Random House, Australia.

Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi. 3rd edition. St. Paul, American

Phytopathological Society.

Hein B. 1989. Liste der Arten und infraspecifischen Taxa von P. Hennings mit Angabe der Typen

in den Herbarien des Botanischen Museums Berlin-Dahlem und des Instituts fiir Allgemeine

Botanik in Hamburg. Englera 10, Botanischer Garten und Botanisches Museum Berlin-

Dahlem.

Hennings P. 1899 [“1900”]. Fungi. 1-38, in: Warburg O (ed.). Monsunia. Beitrage zur Kenntniss

der Vegetation des siid- und ostasiatischen Monsungebietes. Vol. 1. Leipzig, W. Engelmann.

Hernandez JR, Hennen JE 2003. Rust fungi causing galls, witches’ brooms, and other

abnormal plant growths in northwestern Argentina. Mycologia 95: 728-755.

http://dx.doi.org/10.2307/3761948

Hiratsuka N, Chen ZC. 1991. A list of Uredinales collected from Taiwan. Trans mycol Soc Japan

32:3-22,

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y,

Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba, Tsukuba Shuppankai.

Hiratsuka Y, Hiratsuka N. 1980. Morphology of spermogonia and taxonomy of rust fungi. Rep

Tottori Mycol Inst 18: 257-268.

Kobayashi T. 2007. Index of fungi inhabiting woody plants in Japan. Host, distribution and

literature. Tokyo, Zenkoku-Noson-Kyoiku Kyokai Publ. Co., Ltd.

Liu HY, Yang YP, Lu SY, Shih, BL (eds) 2000. Manual of Taiwan vascular plants. Vol. 3. IL.

Spermatophyta. 2. Angiospermae. A. Dicotyledons (continued) (in Chinese). Taipei, Council

of Agriculture Taiwan.

Satake Y, Hara H, Watari S, Tominari T. (eds) 1989. Wild flowers of Japan. Woody plants (in

Japanese). Tokyo, Heibonsha Ltd.

Sawada K. 1918. A new rust fungus parasitic on the cultivated rose (in Japanese with a Latin

diagnosis). Trans Sapporo Nat Hist Soc 7: 36-40.

Sawada K. 1943. Descriptive catalogue of the Formosan fungi. Part IX (in Japanese). Rep Dept Agr

Gov Res Inst Formosa 86: 1-178.

Sawada K. 1944. Descriptive catalogue of the Formosan fungi. Part X (in Japanese). Rep Dept Agr

Gov Res Inst Formosa 87: 1-96.

Shattock RC, Preece TF. 2000. Tranzschel revisited: modern studies of the relatedness of different

rust fungi confirm his law. Mycologist 14: 113-117.

http://dx.doi.org/10.1016/S0269-915.X(00)80086-5

Tai FL. 1979. Sylloge Fungorum Sinicorum. Peking, Science Press, Academia Sinica.

Teodoro NG. 1937. An Enumeration of Philippine Fungi. Techn Bull Dept Agr Comm Manila 4:

1-585.

Texas Invasive Plant and Pest Council 2011. Rosa bracteata, Macartney rose. [retrieved 21 July

2011.] http://www.texasinvasives.org/plant_database/detail. php?symbol=ROBR

Wu Z-y, Raven PH (eds) 2003. Flora of China. Vol. 9. Pittosporaceae through Connaraceae. Beijing,

Science Press & St. Louis, Missouri Botanical Garden.

Zhuang JY. 1983. A provisional list of Uredinales of Fujian Province, China. Acta Mycol Sin

2: 146-158.

MY COTAXON

http://dx.doi.org/10.5248/121.215

Volume 121, pp. 215-223 July-September 2012

New records of smut fungi. 6

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

ABSTRACT — Five species of smut fungi, Leucocintractia scleriae, Sporisorium caledonicum,

Sporisorium paspali-thunbergii, Sporisorium ugandense, and Ustanciosporium scleriicola, are

recorded for the first time from the Democratic Republic of the Congo. Cyperus distans is

a new host of Cintractia limitata in Africa. For Entorrhiza casparyana var. tenuis on Juncus

tenuis, a new combination and status, E. tenuis, are proposed.

KEY worps — taxonomy, Ustilaginomycetes

Introduction

Vanky et al. (2011) published a checklist containing 427 species in 47

genera of smut fungi in Africa and adjacent islands. We add a further five

species to the smut fungi known from the Democratic Republic of the Congo

and report Cyperus distans as a new host of Cintractia limitata in Africa. A

new combination and status, at the species level, are proposed for Entorrhiza

casparyana var. tenuis on Juncus tenuis.

Material & methods

Dried specimens from the mycological collection of the National Botanical Garden

of Belgium (BR) were examined under light (LM) and scanning electron (SEM)

microscopes. For LM observations, spores were mounted in lactophenol solution on

glass slides, gently heated to boiling point to rehydrate the spores, and then cooled.

Spore measurements are given in the form: min-max [mean + 1 standard deviation].

For SEM, spores were attached to specimen holders by double-sided adhesive tape and

coated with gold with an ion sputter. The surface structure of spores was observed at 10

kV and photographed with a JEOL JSM-5510 scanning electron microscope.

The distribution of the smut fungi in Africa is given in accordance with Vanky et al.

(2011) while the general distribution is after Vanky (2011).

216 ... Denchev & Denchev

New records for the Democratic Republic of the Congo

Leucocintractia scleriae (DC.) M. Piepenbr., Begerow & Oberw.,

Mycologia 91: 497, 1999. FIGs 1-2

Sort around all the peduncles of an inflorescence with rudimentary spikelets,

more or less cylindrical, often curved, 5-25 x 2-3 mm wide, when young

covered by a thick peridium, later becoming exposed; spore mass of the mature

sori dark reddish brown, agglutinated, powdery on the surface. Spores slightly

flattened, in plane view orbicular, suborbicular or broadly elliptical, in plane

view 13-16.5 x 11.5-15.5 (14.8 + 0.7 x 13.4 + 0.9) um (n = 50), in side view 9.5-12

uum wide, reddish brown; wall 0.9-1.5 um thick, in LM with connected warts,

forming rows, irregular and incomplete reticulum-like ornaments, or parallel

ridges (especially, in side view); in SEM with well developed ornamentation: in

plane view of coarse, irregular, connected warts, forming short or long rows, or

fusing into irregular meshes, in side view of connected warts forming irregular

meshes (the interspaces often with single or connected warts) or uneven, more

or less parallel ridges, often connected by lower, transverse rows.

SPECIMEN EXAMINED — On Rhynchospora corymbosa (L.) Britton: DEMOCRATIC

REPUBLIC OF THE CONGO, ORIENTALE PROVINCE, Garamba National Park, near

Bagbele, 03°40’ N, 29°00’ E, 24 April 1950, leg. A. Noirfalise, no. 199 (BR 49871,13).

DISTRIBUTION — On Rhynchospora spp. (Cyperaceae), distributed in the

tropics. In Africa known on R. corymbosa and R. spectabilis from Cameroon,

Guinea, Namibia, Sierra Leone, South Africa, and Tanzania.

Sporisorium caledonicum (Pat.) Vanky, Mycotaxon 40: 165, 1991. FIGs 3-4

Sor! destroying the whole inflorescence, when young covered by a greyish

brown peridium, which ruptures irregularly exposing a dark reddish brown

mass of permanent spore balls and several filiform columellae. SPORE BALLS

irregular, broadly ellipsoidal, ellipsoidal or ovoid, 35-85 x 25-50 um, dark

reddish brown, often opaque, composed of tens, rather firmly united spores.

STERILE CELLS absent. Spores dimorphic. Outer spores subglobose, broadly

ellipsoidal or slightly irregular, 9.5-13 x 7.5-11.5 (11.4 + 0.9 x 9.5 + 1.0) um

(n = 50), medium to dark reddish brown; wall 0.7-1.5 um thick, in LM punctate

to finely verruculose, spore profile slightly affected, in SEM punctate to finely

verruculose.

SPECIMEN EXAMINED — On Heteropogon contortus (L.) P. Beauv. ex Roem. & Schult.:

DEMOCRATIC REPUBLIC OF THE CONGO, KINSHASA PROVINCE, Boma, 05°51’

S, 13°03’ E, 1921, leg. Claessens in Vanderyst, sine num. (BR 49827,66).

DISTRIBUTION — On Heteropogon spp. (Poaceae), Africa, South Asia, Australia,

Papua New Guinea, North & South America. In Africa known on H. contortus

from Ethiopia, South Africa, Zambia, and Zimbabwe.

Smuts new to the Democratic Republic of the Congo ... 217

Fics 1-2. Leucocintractia scleriae on Rhynchospora corymbosa (BR 49871,13). Spores in LM and

SEM. Fics 3-4. Sporisorium caledonicum on Heteropogon contortus (BR 49827,66). Spore ball in

LM and SEM. Scale bars: 1, 3, 4= 10 um, 2 = 5 um.

218 ... Denchev & Denchev

Sporisorium paspali-thunbergii (Henn.) Vanky, Publications from the

Herbarium Ustilaginales Vanky 3: 9, 1986. Fics 5-6

Sori destroying the whole inflorescence, up to 10 cm long, 1-2.5 mm

wide, partly hidden by the leaf sheath, when young covered by a thick, greyish

brown peridium which ruptures irregularly exposing a powdery, dark reddish

brown mass of spore balls and spores, and a long, filiform, central columella;

the columella sometimes with short, lateral branches. SpoRE BALLS broadly

ellipsoidal, globose, ovoid or irregular, 35-70 x 30-55 um, dark reddish

brown, composed of eight to tens spores, separating easily. STERILE CELLS

absent. SporEs irregular to subpolygonal, broadly ellipsoidal or subglobose,

sometimes elongated, 11-17 x 9-13.5 (13.9 + 1.3 x 11.6 + 1.2) um (n = 50),

medium reddish brown; wall uneven, thicker at the angles, 0.7-2.0 um thick,

in LM punctate to verruculose on the free surface of the spores, and smooth

on the contact surface, spore profile not affected, in SEM finely echinulate or

verruculose.

SPECIMEN EXAMINED — On Paspalum sp.. DEMOCRATIC REPUBLIC OF THE

CONGO, ORIENTALE PROVINCE, Kisangani, 00°31’ N, 25°11’ E, 19 January 1931, leg.

R.L. Steyaert, no. 330 (BR 49888,30).

DISTRIBUTION — On Paspalum spp. (Poaceae), Africa, South and East Asia,

Philippines, Australia, and Hawaii. In Africa known on P scrobiculatum from

Ethiopia, Malawi, South Africa, and Uganda.

Sporisorium ugandense (Henn.) Vanky, Mycotaxon 91: 250, 2005. Fics 7-8

Sort destroying the distal part of sterile shoots, 4-6 cm long, 0.5-2 mm

wide, partly hidden by the leaf sheath, covered by a greyish peridium which

later ruptures, exposing powdery, dark brown mass of loose spore balls and

spores, and numerous filiform columellae. SpoRE BALLS globose, broadly

ellipsoidal, ovoid or slightly irregular, 30-65 x 25-55 um, medium reddish

brown, composed of fifteen to tens spores, separating easily. STERILE CELLS

absent. Spores subglobose, broadly ellipsoidal, ellipsoidal, ovoid or slightly

irregular, 7.5-12 x 7-9.5 (9.4 + 0.8 x 8.1 + 0.4) um (n = 50), light reddish

brown; wall 0.3-0.5 um thick, in LM punctate to verruculose, spore profile not

affected, in SEM verruculose.

SPECIMEN EXAMINED — On Digitaria abyssinica (A. Rich.) Stapf: DEMOCRATIC

REPUBLIC OF THE CONGO, Tshirumbi, 02°19’ S, 28°47’ E, August 1945, leg.

EL. Hendrickx, no. 2882 (as D. scalarum (Schweinf.) Chiov., BR 49849,88).

DISTRIBUTION — On Digitaria spp. (Poaceae), Africa and Australia. In Africa

known on D. abyssinica from Ethiopia, Kenya, and Uganda.

Smuts new to the Democratic Republic of the Congo ... 219

. i

Fics 5-6. Sporisorium paspali-thunbergii on Paspalum sp. (BR 49888,30). Spore balls and

spores in LM and SEM. Fics 7-8. Sporisorium ugandense on Digitaria abyssinica (BR 49849,88).

Spore balls and spores in LM and SEM. Scale bars: 5, 7 = 10 um, 6, 8 = 5 um.

220 ... Denchev & Denchev

Ustanciosporium scleriicola (Cant.) M. Piepenbr., Nova Hedwigia 70: 352, 2000.

FIGS 9-10

Sori in all flowers of the spikelets; spore mass of the mature sori black,

powdery. Spores single, flattened, in plane view orbicular, suborbicular, broadly

elliptical or slightly irregular, in plane view 16.5-23 x 14.5-20.5 (18.9 + 1.7 x

17.3 + 1.4) um (n = 50), in side view 9.5-13 um wide, medium to dark reddish

brown, no hyaline appendages; wall 1.0-1.9 um thick, in LM foveolate-reticulate,

the ornamentation slightly affecting the spore profile, in SEM foveolate or with

reticulate pattern of low muri and irregular, polygonal meshes.

SPECIMEN EXAMINED — On Scleria melanomphala Kunth: DEMOCRATIC REPUBLIC

OF THE CONGO, Katomia, Marungu, 07°20’ S, 29°40’ E, 28 April 1939, leg. Van den

Brande, no. 120 (as Ustilago sp., BR 49895,37).

DISTRIBUTION — On Scleria melanomphala, S. lagoensis Boeckeler, Scleria sp.

(Cyperaceae), Africa; known from Cameroon, Central African Republic, and

Ethiopia.

Anew host of Cintractia limitata in Africa

Cintractia limitata G.P. Clinton, Proc. Boston Soc. Nat. Hist. 31: 399, 1904. Fics 11-12

Sor! in individual flowers or groups of flowers; spore mass of the mature

sori dark reddish brown, powdery. Sporss flattened, in plane view orbicular,

suborbicular or broadly elliptical, sometimes slightly irregular, in plane view

9.5-13.5 x 7.5-12.5 (11.8 + 1.0 x 10.5 + 0.9) um (n = 50), in side view 6.5-8.5

um wide, light to medium reddish brown, wall 0.6-1.0 um thick; in LM smooth,

in SEM densely punctate of very low warts (often forming small groups or short

rows).

SPECIMEN EXAMINED — On Cyperus distans L.f.: DEMOCRATIC REPUBLIC OF THE

CONGO, Equateur Province, Koli Koli, 00°05’ S, 18°12' E, July 1930, leg. P. Staner, no.

213 (BR 49875,17).

DISTRIBUTION — On Cyperus, Kyllinga, and Mariscus spp. (Cyperaceae), tropics

and subtropics. In Africa known from Cameroon, Congo, Ethiopia, Gabon,

Ghana, Guinea, Ivory Coast, Malawi, Nigeria, Reunion, Sierra Leone, South

Africa, Sudan, Tanzania, Togo, Uganda, Zambia, and Zimbabwe. Cyperus

distans is reported here as a new host of Cintractia limitata in Africa.

New combination and status of Entorrhiza casparyana var. tenuis

Entorrhiza casparyana (Magnus) Lagerh. var. casparyana attacks 17 species

of Juncus (not including J. tenuis), and is distributed in Europe, North America,

Africa, Australia, and New Zealand (Vanky & McKenzie 2002, Denchev &

Minter 2008, Vanky & Shivas 2008, Denchev et al. 2011, Vanky et al. 2011).

Denchev et al. (2007) described a new variety of Entorrhiza casparyana,

var. tenuis, restricted to one host, Juncus tenuis, and recorded only from South

Korea, Austria, Romania, and Costa Rica (Vanky 1985, Piepenbring 2003,

Smuts new to the Democratic Republic of the Congo ... 221

Fics 9-10. Ustanciosporium scleriicola on Scleria melanomphala (BR 49895,37). Spores in LM and

SEM. Fics 11-12. Cintractia limitata on Cyperus distans (BR 49875,17). Spores in LM and SEM.

Scale bars: 9, 11 = 10 um, 10, 12 = 5 um.

222 ... Denchev & Denchev

Denchev 2004 [all as E. casparyana]; Denchev et al. 2007). It differs from var.

casparyana especially by the shorter sori and spores. ‘The sori of var. casparyana

are up to 15 mm in length, whereas those of var. tenuis are only up to 5 mm

long. The spores of var. casparyana are (12—)13.5-23(-32) um long (including

the ornamentations) while those of var. tenuis are 11.5-20(-21.5) um long.

We suggest that Entorrhiza casparyana var. tenuis is morphologically distinct

from E. casparyana var. casparyana and should be recognized at the species

level.

Entorrhiza tenuis (Denchev & H.D. Shin) Denchey, Vanky & T. Denchev,

comb. et stat. nov.

MycoBank MB 564487

= Entorrhiza casparyana var. tenuis Denchev & H.D. Shin, Mycotaxon 100: 74, 2007.

TYPE COLLECTIONS: on Juncus tenuis Willd. KOREA, GANGWON PRov., near

Hoengseong, 37°31'24.82" N, 128°17'27.98" E, 7 September 2006, C.M. Denchev

(holotype, SOMF 26 206); 37°31'24.84" N, 128°17'28.11" E, 7 September 2006, C.M.

Denchev (paratype, SOMF 26 207).

ILLUSTRATIONS: Denchev 2004: 50 (Figs 1-2, as E. casparyana), Denchev et al. 2007: 75

(Figs 1-2, as E. casparyana var. tenuis).

CoMMENTS — Four Entorrhiza species are known on Juncus: E. aschersoniana

(Magnus) Lagerh. (Europe, Central America, New Zealand), E. caricicola

Ferd. & Winge (Europe, New Zealand), E. casparyana, and E. casparyanella

Vanky (New Zealand). Entorrhiza tenuis differs from E. casparyanella in having

tuberculate or verrucose spore ornamentation with coarse tubercules or warts,

whereas E. casparyanella spores have a smooth or undulate outer wall layer (cfr

Vanky 1998: 342-343). Spore shape primarily distinguishes E. aschersoniana

and E. caricicola from E. tenuis, with the mainly broadly ellipsoidal, ellipsoidal

or ovoid (occasionally subglobose) spores with a length/width ratio >1.06

of the first two species contrasting with the globose or subglobose spores of

E. tenuis (length/width ratio 1.04-1.05).

Key to Entorrhiza species on Juncus

1 Spores mainly broadly ellipsoidal, ellipsoidal or ovoid, occasionally

subslobosesfeneth/ width ratio 06<s:.-¢ sores scnee eA attests Raed 2

1* — Spores globose or subglobose, length/width ratio <1.05 .................04. 3

2 Spore wall rugulose-undulate or smooth ....................0.. E. caricicola

2* “Spore swalllverrucose”. fe. os-fag sao setae’ steak shal sas E. aschersoniana

3 Outer wall layer smooth or undulate; spores 10-17 um long ... E. casparyanella

3* Outer wall layer coarsely tuberculate or verrucose, occasionally smooth ...... 4

4 — Spores (12-)13.5-23(-32) um long, sori up to 15mm long ...... E. casparyana

4* Spores 11.5-20(-21.5) um long, sori 1.2-5 mm long ................ E. tenuis

Smuts new to the Democratic Republic of the Congo ... 223

Acknowledgements

We gratefully acknowledge Dr Kalman Vanky (Herbarium Ustilaginales Vanky,

Tubingen, Germany) and Dr Roger G. Shivas (Biosecurity Queensland, Australia) for

critically reading the manuscript and serving as pre-submission reviewers, and the

Curator of the mycological collection of the National Botanical Garden of Belgium (BR)

for the loan of the cited specimens.

Literature cited

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Balcanica 1: 49-50.

Denchev CM, Minter DW. 2008. Entorrhiza casparyana. IMI Descriptions of Fungi and Bacteria.

No. 1761. CAB International, Egham. 5 pp.

Denchev CM, Shin HD, Kim SM. 2007. New records of smut fungi from Korea. 2. — Mycotaxon

100: 73-78.

Denchev CM, Denchev TT, Spooner BM, Helfer S. 2011. New records of smut fungi. 3. Mycotaxon

114: 225-230. http://dx.doi.org/10.5248/114.225

Piepenbring M. 2003. Smut fungi (Ustilaginomycetes p.p. and Microbotryales, Basidiomycota). Flora

Neotropica, Monograph 86. The New York Botanical Garden Press, Bronx, NY.

Vanky K. 1985. Carpathian Ustilaginales. Symbolae Botanicae Upsalienses 24(2): 1-309.

Vanky K. 1998. New Australasian Ustilaginales. Mycotaxon 68: 327-344.

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

1458 pp.

Vanky K, McKenzie EHC. 2002. Smut fungi of New Zealand. Fungi of New Zealand. Vol. 2. Fungal

Diversity Press, Hong Kong.

Vanky K, Shivas RG. 2008. Fungi of Australia: The smut fungi, in Fungi of Australia Series.

