IV ... MYCOTAXON 116

MY COTAXON

VOLUME ONE HUNDRED SIXTEEN — TABLE OF CONTENTS

COVER SECTION

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SUDMUSSIONPTOCCM UTES SLA. Von NST, SSO a ES Le Poot Meat eee ie LANs xi

RESEARCH ARTICLES

First record of Leptomeliola uvariae for South America

Michelline L. Silvério, Maria Auxiliadora Q. Cavalcanti & José L. Bezerra

Notes on the neotype of Tuber taiyuanense

Jin-Zhong Cao, Wei-Dan Chi, Li Fan & Yu Li

Wood-rotting fungi in eastern China 6. Two new species of Antrodia

(Basidiomycota) from Mt. Huangshan, Anhui Province

Bao-Kai Cui, Hai-Jiao Li & Yu-Cheng Dai

Three corticolous species of Lecanora (Lecanoraceae) new to China

Liu-Fu Han, Shou-Yu Guo & Hao Zhang

A new Dothidasteroma species on leaves of Psidium laruotteanum

from the Brazilian Cerrado

C.A. Inacio, R.C. Pereira-Carvalho, E.S.C. Souza & J.C. Dianese

The genus Pythium in Taiwan (2) — an illustrated diagnostic key Hon H. Ho

Lecanora, Phaeophyscia & Rinodina species new to Turkey

Belgin Arslan, Sule Oztiirk & Seyhan Oran

Four new records of Caloplaca (lichenized Ascomycetes) from India

Yogesh Joshi & Dalip K. Upreti

Contribution to the lichen mycota of South Korea Yogesh Joshi, Thi Thuy Nguyen,

Xin Yu Wang, Laszlé Lék6s, Young Jin Koh & Jae-Seoun Hur

Glomeromycetes: three new genera and glomoid species reorganized

Fritz Oehl, Gladstone Alves da Silva, Bruno Tomio Goto, & Ewald Sieverding

Tleodictyon gracile, new to Italy Alessandro Saitta, Riccardo Compagno,

Alfonso la Rosa, Danilo Lopez & Giuseppe Venturella 121

Observations on two rarely collected species of Russula

P. Manimohan & K.P. Deepna Latha 125

Syncephalis clavata (Zoopagales, Zygomycetes), a first record from the neotropics

Roger Fagner Ribeiro Melo, André Luiz C. M. de A. Santiago

& Maria Auxiliadora de Q. Cavalcanti 133

Jahnula aquatica and its anamorph Xylomyces chlamydosporus on submerged

wood in Thailand Somsak Sivichai, Veera Sri-Indrasutdhi & E.B. Gareth Jones 137

Glomus crenatum (Glomeromycetes), a new ornamented species from Cuba

Eduardo Furrazola, Yamir Torres-Arias, Roberto L. Ferrer,

Ricardo A. Herrera, Ricardo L.L. Berbara & Bruno Tomio Goto 143

APRIL-JUNE 2011... V

A new species of Xylaria from China Hai-Xia Ma, Larissa Vasilyeva & Yu Li 151

Dendrophlebia (Agaricomycetes), a new corticioid genus from India

G.S. Dhingra & Priyanka 157

Orbispora gen. nov., ancestral in the Scutellosporaceae (Glomeromycetes)

Fritz Oehl, Danielle Karla Alves da Silva,

Leonor Costa Maia, Natalia Mirelly Ferreira de Sousa,

Helder Elisio Evangelista Vieira & Gladstone Alves da Silva 161

Uncispora sinensis, a new species from China Guang-Zhu Yang, Jun Lu,

Zefen Yu, Keqin Zhang & Min Qiao 171

Puccinia species new to Azad Jammu and Kashmir, Pakistan

N.S. Afshan, A.N. Khalid, A.R. Niazi & S.H. Iqbal 175

Hydnoid basidiomycetes new to Brazil

Alice da Cruz Lima Gerlach & Clarice Loguercio-Leite 183

Phylogenetic relationships and reclassification of Spirosphaera lignicola,

an enigmatic aeroaquatic fungus Herman Voglmayr 191

Spiroplana centripeta gen. & sp. nov., a leaf parasite of Philadelphus and

Deutzia with a remarkable aeroaquatic conidium morphology

Hermann Voglmayr, Mi-Jeong Park, Hyeon-Dong Shin 203

On the variability of spore ornamentation in Laccaria tortilis

(Basidiomycota, Agaricales)

Alfredo Vizzini, Marco Contu, Kuulo Kalamees, Enrico Ercole,

Enzo Musumeci, Gabriel Moreno, J.L. Manjon & P. Alvarado 217

Validation of Kwoniella heveanensis, teleomorph of the basidiomycetous yeast

species Cryptococcus heveanensis

Sheng Sun, Banu Metin, Keisha Findley, Alvaro Fonseca & Joseph Heitman 227

Observations on Inocephalus virescens comb. nov. and Alboleptonia stylophora

from northeastern Queensland David L. Largent & Sandra E. Abell-Davis 231

The family Chaetomiaceae from China 4. Two newly recorded species

of Chaetomium Yunzhong Guo, Guangyu Sun & Mingqi Zhu 247

Saxicolous species of Claudopus (Agaricales, Entolomataceae) from Australia

David L. Largent, Sandra E. Abell-Davis,

Griffin A. Cummings, Kathryn L. Ryan & Sarah E. Bergemann 253

Notes on Trametes (Basidiomycota) in China Hai-Jiao Li & Shuang-Hui He 265

Notes on Amylocorticiellum (Amylocorticiales, Basidiomycota), with some new

combinations Sergio P. Gorjén, Alina G. Greslebin & Mario Rajchenberg 283

Melanoderma microcarpum gen. et sp. nov. (Basidiomycota) from China

Bao-Kai Cui, Chang-Lin Zhao & Yu-Cheng Dai 295

Zygomycetes from “Reserva Bioldgica de Mogi Guacu’, Sao Paulo State, Brazil

José Ivanildo de Souza, Carmen Lidia Amorim Pires-Zottarelli,

Jailson Francisco dos Santos & Janaina Pinheiro Costa 303

A new species and a new record of Herpothallon (lichenized Ascomycota) from India

T.A.M. Jagadeesh Ram & G.P. Sinha 313

Cortinarius xanthodryophilus sp. nov. - a common Phlegmacium under oaks

in California Dimitar Bojantchev & R. Michael Davis 317

VI... MYCOTAXON 116

Zwackhiomyces turcicus sp. nov. (Ascomycota, Xanthopyreniaceae) from Turkey

Mustafa Kocakaya, Mehmet Gokhan Halici & Ahmet Aksoy 329

Arthonia anatolica (Arthoniaceae) sp. nov. on Aspicilia contorta

subsp. hoffmanniana, a new lichenicolous species from Turkey

Mehmet Gokhan Halici & Mehmet Candan

A new species and a new record of Marasmius from China

Chun-Ying Deng, Tai-Hui Li & Bin Song

Two new species of Tuber from China

Li Fan, Jin-Zhong Cao, Yan-Yun Liu & Yu Li

AFLP molecular characterizations of some Saccobolus species

(Ascomycota, Pezizales) Araceli M. Ramos, Isabel E. Cinto,

Luis Franco Tadic & Maria Esther Ranalli

Glomeromycota: two new classes and a new order Fritz Oehl,

Gladstone Alves da Silva, Bruno Tomio Goto,

Leonor Costa Maia & Ewald Sieverding

First report of Morganella compacta (Agaricales, Lycoperdaceae)

from South America Marcos Mateus Barros Barbosa,

Maria Aparecida da Silva, Rhudson Henrique Santos Ferreira da Cruz,

Francisco de Diego Calonge, & Iuri Goulart Baseia

Two new taxa close to Lepiota cristata from China Jun F. Liang & Zhu L. Yang

Two new species of Septobasidium (Septobasidiaceae) from

Gaoligong Mountains in China Chunxia Lu & Lin Guo

Racocetra undulata sp. nov., a new species in the Glomeromycetes

from Taiwan Tzu-Chao Lin & Chiang-Her Yen

Notes on the identity of Chrysothrix populations (Arthoniales, Ascomycota)

containing pinastric acid from southern and central California

Martin Kukwa & Kerry Knudsen

Entoloma mastoideum and E. praegracile — two new species from China

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

New records of corticioid fungi in Turkey

Hasan Hiiseyin Dogan, Mitko Karadelev, Katerina Rusevska & Sinan Aktas

A new species of Hansfordia isolated from the marine brown alga,

Colpomenia sinuosae Xiao-Li Cheng, Kai-Ming Sun, Wei Li,

Tian-Yu Zhang & Chang-Lin Li

Cantharellus in southwestern China: a new species and a new record

Shi-Cheng Shao, Xiao-Fei Tian & Pei-Gui Liu

Passalora papaveris comb. nov. from China

Feng- Yan Zhai, Ying-Lan Guo, Ying-Jie Liu & Yu Li

Cladophialophora pucciniophila, a new hyphomycete parasitizing a rust fungus

Mi-Jeong Park & Hyeon-Dong Shin

Notes on Ceriporia (Basidiomycota, Polyporales) in China

Bi-Si Jia & Bao-Kai Cui

335

341

349

355

365

381

387

395

401

407

413

421

431

437

447

449

457

APRIL-JUNE 2011... VII

BOOK REVIEWS AND NOTICES Else C. Vellinga (EpIToR) 469

MYCOBIOTAS ONLINE

Abstracts of newly posted annotated regional species lists 479

NOMENCLATURE

1. Melbourne approves a new CODE Lorelei L. Norvell 481

2. The Amsterdam Declaration on fungal nomenclature

David L. Hawksworth’, Pedro W. Crous, Scott A. Redhead,

Don R. Reynolds, Robert A. Samson, Keith A. Seifert,

John W. Taylor, Michael J. Wingfield, & 80 signatories 491

3. A critical response to the Amsterdam Declaration

é Walter Gams, Walter Jaklitsch, & 77 signatories 501

4. Letter of concern regarding Props. (117-119) to amend the ICBN

to require pre-publication deposit of nomenclatural information

Paul J. Morris, James A. Macklin, Jim Croft,

Nicky Nicolson & Greg Whitbread 513

NOMENCLATURAL NOVELTIES proposed in volume 116 519

PUBLICATION DATE FOR VOLUME ONE HUNDRED FIFTEEN

MYCOTAXON for JANUARY-MARCH, VOLUME 115 (I-xII + 1-539)

was issued on May 9, 2011

vu ... MYCOTAXON 116

REVIEWERS — VOLUME ONE HUNDRED SIXTEEN

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.

Mehrdad Abbasi

Mohamed Ahmed

Abdel-Wahab

Hakan Alli

Joe Ammirati

Mikhail Andreev

Vladimir Antonin

Andre Aptroot

Hans-Otto Baral

Timothy J. Baroni

Niclas Bergius

Lina Bettucci

José Luiz Bezerra

Janusz Blaszkowski

Uwe Braun

Peter Buchanan

Bart Buyck

Matias J. Cafaro

Paul Francis Cannon

Johan C. Coetzee

Vagner Gularte Cortez

Danny Coyne

Florin Crisan

Yu-Cheng Dai

Eric Danell

Gregorio Delgado

Dennis E. Desjardin

Paul Diederich

Pradeep K. Divakar

John A. Elix

Jack W. Fell

Adam Flakus

Francisco Das Chagas

Oliveira Freire

Walter Gams

Genevieve M. Gates

Masoomeh

Ghobad-Nejhad

Bruno Tomio Goto

Anna Guttova

M. Gokhan Halici

Ian Robert Hall

Nils Hallenberg

Richard T. Hanlin

Shuang-Hui He

Chuanxue Hong

Tom Hsiang

Man-Rong Huang

Semra Ilhan

Paul M. Kirk

Roland Kirschner

Kerry Knudsen

Irmgard Krisai-Greilhuber

Wen-Hsiun Ko

Jana Kocourkova

Richard P. Korf

Cletus P. Kurtzman

James C. Lendemer

Shu- Yan Liu

Laszlé Lék6s

Guozhong Lit

Sandra Lupo

Eric H.C. McKenzie

Andrew S. Methven

Gabriel Moreno

Karen K. Nakasone

Sanjeeva Nayak

Maria Alice Neves

Machiel E. Noordeloos

Lorelei L. Norvell

Fritz Oehl

Shoji Ohga

Ka-Lai Pang

Shaun R. Pennycook

Olinto Liparini Pereira

Ronald H. Petersen

Marcin Piatek

Mario Rajchenberg

Scott A. Redhead

Bernard Rivoire

Jack D. Rogers

Adam W. Rollins

Gary J. Samuels

André Luiz C.M.

de A. Santiago

Ertugrul Sesli

B.M. Sharma

Ewald Sieverding

Danielle Karla

Alves da Silva

Viacheslav Spirin

Steven L. Stephenson

Ave Suija

Evrim Taskin

Atti Tchabi

Goran Thor

Michal TomSovsky

Ludmilla Fritri Untari

Sandra Farto

Botelho Trufem

Aysen Ozdemir Tiirk

Else C. Vellinga

Alfredo Vizzini

Yei-Zeng Wang

Zheng Wang

Anthony J.S. Whalley

Merlin White

Zhu L. Yang

Yi-Jian Yao

Hai-Sheng Yuan

Georgios Zervakis

Meng Zhang

Zhongyi Zhang

Guozhu Zhao

Ru-Yong Zheng

APRIL-JUNE 2011... IX

FROM THE EDITOR-IN-CHIEF

VANISHING WATERMARKS AND SUBSCRIBERSHIP PROTOCOLS — One advantage of an

online subscription is that PDFs are portable, searchable, and take up no room in the

luggage, allowing research to continue even during field expeditions and sabbaticals.

However, electronic publication has one drawback for the publisher: PDFs are too

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as the MycoTaxon FREE PRESS - exacting no page charges and offering the entire

journal online for free — the unfortunate truth is that we do require subscriptions to

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spend thousands of hours corresponding with authors and reviewers, accessioning and

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monthly or Mycotaxon will disappear.

This explains why we decided to use a watermark to remind our subscribers not

to share their online subscriptions with non-subscribers. Unfortunately, the first to

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the watermark in the middle of each page highly distracting. One subscriber wrote to us

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INGENTA I may be a potentially criminal violator of copyright laws who needs a deterrent

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After learning that we had no option to restrict the watermark to the first page, we

decided to remove it entirely. In return, we request our subscribers to honor the same

and non-subscriber alike — to download the many free and open access papers in each

volume paid for by MycoTaxon and its authors.

LATE AGAIN! Despite our best hopes and intentions, preparation for and attendance

of the International Botanical Congress in Melbourne (July 18-30) by the Ep1ror-1n-

CureFr and NOMENCLATURE EpITor interfered with the normal publication schedule,

severely delaying the April-June volume in the process.

The 18" IBC was significant and definitely warranted our attention, as the significant

nomenclatural decisions ratified by the Melbourne Congress show. Among the important

amendments to the CopE summarized in MycoTaxon 116 (pp. 481-490) are valid

electronic publicatiion of names, permission to use English in diagnoses, recognition

of “one fungus : one name” and the radical revision of Art. 59, required registration of

fungal names for valid publication, and a change of title to the INTERNATIONAL CODE OF

NOMENCLATURE FOR ALGAE, FUNGI, AND PLANTS.

With no other major delays anticipated before the end of the year, MycoTaxon still

hopes to publish volumes 117 and 118 in 2011, bringing the journal back on schedule

by January, 2012.

Warm regards, Lorelei Norvell

Mycotaxon Editor-in-Chief

26 August 2011

x ... MYCOTAXON 116

ERRATA FROM PREVIOUS VOLUMES

VOLUME 114

p.343 (bottom line)-344, lines 1-2:

for: However, only two species of Hygrocybe in section Firmae have been described so far

from India: H. alwisii (Berk. & Broome) Pegler from Kerala (Leelavathy et al. 2006), and

H. natarajanii Senthil. & Kumaresan from Karnataka (Senthilarasu et al. 2010).

read: Six species representing Hygrocybe sec. Firmae have been reported from India:

H. hypohaemacta (Vrinda et al. 1996); H. batistae Singer, H. diversicolor (Petch) Pegler,

H. martinicensis Pegler & Fiard (Vrinda et al. 2009); H. alwisii (Berk. & Broome) Pegler

(Leelavathy et al. 2006); and H. natarajanii Senthil. & Kumaresan (Senthilarasu et al.

2010).

p. 349, after bottom line add:

Vrinda, KB, Pradeep CK, Abraham TK. 1996. Agaricales from Western Ghats - III.

Mushroom Research 5: 47-50.

Vrinda KB, Pradeep CK, Varghese SP. 2009. Noteworthy records of Hygrocybe section

Firmae from Western Ghats of Kerala. Mushroom Research 18(1): 1-4.

p.487, line 4in banner for: October-December 2011 read: October-December 2010

p-500, line 4in banner for: October-November 2011 read: October-December 2010

VOLUME 115

p.539, line 22 —_ for: Protoparmeliopsis klauskalbii (Sipman) Senkard., p. 288

read: Protoparmeliopsis klauskalbii (Sipman) Senkard., p. 266

APRIL-JUNE 2011... XI

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MY COTAXON

Volume 116, pp. 1-5 April-June 2011

DOI: 10.5248/116.1

First record of Leptomeliola uvariae for South America

MICHELLINE L. SILVERIO’, MARIA AUXILIADORA Q. CAVALCANTI’

& JOSE L. BEZERRA”

‘Universidade Federal de Pernambuco, Departamento de Micologia,

Av. Prof. Nelson Chaves, s/n°, CEP 50670-901, Recife, PE, Brazil

?Universidade Estadual de Santa Cruz, Rodovia Ilhéus-Itabuna,

Km 16, CEP 45662-900, Ilhéus, BA, Brazil

CORRESPONDENCE TO *: silverio_ml@hotmail.com,

xiliamac@terra.com.br, *jlulabezerra@hotmail.com

ABsTRACT — During a mycological expedition to the “Reserva Ecolégica de Saltinho,

Municipality of Tamandaré, Pernambuco State, Brazil, the ascomycete Leptomeliola uvariae

was found on leaves of the fern Lygodium volubile, parasitizing the mycelium ofa meliolaceous

fungus. This is the first report of Leptomeliola uvariae from South America.

Key worps — Ascomycota, epiphytic fungi, hyperparasite, Atlantic Forest

Introduction

Hohnel (1919) described the genus Leptomeliola Hohn. (Ascomycota,

Parodiopsidaceae) as a hyperparasite of meliolaceous fungi. Of the 14 names

published in Leptomeliola cited by Index Fungorum (2010), five are muscicolous

species (Racovitza 1959) that should probably be excluded from the genus, while

three others should be placed in synonymy (Hughes 1993). The six accepted

species are L. cryptocarpa [= L. anomala], L. cymbisperma [= L. hyalospora, the

type species], L. puberula, L. quercina, L. torta [= L. javensis], and L. uvariae.