Australian Biological Resources Study (ABRS), Canberra & CSIRO Publishing, Melbourne.

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

8: 1-77.

MY COTAXON

http://dx.doi.org/10.5248/121.225

Volume 121, pp. 225-232 July-September 2012

Suillus flavidus and its ectomycorrhizae with

Pinus wallichiana in Pakistan

S. SARWAR*, A.N. KHALID, M. HANIF & A.R. NIAZI

Department of Botany, University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

* CORRESPONDENCE TO: samina_boletus@yahoo.com

Asstract — Suillus flavidus (Boletales, Suillaceae) was found associated with Pinus

wallichiana during a survey of macrofungi from moist coniferous forests of Pakistan. Both

the fruiting body and ectomycorrhizae were characterized morpho-anatomically as well as

by molecular analysis. This fungus is a new record for Pakistan and its ectomycorrhizae with

Pinus wallichiana are described for the first time by molecular analysis.

Key worps —boletes, ITS, mantle, PCR, rDNA

Introduction

Coniferous forests of Pakistan are located at an elevation of 1373 to 3050 m

a.s.l. and are characterized by luxuriant growth of trees such as Abies pindrow,

Cedrus deodara, Picea smithiana, Pinus roxburghii, P. wallichiana, and Taxus

wallichiana. Among these conifers, some deciduous trees and shrubs of

different species also occur (Hussain 1995). Another important feature of these

forests is the high level of rainfall during summer (July-August). High rainfall

and temperature make an environment suitable for the growth of mushrooms.

Most of these fungi form mutualistic symbiotic associations with forest trees

in the form of ectomycorrhizae that facilitate tree growth through enhanced

nutrient absorption and protection of roots from root pathogens (Marx 1991).

Suillus Gray, a genus with approximately 50 species (Kirk et al. 2008), is

characterized by growth under conifers, slimy caps, glandular dots on the stipe,

large pore openings that are often arranged radially, and a partial veil that leaves

a ring around the stipe or tissue hanging from the cap margin (Kuo 2004). Nine

Suillus species have been reported from Pakistan (Ahmad et al. 1997; Razaq

2007; Niazi 2008) associated with different conifers.

During field surveys of Pakistan forests, we collected Suillus mushrooms

under pine trees in order to explore some new and noteworthy species of this

226 ... Sarwar & al.

genus. The aim ofthe present work is to describe the morpho-anatomical features

of the Suillus collections and their ectomycorrhizae with Pinus wallichiana as

well as to confirm their identification by molecular analysis.

Materials & methods

The sampling was carried out in coniferous forests of Pakistan located at an elevation

of around 2200 m during the 2010 rainy season (July-August). The sporocarps and

blocks of soil exactly beneath the fruiting bodies were taken from the rhizosphere

of Pinus wallichiana. The sporocarps were air dried while the soil blocks containing

ectomycorrhizae (ECM) were wrapped in polythene bags and brought to the lab for

further analysis. Field data on basidiocarps (site, habitat, association etc) was noted.

Specimens were studied macroscopically and microscopically in the laboratory following

the methods of Bessette et al. (2000). In the spore dimensions, the first values present the

range of lengths and widths, and the values in parentheses present mean spore lengths

and widths + standard deviations followed Q, + standard deviation, where Q_ is the

mean of Q (= length/width ratio of an individual spore). Other measurements are given

as a range with exceptional values in parentheses. ECM were carefully placed in water to

clean off soil particles and characterized morphologically under the stereomicroscope.

The microscopic description of the ECM follows the terminology of Agerer (1991, 1999).

Mantle views, emanating elements, and illustrations were documented with the help of

TABLE 1: rDNA sequences of Suillus species from Pakistan retrieved from GenBank

for phylogenetic analysis.

NAME ACCESSION NO. IsOLATE No. ORIGIN SOURCE

Suillus bellinii AY898621.1 CCMA-22 Spain Fruitbody

Suillus bovinus AY898623.1 CCMA-67 Spain Fruitbody

Suillus brevipes FJ845440.1 SMI330 Canada Fruitbody

Suillus caerulescens EU486453.1 UBC F16304 Canada Fruitbody

Suillus collinitus AY953421.1 ECM 14 Spain Ectomycorrhiza

DQ440569.1 CCMA-73 Spain Fruitbody

Suillus flavidus — 2C Pakistan Fruitbodies

— 2D Pakistan Fruitbodies

— 2H Pakistan Fruitbodies

— 3H Pakistan Fruitbodies

— H10 Pakistan Fruitbodies

— 10B Pakistan Fruitbodies

— 2H 10H Pakistan Ectomycorrhiza

— 3A 11G Pakistan Ectomycorrhiza

— 8G Pakistan Ectomycorrhiza

AJ971403.1 pl2 UK Ectomycorrhiza

FJ845439.1 SMI206 Canada Fruitbody

Suillus granulatus AY898617.1 CCMA-02 Spain Fruitbody

Suillus grevillei AF347102.1 EL 38-99 Sweden Fruitbody

Suillus lakei DQ367917.1 OUC97024 Canada Fruitbody

Suillus luteus DQ440568.1 CCMA-37 Spain Fruitbody

Suillus mediterraneensis AY935512.1 CCMA-26 Spain Fruitbody

Suillus quiescens GQ249402.1 UC1860306 USA Fruitbody

Suillus variegatus AM086444.1 P17F UK Ectomycorrhiza

AM086446.1 P22 F UK Ectomycorrhiza

Suillus flavidus + Pinus wallichiana ectomycorrhizae (Pakistan) ... 227

a camera lucida. Voucher specimens were deposited in the Herbarium, Department of

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

Molecular analysis

DNA was extracted from dried basidiocarps using Extract N. Amp.TM Plant kit

(SIGMA), and the nrDNA was amplified using fungal specific primers pair ITS3

(5’-GCATCGATGAAGAACGCAGC-3") and ITS6-R (5’-TTCCCGCTTCACTCGCAGT-3’).

Amplification parameters were denaturation at 94°C for 4 min., then 35 cycles of 45 s at

94°C, 45 s at 54°C, and 1 min 30 s at 72°C, and a final extension at 72°C for 2 min. The

purified PCR products were sequenced bidirectionally by Macrogen (South Korea). The

sequence was BLAST searched in GenBank for comparison with available sequences.

To calculate percent identity, similarity, and divergence, selected sequences were aligned

using Clustal W and corrected manually. All ambiguous insertions and deletions were

removed prior to further analyses. Percent Identities (PID) and DNA divergence were

calculated by DNAStar. Maximum Likelihood and Maximum Parsimony criteria were

used to describe the phylogenetic placement with other species included in the present

study. Phylograms were made using Mega5.

Taxonomy

Suillus flavidus (Fr.) J. Presl, WSobecny rostl. 2: 1917 (1846) PLATE 1

PiLEus 3-9 cm wide, convex to hemispherical to nearly plane, sometimes

slightly umbonate at maturity, sometimes margins straight and flaring to slightly

deflexed with whitish remnants of veil, surface viscid to glutinous when wet,

glabrous, yellow to yellowish brown. ConTExtT light yellow, browning when

bruised, not bluing. Stipe 3-10 cm long, 1.5-2 cm thick, nearly equal, cylindrical,

centric and curved, solid, slightly dry, reddish when young, yellow to white

with reddish tinge when mature, whitish glandular dots in some case, whitish

thick band like ring present above centre of stipe, color above ring yellow. PoRE

SURFACE yellow becomes slightly brown upon bruising, adnate and horizontal,

pores angular to irregular, infrequent, about 2 per mm, tubes 3-9 mm deep.

BASIDIOSPORES ellipsoid to fusoid, smooth, yellow, 9-13 x 4-6 um, (11.3 + 1.2

x 5.2 + 0.6; Q = 2.26 + 0.17). Basrp1a cylindric to long clavate, thick walled,

yellowish brown contents visible in Meltzer’s, 1-4 sterigmate, 22-26 x 8-10

um. Cystip1a cylindrical to fusoid-ventricose, brown contents visible, thick

walled, dark brown, 32-34 x 9-10 um. PILEIPELLIs long, cylindrical to slightly

clavate, thick walled, brown, 77-84 x 18-20 um, most terminal cells cylindrical

to clavate, in clusters and separate also, some are globose from above, dark

brown, thick walled, 71-77 x 8-10(-14) um. CHEMICAL REACTIONS pileipellis

reddish in KOH, spores brownish in Meltzer’s, light yellow to honey yellow in

lactic acid. SMELL & TASTE not distinctive. Eprsiuity edible.

MATERIAL EXAMINED: PAKISTAN: KHYBER PAKHTUNKHWA, Ayubia, 2250 m a.s.l.,

under Pinus wallichiana A.B. Jacks., solitary, on ground, 19 July 2010, S. Sarwar (LAH

S.B.#6A).

228 ... Sarwar & al.

Pirate 1. Suillus flavidus. A, B, Sporocarps; C, Basidia; D, Cystidia; E, Terminal cells of Pileipellis;

F, Basidiospores; G, Pileipellis hyphae. Scale bars: A, B = 2.5 cm; C = 8 um; D = 11 um; E = 25 um;

F =6 um; G= 16 um.

Suillus flavidus + Pinus wallichiana ectomycorrhizae (Pakistan) ... 229

PLATE 2. ECM of Suillus flavidus with Pinus wallichiana and its anatomical features. A, Habit;

B, Parenchymatous outer mantle; C, Parenchymatous inner mantle; D, Rhizomorph. Scale bars:

A=1cm;B=9 um; C= 8.5 um; D= 19 um.

Description of ectomycorrhizae of Suillus flavidus

Morphology PLATE 2A

MycorRHIZALSYSTEM found under the fruiting bodies, highly dichotomously

branched, system < 5 mm long, with < 0.8 mm thick main axis, unramified ends

straight, < 2 mm long, < 0.5 mm diam., color of system reddish brown to dull

yellow, older tips dark brown, apices light brown to honey brown, host tissue

visible under the sheath. RHIZOMORPHS frequent and attached at restricted

points, whitish brown.

Anatomy of mantle PLATE 2B,C

Mantle parenchymatous in all layers. OUTER MANTLE parenchymatous

(type L, Agerer 1987-2002); cells angular, 5 x 5 um, honey brown, no matrix

material, no septa and clamps observed. INNER MANTLE parenchymatous, (type

M, Agerer 1987-2002); cells slightly angular, 4 x 4 um, yellowish brown, no

matrix material, no septa and clamps.

Anatomy of emanating elements PLATE 2D

RuHIzOMoRPHS slightly differentiated (type A, Agerer 1991), cells 26 x 7

um, light brown, clamps common, septa less common and only at clamps,

H-shaped anastomoses without clamps present. EMANATING HYPHAE absent.

MATERIAL EXAMINED: PAKISTAN: KHYBER PAKHTUNKHWA, Ayubia, 2250 m a.s.l.,

Ectomycorrhizae Under Suillus flavidus near rhizospere of Pinus wallichiana, 19 July

2010, S. Sarwar (LAH S.B.#6B).

Molecular identification and phylogeny

When ITS-rDNA sequence was submitted for similarity with GenBank

data, it was identified as Suillus flavidus with 98% maximum identity and 100%

230 ... Sarwar & al.

45, Suillus_flaidus 108 (On Pinus watlichiana roots)

Suillus_flavdus_8G

Suillus fawdus_ 2C

|| Suillus_flavidus_3A_11G (On Pinus wallichiana roots)

Suillus_flavdus_2H_10H

Suillus_flavdus_2H (On Pinus walichiana roots)

| Suillus_flavdus_H10

Suillus_flawdus_3H

Suillus_flaidus_20

Suillus. flavdus_FJ845439.1

Suillus_fiavidus_AJ971403,1 (On Pinus sylvestris roots)

Suillus_lakei_DQ367917.1

100 Suillus_caenlescens_EU486453. 1

52 : : Suillus_grevllei_AF347102.4

Suillus_bovinus_AY898623.4

Suillus_variegatus_AMO86444. 7

100 Suillus_vanegatus_AM086446.1

Suillus_quiescens_GOQ249402.1

Suillus_mediterraneensis_AY935512.1

98 | Suillus_collinitus_AY953421.4

| Suillus_collinitus_DQ440569.1

Suillus_bellinii_AYS98621.1

Suillus_granulatus AY898817.1

Suillus_luteus_DQ440568.4

35 Suillus_brevipes. FJ645440.1

Figure 1: Molecular phylogenetic analysis by Maximum Likelihood method. The evolutionary

history was inferred by using the Maximum Likelihood method based on the Poisson correction

model (Zuckerkand] & Pauling 1965). The percentage of trees in which the associated taxa clustered

together is shown next to the branches. Initial tree(s) for the heuristic search were obtained

automatically as follows: when the number of common sites was <100 or less than one fourth of

the total number of sites, the maximum parsimony method was used; otherwise BION] method

with MCL distance matrix was used. The tree is drawn to scale, with branch lengths measured in

the number of substitutions per site. The analysis involved 25 sequences. There were a total of 426

positions in the final dataset. Evolutionary analyses were conducted in MEGA5 (Tamura et al.

2011).

query coverage with S. flavidus (FJ845439.1) from Canada. The phylogenetic

analysis includes 25 sequences from 13 species (TABLE 1). For phylogenetic

analysis, 426 genetic characters were used in aligned datasheet containing

330 conserved sites, 91 variable sites, and 66 parsimony sites. Phylogenetic

analysis also supports the morpho-anatomic and molecular identification.

The phylogram based on maximum likelihood criterion is represented by two

major clades (Fic. 1). Clade I comprises 11 sequences from S. flavidus, S. lakei

(Murrill) A.H. Sm. & Thiers (DQ367912.1), S. caerulescens A.H. Sm. & Thiers

Suillus flavidus + Pinus wallichiana ectomycorrhizae (Pakistan) ... 231

(EU486453.1), and S. grevillei (Klotzsch) Singer (AF347102.1); their clustering

is not highly resolved (bootstrap = 42%). Suillus flavidus, which occupies the

top position in the phylogram, is represented by nine Pakistan sequences (six

from fruitbodies and three from P. wallichiana ectomycorrhizal roots) and two

GenBank sequences. The Pakistan rDNA-ITS sequences shared 100% genetic

similarity with one another and about 98% with S. flavidus FJ845439.1. The

two neighboring S. flavidus sister clades in clade 1 have a different topological

placement due to intraspecific variations within the region sequenced. ‘This

species has a 92.1% genetic similarity with S. lakei DQ367912.1 and 91.6%

with S. caerulescens EU486453.1. Genetic divergence was also measured for

S. flavidus with all the sequences included in the analysis. No genetic divergence

was found among the rDNA-ITS of S. flavidus from Pakistan (Fic. 2), and there

was little genetic divergence (0.5-2.5) compared with S. flavidus FJ845439.1.

Percent Identity

Suillus_flavidus_2C

Suillus_flavidus_2D

Suillus_flavidus_2H

Suillus_flavidus_3H

Suillus_flavidus_H10

Suillus_flavidus_10B.

Suilllus_flavidus_2H_10H

Suillus_flavidus_3A_11G

Suillus_flavidus_8G

Suillus_flavidus_FJ8454239

Suillus_flavidus_AJ971403

Suillus_quiescens_GQ249402

Suillus_collinitus_AY953421

Suillus_collinitus_DO440569

Sulllus_mediterranesnsis_A¥935512

Suillus_grevillei_AF347102

Suillus_luteus_DQ440568

Suillus_granulatus_AY898617

Suillus_lakei_DQ367917

Sulllus_brevipes_FJ845440

Suillus_caerulescens_EU486453

Suillus_variegatus_AM086444

Suillus_variegatus_AM086446

Suillus_bovinus_A¥898623

Sulllus_bellinii_AY898621

Divergence

FIGURE 2: Percent divergence is calculated by comparing sequence pairs in relation to the phylogeny

reconstructed by MegAlign (DNASTAR). Percent similarity compares sequences directly without

accounting for phylogenetic relationships.

Discussion

We report S. flavidus and its ectomycorrhizae with P wallichiana for the first

time from Pakistan. This fungus is characterized by a convex to hemispheric

yellowish pileus with reddish brown spots on the margin and typically small

hanging veil remnants and a stipe with a prominent ring. Spores are smooth

and range from light to dark brown. Suillus flavidus resembles S. lakei, which

differs in a pileus surface covered with dull reddish brown scruffies and a stipe

with reddish streaks and without glandular dots. The similar S. caerulescens also

lacks glandular dots but has occasional appressed hairs on the stipe. Finally,

S. grevillei, which is also resembles S. flavidus, has a somewhat reticulated stipe

that lacks the glandular dots that characterize S. flavidus (Bessette et al. 2000).

232 ... Sarwar & al.

We characterize S. flavidus-P. wallichiana ectomycorrhizae here for the

first time by noting their highly dichotomously branched ramifications with

parenchymatous inner and outer mantle surfaces, slightly differentiated

rhizomorphs, and no emanating hyphae. Different Pakistani ectomycorrhizae

associated with Cedrus deodara have recently been molecularly analyzed (Hanif

et al. 2010) but there is no literature on molecular description of ectomycorrhizae

of boletes from Pakistan. We report S. flavidus as new record for Pakistan and

believe that P. wallichiana is documented here as a new ectomycorrhizal host

for S. flavidus for the first time.

Acknowledgements

The authors would like to thank Higher Education Commission Pakistan for

providing funds to complete this project. We are also thankful to Dr. Jean Michel Savoie

(Institut National de la Recherche Agronomique (INRA), France) and Dr. Najm-u-sehar

Afshan (University of the Punjab, Lahore, Pakistan) for reviewing this manuscript and

providing helpful suggestions.

Literature cited

Agerer R. 1987-2002. Color atlas of ectomycorrhizae. 1-12" delivery, Einhorn Verlag Edward,

Dientenurger. Germany.

Agerer R. 1991. Characterization of ectomycorrhiza. 25-73, in: JR Norris et al. (eds). Techniques

for the study of mycorrhiza. Methods in Microbiology, vol. 23. Academic Press London.

Agerer R. 1999. Never change a functionally successful principle: the evolution of Boletales s.1.

(Hymenomycetes, Basidiomycota) as seen from below ground features. Sendtnera 6: 5-91.

Ahmad §S, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Sultan Ahmad Mycological Society of

Pakistan, Department of Botany, University of Punjab, Quaid-e-Azam Campus, Lahore,

Pakistan. 121 p.

Bessette AE, Roody WC, Bessette AR. 2000. North American boletes: a color guide to the fleshy

pored mushrooms. Syracuse University Press: Syracuse NY.

Hanif M, Khalid AN, Sarwar S. 2012. Additions to the ectomycorrhizae associated with Himalayan

cedar (Cedrus deodara) using rDNA-ITS. International Journal of Agriculture and Biology

14(1): 101-106.

Hussain SS. 1995. Pakistan manual of plant ecology. Mirror Press Ltd. Karachi.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th ed. Wallingford:

CABI.

Kuo M. 2004. The genus Suillus. Retrieved from the MushroomExpert.Com Web site:

http://www.mushroomexpert.com/suillus.html.

Marx DH. 1991. Forest application of ectomycorrhizal fungus Pisolithus tinctorius. The Marcus

Wallenberg Prize, Stockholm. 36 p.

Niazi AR. 2008. Biodiversity of ectomycorrhizas in conifers from Himalayan moist temperate forests

of Pakistan. Ph.D. Thesis, Department of Botany, University of Punjab, Lahore. Pakistan.

Razaq A. 2007. Taxonomic studies on Basidiomycota from northern areas of Pakistan. Ph.D. thesis,

University of Karachi, Karachi.

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. Molecular Biology and Evolution 28(10): 2731-2739.

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

Zuckerkandl E, Pauling L. 1965. Evolutionary divergence and convergence in proteins. 97-166, in:

V Bryson, HJ Vogel (eds). Evolving Genes and Proteins. Academic Press, New York.

MY COTAXON

http://dx.doi.org/10.5248/121.233

Volume 121, pp. 233-253 July-September 2012

Russulaceae of the Pakaraima Mountains of Guyana 2.

New species of Russula and Lactifluus

STEVEN L. MILLER’, M. CATHERINE AIME?, & TERRY W. HENKEL?

‘Department of Botany, University of Wyoming, Laramie, Wyoming 82071

*Department of Plant Pathology & Crop Physiology, Louisiana State University Agricultural Center,

Baton Rouge, Louisiana 70803

*Department of Biological Sciences, Humboldt State University, Arcata, California 95521

*CORRESPONDENCE TO: fungi@uwyo.edu

AxBstRAcT— Morphological and ecological descriptions, illustrations, and taxonomic

discussions are presented for a subiculate species of Lactifluus, L. subiculatus, and three species

of Russula, R. myrmecobroma, R. paxilliformis, and R. gelatinivelata, all newly described from

the Pakaraima Mountains of Guyana. Sequence data confirm relationships of taxa collected

in Guyana within a large Russulaceae database.

Key Worps— basidiomycetes, Basidiomycota, biodiversity, ectomycorrhiza, macrofungi,

neotropics, Russulales

Introduction

A recent paper (Henkel et al. 2011) documented for the first time the

sporocarp diversity of putative or confirmed ectomycorrhizal fungi in lowland

tropical forests of Guyana. In that study, conducted over a ten year period

from 2000 to 2010, sporocarps from 172 species of macrofungi representing

13 families and 25 genera of primarily Agaricomycetes, but also Ascomycota

(Elaphomycetaceae), were collected from plot studies and opportunistic

exploration of the Dicymbe (Fabaceae subfam. Caesalpinioideae) dominated

forests of the Upper Potaro River Basin. Of these families, the Russulaceae

contained the most species, many new to science.

The present communication provides macro-, micromorphological, and

ecological descriptions and illustrations for four new taxa in the Russulaceae

that contributed significantly to the biodiversity in the aforementioned plot

studies (Henkel et al. 2011). Taxonomic discussions are also provided, guided

by sequence analyses that will be detailed elsewhere.

234 ... Miller, Aime & Henkel

Materials & methods

Collecting expeditions were made in the rainy seasons (typically May-July)

2000-2011 to the Upper Potaro River Basin along Guyana’s western border with Brazil,

in the south-central Pakaraima Mountains (general area: 5°05'N 59°58’W). Basidiomata

were examined in the field for fresh characteristics. Color characteristics were coded

according to Kornerup & Wanscher (1981; code noted in parentheses) and described

subjectively. Spore deposits were taken on acetate sheets and examined for fresh color

characteristics. The FeSO, macrochemical test was performed using a large mineral

crystal that was rubbed directly on the stipe, lamellae and flesh. Basidiomata were dried

slowly using large bead silica gel and subsequently placed in re-sealable plastic bags

with small bead silica gel to prevent spoilage in the excessively humid conditions. In

addition, basidiomata in various stages of development were immersed in ethanol for

preservation and for molecular analysis. DNA extraction, sequencing and molecular

analysis were generally those described in Miller et al. (2006).

Microscopic anatomical details were determined from dried basidiomata with an

Olympus BH-2 microscope with bright-field optics and drawings were made with a

camera lucida attachment. For basidiospores at least 20 individuals were observed and

measured per taxon. The acid resistant reaction in basic fuchsin, the cresyl blue reaction,

and the sulfovanillin reaction were prepared according to Singer (1986).

Herbaria designations are after Holmgren et al. (1990) and include: BRG -

University of Guyana, Georgetown; HSU - Humboldt State University; RMS — Solheim

Mycological Herbarium; LSUM - Louisiana State University Bernard Lowy Mycological

Herbarium.

Taxonomy

Lactifluus subiculatus S.L. Mill., Aime & T.W. Henkel, sp. nov. PLaTEs 1-2

MycoBAnk MB 564265

Sed Lactifluus subiculatus atrohervolo pileo maturo et crustoso, versus gracili tomentoso

stipite differt.

Type: Guyana, Pakaraima Mountains, Upper Potaro River Basin, Dicymbe altsonii stand,

22.V.2000, SL Miller 10047 (Holotype, RMS; isotype, BRG; GenBank JQ405654).

EryMo.Loey: subiculatus (Lat.), referring to the dense subiculum from which the

basidiomata arise.

PitEus 0.9-3 cm broad, at first convex, then plane to slightly depressed to

irregularly infundibuliform; margin incurved and regular when young, then

decurved, entire to undulating, obscurely striate at first, then irregularly

subsulcate when older; extreme margin distinctly membranous for up to 1 mm;

pellis finely velutinous to hispidulose or subtomentose when young, then

rugulose and minutely areolate especially at the disk, tacky to slightly lubricous

when moist, light brown (6D6-—8) overall impression, light brown (6D6-—8) to

yellowish brown to golden brown (5D6—8) when young, mottled and streaked

with pale to light orange (5A3-—4) and less frequently with brown (6E-7) with

age, with extreme margin pale orange white. LAMELLAE 2—3.5 mm wide at

Russula & Lactifluus spp. nov. (Guyana) ... 235

PrateE 1. Lactifluus subiculatus habit (Miller 10047). 1. Basidiomata produced on shaggy subiculum

enshrouding base of sapling, x1.8. 2. Stipe showing disrupted crust of pigment and minutely fluted

topography, along with lamella morphology, x2.1. 3-4. Extensive subiculum on boles of large

Dicymbe corymbosa trees. 3. x0.15. 4. Arrow indicates L. subiculatus basidiomata and cluster of

primordia, x0.2.

236 ... Miller, Aime & Henkel

PiaTE 2. Lactifluus subiculatus (Miller 10047), microscopic features. 5. Diagram of a section

of the two-layered pileipellis; the suprapellis is a lampropalisade of thick-walled hairs over a

pseudoparenchymatous subpellis. 6. Elements of the suprapellis. 7. Gloeopleurous hyphae.

8. Basidia. 9. Basidiospores. Scale bars = 10 um.

Russula & Lactifluus spp. nov. (Guyana) ... 237

mid-radius, irregularly adnate to subdecurrent, unequal, close when young

then subdistant, not or occasionally intervenose and forking, thin at first then

moderately thick, rounded, white to pale yellow (4A3—4 to 5A2), unchanging;

edge entire, concolorous; lamellulae of three different lengths. StipE 4-8 x

8—15 mm, cylindrical, equal or tapered to the base, curved, attachment central,

occasionally distinctly fluted, yellowish white to pale creamy tan (4A2 to

5B4—5) above, concolorous with pileus below, subvelutinous toward the base

at first, then this tissue cracking into distinct isolated patches; arising from a

thin, coarse, erect tomentose subiculum. CONTEXT 1-2 mm at mid-radius,

firm, solid in lamellae and stipe, white, unchanging; odor strong fungal; taste

disagreeable. LaTex absent or scant, watery, unchanging. FeSO, quickly dull

green on lamellae and trama. SUBICULUM usually distinct and extensive, shaggy,

off-white to greyish cream, composed of fascicles of hyaline, linear hyphae or

hairs 2-8 um wide, thick-walled (< 2—2.5 um).

BASIDIOSPORES 7.6—9.2(-9.6) x 6.4—7.6 um (Q= 1.18—1.21-1.26), subglobose

to short ellipsoid; ornamentation amyloid, composed of irregularly sized and

shaped warts; warts 0.5—-1.0 um high, usually obtuse, isolated or connected by

fine lines forming a fine partial reticulum; hilar appendix prominent, 3-4 x

1.5—2 um, hyaline; suprahilar plage non amyloid or with faint fine reticulum,

rarely with an isolated amyloid spot. Basip1A clavate, 45-60 x 15-18 um,

4—spored, sterigmata 7-9 x 1-2 um. PsEUDOCysTIDIA 5-10 um wide,

cylindrical or irregularly contorted, rounded or blunt-pointed at the apex, not

emergent, infrequent to abundant, thin-walled; contents refractive granular,

SV-; connected deeply to laticiferous hyphae in the trama with similar contents.

PLEUROCYSTIDIA absent. HYMENOPHORAL TRAMA irregular, composed of

interwoven, hyaline, thin-walled hyphae, which are 3—5 um diam with scattered

inflated cells of 8-10 um wide; lactiferous hyphae scattered, 8-11 um wide with

refractive guttulate contents; sphaerocytes rare or absent; subhymenial layer

not well developed. PILEIPELLIS a sparse to dense lampropalisade; elements of

suprapellis 35—150(-220) x 5-10 um, cylindrical, hair-shaped, septate, with wall

1-2 um thick, frequently with swollen base; subpellis pseudoparenchymatous;

cells isodiametric, obovate, napiform to obpyriform, 10-40 um diam, with

yellowish brown contents in KOH, cylindrical to slightly contorted; all cells

SV-. STIPITIPELLIS similar to pileipellis but hair-like elements less numerous,

surface tissue often nearly occluded by external yellowish brown amorphous

pigment in KOH; SV-.

HABIT, HABITAT, AND DISTRIBUTION — Solitary to fruiting in small numbers

on lower trunks of saplings, larger trees, stumps and other elevated positions,

arising from a shaggy, often deep and extensive, persistent subiculum which

enshrouds living and dead objects above the surface of the ground and which

is often interspersed with or overriding bryophyte growth, subtended by

238 ... Miller, Aime & Henkel

ectomycorrhizal rootlets; found May through early July during wet weather

in forests dominated by Dicymbe corymbosa and D. altsonii. Ectomycorrhizal

associations with the leguminous hosts in Guyana were confirmed with the aid

of molecular analysis (Smith et al. 2011)). Known only from the type locality in

the Upper Potaro Basin of Guyana, and Mabura.

ADDITIONAL SPECIMENS EXAMINED. GUYANA. REGION 8 POTARO-—SIPIRUNI.

Pakaraima Mountains, Upper Potaro River Basin, within 5 km radius of Potaro base camp

located at 5°18'04.8"N 59°54'40.4"W, 710-750 m, 17.V.2000, Miller 10010 (BRG, RMS);

Miller 10021 (BRG, RMS); 28.VII.2000 Henkel 7922 (BRG; HSU). Near Ayanganna

Airstrip, 16.V.2000, Miller 10004 (BRG, RMS); 20.V.2000 Miller 10034 (BRG, RMS).

Dicymbe altsonii stand, 22.V.2000 Miller 10047 (BRG, RMS). Dicymbe Research Plots,

27.V.2000, Miller 10061 (BRG, RMS); 1.V1.2000, Miller 10087 (BRG, RMS); 3.V1.2000,

Miller 10094 (BRG, RMS); 8.VI.2000 Miller 10114 (BRG, RMS). Vicinity of base camp,

28.VII.2000, Henkel 7922 (BRG, HSU); 15.V.2010 Aime 3937 (BRG, LSUM). ~3 km SE

of Potaro base camp near Dicymbe plot 1, 25.V.2001, Henkel 8210 (BRG, HSU). 1 km

SW of base camp near Blackwater point, 10.VII.2010, Henkel 9020 (BRG, HSU). Near

Dicymbe Plot 2, 26.V.2010, Henkel 9229 (BRG, HSU). REGION 10—UPPER DEMERARA-

BERBICE. Mabura Ecological Reserve, Dicymbe altsonii stand, 24.V.2011 Aime 4276

(BRG, LSUM).

COMMENTARY — Previously, three neotropical species of Lactarius that produce

basidiomata from an extensive and well developed subiculum have been

described (Singer 1984; Miller et al. 2002; Miller & Henkel 2004): L. panuoides

Singer, L. brunellus S.L. Mill. et al., and L. multiceps S.L. Mill. et al.. Lactarius

panuoides and L. brunellus are pleurotoid, while L. multiceps has a short, but

occasionally well-developed, eccentric stipe (Miller et al. 2002). The stipe of

the milkcap described here is larger and centrally attached, and the basidioma

stature is typically more agaricoid than pleurotoid. Molecular data confirm that

our species forms ectomycorrhizae with the leguminous hosts D. corymbosa,

D. altsonii, and Aldina insignis in Guyana (Smith et al. 2011).

The infrageneric placement of L. subiculatus is unclear. It is clear that the

species belongs to Lactifluus, which was recently split from Lactarius (Buyck

et al. 2008, 2010; Verbeken et al. 2012). The rDNA molecular phylogenetic

analysis (SL Miller, unpublished) indicates a close relationship with Lactifluus

neotropicus (Singer) Nuytinck, originally described from Trinidad, but collected

also from Guyana (SL Miller, unpublished). The combination of long thick-

walled hairs in the suprapellis arising from an epithelioid subpellis (which both

species share) is characteristic of several Lactifluus groups in both the old- and

new-world tropics, including L. sections Lactariopsis Verbeken, Chamaeleontini

(Verbeken) Verbeken, Pseudogymnocarpi (Verbeken) Verbeken, Phlebonemi (R.

Heim ex Verbeken) Verbeken, and Polysphaerophori (Singer) Verbeken (Pegler

& Fiard 1979, Singer et al. 1983, Verbeken & Walleyn 2010). Singer (Singer

et al. 1983) and Pegler & Fiard (1979) differentiated sections Lactariopsis and

Polysphaerophori by the presence of a partial veil or, more precisely, an annulus

Russula & Lactifluus spp. nov. (Guyana) ... 239

or remnants of a veil on the stipe in sect. Lactariopsis. Similarly, Verbeken &

Walleyn (2010) distinguished sect. Lactariopsis from sect. Chamaeleontini

based on the presence of an annulus in the former. Sect. Polysphaerophori

Singer, erected for South American species (Singer 1973), was used to place the

African “Gymnocarpi” by Verbeken & Walleyn (2010), who rightly recognized

the artificial distinction between several of these groups and maintained them

only for practical reasons.

Macroscopically, the brownish orange colours and pilose to velutinate pileus

surface, and microscopically, the abundant sphaerocytes in the hymenophoral

trama, the thick-walled elements in the pileipellis, and spores with low

ornamentation forming a partial to nearly complete reticulum in L. subiculatus

are consistent with L. sect. Lactariopsis. Although L. subiculatus has not been

observed to form a distinct annulus, the extreme margin does have a distinct

flap of tissue that may be easily interpreted as the remnants of a partial veil.

The other subiculate species, L. multiceps, L. brunellus and L. panuoides,

have not yet been recombined in Lactifluus. They all combine the subiculate

character with a pleurotoid habit and, up to now, all pleurotoid milkcaps that

have been molecularly investigated belong to Lactifluus.

Russula myrmecobroma S.L. Mill., Aime & T.W. Henkel, sp. nov. PLATES 3-4

MycoBank MB 564264

Ab aliis speciebus Russulae stipite cinereo, pileo fusco, deinde irregulariter diaphano ubi

udo et fascianti in centro propriam ordinationem reticulatam, ordinate dispositis striis pro

longitudine aequalibus, lamellis aggregatis et sapore acris differt.

Type: Guyana, Pakaraima Mountains, Upper Potaro River Basin, near Hogback Ridge,

5.V1.2000, SL Miller 10109 (Holotype, RMS; isotype, BRG; GenBank JQ405657).

ETyMOLoGy: myrmecobroma (Gk.), literally ant food, referring to the regular

dismemberment of the basidiomata by ants.

Piteus 3.5—4.5 cm broad, broadly convex at first, then plane and finally

depressed; margin entire when young, then undulating, obscurely striate at

first, then with regularly spaced and sized striations; striations 2 mm long; pellis

dull, dry, smooth at disk, frequently pruinose at mid-radius and at margin,

yellowish brown (5E—F-—8 to 5D-E 5) overall at first, becoming hygrophanous

with irregular reticulate pattern especially halfway the radius and with color

fading to dark blond (5C-—4, 5C-—D-3) to orange grey (5B—2) with irregular

areas of 5B—3—5; disk and margin not hygrophanous and usually remaining

darker. LAMELLAE 1-2 mm broad at mid-radius, adnate, sinuate or slightly

subdecurrent, crowded, forked at stipe and occasionally elsewhere, arising at

different positions relative to the stipe giving uneven appearance, pallid cream

(near 4A—2), becoming spotted with yellow or reddish brown where injured;

lamellulae present, 4—5 ranked; edge entire. StrpE 3—-5.7 x 0.8—-1.1 cm, equal

to subclavate or tapering to the base, centrally attached, moderately distinct

240 ... Miller, Aime & Henkel

! 11.

PLATE 3. Russula myrmecobroma habit. 10. Bisected basidiome (Miller 10109), x1.1.

11. Basidiome (Miller 10161) exhibiting damage by ants (arrow), x1.2.

longitudinal ridges beneath felted subtomentose covering especially below,

tomentum white to pale grey (SA-—B-1), pale orange grey (5B 2-3) beneath,

grey brown where handled and where longitudinal ridges become exposed.

CONTEXT in pileus 2—2.5 mm at mid-radius, stuffed pallid grey cream; context

in stipe cylinder whitish marbled with pale grey, turgid to pliant, pallid cream

to pale grey brown, discoloring pale orange or orange brown where injured;

odor sharp acrid heading toward latex paint; taste strongly acrid; FeSO, quickly

salmon on stipe surface and trama.

BASIDIOSPORES 6.4—7.2(-8) x 5.6—6.8 um (Q = 1.05—1.14-1.17), subglobose

to broadly ellipsoidal; ornamentation reticulate, consisting of low blunt spines,

1-1.3 um high, connected by fine lines or verrucae, strongly but often partially

amyloid; suprahilar plage not distinct to finely reticulate. Basip1A 55-62 x 12-15

um, subclavate to nearly cylindrical, 4—spored; sterigmata stout 7-10 x 2-3 um.

Cystip1aA 80-95 x 15-20 um, subclavate to subfusiform, thin or thick walled,

emergent for ca. 20-30 um, numerous, arising from gloeopleurous elements,

with refringent to crystalline contents, SV+, thick walled lamprocystidia present

but not numerous. MARGINAL CELLS 55-105 x 8-15 um, narrowly subclavate,

tortuous, thin-walled, optically empty, abundant. SUBHYMENIUM distinct, a

gelatinous layer composed of interwoven cylindrical flattened and variously

swollen hyphae of 2—5 um diam. LAMELLAR TRAMA with large sphaerocytes

and scattered hyphae, with gloeopleurous fragments and embedded

dermatocystidia near the pileus. PILEIPELLIs orthochromatic in Cresyl Blue,

two-layered; subpellis gelatinized, forming a dense mat close to the underlying

trama, of tightly interwoven hyphae; hyphae 2-5 um diam, thin-walled,

frequently septate, with scattered strongly refringent gloeopleurous elements

of 5 um diam, frequently terminating with cylindrical to swollen or mucronate

Russula & Lactifluus spp. nov. (Guyana) ... 241

PLaTE 4. Russula myrmecobroma (Miller 10109), microscopic features. 12. Diagram of a

section of the two-layered pileipellis; the suprapellis is a trichodermial palisade over a thin

pseudoparenchymatous subpellis. 13. Elements of the suprapellis. 14. Elements of the stipitipellis.

15. Hymenial cystidia. 16. Marginal cells. 17. Basidia. 18. Basidiospores. Scale bars = 10 um.

242 ... Miller, Aime & Henkel

embedded dermatocystidia; suprapellis composed of 2—5 strongly inflated,

spherical cells, often gradually smaller towards the terminal cell, the latter

cylindrical to narrowly subclavate, ampullaceous, or mucronate, resembling an

epithelium; pileocystidia dispersed, terminal, more or less the same diam. as

other terminal elements, 50-90 x 10-13 um, contents granular-refringent in

KOH. StipiTIPELtis a turf of variously shaped caulocystidia, 100-170 x 9-14

um, these narrowly cylindrical, mucronate, capitate or tortuous.

HABIT, HABITAT, AND DISTRIBUTION — Solitary or fruiting in small numbers

on root mats of the ECM trees Dicymbe corymbosa, D. altsonii, D. jenmanii,

Aldina insignis and Pakaraimaea dipterocarpacea, widespread in the Pakaraima

Mountains but not common; found May through early July during wet weather

and also in December-January. Known from Mabura Hill and the Upper Potaro

and Upper Mazaruni River Basins of Guyana.

ADDITIONAL SPECIMENS EXAMINED: GUYANA. REGION 8 POTARO-—SIPIRUNI.

Pakaraima Mountains, Upper Potaro River Basin, within 15 km radius of Potaro base

camp located at 5°18'04.8"N 59°54'40.4"W, 710-750 m. Vicinity of Potaro Base Camp,

21.V.2000, Miller 10045 (BRG, RMS); 24.V1.2000, Miller 10161 (BRG, RMS). ~3 km

SW of base camp near Dicymbe plot 3, 8.VI.2001 Henkel 8258 (BRG; HSU). 1 km SW

of base camp near Blackwater point, 10.VII.2009, Henkel 9016 (BRG; HSU). 15 km east

of Potaro base camp near Tadang base camp, in mixed Dicymbe corymbosa—Dicymbe

altsonii—Aldina insignis forest, 24.X1I.2009, Henkel 9145 (BRG; HSU). Dicymbe plot 1,

5.VI.2001, Aime 1783 (BRG, LSUM); 15.V.2010, Aime 3935 (BRG, LSUM). Dicymbe

plot 2, 6.VI.2001, Aime 1823 (BRG, LSUM). REGION 7 CuYUNI-MAZARUNI. Pakaraima

Mountains, within 1 km radius of Piegaimah base camp at 5°26'21.3"N 60°04'43.1" W.

Vicinity of base camp, in savanna fringing forest under Pakaraimaea dipterocarpacea

and Dicymbe jenmanii, 24.X11.2010, Henkel 9523 (BRG; HSU). ~1 km SW of base camp,

in stand dominated by Pakaraimaea dipterocarpacea and lacking Dicymbe jenmanii,

26.XII.2010, Henkel 9546 (BRG; HSU). REGION 10—UPpPER DEMERARA-BERBICE.

Mabura Ecological Reserve, Dicymbe altsonii stand 1, 1.VI.2011, Aime 4360 (BRG,

LSUM).