Only two species have previously been recorded from Brazil: L. cryptocarpa

(in Sao Paulo, Amazonas and Rio Grande do Sul States) and L. puberula (in

Pernambuco State) (Batista 1951, Hughes 1993).

Species of the family Meliolaceae are biotrophic parasites, which are usually

foliicolous and occur worldwide on phanerogam and pteridophyte hosts. These

fungi are frequently parasitized by anamorphs and other Ascomycota (Hansford

1946, 1961, Hughes 1993).

Below we describe and illustrate L. uvariae and report its first occurrence in

South America.

2 ... Silvério, Cavalcanti & Bezerra

Materials & methods

The collection site lies in the ‘Reserva Bioldgica de Saltinho (8°43'55"S 35°10'26"W),

situated by PE-60 near Tamandaré Municipality, South Coast of Pernambuco State. The

548-hectare Atlantic Rain Forest reserve has a 12-36°C temperature range and annual

rainfall up to 3000 mm (MUSEU DO UNA 2008).

During February 2010, living leaves of Lygodium volubile with black mildew

symptoms were collected. The leaves were taken to the laboratory of the ‘Departamento

de Micologia da Universidade Federal de Pernambuco’ and examined under the

stereomicroscope. Fruiting bodies were manually scraped, cut, and mounted in PVLG +

cotton blue. Slides were examined under a light microscope Leica DM50 equipped with

a digital camera and fungal structures of taxonomic value were studied. Species were

identified by comparing descriptions of similar ascomycetes parasitizing Meliolaceae

(Hansford 1946, Miller & Arx 1962, Arx & Miiller 1975, Hughes 1993).

In order to isolate Spiropes capensis, conidia were cultured on potato-dextrose-

agar medium (PDA) amended with chloramphenicol 250 ug.ml' for seven days under

ambient laboratory conditions (28 + 2°C, 12 h light and 12 h darkness).

Taxonomy

Leptomeliola uvariae (Rehm) S. Hughes, Mycological Papers 166: 203 (1993).

Fics 1-8

SPECIMEN EXAMINED: BRAZIL. PERNAMBUCO: Tamandaré Municipality, on

Meliolaceae undetermined, on living leaves of Lygodium volubile Sw. (Schizaeaceae), 08

February 2010, Michelline L. Silvério (URM 82258).

COLONIES amphigenous, superficial, composed by subhyaline to pale brown

hyphae, 2.7—4 um thick. PSEUDOTHECIA isolated, brown, globose to subglobose,

190-270 x 170-250 um, ostiolate, setose; setae simple, straight, cylindrical,

obtuse apex, septate, dark brown, 107.5-212.5 x 7.5-8.75 um; peridium

40-45 um thick, composed by many layers of thick-walled angular cells. Asc1

bitunicate, short-stipitate, clavate, 8-spored, 92.5-115 x 35-45 um; paraphyses

absent. Ascosporgs clavate, hyaline becoming brown, smooth, 33-40 x

11-14 um, 3-septate, strongly constricted at the middle septum, two superior

cells broader.

GEOGRAPHICAL DISTRIBUTION: Philippines, Uganda, Ghana, Sierra Leone, Congo(-

Kinshasa), Puerto Rico, Brazil.

Notes: In the same material, conidiophores of Spiropes capensis (Thitm) M.B.

Ellis seemingly developing from the mycelium of L. uvariae were observed;

these were solitary or aggregated, cylindrical, septate, brown to dark brown,

becoming paler at the tip, straight or flexuous, 175-347.5 x 6.25-8.75 um,

with conidial scars; conidia straight, fusiform, mucronate, olivaceous light

brown to brown, smooth, straight, with 3 to 5 transverse pseudosepta, formed

sympodially on the conidiophore apex, 38-54 um long, 7-12 um wide in the

broadest region, 2—4.5 um at the apex and 3-6 um at the base (Fics 9-13).

Leptomeliola uvariae new to South America ... 3

Fic. 1-8: Leptomeliola uvariae— 1. Pseudothecia in colonies on leaves of Lygodium volubile;

2. Vertical section of pseudothecium; 3. Setose pseudothecium; 4. Setae; 5. Asci and ascospores;

6-8. Ascospores. Fic. 9-13: Spiropes capensis— 9. Conidiophores on the leaf surface;

10-11. Conidiophores and young conidium; 12-13. Mature conidia.

4 ... Silvério, Cavalcanti & Bezerra

TABLE 1. Characteristics of known species of Leptomeliola.

SPECIES

am diam.) im)

PSEUDOTHECIA

ASCI

ASCOSPORES

Hosts

REFERENCES*

L. cryptocarpa 150-350: 80-100 x 30-50 x Meliola sp.; Hughes (1993);

(Ellis & globose to 25-35, 10-12, Irene sp. Hansford (1946,

G. Martin) flattened, clavate 3-5septate, as Ph. meliolicola)

S. Hughes hairy to saccate clavate

= L. anomala

L. cymbisperma Minute: Clavate- 27-36 x Asteridiella sp. Hughes (1993);

(Mont.) spherical, pyriform 8.5-10, Hohnel (1919)

S. Hughes +/-setose (as (as L. 3septate,

= L. hyalospora L.hyalospora) —_hyalospora) _ ellipsoid

L. puberula 150-212.5: 47.5-57.5x 25-32.5 x Not available Batista (1951)

Bat. globose, 20-25, 5-7.5,

setose clavate multiseptate,

fusoid

L. quercina Not available Not available 36-43 x Unknown sp. — Hughes (1993)

(Pat.) Hohn. 9-12, (Meliolaceae)

4—5septate,

ellipsoid

to clavate

. 180-260: 80 x 27-29 x Meliola torta Hughes (1993);

L, torta (Doidge) spherical, 20-24, 9-11, Doidge; Sree (1919)

S. Hughes | ees 3. I b

Steaentis almost smooth clavate septate, rene scabra

(as L. javensis) (as L. ellipsoid (Doidge)

javensis) to clavate Doidge

L. uvariae < 280: 100 x 32-44 x Unknown sp. —_ Hughes (1993);

globose to 20-30, 7-14, (Meliolaceae) Hansford (1946,

subglobose, ellipsoid 3-septate, as Ph. meliolicola)

setose to saccate clavate-oblong

*Ph. = Phaeophragmeriella

The presence of pseudothecia and conidiophores apparently on the same

mycelium was also observed by Hansford (1946: 96, as Phaeophragmeriella

meliolae and Helminthosporium capense) and Hughes (1993: 204); however,

molecular and cultural confirmation is needed before this observation can be

accepted as indicating a teleomorph/anamorph connection. Attempts to isolate

the Spiropes conidia on culture media have been unsuccessful.

Leptomeliola uvariae can be easily distinguished from the other species of the

genus based on morphological differences, such as pseudothecial size and

shape; presence or absence of setae on the pseudothecia; size and shape of asci;

septation, shape and size of the ascospores (TABLE 1).

The collection substrate, Lygodium volubile, is a member of a pteridophyte

genus that has a pantropical distribution (Tryon & Tryon 1982, Barros et al.

2010).

Leptomeliola uvariae new to South America... 5

Acknowledgments

The authors are grateful to the Programa de Pés-Graduacao em Biologia de Fungos

for material support, to IBAMA (Pernambuco) for license to collect in the protected

area, to Dr. Iva C. L. Barros and MSc. Keyla Roberta M. S. Souza (PPGBV-UFPE) for

botanical identification, to Dr. Riviane Bellenand for help with literature acquisition

and to Conselho Nacional de Desenvolvimento Cientifico e Tecnoldgico (CNPq) for

financial support.

Literature cited

Arx JA von, Miller E. 1975. A re-evaluation of the bitunicate ascomycetes with keys to families and

genera. Studies in Mycology 9: 1-159.

Barros ICL, Santiago ACP, Pereira AF de N. 2010. Lygodiaceae. In, Lista de Espécies da Flora do

Brasil, Jardim Botanico do Rio de Janeiro. http://floradobrasil.jbrj.gov.br/ 2010/FB091482

(retrieved 2 July 2010).

Batista AC. 1951. Alguns fungos de fumagina de Pernambuco. Mycopathologia et Mycologia

Applicata 5(2-3): 147-172. doi:10.1007/BF02142292

Hansford CG. 1946. The foliicolous ascomycetes, their parasites and associated fungi. Mycological

Papers 15: 1-240.

Hansford CG. 1961. The Meliolineae - a monograph. Sydowia Beiheft 2: 1-806.

Hohnel F von. 1919. Fragmente zur Mykologie - XXIII Mitteilung, Nr. 1154 bis 1188.

Sitzungsberichten der Akademie der Wissenschaften in Wien, Mathem.-naturw. Klasse, Abt.

1,928; 535-62),

Hughes SJ. 1993. Meliolina and its excluded species. Mycological Papers 166: 1-255.

Index Fungorum. 2010. http://www.indexfungorum.org/Names/Names.asp (retrieved 25 June

2010).

Miller E, Arx JA von. 1962. Die Gattungen der didymosporen Pyrenomyceten. Beitrage zur

Kryptogamenflora der Schweiz. Bern 11(2): 1-922.

MUSEU DO UNA. 2008. Reserva Bioldgica de Saltinho. http://www.museudouna.com.br/ saltinho.

htm (retrieved 16 April 16 2008).

Racovitza A. 1959. Diagnoses des espéces nouvelles. Mémoires du Muséum National d’Histoire

Naturelle, série B, Botanique 10: 91-98.

Tryon RM, Tryon AF. 1982. Ferns and allied plants, with special reference to Tropical America.

Springer-Verlag: New York. 857 p.

MY COTAXON

Volume 116, pp. 7-11 April-June 2011

DOI: 10.5248/116.7

Notes on the neotype of Tuber taiyuanense

JIN-ZHONG Cao’ WEI-DAN CHP LI FAN? & Yu LI”

' Institute of Mycology, Jilin Agricultural University, Changchun 130118, China

’ College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

CORRESPONDENCE TO *: fanli@mail.cnu.edu.cn & yuli966@126.com

ABSTRACT — The neotype of Tuber taiyuanense was re-examined and found to contain two

different species. One of these corresponded with the protologue and is here designated as the

neotype of Tuber taiyuanense; the other was identified as T. liaotongense.

KEY worpDs — Ascomycota, neotypification, truffle

Introduction

Tuber taiyuanense, the first record of the genus Tuber in China, was

established by Liu (1985) based on a specimen from Taiyuan, Shanxi province.

Unfortunately, the original type specimen was destroyed in a fire while the

paper was being printed. Later, Wang & Pei (2001) selected a specimen collected

from Beijing as the neotype for this species. Two clearly different descriptions,

both citing the neotype, have since been published by Chen et al. (2005) and

Song (2005). Chen et al. (2005) described the ascospores as “broadly ellipsoid’,

but Song described them as “long ellipsoid” We re-examined the neotype

deposited in HMAS, Beijing, China (HMAS 75888) and found it contained

two completely different species. One labeled HMAS 75888A has ellipsoid

ascospores with spinose-reticulate ornamentation, while the other HMAS

75888B, contains sub-globose to broad ellipsoid ascospores with more or less

regular reticulate ornamentation. From the two elements we have selected the

one that corresponds with the protologue of T’ taiyuanense and re-describe it

and designate it as neotype here.

Methods

Macroscopic characters are described from the rehydrated dried specimens and

microscopic characters from razor-blade sections mounted in 3% KOH (w/v), Melzer’s

reagent, or 0.1% (w/v) cotton blue in lactic acid.

8 ... Cao &al.

For scanning electron microscopy (SEM), spores were scraped from the dried gleba

onto doubled-sided tape, which was then mounted directly onto an SEM stub, coated

with gold-palladium, and examined and photographed with a HITACHI S-4800 SEM.

Measurements were made from 20 ascospores and asci for each specimen using material

mounted in 3% KOH.

Taxonomy

From the original description: “Ascospores brown, 28.4-32.1 x 18.9-24.6 um

(4-spored), 37.8 x 26.5 um (2-spored), spinose-reticulate, spines 3.8-4 um long,

hooked; reticulum 2.8-4.7 um in diameter” and illustrations of T: taiyuanense by

Liu (1985), the HMAS 75888A specimen (Fics. 1-3) is completely compatible

with Liu’s original type material, whereas HMAS 75888B (Fic. 4) is clearly

different. We therefore designate the HMAS 75888A specimen as the neotype.

We identified the other specimen HMAS 75888B as Tuber liaotongense,

a species endemic to China and common in northern China. Below we

re-describe T: taiyuanense and T: liaotongense and provide illustrations of the

HMAS 75888A and 75888B specimens.

Tuber taiyuanense B. Liu, Acta Mycologica Sinica 4: 84 (1985). FIGS.1-3

TYPE SPECIMEN EXAMINED: CHINA. BEIJING, Dongling Mountains, in soil about 5 cm

deep under pines, 20 Aug. 1998, H.A. Wen, X.Q. Zhang & Z. Wang 294 (HMAS 75888A,

neotype designated here).

Ascomarta globose to subglobose, sometimes irregularly lobed, 0.7-1.5 cm in

diam., pale yellow, yellow brown or brown, the surface smooth, or sometimes

with indistinct hairs in the furrows. Odor light when fresh. PERIpIUM

mostly 150-300 um in thickness, composed of two layers. Outer layer

pseudoparenchymatous, composed of subglobose or subangular cells, pale

yellow or light brown, and up to 5-20 um in diam.; hair-like hyphae sometimes

arising from the outermost cell, 75-125 x 2.5-5 um, hyaline, thin-walled, with

blunt tips. Inner layer composed of intricately interwoven, hyaline hyphae up

to 2.0-3.5 um in diam. GLEBA white when young, gray brown or pale brown at

maturity, marbled with large and rare whitish or cream veins. Asci subglobose

to globose, ellipsoid or irregular shaped, 50-90 x 35-70 um, with a short stalk

7-25 x 5-12.5 um, 1-4(-5) spored, irregularly arranged. AscosporEs mostly

ellipsoid, occasionally broad ellipsoid, yellow-brown at maturity, 20-45 x 18-30

um excluding ornamentation, the ornamentation distinctly spinose-reticulate,

with the spines 2.5-5 um long, slightly curving at apex, and basally connected

by an alveolate reticulum, the alveolae regular or irregular, 5-6 sided, mostly

complete and closed, 2.5-5 um in diam., 6-8 across the spore width.

Eco_Locy, DisTRIBUTION — Hypogeous, under Pinus spp. and Quercus spp.

Known from Beijing, Hebei, Hubei, Shanxi and Sichuan in China.

ADDITIONAL SPECIMENS EXAMINED: CHINA. BeyineG, Dongling Mountains, under

Quercus mongolica Fisch. ex Ledeb., alt. ca. 2100 m, 13 Sept. 2003, M.S. Song 091

The neotype of Tuber taiyuanense ... 9

Fics 1-3. Tuber taiyuanense (HMAS 75888A, neotype). 1. Ascocarps. Bar = 1 cm. 2. Ascospores

observed under the light microscope. Bar = 20 um. 3. Ascospores observed under the scanning

electronic microscope. Bar = 20 um.

(HMAS 97109); C.H. Dong 092 (HMAS 97139); B.H. Tang 093 (HMAS 97140). HEBEI

PROVINCE, Xuanhua, 11 Oct. 1988, B.C. Zhang 517 (HMAS 60234, as T. texense). HUBEI

PROVINCE, Shennongjia, under Pinus spp., alt. ca. 1700 m, Sept. 2003, G.Z. Zhao 095

(HMAS 97126). SHANXI PROVINCE, Yangcheng, Manghe, under Quercus spp., alt.

ca. 620-640 m, 14 July 1990, M.C. Chang & Y. Ma 91037 (HMAS 88573); Yuncheng,

Qijiahe forest farm, under Quercus spp., alt. ca. 880 m, 18 Aug. 1990, B. Qiao 91060

(HMAS 88595). SICHUAN PROVINCE, Wenchuan, Wolong national nature reserve, 22

Sept. 1989, B.C. Zhang 622 (HMAS 60241B); B.C. Zhang 616 (HMAS 60238); B.C.

Zhang 619 (HMAS 60240).

Tuber liaotongense Y. Wang, Atti del II Congresso Internazionale sul Tartufo,

Spoleto: 46 (1988). FiG.4

Ascomarta globose to subglobose, sometimes irregularly lobed, with shallow

to deep furrows, 0.4-1.8 cm in diam., pale yellow brown to brown, the surface

smooth, or usually with indistinct hairs in the furrows. Odor light when fresh.

PERIDIUM mostly 150-250 um in thickness, composed of two layers. Outer

10 ... Cao & al.

Fic. 4. Tuber liaotongense (HMAS 75888B). Ascospores observed under the light microscope

(Bar = 20 um) and the scanning electronic microscope (Bar = 10 um).

layer pseudoparenchymatous, composed of subglobose or subangular cells,

pale yellow or light brown, and up to 6-15 um in diam.; hair-like hyphae

sometimes arising from the outermost cell, 20-80 x 2.5-5 um, hyaline, thin-

walled, with blunt tips. Inner layer composed of intricately interwoven, hyaline

hyphae up to 2.5-5 um in diam. GLEBA white when young, pale yellow brown,

brown or deep brown at maturity, marbled with large and rare whitish or

cream veins. Ascr subglobose, ellipsoid or irregular shaped, 50-90 x 30-70

um, with a short stalk 9-25 x 4-10 um, 1-4(-5) spored, irregularly arranged.

Ascosporss subglobose to broad ellipsoid, yellow-brown at maturity, 22-40

x 20-35 um excluding ornamentation, the ornamentation spinose-reticulate,

2.3-3.5 um high, spines short, usually higher than reticulum a little, making

the ornamentation look like a typical reticulum, the alveolae regular, 5-6 sided,

mostly complete and closed, 3.5-5(-8.5) um in diam., (4—) 5-6(-7) across the

spore width.

SPECIMEN EXAMINED: CHINA. BEING, Dongling Mountains, in soil about 5 cm deep

under pines, 20 Aug. 1998, H.A. Wen, X.Q. Zhang & Z. Wang 294 (HMAS 75888B).

COMMENTS — Our study of the specimens deposited in various herbariums

suggests that T: taiyuanense is common in northern China. It can be recognized

by the small, more or less brown colored ascocarps and the typical spinose-

reticulate ascospores. In contrast, the similar species T. huidongense Y. Wang

and T’ umbilicatum Juan Chen & P.G. Liu, both originate from southwestern

China. Tuber huidongense can be easily differentiated from T’ taiyuanense by

the alveolae and ascospore size, which are important characteristics for both the

phylogeny and taxonomy of Tuber (Cao 2010). The alveolae in T: huidongense

are 3-5(-8.5) um in diam and 3-6 across the spore width, are larger than in

T. taiyuanense (2.5-5 um in diam and 6-8 across the spore width). Tuber

umbilicatum has ascospores similar to T: taiyuanense but is clearly differentiated

by its umbilicate ascocarp. In addition, T. huidongense is common and

T:. umbilicatum is not uncommon in southwestern China, whereas T! taiyuanense

has yet to be found there.