COMMENTARY ~ It is relatively difficult to find fresh basidiomata of R. myrmeco-

broma that have not been extensively damaged by ants. While ants will

disarticulate and carry off pieces of a variety of fungal fruiting structures,

they seem to be especially attracted to R. myrmecobroma, and in Guyana only

R. batistae Singer seems to be more attractive. Microscopically both fungi appear

to have relatively primitive traits such as an abundant gloeoplerous system with

embedded dermatocystidia in the pileipellis and abundant macrocystidia of

two types deeply rooted in the hymenium, all strongly positive in sulfovanillin.

Both also have an acrid to strongly acrid taste. It is unknown whether any of

these characteristics serve as an attractant to the ants.

From an ecological standpoint, R. myrmecobroma may be a quite important

mycorrhizal symbiont in Guyana’ Dicymbe forests. In a recent study by Smith

et al. (2011) in the Upper Potaro Basin, R. myrmecobroma was the most

frequently ECM fungus out of 118 taxa recovered by molecular methods on

Russula & Lactifluus spp. nov. (Guyana) ... 243

ECM roots of 19 trees each of three sympatric host species, D. corymbosa,

D. altsonii, and the papilionoid Aldina insignis. In a second study (ME Smith

& TW Henkel, unpublished) investigating mycorrhizal symbionts of the ECM

dipterocarp, Pakaraimaea dipterocarpacea, and the sympatric Dicymbe jenmanii

in the Upper Mazaruni Basin, R. myrmecobroma was the sixth most common

mycobiont recovered among about 50 species of EM fungi associated with 20

trees of each host species, indicating a broad host and distribution range for

the fungus in Guyana. It is interesting to speculate that movement of tissue

pieces by the ants might contribute to the frequency of R. myrmecobroma in

the forest.

Molecular analysis (SL Miller, unpublished) indicates a close relationship

between R. myrmecobroma and R. batistae, despite the fact that macroscopically

they are very different. Russula batistae has irregular, broad, subdistant to

distant lamellae, no lamellulae, and a deeply sulcate pileus margin, while

R. myrmecobroma has regular, rather narrow, crowded lamellae, abundant

lamellulae, and a regularly short-striate margin. Singer erected R. subsect.

Batistinae in the large section Pelliculariae to accommodate R. batistae. ‘This

subsection likely will not suffice as no velar tissue has been observed in

R. myrmecobroma, suggesting a need to refine the description of the subsection.

Buyck (1990a) has shown that R. sect. Pelliculariae is highly heterogeneous

and possibly not monophyletic, indicating the possible need for a new

infrageneric group at the sectional level to accommodate R. batistae and

R. myrmecobroma.

Russula paxilliformis S.L. Mill., Aime & T.W. Henkel, sp. nov. PLATES 5-6

MycoBank MB 564266

Pagina pilosa rupenti in fine squamellas tenues prope marginem, exponenti texturam

pallidam et stipitem velutinum basin versus decrescentem et sine profunde striato

margine.

Type: Guyana, Pakaraima Mountains, Upper Potaro River Basin, vicinity of Potaro

Base Camp, 3.VI.2000, SL Miller 10097 (Holotype, RMS; isotype, BRG; GenBank

JQ405656).

Erymo oey: paxilliformis (Lat.), referring to the pilose pileus with inrolled margin and

coloration reminiscent of Paxillus involutus.

PiLEus 3-8.5 cm broad, convex at first with deeply depressed disk, becoming

plane or uplifted, often with a hole entirely through the disk of the pileus where

ants have eaten away the tissue; margin tightly inrolled around stipe at first

with a frail tomentose partial veil connecting margin to stipe, in age margin

becoming separated, then decurved, and finally plane leaving no annulus

or other trace of a veil, entire, not striate when young, becoming obscurely

striate with age; pellis dull, dry, densely subtomentose to velutinous when

young, concentrically cracking to become areolate, granulose to squamulose,

244 ... Miller, Aime & Henkel

Pirate 5. Russula paxilliformis habit. 19. Top surface of unexpanded pileus showing radial cracking

of pilose pellis (Miller 10097), x1.8. 20. Top surface of expanded pileus showing radial cracking

at the disk and areolate or granulose texture near the margin (Miller 10097), x0.8. 21. Side view

of unexpanded pileus showing tightly inrolled and pale colored margin, and subtomentose and

ornamented stipe (Miller 10097), x1.8. 22. Mature basidiome with partially expanded pileus

showing velar remnants at the margin, and subtomentose stipe (Miller 10070), x0.9.

remaining so especially at the disc; pileus and partial veil light brown to golden

brown (5C-—D 6-7) overall, disk yellowish brown (5D6-—8) when young, with

an orange white subcutis (near 5A—2) showing through, with age areoles (?)

darkening to brown (6E 7-8) with orange white 5A—2 or light brown (near

6D4—5) showing through, disk brown (6—7 E 6—7). LAMELLAE 3—4 mm broad

at mid-radius, adnate to slightly decurrent, close, rarely forking at stipe and

Russula & Lactifluus spp. nov. (Guyana) ... 245

elsewhere, entire, firm when young, soft with age, whitish to cream, discoloring

pale reddish brown where handled; lamellulae absent. Stipe 5.5—8 x 0.9-1.0

cm, cylindrical to tapering to the base when young, strongly tapering to the

base and often flared at the apex in age, usually with a pronounced depression

around stipe apex where tightly inrolled pileus and partial veil were connected,

commonly curving, attachment central, even, highly tomentose to scurfy, with

more or less concentric areolate stretch marks when older, matted to flocculose

where handled; light brown to golden brown (5 C-—5-7 to 5D—5-8), at first

areolae yellow brown (5E-8, with age 5E-6—7), over light brown (5C-—5-6)

ground beneath, then yellowish white to pale creamy tan (4A2 to 5B4—5) above,

light brown to brown (5—6D —6-—7) or dark brown (6F5-6) below. CONTEXT in

pileus 2.5-3 mm at mid-radius, firm at first, then soft, pallid cream; trama in

stipe with distinctive anatomy in longitudinal section, with a discrete separable

cylinder-like plug of hard tissue forming a central core from pileus in upper

two thirds of the stipe, surrounded by firm tissue comprising most of the stipe,

outer cylinder marbled bluish steel gray from base over lower one-half, in

older specimens the central core has frequently been completely eaten away

by ants so that the stipe is hollow; odor strongly fungal, like Scleroderma; taste

disagreeable at first, then slowly acrid; FeSO, quickly salmon on stipe trama.

BASIDIOSPORES (6.8—)7.2—7.46—8 x 6—6.8(-7.2) um (Q = 1.06-1.13-1.27),

subglobose to broadly ellipsoidal; ornamentation reticulate or incompletely

reticulate; ornamentation consisting of partial crests with short lateral

diverticulations and large broadly conical or multiplex blunt spines, 1.6—2.8

um high, connected by fine lines or verrucae, strongly but often partially

amyloid; suprahilar plage moderately large, verruculose, barely decurrent

on apiculus. Bastp1a 50-65 x 12-15 um, subclavate to subcylindrical,

4-spored; sterigmata 8-10 x 1-3 um. CystTip1a 70-95 x 6-13 um, cylindrical

to subfusiform, obtuse, deeply rooting, mostly thin-walled, emergent 30-35

um above basidia, SV+, contents crystalline, numerous. MARGINAL CELLS

65-105 x 10-18 um, cylindrical to subfusiform, SV+, numerous. TRAMA of

lamellae and pileus composed of large well-developed nests of sphaerocytes;

subhymenium not well developed. PILEIPELLIs orthochromatic in Cresyl

Blue, two-layered; subpellis of loosely interwoven hyphae of 4-5 um diam.

with scattered gloeoplerous elements of 5 um diam., pale brownish grey in SV;

suprapellis a dense trichoderm of digitate, septate hyphae; terminal elements

cylindrical, obtuse, often sinuous, 20-60 x 2.5-5 um, occasionally thick-

walled, arising from long chains of spherical to irregularly shaped cells, evenly

brown in KOH, interior walls irregularly thickened and jagged; pileocystidia

25-60 x 3-6 um, narrow cylindrical, obtuse to subcapitate, rare or absent;

all elements SV-, encrustations highly acid resistant in basic fuchsin, dark

purple, abundant. STIPITIPELLIs a dense trichoderm similar to the suprapellis,

246 ... Miller, Aime & Henkel

composed of long chains of cylindrical, spheroidal to irregularly shaped cells,

terminated by brownish ellipsoid or pyriform cells, internal walls irregularly

thickened; cystidia rare or absent.

HABIT, HABITAT, AND DISTRIBUTION — Solitary to widely scattered on root

mats of Dicymbe corymbosa, uncommon; found May through early July during

wet weather. Known only from the type locality in the Upper Potaro Basin of

Guyana.

ADDITIONAL SPECIMENS EXAMINED: GUYANA. REGION 8 POTARO-—SIPIRUNI.

Pakaraima Mountains, Upper Potaro River Basin, within 5 km radius of Potaro base

camp located at 5°18'04.8"N 59°54'40.4"W, 710-750 m. Near Ayanganna Airstrip,

20.V.2000, Miller 10039 (BRG, RMS). Dicymbe Research Plots, 27.V.2000, Miller 10070

(BRG, RMS). Vicinity of Potaro Base Camp, 31.V.2001, Aime 1724 (BRG, LSUM).

~3.5 km SE of base camp near Dicymbe plot 2, 12.V1.2001, Henkel 8270 (BRG; HSU);

19.V.2010 Aime 3974 (BRG, LSUM). Mixed Forest Research Plots MP1, 7.VII.2009,

Henkel 9006 (BRG, HSU). Near Potaro Falls, 4. VII.2002, Aime 2196 (BRG, LSUM).

COMMENTARY — This is an unusual Russula species whose initial encounter

in the low-tech environment of field work in Guyana sparked a great deal of

debate regarding generic placement. The combination of pilose pileus, strongly

inrolled margin, somber yellowish brown to dull brown colors, and tendency for

the lamellae and stipe to discolor brownish, was reminiscent of the temperate

Paxillus involutus. However, close observation of the basidiospores in Melzer’s

reagent, even without a microscope, confirmed the amyloid reaction.

Molecular analysis (SL Miller, unpublished) places R. paxilliformis

consistently near R. matoubensis Pegler. Russula matoubensis is a much

brighter orange but also has a subtomentose (especially when collected fresh

in dry conditions) pileus that becomes finely granular to areolate-disrupted

and squamulose away from the disc, exposing the underlying and much lighter

colored tissue beneath. Microscopically, both fungi also possess a trichodermial

palisade in the pellis, composed of ascendant hyphae, rare dermatocystidia and

interwoven hyphae in the subpellis that are heavily encrusted with material

that is positively acid resistant in basic fuchsin. In both fungi, scalp sections of

the pileus cuticle turn instantly dark brown in KOH. Also, both taxa contain

subpellis cells with irregularly thickened cell walls and lamprocystidia in the

hymenium.

Pegler & Fiard (1979) placed R. matoubensis in R. sect. Decolorantinae

Meltzer & Zvara based on orange pigment, mild taste, white context that

discolors grey, and the presence of dermatocystidia in the pellis. Singer et

al. (1983) acknowledged possible placement in R. subsect. Decolorantes (=

Decolorantinae) but, noting the close resemblance of the anatomical characters

to those of R. fistulosa R. Heim, ultimately placed R. matoubensis into subsect.

Fistulosinae R. Heim ex Sing. Indeed, our molecular analysis consistently

supports R. paxilliformis and R. matoubensis as a sister group to a clade

Russula & Lactifluus spp. nov. (Guyana) ... 247

sve

PiaTE 6. Russula paxilliformis (Miller 10097), microscopic features. 23. Diagram of a section of the

dense trichoderm comprising the suprapellis. 24. Elements of the suprapellis showing long narrow

pileocystidia, catenulate hyphae and acid resistant encrustations. 25. Elements of the stipitipellis.

26. Hymenial cystidia. 27. Marginal cells. 28. Basidia. 29. Basidiospores. Scale bars = 10 um.

248 ... Miller, Aime & Henkel

containing R. ochraceorivulosa Buyck, which Buyck (1994a) placed into subsect.

Pallidorimosinae Buyck, and R. patouillardii Singer, which Buyck (1994b)

placed into subsect. Virescentinae Singer. Morphologically, R. paxilliformis

appears similar to members of the African subsections Brunneofloccosinae

(e.g., R. aurantiofloccosa and R. brunneofloccosa) or Testaceoaurantiacinae (e.g.,

R. testaceoaurantiaca Beeli or R. binganensis Beeli), both described by Buyck

(1990b). The features common to both the South American and African species

are the abundant catenulate chains of spherical to cylindrical highly encrusted

cells in the pellis, the pilose to velutinous pileus that develops a distinctive radial

cracking pattern toward the margin, the basidiospores with fine connectives

between amyloid ornamentation elements and frequently a large amyloid plage,

and flesh or stipe surface that discolors brownish.

Russula gelatinivelata S.L. Mill, Aime & T.W. Henkel, sp. nov. PLATES 7-8

MycoBank MB 564267

Distincto gelatinoso partiali velo circa pileum et inferam partem stipitis juniorum, piloso

rubrobrunneo pileo, et stipite cum rubra pilosa pagina in medio in distinctam areolatam

ordinationem frangatur differt.

Type: Guyana, Pakaraima Mountains, Upper Ireng River Basin, east bank of Ireng River,

1 km downstream from Kurutuik Falls in Dicymbe-dominated slope forest, 15.V.1998,

TW Henkel 6410 (Holotype, HSU; isotype, RMS, BRG; GenBank JQ405655).

EryMo.oey: gelatinivelata (Lat.), referring to the gelatinous partial veil evident in

young, fresh specimens.

PiLEus 5.5 cm broad, convex at first, then broadly convex to slightly depressed;

margin incurved at first, then decurved, entire when young, then frequently

lacerate with age, obscurely striate when young, tuberculate striate when older,

striations 6 mm in length; pellis dull to shiny, with a thick gelatinous partial veil

present when young with material compacted into cavity formed by margin

and stipe, subvelutinous in age, violet brown (11F7—8) with extreme margin

cream (3A—2) when young, then reddish brown (9—10D-—E-8) to brownish

violet (11E6—7) or violet brown (10F—8), disk violet brown 10—11F6-8 or

occasionally with pale yellow centrally, older specimens fading to grayish red

(9C-—D-—5-—6) with disk concolorous to violet brown (10F—8). LAMELLAE 3-3.5

mm broad at mid-radius, adnate to slightly sinuate to subdecurrent, close,

frequently forking at stipe, entire, pale yellow (3A 2-3); lamellulae absent.

STIPE 4.5 x 1.3 cm, equal or more typically tapered to base, attachment central,

even to slightly irregular, viscid to gelatinous over lower one-half when young,

becoming subtomentose above, conspicuously areolate below, with a white to

cream (3A 2-3) zone at apex (I.E. concolorous with lamellae), rarely with a

pinkish tinge, middle portions overlain with grayish- to brownish-red (10D

5-7) areolae frequently in a stretch-mark like pattern over a white or yellowish

white (3A2) ground; the base concolorous with the apex or yellowish white

Russula & Lactifluus spp. nov. (Guyana) ... 249

™m 4233

PiaTE 7. Russula gelatinivelata habit (Henkel 8233). 30. x0.9.

(3A2). CONTEXT in pileus 2-3 mm at mid-radius pliant, white, pale pink

immediately below cuticle, especially at disk; trama in stipe stuffed at maturity,

outer cylinder pale yellow surrounding white central core; odor fragrant like

Avon hand lotion; taste slightly bitter; FeSO, yellow on stipe surface, quickly

salmon red on trama.

BASIDIOSPORES 6.8—7.6 X 6—6.8 (Q = 1.11-1.13), subglobose to broadly

ellipsoid; ornamentation of widely distributed, isolated, acute, narrowly conical

elements, 1.5(—2) um high, amyloid, darkest near the apiculus; suprahilar

plage large and verruculose, not or slightly decurrent on the apiculus. BAsIDIA

(24) 28-32(-36) x 8-13 um, clavate to subcylindrical, 2-, 3-, or 4-spored,

sterigmata long, 6.4—8 x 1—-1.6 um. Cystip1a 75-130 x 15-18 um, subclavate

to cylindrical, obtuse to capitate, thin-walled, some long, arising deep within

the lamella trama, others shorter, arising in hymenium, emergent 10-40 um

above basidia, scattered to numerous, SV-. MARGINAL CELLS 85-95 x 15-20

um, fusiform, mucronate, strongly emergent, mixing with small basidia,

numerous. SUBHYMENIUM well developed of small tightly packed spherical

cells of 10 um diam. LAMELLAR TRAMA composed of many large sphaerocytes,

and nests of relatively small, nearly isodiametrical to globose sphaerocytes,

interwoven with cylindrical hyphae. PILEIPELLIs orthochromatic in Cresyl

Blue, two-layered; subpellis of slightly gelatinized interwoven hyphae of 4—5

uum diam intergrading with discrete regularly spaced rosettes, these often

encrusted with reddish brown pigment in KOH; suprapellis a loose trichoderm

composed of frequently branching spindly hyphae; these 5-12 um diam,

cylindrical to clavate, obtuse and often sinuous and irregularly constricted,

often multi-septate, thin- or exceptionally thick-walled, arising from large

250 ... Miller, Aime & Henkel

PiaTE 8. Russula gelatinivelata (Miller 10066), microscopic features. 31. Diagram of a section of the

two-layered pileipellis; the suprapellis is a trichoderm over a thin pseudoparenchymatous subpellis.

32. Elements of the suprapellis. 33. Elements of the stipitipellis. 34. Hymenial cystidia. 35. Marginal

Cells. 36. Basidia. 37. Basidiospores. Scale bars = 10 um.

Russula & Lactifluus spp. nov. (Guyana) ... 251

inflated spherical cells 2-6 deep that in aggregate resemble an epithelium;

these easily disarticulated in microscopic preparations; cystidia 50-100 um,

long pedicellate, septate, irregularly constricted or swollen at the septum, thin-

or thick-walled, arising deep in the trama, SV-, the contents or interior of the

walls acid resistant in basic fuchsin. STIPITIPELLIS resembling the pileipellis,

composed of branching, septate, digitate to irregularly shaped hyphae that taper

at the apex, and pedicellate, septate, thick-walled hyphae that are cylindrical,

clavate, obtuse and irregularly constricted.

HABIT, HABITAT, AND DISTRIBUTION — Solitary to usually gregarious on

root mats of Dicymbe corymbosa, or in stands of Dicymbe altsonii lacking

D. corymbosa; common and widespread; found May through early July during

wet weather. Known from the Upper Ireng and Upper Potaro Basins of Guyana.

ADDITIONAL SPECIMENS EXAMINED: GUYANA. REGION 8 POTARO-SIPARUNI.

Pakaraima Mountains, Upper Ireng River Basin, elevation 710-850 m. Suruwabaru

Creek, 1 km upstream of juncture with Yuarka Creek, in Dicymbe-dominated riparian

forest, 2.III.1997, Henkel 6269 (BRG; HSU). East bank Ireng River near Sandhill Village,

under Dicymbe altsonii in riparian forest, 1.V.1998, Henkel 6360 (BRG; HSU). East bank

of Sukabi River, 1.5 km upstream from Juncture with Ireng River in Dicymbe corymbosa-

dominated slope forest, 22.V.1998, Henkel 6441 (BRG; HSU). Toe slopes of Mt.

Kukuinang, 3 km east of confluence of Sukabi and Ireng Rivers in Dicymbe corymbosa-

dominated slope forest at 850 m elevation, 23.V.1998, Henkel 6550 (BRG; HSU). East

bank of Ireng River near Sukabi camp, under Dicymbe corymbosa, 27.V.1998, Henkel

6638 (BRG; HSU). ~1 km east of confluence of Sukabi and Ireng Rivers, in slope forest

dominated by Dicymbe altsonii lacking in Dicymbe corymbosa, 27.V.1998, Henkel 6750

(BRG; HSU). Pakaraima Mountains Upper Potaro River Basin, within 5 km radius of

Potaro base camp located at 5°18'04.8"N 59°54'40.4"W, 710-750 m. Vicinity of Potaro

Base Camp, 20.V.2000, Miller 10066 (BRG, RMS). Dicymbe stand on Benny’s ridge near

Potaro River camp, 11.VI.200, Miller 10125 (BRG, RMS). ~3.5 km SE of base camp

near Dicymbe plot 2, 26.V.2010, Henkel 9226 (BRG; HSU). ~3 km SE of base camp

in Dicymbe plot 1, 30.V.2001, Henkel 8233 (BRG; HSU). Vicinity of Dicymbe plot 3,

21.V1.2002, Aime 2096 (LSUM, HSU).

COMMENTARY — Young basidiomata of R. gelatinivelata in moist conditions are

encased in a thick gelatinous layer around the lower portion of the stipe and

pileus that is present as a veil in the space between the stipe and inrolled pileus

margin. The red tomentum and striped areolate patches in the middle portion

of the stipe overlaying yellow ground are remarkable.

Molecular analysis (SL Miller, unpublished) indicates that R. gelatinivelata

has a closer relationship with Russula temperate species than any other species

collected in Guyana. Specifically there appears to be a consistent relationship

with R. nauseosa (Pers.) Fr., R. cessans A. Pearson, and R. laricina Velen., or with

R. aurea Pers. and R. romellii Maire, all temperate taxa that have large, abundant

multiseptate cystidiate hyphal elements in the pellis. In R. gelatinivelata the

walls of these elements, while not encrusted per se, are highly positive for acid

resistant coloration in basic fuchsin. The temperate taxa appear to lack this

252 ... Miller, Aime & Henkel

characteristic, although taxa that have been placed in closely related sections

and subsections (e.g. section Incrustatae Romagn.) are also highly positive for

acid resistant coloration in basic fuchsin, but this is expressed in the form of

abundant encrustations on the exterior of the elements. Russula gelatinivelata,

along with R. nauseosa and R. laricina, has spores with isolated elements,

while R. cessans, R. aurea, and R. romellii have spores with at least a partial

reticulum.

Acknowledgements

The authors thank Drs. Annemieke Verbeken and Kanad Das for providing helpful

reviews of this manuscript. Funding was provided by the National Science Foundation

grant DEB-1050292 to SLM. TWH received funding from the National Science

Foundation DEB—0918591, the Smithsonian Institution’s Biological Diversity of the

Guiana Shield Program, the National Geographic Society's Committee for Research

and Exploration, the Linnaean Society of London, and the Humboldt State University

Foundation. Funding from the Explorer’s Club Exploration and Field Research grant,

a field research gift from W.K. Smith, and NSF DEB—0732968 was provided to MCA.

Research permits were granted by the Guyana Environmental Protection Agency. Dillon

Husbands functioned as Guyanese local counterpart and assisted with field collecting,

descriptions, and specimen processing. Expert field assistance was provided by C.

Andrew, L. Edmund, D. Husbands, V. Joseph, P. Joseph, and L. Williams. Special thanks

to Terry McClean of the Nucleic Acid Exploration Facility at the University of Wyoming

for help with sequencing and analysis. This paper is number 189 in the Smithsonian

Institution's Biological Diversity of the Guiana Shield Program publication series.

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MYCOTAXON

http://dx.doi.org/10.5248/121.255

Volume 121, pp. 255-263 July-September 2012

Tuber in China: T. sinopuberulum and T. vesicoperidium spp. nov.

Li FAN’, JIN-ZHONG CAO? & JIA YU?

' College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

? Institute of Mycology, Jilin Agricultural University, Changchun 130118, China

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

ABSTRACT — Two newspecies are described and illustrated. Tuber sinopuberulum is recognized

by its glabrous peridium and globose to subglobose ascospores. Tuber vesicoperidium is

distinguished by its peridial structure of large, thick-walled swollen cells. The two new species

are supported by molecular studies.

Key worps — Ascomycota, Tuberaceae, truffle

Introduction

The first species of Tuber P. Micheli ex EH. Wigg. to be described in China

was T: taiyuanense B. Liu in 1985 (Liu 1985, Cao et al. 2011). Of the additional

28 species since described, 19 appear endemic to China (TaBLE 1). The other

nine species (T: aestivum Vittad., T. borchii Vittad., T. californicum Harkn.,

T. excavatum Vittad., T: indicum Cooke & Massee, T. lyonii EK. Butters,

T. maculatum Vittad., T. puberulum Berk. & Broome, T: rufum Picco) have

been recorded from India, Europe, or North America (Song 2005; Cao 2010).

In recent years interest in Chinese Tuber species has accelerated partly because

some are edible and are being marketed both at home and abroad. Marketplaces

where locals sell their wild mushrooms have proven to be excellent places to

find new species. Here we describe two new Tuber species, which we discovered

in a Kunming (Yunnan Province, China) mushroom market where locals,

including the Yi people (Wikipedia 2011), offer wild mushrooms for sale.

Materials & methods

Morphological studies

Truffles were collected from the mushroom market in Kunming, China on

20 December 2010. Specimens were deposited in BJTC (Herbarium of Biology

Department, Capital Normal University). Fresh specimens were described both macro-

256 ... Fan, Cao & Yu

SPECIES

T: sinense K. Tao & B. Liu

T. liaotongense Y. Wang

T: gigantosporum Y. Wang & Z.P. Li

T. formosanum H.T. Hu

T. pseudohimalayense G. Moreno et al.

T. pseudoexcavatum Y. Wang et al.

T. liui AS. Xu

T. xizangense A.S. Xu

T. huidongense Y. Wang

T. zhongdianense X.Y. He et al.

T. furfuraceum H.T. Hu & Y. Wang

T. umbilicatum Juan Chen & PG. Liu

T. latisporum Juan Chen & P.G, Liu

TABLE 1. Tuber species described from China since 1985

REFERENCE

Tao et al. 1989

Wang, 1990

Wang & Li 1991

Hu 1992 (from Taiwan)

Moreno et al. 1997

Wang et al. 1998

Xu 1999

Wang & He 2002

He et al. 2004

Hu & Wang 2005 (from Taiwan)

Chen et al. 2006

Chen et al. 2007

Fan et al. 2011

Fan et al. 2012a

Fan et al. 2012b

T. lijiangense L. Fan & J.Z. Cao

T: sinoexcavatum L. Fan & Yu Li

T. polyspermum L., Fan & C.L. Hou

T. sinoalbidum L. Fan & J.Z. Cao

T. microspermum L. Fan & J.Z. Cao

T. microspiculatum L. Fan & Yu Li

and microscopically. Razor-blade sections were mounted in 3% KOH or stained with

Melzer’s reagent, rinsed, and mounted in polyvinyl lactic glycerol to make permanent

slides for archiving with dried specimens. For scanning electron microscopy (SEM),

ascospores were scraped from the dried gleba onto double-sided tape mounted directly

on an SEM stub, coated with gold-palladium, and examined and photographed with a

HITACHI S-4800 SEM.

Molecular methods

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

301, Retsch, Haan, Germany) ina 1.5 ml tube 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 ITS region was amplified with PCR using the

primers ITS1/ITS4 (White et al. 1990). PCR was performed in 50 ul reactions containing

DNA template 2 ul, primer (10 uM) 2 pl each, 2x Master Mix (Tiangen Biotech (Beijing)

Co. Ltd.) 25 ul. PCR reactions were run as follows: an initial denaturation at 95 °C for 3

min, followed by 30 cycles at 95 °C for 2 min, 55 °C for 25 s, 72 °C for 2 min, and a final

extension at 72 °C for 10 min. The PCR products were sent to Invitrogen Biotechnology

Co. Ltd. (Beijing, China) for purifying, sequencing, and editing. Other ITS rDNA

sequence data included in this study were downloaded from GenBank (TABLE 1).

Phylogenetic analyses

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

was manually adjusted with Se-Al v.2.03a (Rambaut 2000). The aligned dataset was

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

Maximum parsimony analysis was conducted using heuristic searches with 1000

replicates of random-addition sequence, tree bisection reconnection (TBR) branch

Tuber spp. nov. (China) ... 257

TABLE 2. Tuber specimens and sequence numbers used in molecular studies

SPECIES ITS LSU

T. sinopuberulum (BJTC FAN157) JQ690073 JQ690070

T. vesicoperidium (BJTC FAN155) JQ690071 JQ690068

(BJTC FAN156) JQ690072 JQ690069

T. borchii HM485343 FJ809799

HM485343

FJ809852

T. californicum DQ974799 AF127120

HM485351 AF159627

HM485346

T. dryophilum Tul. & C. Tul. HM485353 FJ809800

HM485354 FJ809801

T. maculatum EU784428

FM205649

AJ969627

AF106889

T. oligospermum FM205506 AY515306

FM205507

T. puberulum AJ557536

AJ969625

T. sphaerosporum Gilkey FJ809853 FJ809805

FJ809854 FJ809806

HM485390

T. zhongdianense DQ898186

DQ898187

T. panniferum Tul. & C. Tul. FJ809845

FJ809846

T. melanosporum Vittad. AF132501 GU979139

AF106878 FJ809819

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.

Results

Molecular phylogenetics

The maximum parsimony analysis of ITS sequences produced one most

parsimonious tree (Fic. 1) with a length (TL) = 613 steps, consistency index

(CI) = 0.7276, retention index (RI) = 0.8706, homoplasy index (HI) = 0.2724,

and rescaled consistency index (RC) = 0.6335.

The maximum parsimony analysis of LSU sequences produced one most

parsi-monious tree (Fic. 2) with a length (TL) = 380 steps, consistency index (CI)

= 0.8816, retention index (RI) = 0.8308, homoplasy index (HI) = 0.2724, and

rescaled consistency index (RC) = 0.7324.

258 ... Fan, Cao & Yu

100

100)

Tuber dryophilum HM485354

Tuber dryophilum HM485353

Tuber zhongdianense DQ898187

Tuber zhongdianense DQ898186

Tuber vesicoperidium JQ690072

Tuber vesicoperidium JQ690071

Tuber sinopuberlum JQ690073

Tuber oligospermum FM205507

Tuber oligospermum FM205506

Tuber borchii HM485343

Tuber borchii HM485344

Tuber borchii FJ809852

Tuber puberulum AJ557536

Tuber puberulum AJ969625

Tuber sphaerosporum FJ809854

Tuber sphaerosporum HM485390

Tuber sphaerosporum FJ809853

Tuber californicum DQ974799

Tuber californicum HM485346

Tuber californicum HM485351

Tuber maculatum FM205649

Tuber maculatum EU784428

Tuber maculatum AJ969627

Tuber maculatum AF 106889

Tuber melanosporum AF 132501

Tuber melanosporum AF106878

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

Tuber species with pale ascomata and reticulate ascospores, using T: melanosporum as outgroups.

Bootstrap values of more than 70% from 1000 replications are shown above the respective

branches.

Tuber californicum AF127120

Tuber californicum AF 156927

100

Tuber sphaerosporum FJ809806

Tuber sphaerosporum FJ809805

Tuber borchii FJ809799

100

Tuber oligospermum AY515306

Tuber vesicoperidium JQ69006869

Tuber vesicoperidium JQ69006868

Tuber dryophilum FJ809801

Tuber dryophilum FJ809800

Tuber sinopuberulum JQ69006870

Tuber panniferum FJ809846

Tuber panniferum FJ809845

Tuber melanosporum GU979139

Tuber melanosporum FJ809819

Fic. 2. Phylogeny derived from maximum parsimony analysis of LSU nuclear rDNA sequences of

some Tuber species with pale ascomata and reticulate ascospores, additionally two sequences of

Tuber panniferum with spiny ascospores, using T: melanosporum as outgroups. Bootstrap values of

more than 70% from 1000 replications are shown above the respective branches.

Tuber spp. nov. (China) ... 259

The two new species, Tuber sinopuberulum and T: vesicoperidium, group into

one clade with 100% bootstrap support in the ITS-based phylogenetic analyses

(Fic. 1), but in the LSU-based analyses (Fic. 2), they separate into two distinct

clades. Two T. vesicoperidium sequences group in a clade with strong bootstrap

support (99% in both ITS and LSU sequence analyses). Tuber sinopuberulum

and T. vesicoperidium have very similar ascomata and ascospores but very

different peridial structures. Tuber sinopuberulum typically has a two-layered

peridium composed of a pseudoparenchymatous layer and intricate textural

layer, whereas T: vesicoperidium has a one-layered prosenchymatous peridium

with very large and thick-walled swollen cells.

Taxonomy

Tuber sinopuberulum L. Fan & J.Z. Cao, sp. nov. Fig. 3

MycoBank MB 564482

Differs from Tuber puberulum by the absence of hyphae-like hairs on the peridium

surface and by its globose to subglobose ascospores.

Type: China. Yunnan Province, Kunming, from the local mushroom market, 20 Dec.

2010, Jin-zhong Cao 120 (Holotype, BJTC FAN157).

EryMo_oey: sinopuberulum (Lat.), referring to the similarity to the European species,

Tuber puberulum.

ASCOMATA 2 xX 2.5 cm, subglobose or lobed, firm, solid, surface smooth,

glabrous, yellow white to light brown at maturity. Odor slight, not pungent.

PerRIpIuUM 250-350 um thick, two layers; outer layer 80-120 um thick,

pseudoparenchymatous, composed of subangular or subglobose cells 7.5-20

um in diam., with thin or slightly thickened walls, pale yellowish, no distinct

hyphae-like hairs arising from the outermost cells; inner layer composed of

intricately interwoven hyphae, hyaline, thin-walled, branch, septate, 3-5 um

in diam. GLEBA light brown to brown at maturity, marbled with large and

rare, branched white veins continuous with inner peridium. Ascr subglobose,

ellipsoid or irregular, hyaline, thin-walled, 70-110 x 65-100 um, sessile or

short stalk, 1-5 spored. Ascospores mostly globose or subglobose, a few

broad ellipsoid, yellow-brown at maturity, 20-37.5 x 20-32.5 um excluding

ornamentation; ornamentation regularly alveolate reticulum, 2.5-4 um high,

the meshes generally 5-10 across the spore width.

ComMENts — Tuber puberulum from Europe is similar to this new species but

can be easily differentiated by the 100-150 um long hyphal-like hairs densely

coating the peridial surface (Lange, 1956; Pegler et al. 1993) and ascospores

that are not regularly globose.

The European Tuber oligospermum (Tul. & C. Tul.) Trappe and the American

T. californicum might be confused because of their globose ascospores. However,

260 ... Fan, Cao & Yu

Fic. 3. Tuber sinopuberulum (BJTC FAN157, holotype) a. Ascoma. b. Asci and ascospores observed

under light microscope. c. Ascospore observed under SEM.

T. oligospermum is distinguished by a one-layered peridium, and T! californicum

can be separated by its dark colored gleba and puberulent ascomata.

Tuber vesicoperidium (described below) has ascospores similar to T. sino-

puberulum but can be easily differentiated by its peridial structure characterized

by very large and thick-walled swollen cells. The phylogenetic analyses (Fics.

1-2) also show that while T: sinopuberulum groups in a clade with T! vesico-

peridium in the ITS sequence analysis (Fic. 1), they are widely separated in two

different clades in the LSU analysis (Fic. 2), suggesting that they are closely

related but distinct species.

Tuber vesicoperidium L. Fan, sp. nov. Fic. 4

MycoBank MB 564483

Differs from all other Tuber species with globose or subglobose ascospores by its one-

layered peridium of predominantly large, thick-walled, swollen cells.

Type: China. Yunnan Province, Kunming, from the local mushroom market, 20 Dec

2010, Jin-zhong Cao 118 (Holotype, BJTC FAN155).

ETyMOLocy: vesicoperidium (Lat.), referring to the peridium with larger sized swollen

cells.

ASCOMATA 2-4 cm, subglobose or lobed, a few of slight furrows, firm, solid,

surface smooth, glabrous, white to pale yellow, light brown at maturity. Odor

slight, not pungent. PERrIDIUM 550-650 um thick, one layer, prosenchymatous,

composed of intricately interwoven hyphae, hyaline, thin-walled, branched,

septate, 3-7.5 um in diam., distinctly intermixed with large swollen cells of

mostly ellipsoid-shape 50-130 um in diam. with wall 5-7.5 um thick, towards

the inner side of the peridium, the number of the swollen cells less, integrating

with the interwoven hyphae only, more or less parallel to the surface; no

distinct hyphae-like hairs arising from the outermost cells. GLEBA grey brown

to dark brown at maturity, marbled with large and rare, branched, white veins

continuous with peridium. Asci subglobose, ellipsoid or irregular, hyaline, thin-

walled, 65-95 x 50-70 um, sessile, 1-5 spored. Ascospores mostly globose, a

few subglobose, yellow-brown at maturity, 20-35.5 x 20-32.5 um in 2-5 spored

asci and 37.5-45 x 37.5-42.5 um in 1-spored asci excluding ornamentation;

Tuber spp. nov. (China) ... 261

Fic. 4. Tuber vesicoperidium (BJTC FAN155, holotype) a. Ascomaa. b. Peridium with

the large, thick-walled swollen cells. c. Asci and ascospores observed under light

microscope. d. Ascospore observed under SEM.

ornamentation regularly alveolate, 2.5-4 um high, the meshes generally (2-)

3-6(-8) across the spore width.

ADDITIONAL SPECIMEN EXAMINED: CHINA. YUNNAN PROVINCE, Kunming, from the

local mushroom market, 20 Dec 2010, Jin-zhong Cao 119 (BJTC FAN156).

ComMENTs — Tuber vesicoperidium is characterized mainly by its one-layered

peridium and the large, thick-walled, swollen cells occupying nearly all the

peridial tissue, thereby distinguishing it from all other known Tuber species

with globose or subglobose ascospores. Large swollen cells also occur in some

Tuber species with a two-layer peridium and spiny-reticulate ascospores, such

as T: borchii, T. zhongdianense, and T. malacodermum E. Fisch., but with the

swollen cells confined to the outer peridium. Lange (1956), Pegler et al. (1993),

and Chen & Liu (2007) note that the shape and wall thickness of the swollen

cells do not differ greatly from the surrounding cells and they do not normally

exceed 70 um in diam.

ITS (Fic. 1) and LSU (Fic. 2) sequence analyses group the T! vesicoperidium

sequences in a clade with strong support (BP 99 for both analyses), supporting

T. vesicoperidium as a distinct species. In the ITS sequence analysis (Fic. 1), the

two T: vesicoperidium sequences group together with T. sinopuberulum with

strong support (BP 100). This close relationship is reflected by their very similar

262 ... Fan, Cao & Yu

ascomata and ascospores, but the completely different peridial structures also

indicate that the two taxa are morphologically independent.

Acknowledgments

The study was supported by the National Natural Science Foundation of China (Nos.

30770005, 30870008), the Beijing Natural Science Foundation (No. 5072006). We are

grateful to Prof. Wen-Ying Zhuang and Dr. Ian R. Hall for serving as the pre-submission

reviewers.

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Fan L, Cao JZ, Liu YY, Li Y. 2011. Two new species of Tuber from China. Mycotaxon 116: 349-354.

http://dx.doi.org/10.5248/116.349

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from China. Mycotaxon118: 403-418. http://dx.doi.org/10.5248/118.403

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spp. nov. Mycotaxon 119: 391-395. http://dx.doi.org/10.5248/119.391

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Hu HT. 1992. Tuber formosanum sp. nov. and its mycorrhizal associations. Quart. J. Exp. Forest 6:

79-86.

Hu HT, Wang Y. 2005. Tuber furfuraceum sp. nov. from Taiwan. Mycotaxon 93: 155-157.

Huang JY, Hu HT, Shen WC. 2009. Phylogenetic study of two truffles, Tuber formosanum and Tuber

furfuraceum, identified from Taiwan. FEMS Microbiol. Lett. 294: 157-171.

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sp. nov. an Asiatic species commercialized in Spain, similar to the “Perigord” truffle. Mycotaxon

63: 217-224.

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http://dx.doi.org/10.1016/S0007-1536(88)80032-9

MY COTAXON

http://dx.doi.org/10.5248/121.265

Volume 121, pp. 265-273 July-September 2012

Presence of Leucocoprinus cretaceus and L. fragilissimus

in Argentina

N. NIVEIRO'?’, O. POPOFF! & E. ALBERTO?

"Instituto de Botanica del Nordeste,

Sargento Cabral 2131, CC 209 Corrientes Capital, CP 3400, Argentina

?Instituto Tecnologico Chascomus. IIB-INTECH (UNSAM-CONICET),

Cam. Circ. Laguna Km. 6, Chascomus, Buenos Aires, CP 7130, Argentina

* CORRESPONDENCE TO: niconiveiro@hotmail.com

ABSTRACT — Leucocoprinus cretaceus and L. fragilissimus are recorded for the first time in

Argentina. These species are described and illustrated, and a key to the Argentinean species

of this genus is presented.

Key worps — Agaricomycetes, Agaricales, Agaricaceae, Leucocoprineae, taxonomy

Introduction

Leucocoprinus Pat. was considered by Singer (1986) to belong to the

tribe Leucocoprineae Singer in the family Agaricaceae Chevall. The genus is

characterized by a pluteoid habit, convex to umbonate pileus with floccose

pulverulent squamules, a striate-sulcate margin, a very thin context, free

lamellae, and the absence of colour changes with ammonia vapour, a central

stipe with bulbous-clavate base and annulus, and a white to pale pink spore-

print. The basidiospores are smooth, hyaline, thick-walled, with or without

an evident germ pore, dextrinoid, metachromatic in cresyl blue, basidia

are 4-sterigmate, pleurocystidia are either absent or not numerous, and

cheilocystidia are usually abundant. The pileipellis is very variable, formed by

a mixture of different types of cells and hyphae, and generally is a disrupted

epicutis mixed with sphaerocysts or chains of branched/unbranched inflated

hyphae that sometimes form erect, loose fascicles comprising the squamules.