The neotype of Tuber taiyuanense ... 11

Tuber liaotongense, described from Liaoning Province (Wang 1988), is also

very common in China and shares the same distribution area as T: taiyuanense.

Almost all species that have spiny or spinose-reticulate ornamentation on

the spores have ellipsoid spores. The exception is T. liaotongense, which has

subglobose spores. In addition, the spines on T! liaotongense spores are so low

that it is difficult to see them even under high power on an optical microscope,

although they can be clearly seen by SEM photomicrograph (Fic. 4). Molecular

data has also confirmed T. liaotongense to be in the Rufum Group (Song

2005).

Acknowledgments

We are grateful to Dr. Yi-Jian Yao for serving as pre-submission reviewer, and Dr.

Tan R. Hall for editing the manuscript prior to submission. The study was supported by

NSFC (No. 30770005, 30870008)

Literature cited

Cao JZ. 2010. The genus Tuber in China [Dissertation]. Changchun: Jinlin Agricultural University.

86 p.

Chen J, Liu PG, Wang Y. 2005. Tuber umbilicatum, a new species from China, with a key to the

spinose-reticulate spored Tuber species. Mycotaxon 94: 1-6.

Liu B. 1985. New species and new records of hypogeous fungi from China (I). Acta Mycologica

Sinica 4(2): 84-89.

Song MS. 2005. Taxonomic and molecular systematic studies on Tuber in China [Dissertation].

Beijing: Graduate School of Chinese Academy of Sciences. 156 p.

Wang Y. 1988. First report of study on Tuber species from China. Atti del II Congresso Internazionale

sul Tartufo, Spoleto, Nov. 24-27: 45-50.

Wang Z, Pei KQ. 2001. Notes on discomycetes in Dongling Mountains (Beijing). Mycotaxon

79: 307-314.

MY COTAXON

Volume 116, pp. 13-20 DOI: 10.5248/116.13

Wood-rotting fungi in eastern China 6. Two new species of

Antrodia (Basidiomycota) from Mt. Huangshan, Anhui Province

Bao-Kali Cur’, Hat-Jriao Li? & YU-CHENG Dat >

Institute of Microbiology, P.O. Box 61, Beijing Forestry University, Beijing 100083, China

CORRESPONDENCE TO *: ' baokaicui@yahoo.com.cn, * lihaijiao715@126.com &

* yuchengd@yahoo.com

ABSTRACT — Two new polypores, Antrodia bambusicola and A. huangshanensis from

Mt. Huangshan in Anhui Province, eastern China, are described and illustrated. Antrodia

bambusicola is characterized by its annual, resupinate and thin basidiocarps, regular pores

with white to cream or pale buff-yellow pore surface, ellipsoid basidiospores, branched

and septate cystidioles, and its growth on fallen Bambusa spp. Antrodia huangshanensis

has resupinate to rarely effused-reflexed basidiocarps with cream pore surface, larger

pores, branched and flexuous hyphae at dissepimental edges, and cylindrical to allantoid

basidiospores; it was found on a fallen branch of Pinus massoniana, causes a brown rot, and

prefers a dry environment.

Key worps — brown-rot fungi, Fomitopsidaceae, taxonomy

Introduction

Karsten (1879) established the genus Antrodia P. Karst.; its species typicially

have an annual to perennial growth habit, resupinate to effused-reflexed

basidiocarps, a dimitic hyphal system with clamped generative hyphae, hyaline,

thin-walled and usually cylindrical to oblong-ellipsoid basidiospores that are

negative in Melzer’s reagent, and cause brown rot (Bernicchia & Ryvarden 2001,

Gilbertson & Ryvarden 1986, Nufiez & Ryvarden 2001, Ryvarden & Gilbertson

1993). It is a large cosmopolitan genus with more than 45 species (Kirk et al.

2008) of which 20 have been recorded from China (Dai & Niemela 2002, Dai &

Penttila 2006, Dai et al. 2006, 2007a, b, 2009, Du et al. 2009, Li et al. 2008).

Recent studies on diversity of wood-rotting fungi in eastern China have

revealed several new species (Cui & Dai 2008, Cui et al. 2008, Wang et al. 2009).

During a study on wood-rotting fungi from the Mt. Huangshan National Park

(Anhui Province), two species representing the brown rot genus Antrodia were

found that could not be referred to any known species. They are described in

the present paper as A. bambusicola and A. huangshanensis.

14 ...Cui, Li & Dai

Materials & methods

The studied specimens were deposited in the herbarium of Beijing Forestry University

(BJFC). The microscopic procedure follows Du & Cui (2009). In presenting the variation

in the size of the spores, 5% of measurements were excluded from each end of the range,

and given in parentheses. In the text the following abbreviations are used: IKI = Melzer’s

reagent, IKI- = negative in Melzer’s reagent, KOH = 5% potassium hydroxide, CB =

Cotton Blue, CB- = acyanophilous, L = mean spore length (arithmetic average of all

spores), W = mean spore width (arithmetic average of all spores), Q = variation in the

L/W ratios between the specimens studied, n = number of spores measured from given

number of specimens. Sections were studied at magnification up to x1000 using a Nikon

Eclipse E 80i microscope and phase contrast illumination. Drawings were made with

the aid of a drawing tube. Special colour terms follow Anonymous (1969) and Petersen

(1996).

Taxonomy

Antrodia bambusicola Y.C. Dai & B.K. Cui, sp. nov. FIG. 1

MycoBank MB 519531

Carpophorum annuum, resupinatum; facies pororum bubalina vel luteola, pori rotundi vel

angulati, 3 per mm. Systema hypharum dimiticum, hyphae generatoriae fibulatae, hyphae

skeletales contexti 2.2-3.5 um. Sporae hyalinae, ellipsoideae, IKI-, CB-, 5-6 x 3-3.4 um.

Type. — China. Anhui Province, Huangshan County, Mt. Huangshan National Park, on

fallen trunk of Bambusa sp. (Poaceae), 21.X.2010 Dai 11901 (holotype in BJFC).

EryMoLocy — bambusicola (Lat.) refers to growing on bamboo.

FruiTBopy — Basidiocarps annual, resupinate, adnate, corky, without odour or

taste and light in weight when dry, up to 40 cm or more in longest dimension,

5 cm wide, and up to 0.5 mm thick at centre; sterile margin thinning out,

membranous, white, up to 3 mm wide. Pore surface white to cream when fresh,

becoming cream to pale buff-yellow when dry; pores round to angular, about

3 per mm; dissepiments thin, entire. Subiculum very thin to almost lacking,

cream, less than 0.1 mm thick. Tubes concolorous with pore surface, corky, up

to 0.4 mm long.

HyYPHAL STRUCTURE — Hyphal system dimitic; generative hyphae bearing

clamp connections, skeletal hyphae IKI-, CB-; tissue unchanged in KOH.

SUBICULUM — Generative hyphae scanty, hyaline, thin-walled, occasionally

branched, 2-3.3 um in diam; skeletal hyphae dominant, hyaline, distinctly thick-

walled to subsolid, usually unbranched, interwoven, 2.2-3.5 um in diam.

TuBEs — Generative hyphae infrequent, hyaline, thin-walled, occasionally

branched, 1.8-3 um in diam; skeletal hyphae dominant, hyaline, distinctly

thick-walled to almost solid, usually unbranched, some of them at dissepiment

edge occasionally inflated, loosely interwoven, 2-3.8 um in diam. Cystidia

absent; cystidioles present, fusoid, hyaline, mostly thin- walled, occasionally

slightly thick-walled, with a basal clamp connection, usually branched and

Antrodia spp. nov. (China) ... 15

with one to several secondary septum (septa), 16-25 x 4-6 um; basidia clavate,

bearing four sterigmata and a clamp connection at the base, 18-26 x 5-7 um;

basidioles in shape similar to basidia, but slightly smaller.

16 ...Cui, Li & Dai

Spores — Basidiospores ellipsoid, hyaline, thin-walled, smooth, IKI-, CB-,

(4.7-)5-6(-7) x (2.9-)3-3.4(-3.9) um, L = 5.34 um, W = 3.13 um, Q = 1.71

(n=30/1).

TYPE OF ROT — Brown rot.

ADDITIONAL SPECIMEN EXAMINED — CHINA. ANHUI PROVINCE, Huangshan County,

Mt. Huangshan National Park, on fallen trunk of Bambusa sp., 12.X.2004 Dai 6086

(BJFC).

REMARKS — Antrodia bambusicola is characterized by its annual, resupinate

and very thin basidiocarps, regular pores (usually 3 per mm) with white to

cream or pale buff-yellow pore surface, ellipsoid basidiospores (5-6 x 3-3.4

um), presence of branched and septate cystidioles, and by growth on Bambusa

spp.

Morphologically, A. bambusicola is somewhat similar to species of Ceriporia

Donk; however, species of Ceriporia have a monomitic hyphal system with

simple septate generative hyphae, and cause a white rot (Nufez & Ryvarden

2001), while A. bambusicola has a distinct dimitic hyphal system with clamped

generative hyphae, and causes a brown rot.

Antrodia vaillantii (DC.) Ryvarden has similar pores (2-4 per mm) and

basidiospores (5-8 x 3-4 um) as A. bambusicola but differs by having thicker

and separable basidiocarps with a soft-fibrous margin, an absence of cystidioles,

and growth on gymnosperm wood (Gilbertson & Ryvarden 1986).

Antrodia huangshanensis Y.C. Dai & B.K. Cui, sp. nov. Fi. 2

MycoBank MB 519532

Carpophorum annuum, resupinatum vel effuso-reflexum; facies pororum nivea vel cremea;

pori rotundi vel angulati, 1-3 per mm. Systema hypharum dimiticum, hyphae generatoriae

fibulatae, hyphae skeletales contexti 2.4-5.3 um in diam. Sporae hyalinae, cylindricae vel

allantoideae, IKI-, CB-, 5-6.5 x 1.6-2 ym.

Type. — China. Anhui Province, Huangshan County, Mt. Huangshan National Park,

on fallen branch of Pinus massoniana Lamb. (Pinaceae), 11.X.2004 Dai 6082 (holotype

in BJFC).

EryMoLocy — huangshanensis (Lat.) refers to the locality of Mt. Huangshan.

FRuITBODy — Basidiocarps annual, mostly resupinate, rarely effused-reflexed,

tightly attached on the substrate, corky to leathery when fresh, hard corky and

light in weight when dry, without odour or taste, pilei very narrow, projecting

up to 2 mm, resupinate part up to 7 cm long, 3 cm wide, and up to 6 mm thick

at centre. Pileal surface white when fresh, becoming cream when dry, glabrous;

margin acute, sterile margin of resupinate part very narrow to almost lacking.

Pore surface white to cream when fresh, becoming cream to cream-buff upon

drying; pores round to angular, 1-3 per mm; dissepiments thin, mostly entire,

sometimes slightly lacerate. Subiculum cream, corky, thin, ca. 1 mm thick.

Tubes concolorous with pore surface, hard corky, up to 5 mm long.

Antrodia spp. nov. (China) ... 17

s ai

ATR ILEEF SS

eZ -

Fic. 2. Microscopic structures of Antrodia huangshanensis (drawn from the holotype).

a: Basidiospores. b: Basidia and basidioles. c: Hyphae from trama. d: Hyphae from

subiculum.

HYPHAL STRUCTURE — Hyphal system dimitic; generative hyphae bearing

clamp connections, skeletal hyphae IKI-, CB-; tissue unchanged in KOH.

SUBICULUM — Generative hyphae frequent, hyaline, thin- to slightly thick-

walled, occasionally branched, frequently septate with clamp connections

18 ...Cui, Li & Dai

2-5.1 um in diam; skeletal hyphae thick-walled, flexuous, occasionally

branched, interwoven, 2.4—5.3 um in diam.

TuBES — Generative hyphae common, hyaline, thin- to slightly thick-

walled, occasionally branched, 1.7-4.8 um in diam; skeletal hyphae dominant,

thick-walled to subsolid, straight to flexuous, moderate branched, interwoven,

2.2-5 um in diam; hyphae at dissepimental edges usually branched and flexuous.

Cystidia and cystidioles absent; basidia clavate, with four sterigmata and a basal

clamp connection, 14.2-19.7 x 4-6 um, basidioles in shape similar to basidia,

but slightly smaller.

Spores — Basidiospores cylindrical to allantoid, hyaline, thin-walled,

smooth, IKI-, CB-, (4.8-)5-6.5(-7.3) x (1.5-)1.6-2(-2.2) um, L = 5.64 um, W

= 1.84 um, Q = 2.95-3.26 (n=90/3).

TYPE OF ROT — Brown rot.

ADDITIONAL SPECIMENS EXAMINED — CHINA. ANHUI PROVINCE, Huangshan County,

Mt. Huangshan National Park, on fallen branch of Pinus massoniana, 11.X. 2004 Dai

6081 (BJFC); 12.X.2004 Dai 6124 (BJFC).

REMARKS — Antrodia huangshanensis is characterized by its resupinate to rarely

effused-reflexed basidiocarps with white to cream pore surface, larger pores

(1-3 per mm), branched and flexuous hyphae at dissepimental edges, cylindrical

to allantoid basidiospores (5-6.5 x 1.6-2 um), and growth on branches of Pinus

massoniana. A brown rot fungus, it prefers a dry environment.

Morphologically, the new species resembles species of Postia Fr. with an

annual growth habit, cylindrical to allantoid basidiospores, and causing a

brown rot. Postia, however, is characterized by a monomitic hyphal system.

Antrodia sinuosa (Fr.) P. Karst. also produces resupinate to effused

basidiocarps and larger pores (1-3 per mm) but its pale sordid brown pore

surface when dry, sinuous pores with strongly lacerate to split dissepiments

when mature, fusoid cystidioles, and relatively smaller basidiospores (4-6 x

1-2 um, Ryvarden & Gilbertson 1993) distinguish it from A. huangshanensis.

Antrodia hingganensis Y.C. Dai & Penttila from Heilongjiang and Jilin

provinces in northeast China, which may be confused with both A. sinuosa

and A. huangshanensis, is differentiated by smaller pores (3-5 per mm) and

basidiospores (4—-5.4 x 1.1-1.5 um, Dai & Penttila 2006).

Antrodia pseudosinuosa A. Henrici & Ryvarden from England has pores

similar in size (1-2 per mm) to huangshanensis but is distinguished by its

pileate basidiocarps, larger (6-7 x 1.8-2.2 um) basidiospores, and growth on

angiosperm wood (Henrici & Ryvarden 1997).

Antrodia albobrunnea (Romell) Ryvarden produces similarly sized

basidiospores, but is characterized by a brown zone in its subiculum, smaller

(3-5 per mm) pores, and distribution in temperate and boreal forests.

Antrodia spp. nov. (China) ... 19

Antrodia huangshanensis may also be confused with A. ramentacea (Berk.

& Broome) Donk in pore size and growth on Pinus, but the latter species has

bigger basidiospores (9-11 x 4.5-5 um, Ryvarden & Gilbertson 1993).

OTHER SPECIMENS EXAMINED. — Antrodia albobrunnea. FINLAND. Sompion Lappi,

Pelkosenniemi, Luosto, on rotten wood of Picea, 3. VIII.1998 Dai 2747 (IFP); Koillismaa,

Oulanka National Park, on charred wood of Pinus, 19.[X.1997 Dai 2710 (IFP).

— A. hingganensis. CHINA. HEILONGJIANG PROVINCE, Yichun, Fenglin Nature

Reserve, on fallen trunk of Larix, 9.[X.2002 Dai 3748 (IFP & H); on fallen wood of

Picea, 29.VII.2000 Penttila 13011 (H); on fallen trunk of Pinus, 2.VIII.2000 Penttila

13146 (H); JrtIn Province, Antu County, Changbaishan Nature Reserve, on charred

wood of Abies, 28.VII.1993 Dai 817 (IFP & H); on fallen trunk of Pinus, 14.[X.1998 Dai

2909 (IFP & H).

— A. sinuosa. FINLAND. Uusimaa, Kirkkonummi, Jorvas, Finntrask, on fallen

trunk of Pinus, 24.V1.1993 Niemela 5662 & Renvall (H); Sompion Lappi, Pelkosenniemi,

Luosto, Kapusta, on charred wood of Pinus, 14.VIII.1998 Dai 2844 (IFP); Etela- Hame,

Lammi Biological Station, on charred wood of Pinus, 11.1X.1997 Dai 2605 (IFP).

Acknowledgements

We express our gratitude to Drs. Zheng Wang (USA) and Hai-Sheng Yuan (China)

who reviewed the manuscript. The research was financed by the National Natural Science

Foundation of China (Project No. 30900006 and 30910103907) and the Fundamental

Research Funds for the Central Universities.

Literature cited

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Office, London. 1 pp.

Bernicchia A, Ryvarden L. 2001. A new Antrodia species (Coriolaceae, Basidiomycetes). Mycotaxon

79: 57-66.

Cui BK, Dai YC. 2008. Wood-rotting fungi in eastern China 2. A new species of Fomitiporia

(Basidiomycota) from Wanmulin Nature Reserve, Fujian Province. Mycotaxon 105: 343-348.

Cui BK, Yuan HS, Dai YC. 2008. Wood-rotting fungi in eastern China 1. Polypores from Wuyi

Mountains, Fujian Province. Sydowia 60: 25-40.

Dai YC, Niemela T. 2002. Changbai wood-rotting fungi 13. Antrodia sensu lato. Annales Botanici

Fennici 39: 257-265.

Dai Y C, Penttila R. 2006. Polypore diversity of Fenglin Nature Reserve, northeastern China.

Annales Botanici Fennici 43: 81-96.

Dai YC, Yuan HS, He W, Decock C. 2006. Polypores from Beijing area. Mycosystema 25: 368-373.

Dai YC, Cui BK, Huang MY. 2007a. Polypores from eastern Inner Mongolia, northeastern China.

Nova Hedwigia 84: 513-520. doi:10.1127/0029-5035/2007/0084-0513

Dai YC, Cui BK, Yuan HS. 2007b. Notes on polypores from Gansu and Qinghai Province, northwest

China. Cryptogamie Mycologie 28: 177-187.

Dai YC, Yuan HS, Wang HC, Yang F, Wei YL. 2009. Polypores (Basidiomycota) from Qin Mts. in

Shaanxi Province, central China. Annales Botanici Fennici 46: 54-61.

Du P, Cui BK. 2009. Two new species of Megasporoporia (Polyporales, Basidiomycota) from tropical

China. Mycotaxon 110: 131-138. doi:10.5248/110.131

Du P, Cui BK, Wang W. 2009. Wood-rotting fungi in eastern China 3. A species of Antrodia new to

China with notes on the genus. Mycosystema 28: 44-48.

20 ...Cui, Li & Dai

Gilbertson RL, Ryvarden L. 1986. North American polypores 1. Abortiporus - Lindtneria. Fungiflora,

Oslo. 1-433 pp.

Henrici A, Ryvarden L. 1997. Antrodia pseudosinuosa sp. nov. Mycologist 11: 152-154. doi:10.1016/

S0269-915X(97)80088-2

Karsten P. 1879. Symbolae ad mycologiam Fennicam 6. Meddel. Soc. Fauna Flora Fennica 5:

15-46.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s Dictionary of the Fungi

10th. CAB International, Wallingford, Oxon. 771 pp.

Li J, Xiong HX, Dai YC. 2008. Polypores from Shennongjia Nature Reserve in Hubei Province,

Central China. Cryptogamie Mycologie 29: 267-277.

Nufiez M, Ryvarden L. 2001. East Asian polypores 2. Polyporaceae s. lato. Syn. Fungorum 14:

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MY COTAXON

Volume 116, pp. 21-25 April-June 2011

DOI: 10.5248/116.21

Three corticolous species of Lecanora (Lecanoraceae)

new to China

Lru-Fu HAN! SHOU-YU GUO” & HAO ZHANG?

‘College of Life Science, Hebei Normal University, Shijiazhuang 050016, P. R. China

*Key Laboratory of Systematic Mycology and Lichenology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing 100101, P. R. China

*Department of Biology Science, Handan College, Handan 056005, P. R. China

CORRESPONDENCE TO *: hanliufu@163.com, *guosy@im.ac.cn, & zhanghao_55@163.com

ABSTRACT —'hree corticolous species of the lichen genus Lecanora— L. mikuraensis, L.

queenslandica, and L. thysanophora — are reported from China for the first time. General

data on habitats, diagnostic characters, geographic distribution, and habit and anatomical

photos of the newly recorded species are provided.

Key worps —lichenised ascomycetes, Anhui, Hebei, Shaanxi, taxonomy

Introduction

The lichen genus Lecanora Ach. (Lecanoramycetes, Ascomycota) consists of

some 550 species worldwide (Kirk et al. 2008). It is characterized by Lecanora-

type asci, simple ascospores, and crustose thalli. The apothecial margin usually

contains algal cells. The taxa of Lecanora grow on a wide variety of substrata,

such as rocks, soil, bark or wood. In China, Wei (1991) cited 63 species, 11

varieties, and 5 forms, after which an additional 43 species (including 1 new)

were reported (Cao et al. 1995, Abbas et al. 1998, Aptroot & Seaward 1999,

Aptroot & Sparrius 2003, Mamut et al. 2004, 2009, Wang et al. 2007, Han et al.

2009, Lii et al. 2008, 2009a,b,c), bringing to 106 the number of taxa reported

from China.

During our study of Lecanora, we discovered three corticolous species new

to China: L. mikuraensis, L. queenslandica and L. thysanophora, for which we

provide general data on habitats, diagnostic characters, geographic distribution,

brief comments, and habit and anatomical photos.

Materials & methods

The present paper is based on the collections from Sectio Lichenum, Herbarium

Mycologici Academiae Sinicae (HMAS-L) and the Lichen Section of Botanical

22 ... Han, Guo & Zhang

Herbarium, Hebei Normal University (HBNU). Apothecia were observed in free-

hand sections mounted in water. Sections through apothecium were stained by 0.2%

Toluidine blue about 30 min before adding 10% KOH. All specimens were examined

and measured under the dissecting (Motic SMZ-140) and compound (Olympus CH)

microscopes. Apothecial crystals were examined using polarized light under microscope

(Motic PM18). Secondary metabolites were identified using thin layer chromatography

(TLC) (Culberson 1972).

New records from China

Lecanora mikuraensis Miyaw., J. Hatt. Bot. Lab. 64: 296. 1988. Fic. la,d

SPECIMEN EXAMINED: CHINA. Hebei, Mt. Xiaowutai, on bark, 2008/VII/1, Zhang Hao

20080147 (HBNU).

THALLUS crustose, thin to thick, areolate, verrucose to granular, gray, esorediate,

epruinose. PROTHALLUS absent. APOTHECIA lecanorine, abundant, single to

crowded, adnate to constricted at the base, 0.4-1.6 mm in diam., disc yellowish

brown, plane to slightly convex, epruinose; margins prominent, verruculose,

concolorous with thallus. AMPHITHECIUM: cortex thin, or indistinct, algal

dispersed, containing large crystals and small. EPIHYMENIUM not pigmented,

with coarse yellowish crystals soluble in KOH, 13-18 um tall. Hymenrum

hyaline, 50-60 um tall. HypoTHEcIuM hyaline, 75-100 um tall. PARAPHYSES

simple, not thickened or slightly thickened apically. Asci clavate, 8-spored.

Ascosporss ellipsoid, simple, 7.5-12.5 x 4.0-6.0 um. PycniDIA not seen.

CHEMISTRY: K+ yellow; atranorin (TLC).

ComMENts: Lecanora mikuraensis is readily identified by its non-pigmented

epihymenium with coarse yellowish crystals. It is closely related to L. insignis

and L. cinereofusca but differs in lacking “beaded” apothecial margins, pannarin,

and dark brown discs. ‘The species also superficially resembles L. chlarotera and

L. leprosa but lacks gangaleoidin and a pigmented epihymenium.

Lecanora mikuraensis is known previously only from Japan (Miyawaki

1988).

Lecanora queenslandica C. Knight, Syn. Queensland FI. 2, Suppl.: 85. 1888. Fic. 1b,e

SPECIMENS EXAMINED: CHINA. Shaanxi, Mt. Taibai, alt. 3189 m, 2005/VIII/4, Huang

Man-rong 2590 (HMAS-L 070630); alt. 2915 m, 2005/VIH/4, Huang Man-rong 2798

(HMAS-L 070584), Huang Man-rong 2799 (HMAS-L 070585), Huang Man-rong 2598

(HMAS-L 070586), Huang Man-rong 2800 (HMAS-L 070587), Huang Man-rong 2795

(HMAS-L 070592); alt. 2896 m, 2005/VIII/4, Huang Man-rong 2491 (HMAS-L 070591);

alt. 2930 m, 2005/VIII/4, Huang Man-rong 988 (HMAS-L 070652); Ningshan County,

alt. 2100 m, 2005/VII/28, Huang Man-rong 545 (HMAS-L 070571).

THALLUS crustose, coarse, slightly verrucose, pale gray to gray, esorediate,

epruinose. PROTHALLUS absent. APOTHECIA abundant, lecanorine, sessile to

constricted at base, almost single, 0.5-1.4(-2.0) mm in diam.; discs orange-

Lecanora spp. new to China ... 23

lal

Fic. 1. Lecanora species new to China. Habit (a-c; scales = 1 mm): a. L. mikuraensis (Zhang

Hao 20080147, HBNU); b. L. queenslandica (Huang Man-rong 2800, HMAS-L 070587);

c. L. thysanophora (Han Liu-fu 0810042-4, HBNU). Asci (d-e; scales = 10 um): d. L. mikuraensis;

e. L. queenslandica.

brown, epruinose or slightly pruinose, plane; margins smooth, entire,

sometimes slightly crenulate or verruculose, concolorous with thallus.

AMPHITHECIUM: cortex indistinct with algal layer, cortex 25-37 um laterally,

30-40 um basally; algal layer 100-125 um laterally, 100-150 um basally,

containing large crystals insoluble in KOH. EPIHYMENIUM with small crystals

soluble in KOH, pale orange brown, 13-20 um tall. HyYMENIUM hyaline, 50-65

um tall. HypoTHecrium hyaline, 75-100 um tall. PARAPHYsEs not thickened

or slightly thickened apically. Asc1 clavate, 8-spored. Ascospor:s ellipsoid to

broadly ellipsoid, simple, 9.0-12.5 x 6.0-8.0 um.

CHEMISTRY: K+ yellow, C-; atranorin, usnic acid, psoromic acid.

24 ... Han, Guo & Zhang

ComMENTs: Lecanora queenslandica is readily distinguished from other

Lecanora species by the presence of usnic and psoromic acid in the thallus,

the orange brown apothecial discs, and the crystalloid epihymenium. Closely

related to L. chlarotera, it differs in the short ascospores, possession of usnic

and psoromic acids, and absence of gangaleoidin.

Lecanora queenslandica is known previously only from Australia (Lumbsch

1994, 2004).

Lecanora thysanophora R.C. Harris, The Bryologist 103: 790. 2000. Fic. Ic

SPECIMENS EXAMINED: CHINA. Anhui, Jinzhai County, alt. 1450 m, 2008/X/12, Han

Liu-fu & Zhao Hao 0810006 (HBNU)); alt. 1350 m, 2008/X/11, Han Liu-fu 0810042-2

(HBNU).

THALLUS a thin patchy layer of granular soredia, almost leprose, pale green

to yellowish green on surface and with white, conspicuous, webby or fibrous

prothallus developing at the margin. APOTHECIA not seen.

Cuemistry: Thallus K+ yellow; containing atranorin, usnic acid, and an

unidentified substance (TLC).

ComMENtTs: This species is readily distinguished from other Lecanora species

by the powdery thallus with the white fibrous margins, presence of usnic acid

and atranorin. Lecanora thysanophora closely resembles other Lepraria species

(particularly L. impudens), but differs in its powdery thallus with the fibrous

margins.

Brodo (2001) described L. thysanophora from North America, and it is also

recorded from Europe (e.g. Kowalewska et al. 2003, Mrak et al. 2004, Tonsberg

1999, Tonsberg et al. 2001).

Acknowledgements

The project was supported by the Chinese Academy of Sciences (KSCX2-YW-Z-

041), the National Natural Science Foundation of China (No. 30770012), the Natural

Science Foundation of Hebei Province (No. C2008000178) and Doctoral Fund of

Hebei Normal University (L2009B11). The authors would like to thank Prof. Wei Jiang-

Chun (Institute of Microbiology, CAS) for guidance and Ms. Deng Hong (HMAS-L)

for assistance during this study. The authors would like to express their deep thanks

to Dr. André Aptroot (ABL Herbarium, The Netherlands) and Dr. Huang Man-Rong

(Beijing Museum of Natural History, China) for reading the manuscript and serving as

pre-submission reviewers.

Literature cited

Abbas A, Wu JN. 1998. Lichens of Xinjiang. Sci-Tech & Hygiene Publishing House of Xinjiang,

Urumadi. 178 pp.

Aptroot A, Seaward MRD. 1999. Annotated checklist of Hong Kong lichens. Tropical Bryology 17:

57-101.

Aptroot A, Sparrius LB. 2003. New microlichens from Taiwan. Fungal Diversity 14: 1-50.

Lecanora spp. new to China... 25

Brodo IM, Sharnoff SD, Sharnoff S. 2001. Lichens of North America. Yale University Press, New

Haven and London. 795pp.

Cao R, Yong SP, Ma YQ. 1995. Preliminary study of lichens in semi-dry mountains region in

NeiMonggol. Acta Scientiarum Naturalium Universitatis NeiMonggol 26: 587-595.

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.

doi:10.1016/0021-9673(72)80013-X

Han LE, Zhao JC & Guo SY. 2009. Lecanora weii, a new multispored species of Lecanora s. str. from

northeastern China. Mycotaxon 107: 157-161. doi:10.5248/107.157

Kirk PM, Cannon PF, Minter DW, Stalpers JA (eds.). 2008. Ainsworth & Bisby’s dictionary of the

fungi, tenth edition. CABI International, Wallingford. 771 pp.

Kowalewska A, Kukwa M. 2003. Additions to the Polish lichen flora. Graphis Scripta 14: 11-17.

Lit L, Wang CL, Ren Q, Shi XL, Zhao ZT. 2008. The lichen genus Lecanora from Bailong river valley

of Gansu province, China. Mycosystema 27(1): 99-104.

Li L, Ren Q, Wang HY, Zhao ZT. 2009a. New records of four Lecanora species from China.

Mycotaxon 110: 437-441. doi:10.5248/110.437

Li L, Ren Q, Sun LY, Yang F, Zhao ZT. 2009b. Three species of the lichen genus Lecanora new to

China from Bailong River Valley, Gansu Province. Guihaia 29 (3): 311-313.

Lit L, Wang HY, Zhao ZT. 2009c. Five lichens of the genus Lecanora new to China. Mycotaxon 107:

391-396. doi:10.5248/107.391

Lumbsch HT. 1994. Die Lecanora subfusca Gruppe in Australasien. Journal of the Hattori Botanical

Laboratory 77: 1-175.

Lumbsch HT, Elix JA. Lecanora. 12-62, in: McCarthy PM, Mallett K (eds.). 2004. Flora of Australia,

volume 56A, lichens 4. ABRS, CSIRO Australia, Melbourne.

Mamut R, Keyimu A, Abbas A. 2004. New Chinese records of the lichen genus Lecanora Ach.

collected from Khanas Nature Reserve of Xinjiang. Mycosystema 23(1): 167-168.

Mamut R, Tumur A, Xahidin H, Abbas A. 2009. A species of Lecanora new to China. Mycosystema

28(1): 154-156.

Miyawaki H. 1988. Studies on the Lecanora subfusca group in Japan. Journal of the Hattori Botanical

Laboratory 64: 271-326.

Mrak T, Mayrhofer H, Batic F. 2004. Contributions to the lichen flora of Slovenia XI. Lichens from

the vicinity of Lake Bohinj (Julian Alps). Herzogia 17: 107-127.

Tonsberg T. 1999. Lichenes isidiosi et sorediosi crustacei exsiccati. Fascicle 2 (Nos 26-50).

University of Bergen, Bergen. 10 pp.

Tonsberg T, Turk R, Hofmann P. 2001. Notes on the lichen flora of Tyrol (Austria). Nova Hedwigia

72(3-4): 487-497.

Wang CL, Sun LY, Ren Q, Zhao ZT. 2007. A preliminary study of multispored Lecanora Ach. from

Mt. Taibai. Mycosystema 26(1): 46-50.

Wei JC. 1991. An enumeration of lichens in China. International Academic Publishers: Beijing.

278 pp.

MY COTAXON

Volume 116, pp. 27-32 April-June 2011

DOI: 10.5248/116.27

A new Dothidasteroma species on leaves of

Psidium laruotteanum from the Brazilian Cerrado

C.A. INAcIO, R.C. PEREIRA-CARVALHO, E.S.C. SOUZA & J.C. DIANESE,

Departamento de Fitopatologia, Universidade de Brasilia, 70.910-900 Brasilia, D.F., Brazil

*CORRESPONDENCE TO: jcarmine@unb.br

AxBsTRACT— A new Dothidasteroma species found on leaves of Psidium laruotteanum

collected in the Brazilian Cerrado is described, illustrated, and designated as Dothidasteroma

psidii.

KEY woRDS— ascomycetes, foliicolous fungi, Parmulariaceae, fungal taxonomy, Neotropical

mycodiversity

Introduction

Since 1993 over 100 new fungi from the Brazilian Cerrado have been

described (Dianese et al. 1997, Dianese 2000, Inacio & Dianese 1998, 2006),

including major finds among the cercosporoid fungi (Inacio & Dianese 1999a,

Dornelo-Silva et al. 2007, Hernandez-Gutiérrez & Dianese 2008, 2009), two

new genera of rust fungi (Dianese et al. 1993, 1995), three new ascomycete

genera (Dianese et al. 2001, Pereira-Carvalho et al. 2009a, 2010), and eleven

new genera of trichomatous hyphomycetes (Dornelo-Silva & Dianese 2004,

Pereira-Carvalho et al. 2009b).

Foliar ascomycetes are common among the 22,000 specimens deposited in

Herbarium UB, Mycological Colletion, and exsiccates of a myrtaceous host,

Psidium laruotteanum Cambess., revealed a new Dothidasteroma species that is

now described and illustrated.

Materials & methods

Lesions on host leaves were initially observed under a stereomicroscope; samples were

taken for squash preparations and sectioning with a Micron Freezing Microtome, before

being mounted as semi-permanent slides. In most cases the samples were stained with

lacto-glycerol-cotton blue or glycerol-KOH-phloxine B, and the slides sealed with nail

polish, but color observations were done in water preparations. Morphological studies,

measurements, and photomicrography were done with a Leica DM 2500 microscope

28 ... Inacio & al.

coupled with a Leica DFC 490 digital camera connected to a microcomputer. Image

capture, editing, and measurements were made with Leica QWin V3 software. Ascomata

were illustrated also using SEM. The dimensions of all structures were expressed as the

range of sizes.

Taxonomy

Dothidasteroma psidii Inacio, Pereira-Carvalho, E.S.C. Souza & Dianese, sp. nov.

MycosBank MB 518319 FIGS 1-11

Species haec ab Dothidasteroma casuarinae ascis longioribus et ascosporis brevioribus.

Type: Brazil. Distrito Federal, Brasilia, Parque Nacional on leaves of Psidium

laruotteanum Cambess. (Myrtaceae), 13 Sept 1995, leg. M. Sanchez 1205, holotype (UB

Mycol. Coll. 9935).

SYMPTOMS scattered or confluent leaf spots, up to 12 mm diam., mostly

epiphyllous less frequently hypophyllous, elliptical or circular to irregular,

brown to dark brown with pale reddish purple borders; along the midrib

colonies appear amphigenous as elliptical or elongate dark lesions, containing an

aggregation of black conspicuous discoid stromata with a central opening that

splits in a stellar fashion when wet. Mycetrum internal, conspicuous, hyaline,

filamentous; HYPHAE 2-4 um diam. emanating from internal hypostromata

towards the cuticle where stromatic erumpent ascomata are formed. STROMATA

subcuticular, intracuticular to intradermal, sparse to extensive, 5-16 um deep,

mostly one-layered; CELLS brown to dark brown, 4-16 x 5-11 um. ASCOMATA

originating from the immersed stromata, scutate, variable in shape, usually

+ circular, brown to dark brown, confluent, opening by irregular fissures,

measuring 153-790 x 66-194 um in vertical. UPPER SURFACE composed of

pale to dark brown cells 8-13 x 3.5-10 um, that proliferate percurrently in

a dichotomous fashion resulting in textura prismatico-radiata, with a crenate

to laciniate margin; UPPER WALL dense, brown, thick; LOCULEs 70-115 x

183-265 um, single, initially filled by the hamathecium that partially change

into a gelatinous mucus as the asci develop. Asci maturing sequentially, with

young and mature individuals in the same locule; youNG asci obovoid, clavate

to broadly clavate, becoming thick-walled particularly in the upper part; FULL-

SIZE ASCI WITH COLOURLESS SPORES 35-55 x 26-35 um, clavate to broadly

clavate, thick-walled particularly in the upper part, J-, with 8 spores arranged

in two rows or in a cluster; FULL-SIZE ASCI WITH BROWN MATURE SPORES

32-65 x 13-25 um, clavate. AscosporEs 15-21 x 6-8 um, initially colourless,

guttulate, becoming light brown to brown, ellipsoidal to cylindric-ellipsoidal,

thin-walled, smooth, 1-septate, constricted at the septum, LOWER CELLS 6.5-10

x 5-6 um tapering towards the base; UPPER CELL + globose, 8-11 x 5-8 um.