The hymenophoral trama is regular and trabecular and clamp connections

are absent. Leucocoprinus species are solitary to gregarious, terrestrial, and

saprotrophic (Singer 1986, Vellinga 2001, Gimenes 2007, Rother & Borges da

Silveira 2009). The most similar genus, Leucoagaricus Locq. ex Singer, differs

266 ... Niveiro, Popoff & Alberté

mainly by lacking a long striate-sulcate pileal margin, having a thinner context,

and lacking pseudoparaphyses (Pegler 1983, Singer 1986, Vellinga 2001).

Leucocoprinus has a pantropical distribution and is also found growing

in greenhouses of temperate regions (as introduced species) (Vellinga 2004).

The genus comprises approximately 40 species worldwide (Kirk et al. 2008).

In Argentina the specific diversity of this genus is poorly known. Only six

species and two infraspecific varieties have been recorded so far (Raithelhuber

1974, 1987a-c, 1988, 1991, 2004; Singer & Digilio 1952, Spegazzini 1898, 1926;

Wright & Alberté 2002).

This paper resulted from research into the diversity of Agaricales from the

rainforests in northern Argentina. The Atlantic Forest, which stretches along

the Atlantic coast of Brazil with its southern border reaching eastern Paraguay

and northeastern Argentina, is one of the most threatened tropical-subtropical

rainforests in the world (Placi & Di Bitetti 2006) and regarded as one of the 25

biodiversity hotspots for conservation (Myers et al. 2000). The aim of this paper

is to report the results of the study of recent collections of Leucocoprinus from

this habitat.

Materials & methods

The authors recently collected the specimens in protected areas from northern

Argentina. The specimens were macroscopically described according to Largent (1986)

and Lodge et al. (2004). Color names are in accordance with Kornerup & Wanscher

(1978). Microscopic features are described from material mounted in 5% KOH and

phloxine (1%). Meltzer’s reagent, cresyl blue, and cotton blue were used to determine

whether the spores were dextrinoid, metachromatic, and cyanophilic, respectively.

Dimensions were recorded from 20 basidiospores per collection. Standard deviation,

quotient of length/width (Q), and average Q-value (Qm) were calculated for each species

from all spores measured (n). Herbarium abbreviations follow Index Herbariorum

(Thiers 2011). Complete synonymies are not presented; for a complete list see Vellinga

(2010). The collections are deposited in CTES Mycological Herbarium.

Taxonomy

Leucocoprinus cretaceus (Bull.) Locq., Bull. Mens. Soc. Linn. 14: 93 (1945). Fics 1-5

= Agaricus cretaceus Bull., Herb. Fr. 8: tab. 374 (1788).

PrLEus 27-75 mm diam., at first ovoid then convex to broadly conical,

umbonate in maturity, white (1.A1), covered by easily removed, granular

floccose, white squamules, conical to pyramidal on the umbo; margin entire,

sulcate. LAMELLAE white, crowded, free, membranous, 2-4 mm wide, with

lamellulae; margin entire. Stripe 50-90 x 3-7 mm, central, cylindrical, becoming

inflated fusoid towards the base, 15-30 x 8-15 mm, white, glabrescent, fistulose,

with a covering of floccose squamules from the base to the annulus. ANNULUS

present, attached to the upper part of the stipe, simple, membranous, white,

Two Leucocoprinus spp. new to Argentina ... 267

Fic. 1: Leucocoprinus cretaceus (Niveiro & Michlig 1614 CTES)

a: general aspect; b: spores in Meltzer’s reagent.

evanescent. CONTEXT white, thin. SPORE PRINT white.

BASIDIOSPORES 6.8-10 x 5-6.6 um (8.5 + 0.8 x 5.7 + 0.4 um); Q = 1.2-1.7

um; Qm= 1.5, n = 60, ellipsoid, hyaline, smooth, thick-walled, walls 0.6-0.7 um,

with an obvious germ pore, dextrinoid, metachromatic, cyanophilic. BASIDIA

18-30 x 7-11 um, claviform, with 4 sterigmata. PEEUDOPARAPHYSES 11-18 x

6-10 um, claviform. PLEUROCysTIDIA absent. CHEILOCYSTIDIA 38.3-46.6 x

8-16(-20) um, narrowly clavate, lageniform to ventricose, thin-walled, hyaline.

HYMENOPHORAL TRAMA regular, hyphae 3.6-8 um, with smooth, thin walls;

inamyloid. PILEIPELLIS made up of hyaline hyphae with smooth walls and

cylindric terminal elements with excrescences, branched in different shapes

(H, T and Y), 24-45 x 7.5-15 um. STIPITIPELLIs a cutis of narrowly cylindrical,

2-7 um broad hyphae. CLAMP CONNECTIONS absent.

Hasirat (of Argentinean collections) — Solitary or gregarious. On bark of

living gymnosperms (Araucaria angustifolia) and unidentified dicotyledonous

trees in subtropical forest.

268 ... Niveiro, Popoff & Alberté

tod @

Fics 2-5: Leucocoprinus cretaceus (Niveiro & Michlig 1614 CTES)

2: basidia; 3: spores; 4: pileipellis; 5: cheilocystidia. Scale bar = 10 um.

MATERIAL EXAMINED — ARGENTINA, CorRIENTEs: Dpto. MBpurucuyA, Mburucuya

National Park, “Che Roga” trail, 6/[V/2007, leg. Niveiro & Michlig 441 (CTES); Santa

Teresa Ranch, 7/IV/2007, leg. Niveiro & Michlig 480 (CTES). Mistongs: Dpto. GRAL.

BELGRANO, San Antonio Natural Reserve, on Araucaria angustifolia, 26°02'14.2"S

55°47'24.8" W, 526 m, 29/V/2009, leg. Niveiro & Michlig 1614 (CTES).

DISTRIBUTION — Leucoagaricus cretaceus is distributed in tropical regions

and adjacent temperate regions in North America and Europe (Vellinga 2001,

Birkebak 2010). It has been recorded from Africa, Asia, Europe, North America,

and South America (Brazil: Gimenes 2007, Meijer 2006, Rother & Borges da

Silveira 2009, Sobestiansky 2005, Wartchow et al. 2008). This is the first record

of L. cretaceus in Argentina (Misiones and Corrientes Provinces).

COMMENTS — Leucocoprinus cretaceus is easily recognized by its fleshy white

basidiomata covered by floccose squamules that are easily removed when

touched and by a fusoid stipe inflated toward the base. This species is related

to L. squamulosus (Mont.) Pegler from Guyana and Lesser Antilles with similar

basidiospores but which lacks an annulus and an inflated stipe base (Pegler

1983, Dennis 1952, Wartchow et al. 2008).

Dennis (1952, 1970) and Pegler (1977, 1983, 1986) identified tropical

species with a white basidioma and pileus covered by concolorous floccose

Two Leucocoprinus spp. new to Argentina ... 269

squamules as Leucocoprinus cepistipes (Sowerby) Pat. However, Candusso &

Lanzoni (1990) and Vellinga (2001) considered L. cepistipes a separate species

based on the ochraceus to pale brown squamules covering the pileus surface

(Candusso & Lanzoni 1990, Wartchow et al. 2008, Gimenes 2007). Vellinga

(2001), Gimenes (2007), and Rother & Borges da Silveira (2009) explain these

conflicting species concepts.

Leucocoprinus fragilissimus (Ravenel ex Berk. & M.A. Curtis) Pat., Essai Tax.

Hyménomyc.: 171 (1900). Figs 6-8

= Hiatula fragilissima Ravenel ex Berk. & M.A. Curtis,

Ann. Mag. Nat. Hist., Ser. 2, 12: 422 (1853).

PiLEus 20-44 mm diam., conic-campanulate, becoming plane with a small

umbo, even depressed, sulcate-striate to the umbo, white to slightly yellowish

(1.A1-1.A3) when young, becoming whitish and almost translucent except for

a central disk in maturity, umbo pale yellowish-brown (1.A4) with small yellow

squamules, dry. LAMELLAE white, thin, subdistant, free, with lamellulae; margin

entire. STIPE 60-100 x 1-2 mm, equal, fragile, central, light yellow (1.A5), with

scattered small yellowish squamules, fistulose. ANNULUS present, white, small,

membranous, movable, evanescent, attached to the upper to middle part of the

stipe. CONTEXT white, very thin, almost absent, except at the disk, membranous,

subdeliquescent. SPORE PRINT off-white.

BASIDIOSPORES 9-12.7 x 6.5-10 um (11.1 + 0.8 x 8.1 + 0.6 um); Q = 1.1-1.6;

Qm = 1.4, n= 54, broadly ellipsoid to subglobose, smooth, hyaline, thick-walled,

with an obvious germ pore with a hyaline dome, dextrinoid, metachromatic,

cyanophilic. HYMENIAL ELEMENTS not observed. PLEUROCYSTIDIA and

CHEILOCYSTIDIA unobserved. PILEIPELLIs an epithelium with hyaline, globose,

more or less isodiametric elements, 17.2-25.3 um in diameter, thin-walled,

mixed with hyaline, thin, more or less cylindrical hyphae. STrpITIPELLis a cutis

of narrowly cylindrical, 5-12 um broad hyphae. CLAMP CONNECTIONS absent.

HasiraT (of Argentinean collections) — Solitary or gregarious in small

groups. On decomposing dicotyledonous wood and in soil among litter in

subtropical pastures and forests.

MATERIAL EXAMINED — ARGENTINA, CorrRIENTEs: Dpto. MBpurucuyA, Mburucuya

National Park, 7/IV/2007, leg. Niveiro & Michlig 448 (CTES); 8/IV/2007, Santa Maria

Ranch, leg. Niveiro & Michlig 496 (CTES). Mis1ongs: Dpto. GRAL. BELGRANO, San

Anténio Natural Reserve, 26°01'41.4"S 53°47'20.8"W, 322 m, 28/V/2009, leg. Niveiro

& Michlig 1477 (CTES); Dpro. SAN PEDRO, Mocona Provincial Park, 27°09'25.0"S

53°54'07.9"W, 289 m, 25/V/2009, leg. Niveiro & Michlig 1356 (CTES).

DISTRIBUTION — Leucocoprinus fragilissimus has a pantropical distribution

(Pegler 1983). It has been recorded from Africa, Asia, Europe, North America,

and South America (Ecuador: Reid et al. 1981; Brazil: Albuquerque et al. 2006,

Capelari & Maziero 1988, Gimenes 2007, Pegler 1997, Rother & Borges da

270 ... Niveiro, Popoff & Alberté

Fic. 6: Leucocoprinus fragilissimus (Niveiro & Michlig 1356 CTES)

a: general aspect; b: spores in Meltzer’s reagent.

Silveira 2009, Wartchow et al. 2008). This is the first record of this species in

Argentina (Misiones and Corrientes Provinces).

ComMENnts — Leucocoprinus fragilissimus is widely distributed in the tropics

and subtropics (Halling & Mueller 2005). It is characterized by subdeliquescent

basidiomata and a membranous, yellowish-white pileus with small yellow

squamules (Pegler 1983, Rother & Borges da Silveira 2009). Although

the hymenial elements were not observed in our collections due to the

subdeliquescent pileus, the basidiospore shape and size and pileipellis structure

completely agree with those of this species.

According to Pegler (1983) and Smith & Weber (1982), L. fragilissimus

is probably a common pantropical species but rarely collected owing to the

extremely delicate basidiomata, which make field collection difficult. The

presence of cheilocystidia in this species is rarely mentioned. According to

Smith & Weber (1982), the cheilocystidia are 13-25(-36) x 9-15(-20) um in

size.

Two Leucocoprinus spp. new to Argentina ... 271

“J

Fics 7-8: Leucocoprinus fragilissimus (Niveiro & Michlig 1356 CTES)

7: spores; 8: pileipellis. Scale bar = 10 um.

Leucocoprinus tenellus Pegler [nom. illegit., non (Boud.) Locq.] from the

Lesser Antilles and L. thoenii Heinem. from Zaire are also species with thin

fragile basidiomata. Their habit and spore shape also resemble L. fragilissimus,

but L. tenellus and L. thoenii differ in the absence of any yellow pigments (Pegler

1983). Leucocoprinus brunneoluteus Capelari & Gimenes from Brazil also

has fragile basidiomata but differs from L. fragilissimus in its more yellowish

coloration and the presence of brown umbo and squamules (Capelari &

Gimenes 2004, Rother & Borges da Silveira 2009).

Key to species of Leucocoprinus in Argentina

la. Basidiomata yellow or yellowish-white ......... 0... cece eee eee eee eee 2

1b. Basidiomata white, brownish to pinkish-orange (i.e. non yellowish) ............. 4

2a. Pileus extremely thin and subdeliquescent, white with bright yellow

siria Madpressed-oquaniules a-, s.r. Pots ed oances PSUs tae dd alot L. fragilissimus

2b. Pileus fleshy, pale yellow to sulphur yellow, covered by loose,

SCatbereds ALOCCOSE SCAT UIES: 2 Poms 2 tas esa aaa nnte ty ates ony oe tes wl BS al 3

3a. Pileipellis made up of elongated elements; spores 7-10 x 5-7 um,

WITH AP ODWIOUS COT POLEN, ce. ion arctica, teat lan tte 'asar Seaahesenemat atime mane L. birnbaumii

3b. Pileipellis made up of globose elements; spores 5-7.5 x 3-5 um,

WithOUPAN-ObVIOUS {ser POLE 2 Mee iced seatiye cece, ge sera cese Sioa eget cea» L. straminellus

4a. Pileus pinkish-orange; stipe with reddish, greenish, or bluish tinges;

SPOKes +d S=A Sy DROAT | sakes fake ng Hanon lyse Clelnes 4 shetstir th ots Saco L. fibrillosus

4b. Pileus white to yellowish-brown, pale brown or greyish; stipe whitish,

without reddish, greenish or bluish tinges; spores 5-8 um in broad. ........... 5

5a. Pileus 15-25 mm in diam., hemispheric to convex, pale brown with

ANU BERER COMET ca react va atacticvaohe Lavaaats, 0k shy teen cegtbicwa dee Mavksshady Aa nq aa ee as L. acer

5b. Pileus 20-75 mm diam., conic-convex, campanulate, umbonate in maturity,

white: greyish to vellowish DrOWwHE:, toaacton tetas cootok ee tok eater shad ark chased s 6

272 ... Niveiro, Popoff & Alberté

6a. Pileus covered by ochraceus squamules, darker in the center;

stipe greyish white to yellowish, smooth or covered by

SWALLENE CS CPATATUL SS sasu<e arits gird setiny = opie et wasiehe getienee rains Laas ces L. cepistipes

6b. Pileus covered by white, floccose squamules; stipe covered by white

floccose squamules from the annulus to the base .................. L. cretaceus

Acknowledgments

The authors wish to thank J. Birkebak and M. Capelari for the critical revision

of the manuscript, and E.C. Vellinga for the corrections of a previous version of this

manuscript. This research was made possible by support from Reserva de Biosfera

Yaboty, MERNRyT - Proyecto Regional Araucaria XXI, Bosque Atlantico - AECID;

Myndel Botanical Foundation; SaCyT (UNNE) and the Argentine National Research

Council (CONICET).

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http://dx.doi.org/10.5248/112.83

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Capelari M, Maziero R. 1988. Fungos macroscépicos do estado de Rondo6nia, regiao dos Rios Jaru

e Ji-Parana. Hoehnea 15: 28-36.

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MY COTAXON

http://dx.doi.org/10.5248/121.275

Volume 121, pp. 275-280 July-September 2012

Two new species of Lasionectria (Bionectriaceae, Hypocreales)

from Guadeloupe and Martinique (French West Indies)

CHRISTIAN LECHAT? & JACQUES FOURNIER?

'Ascofrance, 64 route de Chizé, F-79360 Villiers en Bois, France

*Las Muros, F-09420 Rimont

*CORRESPONDENCE TO: lechat@ascofrance.fr

ABSTRACT —Lasionectria marigotensis sp. nov. on decaying leaves of Cocos nucifera (Arecaceae)

in Guadeloupe and L. martinicensis sp. nov. on dead stems of Passiflora sp. (Passifloraceae) in

Martinique are described and illustrated. The acremonium-like asexual state was obtained in

culture for both species. An updated key to the species of Lasionectria is provided.

KEY worps —Ascomycota, neotropics, palm fungi, taxonomy

Introduction

The genus Lasionectria (Sacc.) Cooke is based on the lectotype L. mantuana

(Sacc.) Cooke, designated by Clements & Shear (1931). The ascomata of

Lasionectria are yellow, pale orange, red-orange or dark brown, and do not

obviously change colour in 3% KOH or lactic acid; thus Lasionectria belongs to

the Bionectriaceae Samuels & Rossman as defined by Rossman et al. (1999). The

genus is distinguished from other genera in the Bionectriaceae by the ascomatal

wall over 20 um thick and composed of thick-walled cells with a small lumen

and hairs scattered over the ascomatal surface. Hairs may be stiff or flexuous,

solitary or fasciculate but do not form a distinct apical crown. The most similar

genus is Ijuhya Starback, which differs mainly in having ascomata with a wall

less than 20 um thick and typically with a discoid flattened apex fringed by

triangular fascicles of hairs, or rarely without hairs (Rossman et al. 1999). Both

genera have acremonium-like anamorphs in culture and occur on various

woody or herbaceous substrates.

In the course of an ongoing research program on the fungal diversity of

Lesser Antilles, «Les champignons des Petites Antilles; diversité, écologie,

protection», conducted by Prof. R. Courtecuisse (Courtecuisse 2006),

two bionectriaceous ascomycetes were collected that permitted a detailed

276 ... Lechat & Fournier

morphological characterization and successful single ascospore isolations.

Based on perithecia not changing color in 3% KOH or lactic acid, ascomatal

wall composed of two regions with the outer region composed of thick-walled

cells with a small lumen, presence of hairs on the surface and comparison with

known species in the genus, these specimens were determined to represent

previously undescribed species of Lasionectria. Both specimens were cultured

from single ascospores and produced acremonium-like asexual state.

Materials & methods

Specimens were examined using the methods described by Rossman et al. (1999).

Microscopic observations and measurements were made in water and the ascospore

ornamentation was observed in cotton blue in lactic acid. Cultures were made from

single ascospores that were isolated on PDA (Difco™ Potato Dextrose Agar). The

holotype specimens are deposited in LIP herbarium (Lille) and cultures at CBS.

Taxonomy

Lasionectria marigotensis Lechat & J. Fourn., sp. nov. PLATE 1a-f

MycoBank MB 564150

Differs from known Lasionectria species in white to pale orange ascomata with erect

non-fasciculate hairs evenly scattered on surface and relatively small, smooth-walled

ascospores.

Type: French West Indies, Guadeloupe, Vieux Habitants, Marigot, [Anse a la barque,

on decaying leaf of Cocos nucifera L. (Arecaceae), 3 Aug. 2011, leg. Christian Lechat

CLLGUAD 11002 (Holotype, LIP; ex-type culture CBS131606).

Erymo.ocy: The epithet is derived from Marigot, the locality where this species was

collected.

PERITHECIA gregarious, superficial, subglobose, (130-—)160-180(-200) um high

x (120-)130-150(-180) um diam. ("= 170 x 145 um, n = 10), white to pale

orange, collapsing cupulate, shining when dry, not changing color in 3% KOH

or lactic acid, hairs erect to flexuous covering perithecial surface. PERITHECIAL

APEX conical, composed of a palisade of cylindrical cells rounded at tip. Harrs

14-47 um long, 3-3.5(-4) um wide, hyaline, cylindrical, thick-walled (1 um),

rounded at tip, aseptate. PERITHECIAL WALL 20-25(-30) um thick, composed

of two regions: outer region 15-20 um wide, of globose to ellipsoidal 2.5-7

x 2-4.5 um cells, with pale yellow walls 1-1.5 um thick, each with a small

lumen; inner region 5-10 um wide, of elongate, flattened cells 4-11 x 1.5-2.5

um, with hyaline walls 0.5-0.8 um thick. Asci evanescent (30-)40-48(-53) x

(7-)8-9(-10) um (™ = 44.5 x 8.7 um, n=20), clavate, apices flattened, without

ring, with eight biseriate ascospores completely filling each ascus. ASCOSPORES

(9-)10-12.5(-13.5) x 3-3.5um (™ = 11.6 x 3.2 um, n=30), fusiform, slightly

curved, 1-septate, hyaline, smooth.

Lasionectria spp. nov. (French West Indies) ... 277

PLaTE 1. Lasionectria marigotensis (holotype). a. Perithecia. b. Hairs. c. Culture. d. Perithecium

in water. e. Ascospores. f. Median section of perithecial wall. Lasionectria martinicensis

(holotype). g—-h. Perithecium. i. Ascus and ascospores. j. Median section of perithecial wall.

k. Striations dehiscing from the ascospores. L. Culture. Scale bars: a, g, h = 100 um; b = 30 um;

d = 50 um; e, i, k = 10 um; f, j = 20 um.