HAMATHECIUM of filiform branched paraphyses initially filling the locules, but

mostly evanescing as the asci develop, colourless, septate, thin-walled, slightly

bulbous at the tip, 1-3 um diam.

Dothidasteroma psidii sp. nov. (Brazil) ... 29

Fics. 1-6. Dothidasteroma psidii on Psidium laruotteanum. 1. Host leaves showing adaxially (left)

well developed circular sometimes coalescent ascomata along the leaf ribs, and linear disposition of

small ascomata mostly on top of the midrib on the abaxial (right) surface (bar = 1 cm). 2. A group

of ascomata seen in SEM (bar = 300 um). 3. Squash preparation showing an ascoma releasing the

asci under pressure (bar = 100 um). 4. Upper surface of a circular ascoma showing a radiate texture

with irregular border with a clearer central area where ascomata usually split open in a stellar fashion

(bar = 30 um). 5. Detailed view of the radiate texture of an ascoma showing the dichotomous

proliferation of the upper wall components resulting in a crenate to laciniate margin (bar = 10 um).

6. Disposition of the asci within an ascoma (bar = 20 um).

30 ... Inacio & al.

Fics. 7-11. Dothidasteroma psidii on Psidium laruotteanum.7. Anascoma (A) witha layer of

intra-cuticular dark hypostroma (arrow) showing hyphal connections with host epidermal

cells (bar = 10 um). 8. Immersed hypostroma growing through the epidermis (arrows) to

originate two ascomata (A) (bar = 5 um). 9. Brown ascospores released among paraphyses

(bar = 20 um). 10-11. Brown mature ascospores (bars = 10 and 5 um, respectively).

ADDITIONAL SPECIMENS EXAMINED: BRAZIL. BRAS{L1a: Parque Nacional on living

leaves of Psidium laruotteanum 20 Feb 1999, leg Z. M. Chaves 220 (UB Mycol. Coll.

10.256). Parque Nacional on living leaves of P laruotteanum, 15 Feb 1999, leg. Z. M.

Chaves 212 (UB Mycol. Coll. 10.127). Parque Nacional on living leaves of P laruotteanum,

27 Sept 1995, leg C. M. Leadebal 44 (UB Mycol. Coll.10.138). Lago Norte; on leaves of

P. laruotteanum, 11 Nov 1992, leg. J. C. Dianese 629 (UB Mycol. Coll. 2752).

ComMENT: Hohnel (1909) erected the genus Dothidasteroma for fungi with

“subcuticular subiculum forming a 1-cell layer, superficial, with rather flat

stromata, with locules mostly irregular, opening by fissures; asci 8-spored

with 1-septate ascospores.” Among the genera of the Parmulariaceae close

to Dothidasteroma is Aulacostroma, which shows superficial mycelium with

bulbous apressoria, contrasting with Dothidasteroma, which lacks superficial

hyphae or hyphal strands. A recent new monotypic genus, Mintera (type species

M. reticulata (Starback) Inacio & P.F. Cannon) also produces an effuse brown

internal stroma connected to the superficial ascomata with small peg-like

columns, but differs from Dothidasteroma mainly by producing a superficial

appressoriate mycelium and ascomata with radiating locules (Inacio & Cannon

2003).

Dothidasteroma species are primarily diagnosed by the presence of

subcuticular effuse brown stromata, which give rise to superficial stromatic

Dothidasteroma psidii sp. nov. (Brazil) ... 31

ascomata. The immersed stromata remain connected to the mature ascomata

by single hyphae, stromatic columns, bundles of hyphae, or peg-like filaments

(Inacio & Cannon 2008). In Asterinaceae three genera (Dothidasteromella,

Macowaniella, and Echidnodes) show immersed stromatic structures similar

to those of Dothidasteroma, although all three clearly differ in forming a

well developed hyphopodiate superficial mycelium (Inacio & Cannon 2008).

Dothidasteroma psidii has been erroneously reported earlier from Brasilia

(Inacio & Dianese 1999b) as a Polycyclinopsis species (Pereira-Carvalho et al.

2009c).

Our report constitutes a first record of the genus on a myrtaceous host and

also represents the first Dothidasteroma species from South America (previously

known only from Australia, Philippines, and Sri Lanka; Inacio & Cannon 2008,

Miller & Arx 1962, Swart 1988, Sydow & Sydow 1917).

As recently revised by Inacio & Cannon (2008), Dothidasteroma contains

three species—the type D. maculosum (Berk. & Broome) Hohn. on Pterygota

alata (Sterculiaceae), D. casuarinae H.J. Swart on Casuarina luehmannii

(Casuarinaceae), and D. dipteridis (Syd. & P. Syd.) Arx on Dipteris conjugata

(Dipteridaceae). The new species differs from all three, and can be segregated

from them, as follows:

La ASEOSDOTES VETRUCOSE «5.2 cteeuek, ste teuek, ste teu n eaten aeate ten eactet ona a wh ee et a 2

LD: SA SCOSPOTES STOOL y Faas. 2 peste! e oeeee al 2 od Wena ees Ae Gaels we wa 3

2a. Asci 35-55 x 26-35 um, spores 18-26 x 8-10 um, on Dipteris........ D. dipteridis

2b. Asci 55-76 x 25-45 um, spores 19-40 x 11-19 um, on Pterygota ...D. maculosum

3a. Ascomata connecting to the host by hyphal bundles, asci 30 um long, ovoid;

ascospores light brown, upper cells 15-19 x 10-15 um, lower cells

P= TD B Galas OTS ASU APT tale ean d Hay. Ones PH wie F week ge D. casuarinae

3a. Ascomata connecting to the host by separated hyphal strands, asci 32-65 x

13-25 um, obclavate to elliptical; ascospores 15-21 x 5-8 um brown,

upper cells 8-11 x 5-8 um, lower cell + globose, 6.5-10 x 5-6 um ..... D. psidii

Acknowledgements

The authors thank CNPq-Brasil and Fundag¢ao Banco do Brasil for financial support,

and Prof. Mariza Sanchez for assistance with the herbarium work. Thanks are also given

to Drs. Richard Hanlin and Paul Cannon for pre-submisson reviews leading to the

improvement of our manuscript. The senior author also thanks FAPDF and FINATEC

for grants.

Literature cited

Dianese JC. 2000. Micodiversidade associada a plantas do Cerrado. 109-115, in: Cavalcanti

TB, Walter BMT, Eds. Topicos atuais em Botanica. Brasilia, Brazil: Soc. Bras. Botanica /

EMBRAPA.

Dianese JC, Inacio CA, Dornelo-Silva D. 2001. Wilmia, a new genus of phaeosphaeriaceous

ascomycetes on Memora pedunculata in central Brazil. Mycologia 93: 1014-1018.

32 ... Inacio & al.

Dianese JC, Medeiros RB, Santos LTP, Furlanetto C, Sanchez M, Dianese AC. 1993. Batistopsora

gen. nov. and new Phakopsora, Ravenelia, Cerotelium, and Skierka species from the Brazilian

cerrado. Fitopatologia Brasileira 18: 436-450.

Dianese JC, Santos LTP, Medeiros RB. 1995. Kimuromyces cerradensis gen. et sp. nov. the rust of

Gongalo Alves. Fitopatologia Brasileira 20: 251-255.

Dianese JC, Medeiros RB, Santos LT. 1997. Biodiversity of microfungi found on native plants of

the Brazilian cerrado. 367-417, in: Hyde KD (ed.). Biodiversity of Tropical Microfungi. Hong

Kong: Hong Kong University Press.

Dornelo-Silva D, Dianese JC. 2004. New hyphomycete genera on Qualea species from the Brazilian

cerrado Mycologia 96: 879-884.

Dornelo-Silva D, Pereira-Carvalho RC, Dianese JC. 2007. New Stenella and Parastenella species

from the Brazilian cerrado. Mycologia 99: 753-764. doi: 10.3852

Hernandez-Gutiérrez A, Dianese JC. 2008. New cercosporoid fungi from the Brazilian cerrado 1.

Species on hosts of the families Anacardiaceae. Mycotaxon 106: 41-63.

Hernandez-Gutiérrez A, Dianese JC. 2009. New cercosporoid fungi from the Brazilian cerrado 2.

Species on hosts of the subfamilies Caesalpinioideae, Faboideae and Mimosoideae (Leguminosae

s. lat.). Mycotaxon 107: 1-24.

Hoéhnel FXR. Von. 1909. Fragmente zur Mykologie (viii. Mitteilung, nr. 407 bis 467).

Sitzungsberichten der kaiserliche Akademie der Wissenschaften in Wien, mathematische-

naturwissenschaftliche Kasse, abt. 118: 1461-1552.

Inacio CA, Cannon PF. 2003. Viegasella and Mintera, two new genera of Parmulariaceae

(Ascomycota), with notes on the species referred to Schneepia. Mycological Research 107:

82-92. doi:10.1017/S0953756202007013

Inacio CA, Cannon PF. 2008. The genera of the Parmulariaceae. CBS Biodiversity Series vol. 8.

Utrecht, Netherlands: CBS Biodiversity Centre. 196 p.

Inacio CA, Dianese JC. 1998. Foliicolous fungi on Tabebuia species. Mycological Research 102:

695-708. doi:10.1017/S0953756297005856

Inacio CA, Dianese JC. 1999a. A new Mycovellosiella species on Myracrodruon urundeuva.

Mycotaxon 72: 25-263.

Inacio CA, Dianese JC.1999b. Some Ascomycetes from Parque Nacional, Brasilia, DE Fitopatologia

Brasileira (Suplement) 24: 291.

Inacio CA, Dianese JC. 2006. Foliicolous fungi on Tabebuia species from the Cerrado. Mycological

Progress 5: 120-127. doi:10.1007/s11557-006-0507-8

Miller E, Arx JA von. 1962. Die Gattungen der didymosporen Pyrenomyceten. Beitrage zur

Kryptogamenflora der Schweiz 11(2): 1-922.

Pereira-Carvalho RC, Dornelo-Silva D, Inacio CA, Dianese JC. 2009a. Chaetothyriomyces: a new

genus in family Chaetothyriaceae. Mycotaxon 107: 483-488. doi:10.5248/107.483

Pereira-Carvalho RC, Inacio CA, Dianese JC. 2009c. New Polyciclynopsis (Ascomycota) species

from the Brazilian Cerrado. Inoculum 60(2): 35.

Pereira-Carvalho RC, Sepulveda-Chavera G, Armando EA, Dianese JC. 2009b. An overlooked

source of fungal diversity: novel hyphomycete genera on trichomes of cerrado plants.

Mycological Research 113: 261-274. doi:10.1016/j.mycres.2008.11.005

Pereira-Carvalho RC, Inacio CA, Dianese JC. 2010. Plurispermiopsis: a new capnodiaceous genus

from the Brazilian Cerrado. Mycologia 102: 1163- 1166. doi: 10.3852/09-253

Swart HJ. 1988. Australian leaf-inhabiting fungi. X. Two interesting Parmulariaceae. Transactions

of the British Mycological Society 91: 581-585. doi:10.1016/S0007-1536(88)80030-5

Sydow H, Sydow P. 1917. Beitrag zur Kenntnis der Pilzflora der Philippinen Inseln. Annales

Mycologici 15: 165-268.

MY COTAXON

Volume 116, pp. 33-47 April-June 2011

DOI: 10.5248/116.33

The genus Pythium in Taiwan (2) —an Illustrated diagnostic key

Hon H. Ho

Department of Biology, State University of New York, 1 Hawk Dr., New Paltz, NY 12561, USA

CORRESPONDENCE TO: hoh@newpaltz.edu

ABSTRACT — A diagnostic key has been developed to facilitate the identification of 48 species of

Pythium recorded in Taiwan so far. Photomicrographs are provided for the 10 most common

or economically important species to highlight their major morphological characteristics.

Key worps — Pythiaceae, oomycetes, Chromista, Straminopila

Introduction

In a review of the genus Pythium in Taiwan, Ho (2009) reported that a total

of 48 Pythium species have been recorded. ‘This is the second part of the paper,

presenting a diagnostic key to all these species. Although taxonomic keys to

Chinese Pythium species are already available (Yu & Ma 1989, Yu 1998), a

similar key is lacking for Taiwan. Considering the importance of the genus in

the ecosystem (Plaats-Niterink 1981), plant pathology (Hendrix & Campbell

1973), fish diseases (Khulbe 2009), the potential benefits to human beings as

agents biological controls of soil-borne diseases (Jones 1995) and mosquitoes

(Su et al. 2001), and as a source of enzymes useful in medicine, food industry

and the production of biofuel (Stredansky et al. 2000, Mendez-Vilas 2009),

there is a need for a diagnostic key to all species of Pythium in Taiwan to assist

mycologists, plant pathologists, and other scientists, which is presented below.

Photomicrographs of the ten most common or economically important species

are also provided to facilitate identification.

Materials & methods

Morphological characters used traditionally to classify Pythium species are limited

and often variable and overlapping. These include the characteristics of the oogonia,

oospores, antheridia, sporangia, and hyphae as well as maximum growth temperatures.

Moreover, there has been controversy and confusion regarding the terminology of some

important taxonomic criteria.

For instance, traditionally, an oospore is considered plerotic if it fills up or nearly

fills up the space of the oogonium that produces it (Middleton 1943, Waterhouse 1967,

34 ... Ho

Plaats-Niterink 1981). More recently, Dick (1990) devised a mathematical formula to

calculate the aplerotic index showing the percentage of the oogonial volume occupied

by the oospore where an oospore is considered plerotic when the aplerotic index

exceeds 70% and aplerotic when the aplerotic index is less than 60%. I have opted for

the traditional concept in using the terms plerotic and aplerotic in this key.

There are some species that form sporangia-like structures but no zoospores.

These structures have been termed “sporangia” (Middleton 1943), “hyphal bodies”

(Waterhouse 1967, Dick 1990), or “hyphal swellings” (Plaats-Niterink 1981). In this key,

“zoosporangia’ refers to sporangia that produce zoospores and “hyphal swellings” refer

to sporangia-like swellings producing no zoospores.

A diagnostic key to the species of Pythium in Taiwan

ka. Ooronia produced sinsle-cultireser : (05 Putts tte tee RS ES ER Be 2

1b. Oogonia not or rarely produced in single cultures .....................0000. 27

2a.(1a). Oogonial wall ornamented with projections ............. 0.0 cece eee eee 3

2beOogontial wall’SmiOGbar dt. wevcn, Rages andy Bomaly Bees Meus MeO ene eee 10

3a:(2a)s Projectionsmnumerous, evetily-distribileds .. 9. 56h. 5 witha cote sere 4 aa 4

3b. Projections 0-5 per oogonium, unevenly distributed....... P. irregulare Buisman

Aan 5 a1) AOOSPOnAai Ola a DSM. occ clei Maca mled wonton ote cai a ocean deal a trlonid 5 tant ote 5

ADA ZOOSPOTANS 1a PEESETIL 1 oem tissue, aes Mon faethe arn cba wince beret ea be ral aya 6

5a.(4a). Projections digitate ............. P. spinosum Sawada var. spinosum (Fic. 1)

5b.-Projections conical, acute ......4. 0.544.068 00. P. hydnosporum (Mont.) J. Schrot.

6a.(4a). Zoosporangia proliferating internally; chlamydospores large,

aVeDOMM CHAT so. eosiis eae, boas P. dimorphum EF. Hendrix & W.A. Campb.

6b. Zoosporangia non-proliferating; chlamydospores absent ..................00. ve

7a.(6b). Projections conical, acute; antheridia often absent ... P. oligandrum Drechsler

7b. Projections conical, sub-acute; antheridia present ................... 00 eee eee 8

7c. Projections conical, obtuse; zoosporangia lobulate ....... P. periplocum Drechsler

7d. Projections conical, obtuse or mammiform; zoosporangia not lobulate ......... 9

8a.(7b). Projections mostly 2-3 um long; zoosporangia often forming discrete

complexes of subglobose and inflated elements

Pact cate PSE Ra, Tee fei Ce P. acanthicum Drechsler (Fic. 2)

8b. Projections mostly 3-5 um long; zoosporangia ellipsoidal to elongated

ois es chim oes Acti OS ee oh oie Os Rh ae al P. helicandrum Drechsler

9a.(7d). Projections mostly 2-6 um long; zoosporangia globose to ovoid

Hacia ny Buchad achaa dana -dreniea dritgcdea dessa irene ee teh P. mamillatum Meurs.

9b. Projections mostly 5-8 um long; zoosporangia subglobose to broadly ovoid

(oogonia large, av. 53 um diam).................... P. polymastum Drechsler

10a(26). Zoosporangia filamentons 00. "oo is oh ea et ane eae ees See 11

LOD: ZOOSPOran Gta OD UAE pee om Ath, ete sea de secede ried weed geod etal watts» 15

LOE ZGOsporanciaslODOSe tO: OVOIds retin ges arabe odious tale ht ado d ate aes Seite I 20

10d. Zoosporangia absent (hyphal bodies lobulate) ......... P. tardicrescens Vanterp.

Key to Pythium species of Taiwan ... 35

11a.(10a). Oogonia <15 um diam ....... P. sukuiense W.H. Ko, S.Y. Wang & PJ. Ann

EIB CGogoniats: | Siem, Gating «mAs, a Merce) iN fd Pets tl baseline tau ear yedee, 12

12a.(11b). Oospore wall pale lilacin color .................... P. coloratum Vaartja

12b. Oospore wall uncolored or not pale lilacin color .....................0000. 13

13a.(12b). Oospores plerotic; antheridia stalked .......... P. monospermum Pringsh.

13b. Oospores aplerotic; antheridia stalked ........ 0... eee eee ee eee eee 14

13c. Oospores aplerotic to nearly plerotic; antheridia with short stalks or sessile

PT ER i ei Rae OMe ee oe at oe P. dissotocum Drechsler (Fic. 3)

14a.(13b). Antheridia often several from the same stalk, 1-4 antheridia per oogonium,

terminal orantercalatys 34s. +94,ty-eadinaer 3 diner F4 Raber 40H P. adhaerens Sparrow

14b. Antheridia usually one per stalk; 1(-2) per oogonium,

Mmostly-teriiitialy 235 ci. 2.26 £7. Ge 2 Gee hon lw ch lew oe Ne P. gracile Schenk

-Sa-Ch0b).-Antheridiatottentintercalary. Loch re h seth epee eee Tae 16

1SbAntheridiadsuallymet-imntercalar ys sesh sh stete yaaa eatin este eee one eueya tae 17

16a.(15a). Oogonial stalks straight ....... P. aphanidermatum (Edson) Fitzp. (Fic. 4)

16b. Oogonial stalks mostly curved towards the antheridia P. deliense Meurs. (Fic. 5)

EAE Bs QOSpOrest Per Ol Gs. 24 eass-g a cesiss-oa osaielen patiate tales re satiate Gatien giieiie 0 Blt ine 18

T/ be DOspOresraPle LOU cray g.:ricteg tt sveg trbesneoa gets petea drhog oa trea dog dope deep ceed pees eis oe 19

18a.(17a). Antheridia mostly diclinous; 15-20 antheridia per oogonium

Mats Reid AaP IML oA tL LU ELL Bi Al P. arrhenomanes Drechsler

18b. Antheridia mostly monoclinous, 1-2 antheridia per oogonium

scat lea eine mete Arts RA eee Aye AMO P. torulosum Coker & P. Patt.