278 ... Lechat & Fournier

In Cutrure: After three weeks at 25°C on Difco PDA containing 5 mg/

L streptomycin, colony 5-7.5 cm diam, without coloration of the medium,

mycelium white to greyish, producing an Acremonium-like anamorph at white

margin of colony, conidiophores monophialidic, 18-34 um long, 2-2.5 um

diam with 1-septum at base, finely spinulose, arising from smooth hyphae 2-2.8

um diam, conidia cylindrical to widely ellipsoidal (3.5-)4—6(-6.5) x 2-2.8 um

("=5.4 x 2.5 um, n = 30), hyaline, smooth, non-septate, with a basal abscission

scar.

Lasionectria martinicensis Lechat & J. Fourn., sp. nov. PLATE 1g-]

MycoBank MB 563442

Differs from known Lasionectria species in pale orange ascomata with erect non-

fasciculate hairs evenly scattered on upper half and striate ascospores.

Type: French West Indies, Martinique, Schoelcher, Riviere Case Navire, on dead stems

of Passiflora sp. (Passifloraceae), 28 Aug. 2010, leg. Christian Lechat CLLMARO085

(Holotype, LIP; ex-type culture CBS129746).

ErymMo.ocy: The epithet is derived from the name of the island where this species was

collected.

PERITHECIA Solitary, superficial, subglobose to obpyriform, (200-—)220-270

(-285) um high x (160-)180-250(-270) um diam. ("= 250 x 220 um, n = 10),

pale orange to orange, collapsing laterally, not changing color in 3% KOH

or lactic acid, hairs erect scattered over the upper half of the perithecium.

PERITHECIAL APEX conical composed of palisades of cylindrical to narrowly

clavate cells. Hairs 40-75 um long, 6-7 um wide at base, attenuated at tip,

hyaline, cylindrical, thick-walled (1-1.7 um), rounded or acute at tip, septate.

PERITHECIAL WALL 25-35 um thick, composed of two regions: outer region

15-20 um wide, of globose to ellipsoidal 3.5-7 x 2-6 um cells, with pale

yellow walls 1.2-1.7 um thick, each with a small lumen; inner region 8-15 um

wide, of elongate, flattened cells 8-19 x 2.5-3.5 um, with hyaline walls 0.5-1

um thick. Ascr (65-)70-75(-80) x (8-)9-11(-12) um (™ = 72.5 x 10.5 um,

n = 20), clavate, apices rounded, without ring, with eight biseriate ascospores.

Ascosporss (14-)16-20(-22) x 3-3.7um (™= 18.9 x 3.4 um, n = 30), fusiform,

1-septate, hyaline, striate with striations finely verrucose and dehiscing from

ascospore when slightly crushed under the cover slip.

In Cutture: After two weeks at 25°C on Difco PDA containing 5mg/L

streptomycin, colony 3-3.5 cm diam, spreading a reddish brown coloration

in medium, mycelium white to pale yellow, producing an Acremonium-like

anamorph at margin of colony, conidiophores monophialidic, 42-55 um long,

3-4 um diam with 1l-septum at base, arising from smooth hyphae 2.5-3.8

um diam, conidia cylindrical to widely ellipsoidal (3.5-)4-6.5(-7) x 2- 4 um

™= 6.1 x 3.2 um, n = 30), hyaline, smooth, non-septate, with a basal abscission

scar.

Lasionectria spp. nov. (French West Indies) ... 279

Dichotomous key to species of Lasionectria

I ASCOspores:sthiated 1.3 ahi. g dikes ddihate ddihared dewey debeced diners d yan’ dngpewid gees 2

LCASCOSDOLESES TOOLING # Piast NIH IS AIM Teh MI tah AINE MUA IAAL RATE ETRE OREN A 5

2. Habitat aquatic; ascomatal wall yellow; ascospores 13-17 x 4.5-6 um... L. fournieri

2. Habitat terrestrial; ascomatal wall pale orange to orange red or orange brown ....... 3

SF Ascospores SLI 6 S=4UMO os hada bade ee edhe eid eds L. vulpina

SPASCOSPORCS LAT OCT, hn eka hon Me chen Min dich Wiel SSR ISO RR TAN le heer 4

4. Ascospores two-celled, 16-20 x 3-3.7 UM ........... eee eee ee eee L. martinicensis

4. Ascospores one-celled, 22.8-30 x 7.5-10.5 UM ........ eee eee eee L. calamicola

. Ascomata with scattered hairs 14-47 um long, evenly distributed,

not bound in fascicles; ascospores 10-12.5 x 3-3.5 um; on palm . . L. marigotensis

5. Ascomata with both scattered and fasciculate hairs; on other plant ............... 6

6. Ascomata with scattered hairs 12-18 um long and fasciculate hairs 30-36 um

long around the apex; ascospores 8.5-9.5 x 3-3.5 um; on wood..... L. mantuana

6. Ascomata with hairs up to 100 um long; ascospores 11-15 x 3-4 um;

ONAL RACE OUS:BUCTIS S069 dis aee g dibateg dinber-g dinberg dinaerg dinberg Ain bend thers L. sylvana

Discussion

The two species described herein meet all the key features of Lasionectria

as defined by Rossman et al. (1999). They differ from the most closely

morphologically related genus Ijuhya by ascomata lacking a discoid apex and

an apical crown of fasciculate hairs and by having an ascomatal wall more than

20 um thick, composed of globose thick-walled cells in the outer region.

Lasionectria marigotensis can be recognized by the combination of white

to pale orange ascomata with scattered erect hairs evenly distributed on the

ascomatal surface and the relatively small, smooth-walled ascospores. Its

occurrence on palm might be significant if further collections confirm a host

preference or specificity. Another Lasionectria species reported from palm is

L. calamicola J. Frohl. & K.D. Hyde, known from two collections from Australia

and Brunei Darussalam (Frohlich & Hyde 2000). This taxon clearly differs from

L. marigotensis in having orange brown ascomata bearing hairs arranged in

fascicles and one-celled ascospores with striate walls.

Lasionectria martinicensis is unambiguously distinguished from other

species based on its large ascospores with a conspicuously striate perispore

easily loosening from the epispore. Other species of Lasionectria known to have

striate ascospores are L. calamicola, L. fournieri Lechat, and L. vulpina (Cooke)

Rossman & Samuels. The former differs in having one-celled ascospores

22.8-30 x 7.5-10.5 um and in occurring on palm. Lasionectria fournieri occurs

exclusively on submerged wood and its ascospores are markedly wider than

those of L. martinicensis, while L. vulpina differs in having smaller ascospores

8-11 x 3-4 um.

280 ... Lechat & Fournier

Acknowledgments

The authors gratefully acknowledge Prof. Régis Courtecuisse (Laboratoire des

sciences végétales et fongiques, Faculté des sciences pharmaceutiques et biologiques,

Université de Lille 2, France) for having initiated, organized and carried out this

ambitious project of exploration of the fungal diversity of French West Indies and for

having associated us to this survey. We also thank Luc Legendre (DREAL of Guadeloupe)

and Jean Baptiste Schneider (Office National des Foréts of Martinique) for contributing

to funding the field work and the Société mycologique de France (S.M.E) for help with

organizing this research program. Felix Lurel (ACED) and Jean Pierre Fiard are thanked

for their scientific expertise and assistance in field work in Guadeloupe and Martinique

respectively. Dr Amy Rossman, Systematic Mycology & Microbiology Laboratory,

USDA-ARS, 10300 Baltimore Ave., Beltsville, Maryland, 20705 USA. and Dr Rosalind

Lowen P.O. Box 1223, Lincoln, NH 03251 USA are warmly thanked for their invaluable

scientific help and for having substantially improved this paper by their suggestions.

Literature cited

Clements FE, Shear CL. 1931. The genera of fungi, 2"¢ edition. HW Wilson, New York. 496 p.

Courtecuisse R. 2006. Liste préliminaire des Fungi recensés dans les Iles francaises des Petites

Antilles: Martinique, Guadeloupe et dépendances. 1. Basidiomycetes lamellés et affines

(Agaricomycetideae s.|.). Doc. Mycol. 34(133-134): 81-140.

Frohlich J, Hyde KD. 2000. Palm microfungi. Fungal Diversity Research Series 3: 1-393.

Lechat C. 2008. Lasionectria fournieri sp. nov. et son anamorphe Acremonium. Bull. Soc. mycol. Fr.

124(1-2) : 1-5.

Rossman AY, Samuels GJ, Rogerson CT, Lowen R. 1999. Genera of Bionectriaceae, Hypocreaceae

and Nectriaceae (Hypocreales, Ascomycetes). Studies in Mycology 42: 1-248.

MY COTAXON

http://dx.doi.org/10.5248/121.281

Volume 121, pp. 281-284 July-September 2012

Steccherinum straminellum, a new record for Italy

ALESSANDRO SAITTA’ & IRENEIA MELO?

"Dipartimento di Biologia Ambientale e Biodiversita, Universita di Palermo,

Via Archirafi 38, I-90123 Palermo, Italy

?Jardim Botanico (MNHN), CBA/FCUL, Universidade de Lisboa,

Rua da Escola Politécnica 58, 1250-102 Lisboa, Portugal

* CORRESPONDENCE TO: alessandro.saitta@unipa. it

ABsTRACT — The first Italian record of Steccherinum straminellum is reported from Sicily.

This collection represents the first record on wood of broadleaved trees. A description and

line drawing of the species based on the Sicilian specimens are provided along with notes on

the taxonomy, ecology, and distribution of this rare taxon.

Key worps —Aphyllophorales, Corticiaceae, wood-inhabiting fungi

Introduction

Knowledge of fungal diversity in Italy has increased remarkably over the

last five years (Venturella et al. 2011). Of the 1582 wood-decay fungi recorded

in Italy, 1241 represent basidiomycetes, including 770 aphyllophoroid fungi

(Saitta et al. 2011). Seventeen species of Steccherinum Gray are known from

Italy (Bernicchia & Gorjén 2010).

Steccherinum is characterized by odontoid hymenophore, encrusted

pseudocystidia, and smooth thin-walled inamyloid basidiospores. Recently, the

genus has been included in the family Meruliaceae along with the poroid genus

Junghuhnia (Larsson 2007). Steccherinum straminellum is a very rare species

previously collected only in Portugal (Bernicchia & Gorjon 2010, Melo 1995).

Originally described as Odontia straminella Bres. (Bresadola 1902), the species

was transferred to Steccherinum seventeen years ago (Melo 1995).

Materials & methods

The basidiomata were identified while fresh and microscopic features were observed

in Melzer’s reagent and solution of potassium hydroxide (KOH 5%) using a Leica

microscope DMLB. The spore measurements were based on 50 observations for each

specimen. The nomenclature is referred to MycoBank (http://www.mycobank.org).

Locality coordinates were derived from the 1:50.000 scale edition of the Official Map of

282 ... Saitta & Melo

the Italian State (I.G.M.I.), following the methodology proposed by Padovan (1994). The

collected specimens are preserved in the Herbarium Mediterraneum Panormitanum,

Palermo (PAL).

Taxonomy

Steccherinum straminellum (Bres.) Melo, Mycotaxon 54: 126. 1995. Fics 1-2

ee as

Fic. 1. Steccherinum straminellum, a. spores (PAL 02047 Mic);

b. basidia (PAL 02047 Mic); c. section through basidiome (PAL 02050 Mic);

d. section through apical part of a tooth (PAL 02050 Mic).

Steccherinum straminellum, new to Italy ... 283

Fic. 2. Steccherinum straminellum basidiome on fallen branch of Quercus ilex.

Basidiome annual, resupinate, effused, adnate, broadly attached, rhizomorphs

well developed, pileus surface white to pale-ochraceous, margin fimbriate,

hymenophore odontioid to hydnoid, aculei up to 0.5 mm long and up 0.2 mm

broad. Hyphal system dimitic, generative hyphae hyaline, thin-walled, 2.3-4.2

uum wide.

Pseudocystidia numerous, cylindrical with obtuse apex, strongly incrusted in

theapical part, 7-12 um wide, distributed along theaculei, sometimes protruding,

thick-walled, becoming thin-walled towards apex. Basidia subclavate, 15-17 x

4.5-5.5 um, with 4 sterigmata and basal clamps. Basidiospores 3.5-4(-4.5) x

2-2.2 um, smooth, ellipsoid to subcylindrical, thin-walled, inamyloid.

SPECIMENS EXAMINED: ITALY, SiciLy, PALERMO, Monte Petroso, I.G.M.I. 594123, 560

m, woodland dominated by Quercus ilex L., on fallen branches and trunk of Q. ilex, 03

Oct 2010, coll. A. Saitta (PAL 02047 Mic); 08 Mar 2011, coll. A. Saitta (PAL 02050 Mic);

Casa Lo Curto, I.G.M.I. 609122, 720 m, mixed wood of Q. suber L. and Q. virgiliana

(Ten.) Ten., on fallen branches of Q. virgiliana, 06 May 2010, coll. A. Saitta (PAL 02024

Mic).

Discussion

Steccherinum is represented by eighteen taxa in Europe, eight of which

(including our new record) are currently reported for Italy: S. bourdotii Saliba

& A. David, S. fimbriatum (Pers.) J. Erikss., S. litschaueri (Bourdot & Galzin)

J. Erikss., S. ochraceum (Pers.) Gray, S. oreophilum Lindsey & Gilb., S. robustius

(J. Erikss. & S. Lundell) J. Erikss., S. straminellum, and S. subcrinale (Peck)

Ryvarden (Bernicchia & Gorjén 2010; this paper). Steccherinum oreophilum,

S. robustius, and S. subcrinale are very rare in Italy (Bernicchia & Gorjon

2010).

284 ... Saitta & Melo

The morphological features of S. straminellum are very close to S. litschaueri.

The two species differ only in the filamentous margin and the length of aculei

that can reach 1 mm in S. litschaueri. Moreover, S. litschaueri has thinner

cystidia and subcylindrical to cylindrical, up to 5.5 um long spores (Eriksson

et al. 1984).

Previously S. straminellum had been collected only on conifers (Bresadola

1902; Melo 1995), but now it has been found growing on fallen branches of

hardwoods, Quercus ilex and Q. virgiliana, in two localities of Sicily (southern

Italy).

Based on this new finding, S. straminellum has a restricted southern

European geographical distribution (Sicily in southern Italy, Portugal). In Sicily,

S. ochraceum is the only common Steccherinum species. Our collection shows

again that oak branches and trunks seem to be the preferred Steccherinum

substrate in Sicily (Saitta et al. 2004).

Acknowledgments

The authors are grateful to Dr. Cristiano Losi for critical comments for supporting

herbarium and bibliographic research. The authors wish to thank Dr. Vladimir Antonin

(Brno, Czech Republic) and Dr. Leho Tedersoo (Tartu, Estonia) for critically reviewing

the manuscript. Thanks are also due to Mrs. Stefania Paglialunga for the drawing of

microscopic characters.

Literature cited

Bernicchia A, Gorjén SP. 2010. Corticiaceae s.]. Fungi Europaei. Vol. 12. Ed. Candusso, Alassio

(Italy). 631 p.

Bresadola D. G. 1902. Mycetes lusitani novi lecti a Cl. Camillo Torrend s. j. Atti Imper. Regia Accad.

Rovereto, Ser. 3, 8(2): 128-133.

Eriksson J, Hjortstam K, Ryvarden L. 1984. The Corticiaceae of North Europe. Vol 7. Fungiflora,

Oslo. pp. 1281-1449.

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59=69.

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Fungal biodiversity and in situ conservation in Italy. Plant Biosystems 145(4): 950-957.

http://dx.doi.org/10.1080/11263504/2011.633115

MYCOTAXON

http://dx.doi.org/10.5248/121.285

Volume 121, pp. 285-289 July-September 2012

A new species of Inonotus (Basidiomycotina, Hymenochaetales)

from tropical Yunnan, China

HAt-JIAo LI & SHUANG-Hu!I HE*

Institute of Microbiology, P.O. Box 61, Beijing Forestry University, Beijing 100083, China

* CORRESPONDENCE TO: heshh1981@yahoo.cn

ABSTRACT — Inonotus puerensis sp. nov. is reported from tropical China. It is characterized

by having a perennial habit, duplex context, a monomitic hyphal structure, presence of both

hyphoid setae and hymenial setae, and coloured basidiospores. Detailed descriptions with

illustration are provided for the new species, and its relationships with similar species are

discussed.

Key worps — Hymenochaetaceae, taxonomy, white rot, wood-inhabiting fungi

Introduction

Yunnan Province is located in southwestern China and very rich in fungal

diversity because of its abundant and diverse vegetation. Although around

300 wood-inhabiting fungi have been recorded in this area (Dai 2011, 2012),

including several recently published new polypore species (Wei et al. 2006; Yuan

& Dai 2008; Cui et al. 2009, 2011; Dai & Korhonen 2009; Dai & Yuan 2009;

Dai 2010), many taxa are still undescribed, especially the tropical species. In

2011 we surveyed wood-inhabiting fungi in tropical Yunnan and collected an

unknown species of Inonotus P. Karst. The species has a perennial habit, duplex

context, a monomitic hyphal structure, both hyphoid setae and hymenial setae,

and coloured basidiospores. Since Ryvarden’s (2005) monograph on Inonotus

was published, more new species have been described (Balezi & Decock 2009;

Dai 2010; Wu et al. 2012). After checking all existing names, we found none

matching all the above characters. Therefore, we propose here a new species,

Inonotus puerensis.

Materials & methods

Voucher specimens are deposited in the herbarium of Beijing Forestry University

(BJFC). The specimens were examined microscopically according to Cui et al. (2011).

Abbreviations include: IKI = Melzer’s reagent, KOH = 5% potassium hydroxide,

286 ... Li & He

CB = Cotton Blue, IKI- = inamyloid and nondextrinoid, CB- = acyanophilous, L = mean

spore length (arithmetical average of all spores), W = mean spore width (arithmetical

average of all spores), Q = variation in the L/W ratios between the specimens studied

(quotient of the mean spore length and the mean spore width of each specimen),

n = the number of spores measured from given number of specimens. In spore and seta

size ranges, the 5% of the measurements excluded from each end of the range are given

in parentheses. Special color terms follow Petersen (1996).

Taxonomy

Inonotus puerensis Hai J. Li & S.H. He, sp. nov. Fic. 1

MycoBank MB 564891

Differs from other Inonotus species by its perennial habit, duplex context, monomitic

hyphal structure, presence of both hyphoid setae and hymenial setae, and coloured

basidiospores.

Type: China. Yunnan Province, Puer, Caiyanghe Nature Reserve, on fallen angiosperm

trunk, 6.VI.2011 Dai 12241 (holotype, BJFC).

ETyMOLocy: puerensis (Lat.): referring to the locality name of Puer, Yunnan Province,

China.

FRUITBODY: Basidiocarps perennial, pileate, solitary, without odour or taste and

hard corky when fresh, becoming woody hard upon drying. Pilei applanate,

projecting up to 3 cm long, 4.5 cm wide and 6 mm thick at base. Pileal surface

fuscous when dry, velutinate to tomentum, concentrically sulcate and zoned;

margin yellowish brown, obtuse. Pore surface cinnamon when fresh, become

dark brown when bruised, clay-buff when dry, glancing; sterile margin distinct,

yellowish brown, up to 2 mm wide, bearing abundant hyphoid setae (by lens);

pores circular to angular, 7-9 per mm; dissepiments thin, entire, matted.

Context cinnamon, up to 2 mm thick, woody hard, duplex, with a black zone

between upper velutinate or tomentum and lower context; lower context woody

hard, up to 1.5 mm thick, upper velutinate or tomentum up to 0.5 mm thick; a

thick dark brown line present between context and tubes. Tubes concolorous

with pore surface, woody hard, distinctly stratified, up to 4 mm long.

HYPHAL STRUCTURE: Hyphal system monomitic, generative hyphae simple

septate, IKI-, CB-; tissue darkening but otherwise unchanged in KOH.

CONTEXT: Generative hyphae yellowish to brownish, thick-walled with a

wide lumen, more or less straight, unbranched, frequently septate, interwoven,

3.2-4.7 um in diam.; hyphoid setae prominent, not dominant, dark brown,

distinctly thick-walled with a narrow lumen, some parts subsolid, straight,

tapering to apex, 240-400 x 10-13 um; hyphae in the black zone dark brown,

distinctly thick-walled with a narrow lumen, dark brown, flexuous, strongly

agelutinate, interwoven, 3-4 um in diam.; hyphae in the tomentum brown,

thick-walled with a narrow to wide lumen, agglutinate, interwoven, 3-6 um

in diam.

Inonotus puerensis sp. nov. (China) ... 287

Le aa

ae Wy ae A

< Sn

10 ym

Fic. 1. Microscopic structures of Inonotus puerensis (holotype).

a: Basidiospores. b: Basidia and basidioles. c: Hymenial setae. d: Hyphoid setae from context.

e: Hyphoid setae from trama. f: Hyphae from trama. g: Hyphae from context.