18c. Antheridia mostly monoclinous; 1-6 antheridia per oogonium

Eth uaa Naa wea Weenie, Heratee teresa Wows Vi Merk ee Mewes P. graminicola Subraman.

19a.(17b). Maximum growth temperature >40°C ... P. myriotylum Drechsler (Fic. 6)

19b. Maximum growth temperature <40°C; antheridial branch

entwining oogonial stalk.................0... P. volutum Vanterp. & Truscott

19c. Maximum growth temperature <40°C; antheridial branch

not entwining oogonial stalk ...................0.. P. aristosporum Vanterp.

20a.(10c). Zoosporangia non-proliferating or occasionally proliferating internally;

OOS Onis BOA TAN says cet cusp aeons awn thcns Ma Biaya ee toon eas As donee 21

20b. Zoosporangia proliferating internally; oogonia >30 um diam ............... 26

20c. Zoosporangia absent; oogonia <30 um diam .... 6... eee eee

Vigeerhtas ceMlane cablen tetera aeoten aaa ee P. ultimum Trow var. ultimum (Fic. 7)

21a.(20a)..Oospores plerotie soe... a. cs ea stews ana saa aaa oa gol oe as 22

ZB OOSPOLEsraPlCTOLIGL 0.2% Patt Poh 2e Fo MM! UNM! UML! USER LAM! 1 FN ee Ee 23

22a.(21a). Antheridia hypogynous or monoclinous, oogonia often in chain

Foes dh ok te Site be Soke Pa eigt PL) OF ORE Fay BS gee A, P. rostratum E.J. Butler

22b. Antheridia absent or monoclinous; oogonia often in chain

Pare ee a eres et a en Sh Mon ee eA P. salpingophorum Drechsler

22c. Antheridia diclinous or monoclinous; oogonia not in chain .. P. pleroticum T. Ité

230 OED) anit heridiy Belleghaped er ut90 F349 Fuh 8th 8 oY RE BR Nh ORLY 24

23h. Antheridiasnot belloshaped o0 2. ue <p Mey lose eee sey beng hs See ee FS 25

36 ... Ho

24a.(23a). Sporangia globose, pyriform or ovoid, occasionally proliferating internally;

irregularly elongated antheridia also present ......... P. vexans de Bary (Fie. 8)

24b. Sporangia various shaped, non-proliferating; elongated antheridia

POPPRESCHIL Raa se hotly. orate eu ca Tet cna ols P. perplexum H. Kouyeas & Theoh.

25a.(23b). Antheridia mostly hypogynous; sporangia and/or oogonia mostly

intercalary and occasionally catenulate ................. P. pulchrum Minden

25b. Antheridia variable, mostly monoclinous, sessile but occasionally

hypogynous; oogonia often intercalary or catenulate; sporangia terminal

OP AMPLE CATAL * Areas actad ott ecto chcte ootad dehesghad dub fubod deh eben stchs P. paroecandrum Drechsler

25c. Antheridia mostly monoclinous, stalked; oogonia terminal and occasionally

intercalary; sporangia terminal or intercalary ......... P. debaryanum R. Hesse

26a.(20b). Antheridia smooth in contour; oogonia terminal; maximum growth

Lelmperatiie SANG sores oie Weed Lai aie alae aay P. helicoides Drechsler (Fic. 9)

26b. Antheridia often wavy in contour; oogonia terminal; maximum growth

LEIP ETAL TE, AAO Ea I en RA iene es lesa d ag iectt a ih P. oedichilum Drechsler

26c. Antheridia often wavy in contour; oogonia intercalary or lateral

a tb sk RM eve We Hava Ree Mewante nN cua Bath Gna se Merk 8a Wows tN end P. palingenes Drechsler

27a.(1b). Zoosporangia non-proliferating, non-papillate....................000. 28

27b. Zoosporangia proliferating internally, papillate .......................000. 31

ZLCPLOOSPOLANGIA-ADGE NIG tescsnbad tet skal ets snbad ch shad dude geben hy pchag di Aken chche setiucd ch beau dri psa Se

28a. (27a). Zoosporangia filamentous ........... P. afertile Kanouse & T. Humphrey

28b. Zoosporangia spherical, ovoid to cylindrical ......... 0... cee eee eee 29

28e-Zoo0sporangialobulates, 2.2 x opus xe ce ew oe nee oe En ee Gees ole ch alc OS 30

29a.(28b). Zoosporangia terminal, spherical, catenulate in water, deciduous,

ZOOSPONCS-HALC so. hace fee eayl Lee ecgs be laee eas eae P. intermedium de Bary

29b. Zoosporangia terminal or intercalary, spherical, pyriform to cylindrical;

exit tube-verylone,upto150 ums irk ett rs P. elongatum V.D. Matthews

30a.(28c). Spherical hyphal swellings (10-20 um) catenulate in water,

HOPOSCIMUOUE So + isa + ieas 7 teas oo ease tlw P. catenulatum V.D. Matthews

30b. Toruloid hyphal elements present; zoospores rare .... P. inflatum V.D. Matthews

31a.(27b). Zoosporangia ellipsoidal, up to more than100 um long, terminal

On Sporangiophore..... <.8u wl. kaw cs wee eae Nees P. undulatum H.E. Petersen

31b. Zoosporangia globose, pyriform to asymmetrically utriform, often

transversely attached to sporangiophore ............. P. marsipium Drechsler

31c. Zoosporangia globose to ovoid, terminal on sporangiophore ................ 32

32a.(31c). Zoosporangia av. 25 um diam; hyphal swellings catenulate

ey WALDEN tein one Min 5 LAMAR s heahe due chy P. carolinianum V.D. Matthews

32b. Zoosporangia av. 30 um diam .................. 000 eee P. middletonii Sparrow

33a.(27c). Hyphal swellings large, globose, mostly 30-40(-49) um diam and

{R23 © AULD GPE Uae veh rt Ey ON AOR eh aoe een P. splendens Hans Braun (Fic. 10)

33b. Hyphal swellings globose to ovoid, mostly 25-30 um diam, intercalary or

RCE TTATIT AL, cer Comet Merve, aM ae a tla P. sylvaticum W.A. Campb. & EE Hendrix

Key to Pythium species of Taiwan ... 37

Discussion

I have not attempted to address the difficult problem of speciation in the

genus Pythium in the above key and all species recorded in Taiwan have been

accepted without prejudice. Although Plaats-Niterink (1981) considered

P. debaryanum to be conspecific with P ultimum var. ultimum, and P. gracile

with P monospermum, I have accepted the thesis of Waterhouse (1976) and Dick

(1990) that they are recognizably distinct taxa. Similarly, P. afertile is retained

in this key, as in the keys developed by Waterhouse (1967) and Dick (1990),

despite the fact that it was rejected by Plaats-Niterink (1981). The preparation

of a traditional diagnostic key for all species of Pythium in Taiwan is only the

first step in the study of the taxonomy of the genus on the island. Hopefully,

in the future, with a combination of morphological studies and the utilization

of modern molecular biology techniques, a clearer picture of the speciation of

Pythium in Taiwan will emerge.

Acknowledgements

Thanks are due to Drs. W.H. Ko and C.X. Hong for reviewing the manuscript, the

National Science Council of Taiwan for financial support, to Dr. H.S. Chang of Institute

of Botany, Academic Sinica, Taipei for providing space, equipment and technical

assistance in the study of isolates of Pythium species from Taiwan, and to Dr. Shaun

Pennycook for his meticulous and helpful editing.

Literature cited

Dick MW. 1990. Keys to Pythium. College of Estate Management, Whiteknights, Reading.

Ho HH. 2009. The genus Pythium in Taiwan, China (1) — a synoptic review. Front. Biol. China

4(1): 15-28. doi:10.1007/s11515-009-0009-6

Hsieh HJ. 1976. Studies on genera Pythium and Pythiogeton in Taiwan. MS Thesis. Taipei: Taiwan

National University (in Chinese).

Jones EE. 1995. Comparative physiology and behavior of the mycoparasites Pythium acanthophoron,

P. oligandrum and P. mycoparasiticum. Biocontrol Science & Technology 5(1): 27-40.

doi: 10.1080/0958319550039990

Khulbe RD. 2009. Pathogenicity of some species of Pythium Pringsheim on certain fresh water

temperate fishes. Mycoses 26(5): 273-275. doi: 10.1111/j.1439-0507

Mendez-Vilas A. (ed.) 2009. Current research topics in applied microbiology and microbial

biotechnology. World Scientific, Singapore.

Middleton JT. 1943. The taxonomy, host range and geographical distribution of the genus Pythium.

Memoirs of the Torrey Botanical Club 20: 1-171.

Plaats-Niterink AJ van der. 1981. Monograph of the genus Pythium. Stud. Mycol 21: 1-242.

Stredansky M, Conti E, Salaris A. 2000. Production of polyunsaturated fatty acids by Pythium

ultimum in solid-state cultivation. J. Industrial Microbiology 26(2-4): 304-307. doi:10.1016/

SO141-0229(99)00146-5

Su XQ, Zhou FH, Guo Q, Huang J, Chen TX. 2001. A report on a mosquito-killing fungus, Pythium

carolinianum. Fungal Diversity 7: 129-133.

Yu YN. (ed.) 1998. Flora Fungorum Sinicorum Vol. 6. Peronosporales. Science Press, Beijing (in

Chinese).

Yu YN, Ma GZ. 1989. The genus Pythium in China. Mycosystema 2: 1-110.

Fic. 1. Pythium spinosum var. spinosum. A. appressorium; B. digitate appendages on an

intercalary hyphal swelling; C. intercalary hyphal swelling; D, E. terminal hyphal swellings; F-J.

ornamented terminal oogonia with stalked antheridia, monoclinous in F, G, and diclinous in H-J;

K-P. ornamented intercalary oogonia with antheridia stalked and monoclinous in K-M but

hypogynous in N-P; Q, R. oogonia with plerotic oospores. Scale bar: 20 um (all at the same

magnification)

Key to Pythium species of Taiwan ... 39

Fic. 2. Pythium acanthicum. A, B. subglobose sporangial complex; C, D. terminal oogonia with

monoclinous antheridia; oogonia with short, subacute conical projections and plerotic oospores.

Scale bar: 20 um (all at the same magnification)

Fic. 3. Pythium dissotocum. A, B. hyphal tip forming filamentous sporangium; C. zoospores in

a vesicle at the tip of an exit tube; D. Empty cyst with an exit tube after the cyst has germinated

by repeated emergence of zoospore; E. Cyst germinates by a germ tube; F-J. oogonia with short

stalked monoclinous antheridia; K, L. oogonia with aplerotic to nearly plerotic oospores Scale bar:

20 um (all at the same magnification except A)

Key to Pythium species of Taiwan ... 41

Fic. 4. Pythium aphanidermatum. A, B. lobulate sporangia, sporangium empty with long exit tube

in B after zoospore release; C-D. cyst germination by germ tube(s); E. Empty cyst with two exit

tubes after germination by repeated emergence of zoospore; F. appressorium; G. oogonium with a

terminal, stalked monoclinous antheridium; H-J. oogonia with intercalary antheridia; K. oospore

with aplerotic oospore. Scale bar: 20 um (all at the same magnification except C and K)

42... Ho

Fic. 5. Pythium deliense. A, B. lobulate sporangia, sporangium empty with long exit tube in B.

C, D. curving of oogonial stalk toward the antheridium (terminal in C and intercalary in D);

E-G. oogonia with aplerotic oospores and antheridia (terminal and diclinous in E but intercalary

in F and G). Scale bar: 20 um (all at the same magnification)

Key to Pythium species of Taiwan ... 43

Fic. 6. Pythium myriotylum. A-C. appressoria, conspicuously broom-like in C; D, E. lobulate

sporangia, empty sporangium with long exit tube in E; F-K. oogonia surrounded by club- or crook-

shaped mostly diclinous antheridia; L, M. mature oogonia with mostly aplerotic oospores in M and

an occasional plerotic oospore in L. Scale bar: 20 um (all at the same magnification, except A)

44 ...Ho

Fic. 7. Pythium ultimum var. ultimum. A. appressorium; B-F. hyphal swellings; G-I. terminal

oogonia with monoclinous sessile antheridia attached to the base of oogonia; J-L. intercalary

oogonia with monoclinous antheridia, sessile in J, on short stalk in K and hypogynous in L;

M-Q. Mature terminal oogonia with thick-walled aplerotic oospore; S, T. intercalary oogonia with

developing aplerotic oospores and monoclinous antheridia, sessile in R; on short stalk in S and

hypogynous in T. Scale bar: 20 um (all at the same magnification except A)

Key to Pythium species of Taiwan ... 45

Fic. 8. Pythium vexans. A. appressorium; B-D. terminal sporangia , empty sporangia with short

exit tubes in C and D; E. an empty sporangium proliferating internally; F. intercalary sporangium;

G-K. developing oogonia with antheridia campanulate in H, I and J but irregularly elongated in

G and K; L-O. mature oogonia with mostly aplerotic oospores in L and M but occasionally nearly

plerotic oospores in N and O. Scale bar: 20 um (all at the same magnification)

46 ... Ho

Fic. 9. Pythium helicoides. A. sporangiophore with cymose branching; B-D. terminal papillate

sporangia showing internal proliferation in C and D; E-G. developing oogonia with antheridial

stalk coiling around the oogonial stalk at the base of the oogonium; H, I. Mature oogonia with large

elongated antheridia applied close to the oogonium in H with thick-walled aplerotic oospores Scale

bar: 20 um (all at the same magnification except A)

Key to Pythium species of Taiwan ... 47

Fic. 10. Pythium splendens. A-C. hyphal swellings terminal in A and B but intercalary in C;

D. hyphal swelling germinate by germ tube; E. developing terminal oogonium with diclinous

antheridium; F-L. mature oogonia with aplerotic oospores and club- or crook-shaped antheridia.

Scale bar: 20 um (all at the same magnification)

MY COTAXON

Volume 116, pp. 49-52 April-June 2011

DOI: 10.5248/116.49

Lecanora, Phaeophyscia and Rinodina species new to Turkey

BELGIN ARSLAN, SULE OztTuUrRK & SEYHAN ORAN’

Department of Biology, Faculty of Arts & Sciences, Uludag University, 16059, Bursa, Turkey

CORRESPONDENCE TO *: seyhana@uludag.edu.tr

ABSTRACT — Four lichenized fungi (Lecanora jamesii, L. juniperina, Phaeophyscia hirsuta,

and Rinodina orculata) are reported for the first time from Turkey. Comments on their

habitat, substrate, and morphological and anatomical features are provided.

Key worps — Ascomycota, Bursa, Canakkale, lichens

Introduction

Lichenological studies in Turkey have rapidly increased in recent years.

Prior to 2004, 361 papers referring to lichens were published (John 2004).

Since then, many floristic studies have added several new records to the lichen

biodiversity of Turkey (e.g. Yazici1 & Aslan 2005, Giiveng et al. 2006, John &

Turk 2006, Halici et al. 2007, Yavuz & Cobanoglu 2007, Candan & Tiirk 2008,

Halici & Aksoy 2009, Kocakaya et al. 2009, Kinalioglu 2009, 2010). In spite of

the increase in the number of studies, additional research is needed in some

provinces where lichen biodiversity is still not sufficiently understood. ‘This

paper contributes further knowledge of the lichenized fungi in Turkey.

Materials & methods

Vouchers are stored in BULU (Herbarium of Uludag University, Sciences and Arts

Faculty, Bursa, Turkey) and their accession numbers are given in parenthesis with the

locality information. The specimens were examined with stereomicroscope (Prior), and

light microscope (Kriiss). Anatomical sections were examined in water, 10% KOH, and

Lugol’s iodine solution. Ascospore measurements were carried out in water and the

extreme values outside the main range are given in parenthesis.

Species recorded

Lecanora jamesii J.R. Laundon 1963 Fic. 1

SPECIMEN EXAMINED—TURKEY. CANAKKKALE: Bayramic; Kazdagi Mountain,

Dalaksuyu place, beech forest, 39°46N, 26°58E, alt. 1300 m, on Fagus orientalis Lipsky

(Fagaceae), 18 Aug. 2005, leg. S. Oran, det. S. Oran (BULU 13785).

50 ... Arslan, Oztiirk & Oran

Thallus to 2-3 cm in diam., forming continuous, smooth or granular patches,

grey; prothallus when present black; soredia arising in + circular soralia, to

1 mm diam, farinose, pale yellow (to whitish on storage). Apothecia absent.

Thallus P-, K-, C-, soralia sometimes weakly K+ yellowish.

On smooth bark of deciduous trees, most frequently Salix in damp situations,

rarely on wood or siliceous rock; locally abundant (Purvis et al. 1992). In our

study area, L. jamesii was found in a forest on smooth bark of Fagus orientalis.

Lecanora jamesii has been reported from western Europe and Chile (Purvis

et al. 1992), and Iran (Valadbeigi & Sipman 2010). Clauzade & Roux (1985) and

Purvis et al. (1992) describe the species in detail.

Lecanora juniperina Sliwa 2004 Fic. 2

SPECIMEN EXAMINED— TURKEY. Bursa: Inegdl; road of Hilmiye-Oylat, 1 km from

Hilmiye, rocky area, 39°56N, 29°35E, alt. 685 m, on Juniperus sp. (Cupressaceae),

19 Sept. 2004, leg. B. Arslan, det. $. Oztiirk (BULU 9959).

Thallus granulose to areolate-rimose, grayish. Apothecia 0.3-1.2 mm in

diam.; disc pale, sometimes yellow, usually pruinose, smooth; margin usually

prominent, crenulate to dentate, paler than the disc; epihymenium granular,

granules not soluble in K and N. Ascospores 9.0-13.5 x 4.5-6.0(-7.5) um,

ellipsoid.

Lecanora juniperina grows on tree bark (Juniperus and Quercus) and wood

at intermediate elevations (Nash et al. 2004). Similar to the literature, our

specimen was found on bark of Juniperus sp. in rocky area and was noted for its

small thallus with very crowded and heavy pruinose apothecia.

Nash et al. (2004) and Sliwa (2007), who also describe the species in detail,

recorded L. juniperina from North America. Valadbeigi & Sipman (2010) also

report the species from Iran.

Phaeophyscia hirsuta (Mereschk.) Essl. 1978 Fic. 3

SPECIMEN EXAMINED—TURKEY. Bursa: Inegél; road of Cayyaka-Littfiye, 1 km from

Cayyaka, oak woodland, 39°59N, 29°28E, alt. 489 m, on Juglans sp. (Juglandaceae),

06 Jun. 2004, leg. B. Arslan, det. §. Oztiirk (BULU 9601).

Thallus foliose, up to 3(-4) cm in diam., often orbicular; lobes 0.5-1(-1.5) mm

broad, upper surface gray to gray-brown or brown, with cortical hairs near the

lobe ends, sorediate; soralia labriform; lower surface black; rhizines simple,

black. Apothecia numerous, up to 2 mm in diam, sessile, margin usually with

cortical hairs.

Phaeophyscia hirsuta grows mostly on bark but also commonly on rock

(Nash et al. 2004). Our specimen was collected from a trunk of Juglans sp. in a

protected forest area.

Clauzade & Roux (1985), Nash et al. (2004), and Liska et al. (2008) have

reported P. hirsuta from North America, Europe, and Africa. The first two

papers describe the species in detail.

Lichens new to Turkey... 51

ene oe

Fic. 1. Lecanora jamesii (BULU 13785). Fic. 2. Lecanora juniperina (BULU 9959).

Fic. 3. Phaeophyscia hirsuta (BULU 9601). Fic. 4. Rinodina orculata (BULU 9818).

Rinodina orculata Poelt & M. Steiner 1970 Fic. 4

SPECIMEN EXAMINED—TURKEY. Bursa: Ineg6l; road of Kursunlu-Giizelyurt, 3 km

from Kursunlu, oak woodland, 40°02N, 29°42E, alt. 575 m, on Quercus sp., 20 Jun. 2004,

leg. B. Arslan, det. B. Arslan (BULU 9818).

Thallus crustose, usually effuse and discontinuous, composed of small

verrucae, whitish, yellowish-brown to reddish-brown. Apothecia lecanorine,

adnate to sessile, 0.2-0.4(-0.6) mm in diam., thalline margin thick, persistent.

Apothecial cortex up to 20 um, I+ blue; disc brown, plane; epihymenium red-

brown. Ascospores 12-15(-17) x 6-8 um, Physconia-type.

Rinodina orculata occurs at high altitudes in coniferous forests, where it

grows mainly on small coniferous twigs. Our lichen sample, which was found

on Quercus bark in a broad-leaved forest, was diagnosed by its numerous and

round apothecia with dark brown discs.

52... Arslan, Oztiirk & Oran

This is a montane species with a distribution centered in the European Alps

where it mostly grows on Rhododendron spp. Giralt (2001), who describes

the species in detail, has reported the species from the Iberian Pyrenees and

Portugal; R. orculata has also been recorded in the Czech Republic (Liska et

al. 2008).

Acknowledgements

We would like to thank Aysen Tiirk (Eskisehir, Turkey) and M. Gokhan Halici

(Kayseri, Turkey) for reviewing this paper and Dr. Harrie Sipman (Berlin) for checking

the specimens and providing valuable suggestions for the manuscript. Collection of

Lecanora jamesii was made possible by project (2006/63), funded by the Unit of Scientific

Research Projects, Uludag University.

Literature cited

Candan M, Ozdemir Tiirk A. 2008. Lichens of Malatya, Elazig and Adiyaman provinces (Turkey).

Mycotaxon 105: 19-22.

Clauzade G, Roux C. 1985. Likenoj de Okcidenta Etropo. Ilustrita Determinlibro. Royan. Bulletin

de la Société Botanique du Centre-Ouest Nouvelle série-Numéro Spécial.

Giralt M. 2001. The lichen genera Rinodina and Rinodinella (lichenized Ascomycetes, Physciaceae)

in the Iberian Peninsula. Bibliotheca Lichenologica, Band 79, Stuttgart.

Gitveng $, Oztiirk $, Aydin S. 2006. Contributions to the lichen flora of Kastamonu and Sinop

provinces in Turkey. Nova Hedwigia 83: 67-98. doi:10.1127/0029-5035/2006/0083-0067

Halici MG, Aksoy A. 2009. Lichenized and lichenicolous fungi of Aladaglar National Park (Nigde,

Kayseri and Adana Provinces) in Turkey. Turkish Journal of Botany 33: 169-189. doi:10.3906/

bot-0810-14

Halic1 MG, Candan M, Ozdemir Tiirk A. 2007. New records of lichenicolous and lichenized fungi

from Turkey. Mycotaxon 100: 255-260.

John V. 2004. Lichenological studies in Turkey and their relevance to environmental interpretation.

Abstract book, XI OPTIMA meeting, 5.-11.9.2004 Belgrad: 45.

John V, Turk A. 2006. Species/area curves for lichens on gypsum in Turkey. Mycologia Balcanica

3: 55-60.

Kinahioglu K. 2009. Lichens from the Amasya, Corum, and Tokat regions of Turkey. Mycotaxon

109: 181-184.

Kinahoglu K. 2010. Lichens of Ordu Province, Turkey. Mycotaxon 112: 357-360.

doi:10.5248112.357

Kocakaya M, Halici MG, Aksoy A. 2009. Lichens and Lichenicolous Fungi of Kizildag (Derebucak,

Konya). Turkish Journal of Botany 33: 105-112. doi:10.3906/bot-0810-2

Liska J, Palica Z, Slavikova §. 2008. Checklist and Red List of lichens of the Czech Republic. Preslia

80: 151-182.

Nash III TH, Ryan BD, Diederich P, Gries C, Bungartz F. 2004. Lichen Flora of the Greater Sonoran

Desert Region, Volume 2. Tempe, Arizona State University.

Purvis OW, Coppins BJ, Hawksworth DL, James PW, Moore DM. (Eds.) 1992. The lichen flora of

Great Britain and Ireland. London, Natural History Museum Publications.

Sliwa L. 2007. A revision of the Lecanora dispersa complex in North America. Polish Botanical

Journal 52(1): 1-70.

Valadbeigi T, Sipman HJM. 2010. New records of lichens and lichenicolous fungi from Iran and

their biogeographical significance. Mycotaxon 113: 191-194. doi: 10.5248/113.191

Yavuz M. Cobanoglu G. 2007. Lichen flora of Pamukkale (Hierapolis) Turkey. Pakistan Journal of

Biological Sciences 10(17): 2998-3001.

Yazici K, Aslan A. 2005. Six new lichen records from Turkey. Mycotaxon 93: 359-363.

MY COTAXON

Volume 116, pp. 53-60 April-June 2011

DOI: 10.5248/116.53

Four new records of Caloplaca (lichenized Ascomycetes) from India

YOGESH JOSHI’ & DALIP K. UpRetr’

"Department of Botany, S.S.J. Campus, Almora 263601, Uttarakhand, India

’Lichenology Laboratory, National Botanical Research Institute,

Rana Pratap Marg, Lucknow-226001, Uttar Pradesh, India

CORRESPONDENCE TO *: *? dryogeshcalo@gmail.com, ? upretidk@rediffmail.com

ABSTRACT — Detailed taxonomic descriptions are presented for four species of the lichen

genus Caloplaca, newly reported from India. Caloplaca litoricola (also newly reported from

Asia) is a littoral species, recorded only from coastal regions in India, while C. chalybaea,

C. lypera, and C. maura are recorded only from inland areas.

Key worps — Chalybaeae, Chrysophorae, Conversae, Kamczatica, Ochrotropae, Sideritis,

Teloschistaceae

Introduction

Until 2009, numerous floristic or revisionary papers dealing with either the

lichen family Teloschistaceae Zahlbr. or the genera Caloplaca Th. Fr., Ioplaca

Poelt, and Xanthoria (Fr.) Th. Fr. from India were published (Acharius 1810,

1814; Rasénen 1950, 1952; Awasthi 1963, 1965, 1986, 1988, 1991, 2000, 2007;

Singh 1964; Poelt 1977; Singh & Awasthi 1978; Singh 1981; Poelt & Hinteregger

1993; Singh & Sinha 1994; Sinha & Singh 2005; Joshi & Upreti 2006, 2007a,b,

2008; Joshi et al. 2008, 2009). Addtionally, Joshi revised the Teloschistaceae

from India in an unpublished thesis (Joshi 2008), describing 78 teloschistacean

species, of which 67 represent Caloplaca. In this thesis, he cited C. chalybaea,

C. litoricola, C. lypera, and C. maura as occurring in India. Since these species

have not been included in any published accounts of the Indian lichen biota,

we provide detailed taxonomic descriptions, chemical and ecological data,

and comments for each species to expand the knowledge of lichen diversity in

India.

Materials & methods

The study is based on lichen specimens deposited in the herbarium of LWG (including

LWG-LWU). Specimens were examined using standard microscopical techniques and

were hand-sectioned under Digi-Zoom binocular. All measurements were made on

54 ... Joshi & Upreti

material mounted in water; lactophenol cotton blue (LCB) was used only as a stain.

Spot test reactions were carried out on hand-sections of thalli and apothecia under

Nikon Eclipse 400 compound microscope. The K+ purple reaction of the epihymenium

should not be confused with K+ red reaction, as the former represents lack of parietin

complex in the taxa. Species with K+ red reaction will always have parietin complex.

Measurements of ascospores were made at x400 magnification mounted in water; only

free ascospores lying outside the asci were measured. The number of microscopical

measurements (n) taken is noted at the end of the pertinent description for each species.

Secondary metabolites were identified by TLC as described by White & James (1985)

using solvent system C. HCl was used to detect the nature of rock (bubbles indicate

calcareous rock).

Taxonomy

Caloplaca chalybaea (Fr.) Mill. Arg., Mém. Soc. Phys. Hist. nat. Genéve 16: 388,

1862.

SPECIMENS EXAMINED: INDIA. Mapuya PRADESH: Anuppur district, Amarkantak, 10

km before Kabir Chabutra (Attaria), alt. 510 m, on quartz rocks, 22 March 2004, Upreti,

Nayaka & Satya 04-002372, 04-002359/B (LWG); Hoshangabad district, Pachmarhi, on

way to little fall, alt. ca. 1050 m, on rock, 24 January 1973, S.R. Singh 73.135 (LWG).

TAMIL Napv: Chennai, Pulboli, LHS, alt. 1030 m, on rock, 25 July 2000, Hariharan &

Balaji CS12a (MSSRF).

THALLUS crustose, saxicolous, appearing as spotted grayish patch on a dark black

hypothallus, areolate to coarsely rimose-areolate, olivaceous gray to brownish-

gray to lead gray to pale gray. CoRTEX paraplectenchymatous. MEDULLA white.

PROTHALLUs black, dendritic.

APOTHECIA numerous, scattered to + clustered, immersed in the thallus

at first, later on sessile, round, 0.2-0.3 mm in diam.; disc brownish-red to

brownish-black to black, flat to subconvex; proper margin thin to moderate,

smooth, entire, persistent, concolorous or slightly darker than disc, glossy, in

cross-section of indistinct to small oval cells; thalline margin thin, at maturity +

restricted to the base of apothecia, concolorous with the thallus. EPtHyMENIUM

brownish to golden-brown. HyMENtvuM hyaline, 40-60 um high. HyPOTHECIUM

hyaline, of paraplectenchymatous cells, impregnated with crystals, oil globules

absent. AMPHITHECIUM with algae. PARAPHYSES simple to furcated, with

upper 1-3 cells swollen. Ascr 8-spored, spores polarilocular, ellipsoid, (6-)8-

14 x 3-6 um, isthmus 1-3 um. Pycnip1A numerous, 1-3 per areolae, ostiole

brownish-black to black, conidia bacilliform, 1-2 x 0.5-0.7 um. [n = 20 for all

measurements.] For further descriptions see Wunder (1974).

CHEMISTRY — Spot test reactions: Thallus K+ yellow, C-. Medulla K-, C-.

Apothecial disc K+ purple or K-. Epihymenium K+ faint purple. Secondary

metabolites: atranorin.

ECOLOGY AND DISTRIBUTION — Caloplaca chalybaea was found growing

over boulders (quartzite) in tropical regions between 510-1050 m. It was

Caloplaca spp. new to India... 55

accompanied by Caloplaca subsoluta (Nyl.) Zahlbr. and C. cinnabarina (Ach.)

Zahlbr.

Wade (1965) and Wunder (1974) reported this species growing over dolomite

and calcareous rocks from Central Europe. Later, Fletcher & Laundon (2009)

reported this species growing over hard limestone, natural boulders, buildings,

and chest tombs from Europe, Macaronesia, Asia, and Africa. Wetmore (1994)

reported it from North and Central America. The present study extends its

distribution in India

REFERENCE SPECIMEN EXAMINED: FRANCE. Vauc use: Lagarde [inter Rustrel et

saint Christol], prope Le Buis, alt. 1000 m.s.m., 23 April 1965, ad saxa calcarea, leg.

G. Clauzade, J. Lambinon et A. Vézda 399 (A. Vézda: Lichenes selecti exsiccati).

REMARKS — According to Hansen et al. (1987) and Poelt & Hinteregger (1993),

C. chalybaea belongs to Chalybaeae group, comprising species characterized

by a crustose to poorly effigurate grayish thallus, brown to black apothecial

disc without anthraquinone crystals but with diffused gray-violet pigments,

and widely ellipsoid spores with a tendency of reduced septum. Later Wetmore

(1994) placed this species in a group of taxa characterized by brown or

black-fruited apothecial disc, K-/K+ yellow thallus, and K-/K+ purplish

epihymenium. This rather large group includes species representing mostly all

ecological groups, e.g., corticolous, lignicolous, and saxicolous (Massalongo

1853; Wetmore 1994). Five other members of this group previously reported

from India (Singh & Awasthi 1978; Poelt & Hinteregger 1993; Joshi & Upreti

2007a; Joshi et al. 2009) include three corticolous species — C. atrosanguinea

(G. Merr.) ILM. Lamb, C. pollinii (A. Massal.) Jatta, C. rinodinopsis Poelt

& Hinter. — and two saxicolous species — C. transcaspica (Nyl.) Zahlbr.,

C. variabilis (Pers.) Mull. Arg.

Caloplaca chalybaea is characterized by a crustose areolate to coarsely

rimose-areolate grayish thallus with a dendritic black prothallus, numerous

small apothecia, a brownish-red to blackish apothecial disc with a concolorous

proper margin, and numerous pycnidia with brownish-black to black ostioles.

Superficially it resembles Aspicilia in habit but is easily distinguished by the

polarilocular spores and K+ purple epihymenium. It is close to C. atroflava

(Turner) Mong., which differs from it in having a dark gray to black, cracked

areolate thallus with a brownish-orange apothecial disc lacking a thalline

margin and broadly ellipsoid spores. Another related species, C. concilians

(Nyl.) H. Olivier, is differentiated by bigger apothecia, branched paraphyses,

and lack of dendritic prothallus.

Caloplaca litoricola Brodo, Bryologist 87: 98, 1984.

SPECIMEN EXAMINED: INDIA. ANDHRA PRADESH: Vishakhapatnam, along sea coast,

on rock, 07 March 1986, D.D. Awasthi, G. Awasthi, R. Mathur & P. Srivastava 86.272

(LWG-LWU).

56 ... Joshi & Upreti

THALLUS crustose, saxicolous, thin, continuous or areolate, whitish-gray.

Cortex cellular and thin. MEDULLA white. PROTHALLUS absent.

APOTHECIA numerous, scattered, sessile, round to angular owing to pressure,

0.2-0.6(-0.9) mm in diam.; disc orange-yellow to dark orange, flat to convex;

proper margin thick, smooth to wavy, entire, persistent, raised above the level

of the disc, black, glossy, in cross-section of radiating elongated hyphal cells,

outer surface aeruginose pigmented; thalline margin absent. EPIHYMENIUM

golden-brown. HyMENtIvuM hyaline, 70-90 um high. HypoTHeEcrum hyaline or

pale brown, of isodiametric cells, oil globules present. AMPHITHECIUM without

algal cells. PARAPHYSES simple, rarely branched, without swollen tips or with

1-3 slightly swollen cells, upper portion covered by epipsamma. Asc1 8-spored,

spores polarilocular, oval to ellipsoid, 11-16 x 6-9(-10) um, isthmus 5-6 um.

Pycnip1A immersed, ostiole black, conidia 2.5-3.2 x 1-1.5 um. [n = 10 for all

measurements.]For further descriptions see Brodo (1984).

CHEMISTRY — Spot test reactions: Thallus and medulla K-, C-. Apothecial

disc and epihymenium K+ red, margin K- (remaining aeruginose). Secondary

metabolites: parietin.

ECOLOGY AND DISTRIBUTION — Caloplaca litoricola was found growing

on siliceous rocks along the shoreline of Vishakhapatnam district, Andhra

Pradesh.

Benton et al. (1977) first reported this species (as Caloplaca sp.) from

Bamfield Marine Station, Vancouver Island (North America). Brodo (1984)

cited C. litoricola from the Queen Charlotte Islands and Long Beach (Ucluelet)

on Vancouver Island’s west coast, and Arup (1995ab) and Wetmore (1996)

reported it growing over non-calcareous maritime rocks from Washington to

Alaska. Our report represents the first for Asia, where it is thus far known only

from India’s east coast.

REFERENCE SPECIMEN EXAMINED CANADA. BRITISH COLUMBIA: Queen Charlotte

Islands: Graham Island, Tow Hill, alt. 54°04'N, 131°47'W, on shoreline rock, 16 June

1967, ILM. Brodo & M.J. Shchepanek 9905 (Isotype-CANL).

REMARKS — Wetmore (1996) placed this C. litoricola in the Sideritis group,

which consists chiefly of saxicolous species with gray, whitish, or brown

thalli that lack anthraquinones and red or orange anthraquinone-containing

apothecia. Sechting (2004), however, referred it to the Kamczatica group,

which comprises both corticolous and saxicolous species with grayish thalli

lacking anthraquinones and black-margined yellowish orange anthraquinone-

containing apothecial discs, but which differs from the Sideritis group in lacking

algal cells in amphithecium. Recently Wetmore (2007) also refers the species to

the Kamczatica group. Poelt & Wunder (1967) earlier considered the presence

or absence of algae in amphithecium a variable character. Molecular analysis

may help settle this taxonomic problem in the future.

Caloplaca spp. new to India... 57

In India, this group is represented by only two species, C. litoricola and

C. tropica Y. Joshi & Upreti. Joshi & Upreti (2007a) initially described C. tropica

under Sideritis group, but later (Joshi 2008) placed it along with C. litoricola

under Kamczatica group.

Caloplaca litoricola is characterized by whitish-gray continuous or areolate

thallus, orange-yellow to dark orange apothecia with black proper margins,

proper exciple of radiating elongated hyphal cells, oval to ellipsoid 11-16 x 6-9

um spores, and littoral distribution. It is close to Caloplaca tropica, which differs

in having indistinct thallus, narrowly ellipsoid spores (9-12 x 1.5-2 um), anda

distribution restricted to inland localities of central India.

Caloplaca lypera Poelt & Hinter., Biblioth. Lichenol. 50: 159, 1993.

SPECIMENS EXAMINED: INDIA. UTTARAKHAND: Bageshwar district, Phurkia

Dakbunglow, alt. 3450 m, on stones, 10 June 1970, D.D. Awasthi, 7676 (LWG-AWAS);

Pithoragarh district, Munsiyari, alt. 2200 m, on rocks, 17 November 2006, Y. Joshi & R.