TuBEs: Generative hyphae hyaline to yellowish brown, thin- to thick-walled,

rarely branched, frequently septate, straight, subparallel along the tubes, 2.5-3.9

um in diam; hyphoid setae not dominant, prominent, dark brown, distinctly

thick-walled with a narrow lumen, some parts subsolid, straight, tapering to

apex, 90-250 x 8-12 um. Hymenial setae frequent, subulate, tapering to apex,

dark brown, thick-walled, 25-42 x 7-11 um; cystidia and cystidioles absent;

basidia barrel-shaped, bearing four sterigmata and a simple septum at the base,

9-11 x 5-6 um; basidioles in shape similar to basidia, but slightly smaller.

288 ... Li & He

Spores: Basidiospores broadly ellipsoid, yellowish to golden yellow, thick-

walled, smooth, IKI-, CB-, (4.4-)4.5-5(-5.2) x (3.6-)3.7-4(-4.2) um, L = 4.78

um, W = 3.9 um, Q = 1.23 (n = 30/1).

TYPE OF ROT: White rot.

REMARKS: Inonotus puerensis is unique in the genus for its perennial habit,

duplex context, monomitic hyphal structure, presence of both hyphoid and

hymenial setae, and coloured basidiospores.

Macro-morphologically Inonotus puerensis resembles members of the

I. baumii complex by sharing pileate and perennial basidiocarps, abundant

hymenial setae, and thick-walled yellowish basidiospores (Dai 2010). However,

I. puerensis has a monomitic hyphal structure and hyphoid setae, while species

in the I. baumii complex have a dimitic tramal structure and lack hyphoid

setae.

Microscopically Inonotus puerensis is similar to I. indurescens Y.C. Dai, also

described from Yunnan Province (Dai & Zhou 2000). Both have a monomitic

hyphal structure, hyphoid setae, and yellowish spores, but I. indurescens is an

annual species with larger pores (5-7 per mm) and dentate dissepiments. In

addition, its hyphoid setae occur in the trama and only sometimes curve and

penetrate into hymenium (Dai & Zhou 2000).

Inonotus pseudoglomeratus Ryvarden, described from South America

(Ryvarden 2002), resembles I. puerensis in pileate basidiocarps, presence of

both hyphoid setae and hymenial setae, and coloured basidiospores. However,

I. pseudoglomeratus is an annual species with larger pores (4-6 per mm), shorter

hymenial setae (15-25 x 6-10 um), and bigger basidiospores (5-6 x 4-4.3 um,

Ryvarden 2002).

Acknowledgments

The authors would like to express their deep thanks to Prof. Yu-Cheng Dai (Institute

of Applied Ecology, Chinese Academy of Sciences) and Dr. Xiao-Yong Liu (Institute of

Microbiology, Chinese Academy of Sciences) for serving as pre-submission reviewers.

This study was supported by the Fundamental Research Funds for the Central

Universities (Project No. YX2010-22) and the National Natural Science Foundation of

China (Project Nos. 31070022 and 31000006).

Literature cited

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undescribed species from the Rwenzori Mountain range. Cryptogamie Mycol 30: 225-232.

Cui BK, Dai YC, Bao HY. 2009. Wood-inhabiting fungi in southern China 3. A new

species of Phellinus (Hymenochaetales) from tropical China. Mycotaxon 110: 125-130.

http://dx.doi.org/10.5248/110.125

Cui BK, Du P, Dai YC. 2011. Three new species of Inonotus (Basidiomycota, Hymenochaetaceae)

from China. Mycol Prog 10: 107-114. http://dx.doi.org/10.1007/s11557-010-0681-6

Inonotus puerensis sp. nov. (China) ... 289

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Divers 45: 131-343.

http://dx.doi.org/10.1007/s13225-010-0066-9

Dai YC. 2011. A revised checklist of corticioid and hydnoid fungi in China for 2010. Mycoscience

52: 69-79. http://dx.doi.org/10.1007/s10267-010-0068-1

Dai YC. 2012. Polypore diversity in China with an annotated checklist of Chinese polypores.

Mycoscience 53: 49-80. http://dx.doi.org/10.1007/s10267-011-0134-3

Dai YC, Korhonen K. 2009. Heterobasidion australe, a new polypore derived from the Heterobasidion

insulare complex. Mycoscience 50: 353-356. http://dx.doi.org/10.1007/s10267-009-0491-3

Dai YC, Cui BK, Yuan HS. 2009. Trichaptum (Basidiomycota, Hymenochaetales) from China with a

description of three new species. Mycol Prog 8: 281-287.

http://dx.doi.org/10.1007/s11557-009-0598-0

Dai YC, Zhou TX. 2000. A new species of Inonotus (Basidiomycotina) from Yunnan, southern

China. Mycotaxon 74: 331-335.

Ryvarden L. 2002. Studies in neotropical polypores 17. New neotropical Inonotus species. Synopsis

Fungorum 15: 70-80.

Ryvarden L. 2005. The genus Inonotus, a synopsis. Synopsis Fungorum 21: 1-149.

Wei YL, Dai YC. 2006. Three new species of Postia (Aphyllophorales, Basidiomycota) from China.

Fungal Divers 23: 391-402.

Wu SH, Dai YC, Hattori T, Yu TW, Wang DM, Parmasto E, Chang HY, Shih SY. 2012. Species

clarification for the medicinally valuable ‘sanghuang’ mushroom. Botanical Studies 53:

135-149.

Yuan HS, Dai YC. 2008. Polypores from northern and central Yunnan Province, Southwestern

China. Sydowia 60: 147-159.

MY COTAXON

http://dx.doi.org/10.5248/121.291

Volume 121, pp. 291-296 July-September 2012

A new species of Grammothelopsis

(Polyporales, Basidiomycota) from southern China

CHANG-LIN ZHAO & BAO-Kal Cul’

Institute of Microbiology, P.O. Box 61, Beijing Forestry University, Beijing 100083, China

* CORRESPONDENCE TO: baokaicui@yahoo.com.cn

ABSTRACT — A new polypore, Grammothelopsis subtropica, is described and illustrated

based on three specimens collected in Guangdong and Hunan provinces, southern China.

Macroscopically, the new species is characterized by an annual growth habit, resupinate

basidiocarps with cream pore surface, and large pores (1-2 per mm); microscopically, it

has a dimitic hyphal system with strongly dextrinoid and cyanophilous skeletal hyphae, and

its basidiospores are ellipsoid to oblong-ellipsoid, thick-walled, slightly dextrinoid, slightly

cyanophilous, and 12.7-15.2 x 4.9-5.9 um. In addition, dendrohyphidia are abundant

in dissepiments. An identification key to the worldwide species of Grammothelopsis is

provided.

Key worps — lignicolous and poroid fungi, Polyporaceae, taxonomy

Introduction

Grammothelopsis Julich was established by Julich (1982) and typified by

G. macrospora (Ryvarden) Julich. It is characterized by resupinate to effused

basidiocarps, shallow irregular pores, and large thick-walled variably dextrinoid

basidiospores. Grammothelopsis species occur mostly in tropical Africa and

America (Robledo & Ryvarden 2007, Ryvarden & de Meijer 2002), with one

species recently described from tropical China (Dai et al. 2011).

As a result of recent surveys of the diversity of wood-inhabiting fungi in

southern China, many new fungal species have been described from tropical

and subtropical areas of the country (Cui & Dai 2008, 2011; Cui et al. 2009,

2010, 2011; Dai 2012a; Dai & Korhonen 2009; Dai et al. 2003, 2004, 2009, 2010,

2011; Du & Cui 2009; Jia & Cui 2011; Li & Cui 2010; Ma et al. 2011; Yuan

2011; Yuan & Dai 2008a,b; Zhang et al. 2012; Zhou & Dai 2012). However, the

diversity and richness of tropical polypores in China are still not well known

(Dai 2012b), and there are still many unidentified specimens from tropical

China. During examination of southern Chinese polypore collections, an

292 ... Zhao & Cui

additional undescribed Grammothelopsis species was found, which is described

and illustrated in the present paper.

Materials & methods

The studied specimens are deposited at the herbaria of the Institute of Microbiology,

Beijing Forestry University (BJFC) and the Institute of Applied Ecology, Chinese

Academy of Sciences (IFP). The microscopic routine used in the study follows Dai

(2010). Sections were studied at magnifications up to x1000 using a Nikon Eclipse E

80i microscope and phase contrast illumination. Drawings were made with the aid

of a drawing tube. Microscopic features, measurements, and drawings were made

from slide preparations stained with Cotton Blue and Melzer’s reagent. Spores were

measured from sections cut from the tubes. In presenting the spore size variation, the

5% of measurements excluded from each end of the range are given in parentheses.

Abbreviations include: IKI = Melzer’s reagent, KOH = 5% potassium hydroxide,

CB = Cotton Blue, CB+ = cyanophilous, L = mean spore length (arithmetic average of

all spores), W = mean spore width (arithmetic average of all spores), Q = variation in the

L/W ratios between the specimens studied, n = number of spores measured from given

number of specimens. Special color terms follow Petersen (1996).

Taxonomy

Grammothelopsis subtropica B.K. Cui & C.L. Zhao, sp. nov. FIG. 1

MycoBank MB 564797

Differs from other Grammothelopsis species by an annual growth habit, resupinate

basidiocarps with creamy shallow large pores (1-2 per mm), ellipsoid to oblong ellipsoid,

slightly dextrinoid and slightly cyanophilous basidiospores, presence of dendrohyphidia,

and absence of hyphal pegs.

Type: China, Guangdong Province, Fengkai County, Heishiding Nature Reserve, on

fallen angiosperm branch, 1.VII.2010, Cui 9035 (holotype, BJFC).

EryMo_oey: subtropica (Lat.): refers to the species being distributed in the subtropics.

FRuITBODY — Basidiocarps annual, resupinate, adnate, soft corky, without

odor or taste when fresh, becoming corky upon drying, < 7.5 cm long, 1.6 cm

wide, 0.7 mm thick at center. Pore surface white to cream when fresh, cream

upon drying; pores round to angular, 1-2 per mm; dissepiments thin, entire.

Sterile margin narrow, white, < 1 mm wide. Subiculum cream, thin, ca. 0.2 mm

thick. Tubes concolorous with pore surface, corky, < 0.5 mm long.

HyPHAL STRUCTURE — Hyphal system dimitic; generative hyphae with

clamp connections; skeletal hyphae strongly dextrinoid in Melzer’s reagent,

CB+, tissues unchanged in KOH.

SUBICULUM — Generative hyphae infrequent, hyaline, thin-walled, 2.1-2.5

um in diam; skeletal hyphae dominant, hyaline, thick-walled with a wide to

narrow lumen, frequently branched, flexuous, interwoven, 2.3-2.8 um in

diam.

Grammothelopsis subtropica sp. nov. (China) ... 293

Lj y J =, IF

SOK

\¢ .

SS <i A|

~~ Z YJ\ "GZ ig

f SS SG NZ

\S ! Z Za \ \ él \ \ \)

VRS TN

“Tl

FiGuRE 1. Grammothelopsis subtropica (holotype) microscopic structures.

a: Basidiospores. b: Basidia and basidioles. c: Cystidioles.

d: Dendrohyphidia. e: Hyphae from trama. f: Hyphae from subiculum.

294 ... Zhao & Cui

TuBES — Generative hyphae infrequent, hyaline, thin-walled, 2-2.3 um

in diam; skeletal hyphae dominant, hyaline, thick-walled with a wide to

narrow lumen, frequently branched, flexuous, interwoven, 2-2.5 um in diam.

Dendrohyphidia abundant in dissepiments, hyaline, thin-walled, < 45 um long.

Cystidia absent, fusoid cystidioles present, hyaline, thin-walled, 28.1-36 x

4.2-6.4 um; basidia clavate to pear-shaped, with four sterigmata and a basal

clamp connection, 36.5-39.1 x 8.9-9.8 um; basidioles dominant, mostly pear-

shaped, slightly smaller than basidia.

Spores — Basidiospores ellipsoid to oblong-ellipsoid, hyaline, thick-walled,

smooth, slightly dextrinoid in Melzer’s reagent, weakly CB+, (11.2-)12.7-

15.2(-16) x (4.7-)4.9-5.9(-6.1) um, L = 13.7 um, W = 5.3 um, Q = 2.4-2.7 (n

= 90/3).

ADDITIONAL SPECIMENS EXAMINED: CHINA. GUANGDONG PROVINCE, FENGKAI

County, Heishiding Nature Reserve, on fallen angiosperm branch, 1.VII.2010, Cui

9041 (BJFC); HUNAN PROVINCE, YIZHANG County, Mangshan Nature Reserve, on

fallen angiosperm trunk, 24.V1.2007, Li 1662 (IFP).

Discussion

Six species have been recorded in Grammothelopsis: G. bambusicola

Ryvarden & de Meijer, G. incrustata A. David & Rajchenb., G. neotropica

Robledo & Ryvarden, and G. puiggarii (Speg.) Rajchenb. & J.E. Wright were

found in Central and South America (David & Rajchenberg 1985; Rajchenberg

& Wright 1987; Robledo & Ryvarden 2007; Ryvarden & de Meijer 2002),

G. macrospora in tropical Africa (Ryvarden & Johansen 1980), and G. asiatica

Y.C. Dai & B.K. Cui in tropical China (Dai et al. 2011).

Grammothelopsis bambusicola has a dimitic hyphal system with strongly

dextrinoid skeletal hyphae and presence of dendrohyphidia. However, it differs

from G. subtropica in having smaller pores (4 per mm) and strongly dextrinoid,

wider basidiospores (11-13.5 x 7.8-9 um: measured from type specimen by

Dai et al. 2011).

Grammothelopsis macrospora may be confused with G. subtropica due to

its resupinate basidiocarps with larger pores (1-2 per mm) and presence of

dendrohyphidia, but it is distinguished from G. subtropica by its non-dextrinoid

and unbranched skeletal hyphae. In addition, its basidiospores are strongly

dextrinoid and larger (15-20 x 7.5-11 um: Robledo & Ryvarden 2007).

Grammothelopsis incrustata and G. neotropica also have resupinate

basidiocarps and a dimitic hyphal system, but both species have smaller

pores (3-4 per mm), non-dextrinoid skeletal hyphae, and distinctly larger

basidiospores (G. incrustata, 16-22 x 6-8 um; G. neotropica, 18-20 x 7-8 um:

Robledo & Ryvarden 2007).

Grammothelopsis puiggarii shares with G. subtropica larger pores (1-2 per

mm) and a dimitic hyphal system with strongly dextrinoid skeletal hyphae, but

Grammothelopsis subtropica sp. nov. (China) ... 295

G. puiggarii has larger basidiospores (17-20 x 10-12 um) and lacks dendro-

hyphidia (Robledo & Ryvarden 2007).

Grammothelopsis asiatica resembles G. subtropica in resupinate basidiocarps,

a dimitic hyphal system, and similar basidiospores (10.5-13 x 5.4-6 um).

However, G. asiatica has smaller pores (3-4 per mm), its skeletal hyphae and

basidiospores are non-dextrinoid, and it lacks dendrohyphidia (Dai et al.

2011).

Key to the species of Grammothelopsis

HeWMend toy pHidis PLesenil. nz, Mewatp. tesa tps ewer Mame eu taawa te awn teawn maar team 2

Lendrohyphidia'absent=. 112s (clear ro tags aoe oka oes (cP achat tay iad tude 4

2s Pores Baa Perini es 2s eve ey Seis nn Mes x5 Mela Seneinys Selene Wola 03 Baur G. bambusicola

Ze OES Le CE ELEM shee Steet er Sindy sed seine! S 9 ina! S oy oes attest ptatnsd attention ese ated 3

3. Basidiospores >15 um long, hyphal pegs present .................. G. macrospora

3. Basidiospores <15 um long, hyphal pegs absent..................... G. subtropica

4oGenerative hyphae simple-septater <M tinct s Peele e smaies sensi ya G. incrustata

4. Generative hyphae with clamp COnNecti ons. 3:5. 2c ce ce 8 eco a ore wee sole a 6

5. Pores 1-2 per mm, basidiospores strongly dextrinoid................. G. puiggarii

5: Pores'3—4 per mim, basidiosporés non-dextrinoid:: » sa... samaed mance y Sete ath wales 6

GL Basidi@spores: LS i LONG 2 asi0 a5tasin, a ecatcses raat cesar Sas a G. neotropica

6 Basidiospores <5 (id VO as acu ockeg dei wckeg dh pkeg chek phere doshas dondros. tucnoans G. asiatica

Acknowledgments

We express our gratitude to Drs. Tatiana B. Gibertoni (Brazil) and Hai-Sheng Yuan

(China), who reviewed the manuscript. The research is financed by the Fundamental

Research Funds for the Central Universities (Project No. BLYJ201205) and the National

Natural Science Foundation of China (Project Nos. 30900006 and 31093440).

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MY COTAXON

http://dx.doi.org/10.5248/121.297

Volume 121, pp. 297-304 July-September 2012

Two new species of white truffle from China

Li FAN” & JIN-ZHONG CAO?

' College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

? Institute of Mycology, Jilin Agricultural University, Changchun 130118, China

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

ABSTRACT — Two new white truffle species from China are described and illustrated. Tuber

pseudomagnatum is characterized by yellow white ascomata with blackish gleba and small

elliptic ascospores with a large meshed reticulum. Tuber liyuanum is separated from other

species by its white ascomata, brown gleba, and elliptic to long elliptic ascospores with a

regular reticulum. ITS based sequence analyses support the erection of the two new species.

KEY worps — Ascomycota, Tuberaceae, taxonomy

Introduction

Southwestern China is a place where new truffle species are being found ona

regular basis. Recently, we were sent 4 kilos of fresh white truffles from Yunnan

Province by someone who has been involved in the Chinese truffle business

for more than a decade. ‘The truffles gave off a not unpleasant, strong, pungent

aroma, but after boiling in 3% salt-water solution, they developed a pleasant

flavor. These truffles could be divided into two different groups according to

the exterior characteristics of ascomata. One was yellow white and tasted a

little sweet, while the other was pure white (at least at the furrows) and tasted

sweet. They resembled several known white truffles, such as Tuber latisporum

Juan Chen & P.G. Liu (Chen & Liu 2007) from China, T: magnatum Picco and

T. borchii Vittad. from Europe (Riousset et al. 2001), and T. oregonense Trappe et

al. and T’ gibbosum Harkn. from North America (Bonito et al. 2010). However,

detailed morphological observation and DNA analysis revealed them as new

species, which we describe here.

Materials & methods

Morphological studies

The truffles were collected under conifers or mixed woodlands in Huize county,

Yunnan Province, China. Macroscopic characters were described both from fresh and

rehydrated dried specimens, and microscopic characters from razor-blade sections

298 ... Fan & Cao

mounted in 3% KOH (w/v), Melzer’s reagent and 0.1% (w/v) cotton blue in lactic acid.

For scanning electron microscopy (SEM), spores were scraped from the dried host

gleba onto doubled-sided tape, which was mounted directly on an SEM stub and coated

with gold-palladium. The treated materials were examined and photographed with a

HITACHI S-4800 SEM. The specimens were deposited in BJTC (Herbarium of Biology

Department, Capital Normal University, Beijing, China).

Molecular methods

Samples from herbarium material 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 then extracted using

the PeqLab E.Z.N.A._ Fungal DNA kit following the manufacturer’s protocol. The ITS

regions were amplified with PCR using the primers ITS1/ITS4 (White et al. 1990). PCR

was performed in 50 ul reactions containing DNA template 2 ul, primer (10 uM) 2 ul

each, and 2 x Master Mix (Tiangen Biotech (Beijing) Co. Ltd.) 25 ul. PCR reactions

were run as follows: an initial denaturation at 95 °C for 3 min, followed by 30 cycles

at 95 °C for 2 min, 55 °C for 25 s, 72 °C for 2 min and a final extension at 72 °C for 10

min. The PCR products were sent to Invitrogen Biotechnology Co. Ltd. (Beijing, China)

for purifying, sequencing, and editing. The other ITS rDNA sequences included in this

study (TABLE 1) were downloaded from GenBank.

TABLE 1. Tuber ITS rDNA sequences used in study.

SPECIES (VOUCHER SPECIMENS) ITS

T. borchii HM485344

HM485343

T. dryophilum Tul. & C. Tul. HM485353

HM485354

T. gibbosum FJ809868

T. latisporum DQ898185

DQ898183

T. liui AS. Xu DQ898182

T. liyuanum (BJTC FAN 162, holotype) JQ771191

(BJTC FAN 187) JQ771193

T. maculatum Vittad. FM205649

EU784428

T. magnatum AJ586308

EU807975

T. melanosporum Vittad. AF132501

AF106878

T. oligospermum (Tul. & C. Tul.) Trappe FM205507

FM205506

T. oregonense FJ809881

FJ809882

FJ809870

T. puberulum Berk. & Broome AJ969626

AJ969625

T. pseudomagnatum (BJTC FAN 163, holotype) JQ771192

T. scruposum R. Hesse DQ011846

DQ011845

T. zhongdianense X.Y. He et al. DQ898187

DQ898186

Tuber pseudomagnatum & T. liyuanum spp. nov. (China) ... 299