Bajpai 06-006276, 06-007008 (LWG); alt. 2500 m, on rocks, 16 November 2006, Y. Joshi

& R. Bajpai 06-006234 (LWG); Khaliya top, alt. 2700-3000 m, on rocks, 17 November

2006, Y. Joshi & R. Bajpai 06-006273 (LWG).

THALLUS crustose, saxicolous, determinate, irregular to + orbicular, zonate,

cracked areolate, gray to ash-gray to ochraceous-gray, glossy. CORTEX

paraplectenchymatous. MEDULLA yellow. PROTHALLUS generally absent but

black when present.

APOTHECIA numerous, scattered to clustered, restricted to the central parts

of the thallus, sessile, initially round, later on angular owing to pressure from

neighboring apothecia, 0.2-0.7(-1.2) mm in diam.; disc reddish-brown to

mostly olivaceous black, flat; proper margin thin, entire, persistent, smooth

to flexuose, dirty brownish-red, paler than disc, in cross-section of slightly

elongated cells to + paraplectenchymatous, broadly fan-shaped; thalline

margin thick, entire, persistent, smooth to flexuose, yellowish to muddy orange

or rarely thallus-coloured. EprHyMENIUM yellowish-orange to brownish-

orange, with fine granular epipsamma. HyMENIvuM hyaline, ca. 100 um high.

HyporuHecium hyaline, of indistinct cells, with oil globules. AMPHITHECIUM

with algae. PARAPHYSES thin, simple, rarely branched, with clavate apical cells

surrounded by epipsamma. Asci 8-spored, spores polarilocular, ellipsoid

to broadly ellipsoid, 15-17 x 7-9 um, isthmus 4.5-6 um. PYCNIDIA present,

numerous, ostiole brown to brownish-red, conidia bacilliform, 3-4 x 1-1.5 um.

[n = 20 for all measurements.] For further descriptions see Poelt & Hinteregger

(1993).

CHEMISTRY — Spot test reactions: Thallus K- to K+ faint red in some

regions, C-. Medulla, apothecial disc and epihymenium K+ red, C-. Secondary

metabolites: parietin.

ECOLOGY AND DISTRIBUTION — Caloplaca lypera is found growing along

roadside on the vertical faces of rocks between elevations of 2200-3450 m in

58 ... Joshi & Upreti

temperate regions of Central Himalaya. Poelt & Hinteregger (1993) reported

this species growing over hard, lime free siliceous rocks at 2700 m in the

Langtang area of Nepal. The present study extends its distribution in India.

REFERENCE SPECIMEN EXAMINED: NEPAL. LANGTANG AREA: On way Khangjung to

Sangsa (E of Khangjung), towards Pang Sang Lekh, pastures, rocks, open woods, alt.

2700 m, on rock, 28 August 1986, J. Poelt 000279609 (Holotype-GZU).

REMARKS —Poelt & Hinteregger (1993) place C. lypera in the Chrysophorae

group, characterized by crustose to lobate, gray to olive-brown thalli with

partially or completely brownish-black apothecia and widely ellipsoid spores.

Members of this group colonize lime-poor rocks. So far it is the only known

representative of this group from India. The Ochrotropae group (Poelt &

Hinteregger 1993) is similar enough to the Chrysophorae group, that molecular

analyses are needed to settle the taxonomy.

Caloplaca lypera is mainly characterized by an irregularly cracked areolate,

grayish or ochraceous coloured thallus with a yellow medulla and a reddish-

brown to olivaceous-black apothecial disc surrounded by yellow thalline

margin. Outwardly it resembles Lecanora luteomarginata Nayaka et al., but

differs in having yellow medulla and polarilocular spores.

Caloplaca maura Poelt & Hinter., Biblioth. Lichenol. 50: 161, 1993.

SPECIMEN EXAMINED: INDIA. Srxxim: North Sikkim, Giagaon, above Thanngu, alt.

4600 m, on detritus, 13 August 2004, D.K. Upreti, S. Chatterjee & P.K. Divakar 04-

004004 (LWG).

THALLUS crustose, muscicolous/terricolous, often coalescing with other thalli

to cover large areas, indistinct to squamulose, squamules small, gray. CORTEX

paraplectenchymatous. MEDULLA white. PROTHALLUS absent.

APOTHECIA numerous, scattered to clustered, sessile to constricted at base,

round, 0.2-0.3(-0.5) mm in diam.; disc brownish-black to black, sometimes

covered with yellowish-brown to yellowish-orange granules (epipsamma), flat;

proper margin thick, smooth, persistent, entire, raised above the level of disc

to + flush, black, in cross-section of elongated radiating cells; thalline margin

+ present, thin, smooth to crenulate, concolorous to thallus. EPIHYMENIUM

golden to golden-brown. HymMEnivu hyaline, 70-100 um high. HyPOTHECIUM

hyaline, thin, of indistinct cells, without oil globules. AMpHITHECIUM with

algae, algal layer continuous below exciple, outer surface aeruginose pigmented.

PARAPHYSES in upper region are repeatedly branched, end cells club-shaped to

capitate. Asci 8-spored, spores polarilocular, broadly ellipsoid, 12-18 x 6-10

um, isthmus 3-5 um. Pycnip1a not seen. [n = 10 for all measurements.] For

further descriptions see Poelt & Hinteregger (1993).

CHEMISTRY — Spot test reactions: Thallus and medulla K-, C-. Apothecial

disc and epihymenium K+ red, C-. Secondary metabolites: parietin.

Caloplaca spp. new to India ... 59

ECOLOGY AND DISTRIBUTION — ‘The present study extends its distribution

in India where it is found growing on detritus in alpine regions at 4600 m in

the eastern Himalaya. Poelt & Hinteregger (1993) reported this species growing

over wood of Kobresia pygmaea in the Khumbu region of Nepal.

REMARKS —Poelt & Hinteregger (1993) place C. maura in the Conversae group,

characterized by crustose to areolate, + dark gray thalli and small, blackish or

black apothecial discs with concolorous margins. Most species in this group

prefer rocky habitats, but some grow over bark, moss, and detritus. Caloplaca

rinodinopsis, a corticolous species, is the other known representative of this

group from India.

Caloplaca maura is characterized by an indistinct to squamulose gray thallus,

apothecia with brownish-red to black discs + covered by yellowish granules.

A subtemperate distribution, orange to orange-brown apothecial disc, and

non-thalline margin separate the closely related muscicolous C. cerina var.

muscorum (A. Massal.) Jatta from C. maura.

Acknowledgments

The authors are thankful to Drs M. Andreev, L. Lékés, and P.K. Divakar for reviewing

the manuscript and providing valuable comments.

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Lichenologica 3: 1-186.

MY COTAXON

Volume 116, pp. 61-74 April-June 2011

DOI: 10.5248/116.61

Contribution to the lichen mycota of South Korea

YOGESH JOSHI’, THI THUY NGUYEN’, XIN YU WANG’,

LAszL6 L6xK6s?, YOUNG JIN KOH? & JAE-SEOUN Hur”

' Department of Botany. S.S.J. Campus, Almora 263601, Uttarakhand, India

? Korean Lichen Research Institute, Sunchon National University, Sunchon 540-742, South Korea

> Botanical Department of the Hungarian Natural History Museum,

Budapest, Pf. 222 H-1476, Hungary

CORRESPONDENCE TO *: " *yogesh36953@rediffmail.com,

?jshur1@sunchon.ac.kr, *lokos@bot.nhmus.hu

ABSTRACT — The present paper briefly describes 14 subtropical to temperate lichen species

new to South Korea. Among the genera Bacidia, Cresponea, Diploschistes, Fissurina, Fuscidea,

Micarea, Mycoblastus, Phyllopsora, Sarcogyne, Scoliciosporum, and Toninia reported for the

first time from South Korea, Fuscidea recensa var. arcuatula, Micarea elachista, Sarcogyne

privigna, and Toninia cinereovirens also represent new records for East Asia, including China

and Japan.

KEY worps — geographical distribution, lichen-forming fungi, taxonomy

Introduction

In common with other areas of East Asian regions, the lichen flora of South

Korea (34-38°N, 126-130°E) is little known. Hur et al. (2005) provided an

annotated checklist of the lichenized fungi of South Korea based on critical

review of archival manuscripts and published literature records and reported

510 species belonging to 113 genera. Further examination of herbarium material

lodged at Korean Lichen Research Institute (KoLRI) and recent collections

made by the authors have revealed 11 genera and 14 species that have not

been previously reported from this country. These species are in addition to

those reported earlier by Joshi et al. (2009a-c, 2010a-h). It is expected that

future collections will add more genera and species to the lichen biota of South

Korea.

The genera Bacidia De Not., Cresponea Egea & Torrente, Diploschistes

Norman, Fissurina Feé, Fuscidea V. Wirth & Vézda, Micarea Fr., Mycoblastus

Norman, Phyllopsora Mill. Arg., Sarcogyne Flot., Scoliciosporum A. Massal. and

62 ... Joshi & al.

Toninia A. Massal. are reported for the first time in South Korea. Cresponea

proximata, Diploschistes scruposus, Fuscidea recensa var. arcuatula, and Toninia

cinereovirens were collected from the maritime regions of South Korea.

The intent of this work is to provide users with the geographical distribution

of lichens in South Korea, expand the knowledge of lichen diversity in East

Asia including China and Japan, and add some new records to the South

Korean lichen checklist of Hur et al. (2005). Brief taxonomic descriptions,

comments, and photographs of each species are provided along with chemical

and ecological data.

Materials & methods

The study is based on lichen specimens lodged in herbarium of Korean Lichen

Research Institute (KoLRI), Sunchon National University, South Korea, as well as

fresh samples collected during recent field trips in various parts of the country.

Specimens were examined using standard microscopic techniques and hand-sectioned

under NIKON C-PS 1068908 dissecting microscope. All measurements were taken

from material mounted in water; lactophenol cotton blue was used only as a stain.

For characters such as size of the thallus, ascomata and thickness of the hymenium,

subhymenium, hypothecium, exciple, ascospores dimension, ten measurements were

recorded per specimen. Ascospore dimensions are generally presented as smallest mean

recorded to the largest mean. Spot test reactions were carried out on hand sections of

thalli and apothecia under the microscope OLYMPUS BX 50. Secondary metabolites

were identified by TLC as described by Orange et al. (2010) using solvent system C and

EA (used for the separation of lecanoric and gyrophoric acids). HCl was used to detect

the nature of rock (bubbles indicate calcareous rock).

Taxonomy

Bacidia arceutina (Ach.) Rehm & Arnold, Verh. zool.-bot. Ges. Wien 19: 624, 1869

Fic. lA

= Lecidea luteola var. arceutina Ach., Method. Lich.: 61, 1803

SPECIMENS EXAMINED —- SOUTH KOREA: Gyeongbuk Prov.: MUNGYEONG Co., Mt.

Juhul, 36°46'30.1"N, 128°06'12.6"E, alt. 788 m, on bark, 29 February 2004, J.-S. Hur

040131 (KoLRI); Mt. Joryeong, 37°48'27"N, 128°03'32"E, alt. 500 m, on bark, 10 July

2008, J.-S. Hur 080306 (KoLRI); Kangwon Prov.: PYEONGCHANG Co., Mt. Hwangbyong,

37°44'44.3"N, 128°37'30.4"E, alt. 779 m, on bark, 14 July 2008, J.-S. Hur 080380

(KoLRI); 37°44'47.6"N, 128°37'31.5"E, alt. 772 m, on bark, 14 July 2008, J.-S. Hur,

080416 (KoLRI); 37°44'52.3"N, 128°37'33.2"E, alt. 751 m, on bark, 14 July 2008, J.-S.

Hur 080404 (KoLRI); Mt. Seokpyeong, 37°34'57.3"N, 128°52'27.1"E, alt. 870 m, on bark,

24 May 2008, J.-S. Hur 080229 (KoLRI); Jeonnam Prov.: Hwasun Co., Doam-myeon,

Caecho-ri, Mt. Cheonbul, Unju-sa, 34°55'13.2"N, 126°52'51.7"E, alt. 70 m, on bark, 04

September 2005, J.-S. Hur 050381 (KoLRI).

BRIEF DESCRIPTION — ‘Thallus corticolous, determinate, thin, continuous,

smooth to minutely granular-warted, pale green-gray. Prothallus absent.

Apothecia numerous, 0.4-0.8 mm diam., at first plane and marginate but

Lichens of South Korea ... 63

often becoming convex and immarginate, pale brown. Proper exciple without

crystals. Rim pigmented in a distinct zone, dark yellow-brown to brown in

upper part, downwards changing to pale yellow or colourless. Epithecium

yellowish-brown, K-, N-. Hymenium colourless. Hypothecium straw or

yellow-brown, K+ intensifying yellow. Spores hyaline, acicular, 45-70 x 1.3-2.5

um, 3-7 septate.

CHEMISTRY — Spot test reactions: thallus and apothecia K-, C-, KC-, P-,

N-. Secondary metabolites: none detected.

ECOLOGY & DISTRIBUTION — ‘The collections were found growing over

Quercus bark between elevations of 70-870 m. Known also from Europe,

Africa, Asia, and North America (Smith et al. 2009).

REMARKS — Bacidia schweinitzii and B. subincompta (also reported here as

new records) are the only other two Bacidia species known from South Korea.

They differ in having bluish-green epithecium that is always N+ purple violet.

The bluish-green epithecium observed in some specimens of B. arceutina may

lead to confusion with the former two species, but can be easily separated from

them in having colourless to straw or yellow-brown hypothecium which is K+

intensifying yellow. For further descriptions see Ekman (1996).

Bacidia schweinitzii (Fr. ex E. Michener) A. Schneid, Guide Study Lich.:

110, 1898 Fie. 1B

= Biatora schweinitzii Fr. ex E. Michener, Flora Cestrica, ed. 3: 447, 1853

SPECIMENS EXAMINED —- SOUTH KOREA: Gyeongbuk Prov.: MUNGYEONG Co., Mt.

Joryeong, 37°48'27"N, 128°03'32"E, alt. 500 m, on bark, 10 July 2008, J.-S. Hur 080329

(KoLRD); Jeju Island, Mt. Halla 33°26'04.4"N, 126°34'01.7"E, alt. 530 m, on bark, 29

August 2004, J.-S. Hur 040876-1 (KoLRI); Kangwon Prov.: PYEONGCHANG Co., Mt.

Hwangbyong, 37°44'41.3"N, 128°37'31.0"E, alt. 630 m, on bark, 14 July 2008, J.-S. Hur

080364 (KoLRI).

BRIEF DESCRIPTION — ‘Thallus corticolous, indeterminate, thin, of discrete

granules or continuously granular, gray to green-gray. Granules + globose.

Prothallus absent. Apothecia numerous, 0.3-0.8 mm diam., at first plane but

later becoming convex, solitary, orange-brown to blue-black to pure black.

Proper exciple with radiating clusters of crystals. Rim colourless to dark brown,

often bluish-green near the hymenium. Epithecium blue-green, K-, N+ purple

violet. Hymenium colourless. Hypothecium orange-brown to dark brown.

Spores hyaline, acicular, 40-68 x 2.5-3 um, 3-7 septate.

CHEMISTRY — Spot test reactions: thallus and apothecia K-, C-, KC-, P-,

N-. Secondary metabolites: none detected.

ECOLOGY & DISTRIBUTION — The collections were found growing over

Quercus bark between 500-630 m. Known also from Eastern Asia (China,

Japan), and North America (Ekman 1996).

64 ... Joshi & al.

REMARKS — The brown hypothecium of B. schweinitzii, which gradually

merges with dark coloured proper exciple near the base, separates it from

B. subincompta, where the hypothecium forms a + distinct zone. Further,

yellowish to orange-brown hypothecial region is K+ intensifying in

B. schweinitzii, and lacking in B. subincompta. The colourless or pale straw-

coloured hypothecium of B. arceutina separates it from B. schweinitzii. For

further descriptions see Ekman (1996).

Bacidia subincompta (Nyl.) Arnold, Flora, Jena 53: 472, 1870 Fic. 1C

= Lecidea subincompta Nyl., Flora, Jena 48: 147, 1865

SPECIMENS EXAMINED — SOUTH KOREA: Kangwon Prov.: PYEONGCHANG Co., Mt.

Hwangbyong, 37°44'52.3"N, 128°37'33.2"E, alt. 751 m, on bark, 14 July 2008, J.-S. Hur

080404 (KoLRI); 37°44'53.0"N, 128°37'31.9"E, alt. 776 m, on bark, 14 July 2008, J.-S.

Hur 080455 (KoLRI); Jeju Island, Mt. Halla 33°23'18.1"N, 126°29'345.1"E, alt. 975 m,

on bark, 27 August 2004, J.-S. Hur 040689 (KoLRI); Gyeongbuk Prov.:: MUNGYEONG

Co., Mt. Joryeong, 37°48'27"N, 128°03'32"E, alt. 500 m, on bark, 10 July 2008, J.-S. Hur

080301 (KoLRI).

BRIEF DESCRIPTION — Thallus corticolous, determinate, thin to thick, either

discontinuous, of discrete low convex areoles, or continuous, without cracks,

gray to gray-green to greenish-brown. Prothallus absent. Apothecia numerous,

0.2-1.1 mm diam.., at first plane but later becoming convex, solitary to +clumped,

blue-black to pure black. Proper exciple without crystals. Rim colourless

to red-brown to black-brown, often with a bluish-green tinge in upper part.

Epithecium green to bluish-green, K-, N+ purple violet. Hymenium colourless.

Hypothecium dark red-brown, K+ purple in upper part, pale red-brown below.

Spores hyaline, fusiform, bacilliform or acicular, 25-32 x 2-3 um, 3-7 septate.

CHEMISTRY — Spot test reactions: thallus and apothecia K-, C-, KC-, P-,

N-. Secondary metabolites: none detected.

ECOLOGY & DISTRIBUTION — The collections were found growing over

Quercus bark at 500-975 m. Known also from Africa, Asia, Europe, Macaronesia,

and North America (Smith et al. 2009).

REMARKS — For comparisons, see B. arceutina and B. schweinitzii above. For

further descriptions see Ekman (1996).

Cresponea proximata (Nyl.) Egea & Torrente, Mycotaxon 48: 328, 1993 Fic. 1D

= Lecidea proximata Nyl., Ann. Sci. Nat. Bot., sér. 4, 19: 356, 1863

SPECIMENS EXAMINED —- SOUTH KOREA: Jeonnam Prov.: YEOsU CITY, Geomun Island,

34°00'38.5"N, 127°19'10.8"E, alt. 46 m, on Camellia japonica bark, 24 March 2007, J.-S.

Hur 070128, 070130 (KoLRI); 34°00'37.1"N, 127°19'13.0"E, alt. 21 m, on bark, 24 March

2007, J.-S. Hur 070131 (KoLRI); WANDo Co., Bogil-myeon, Bogil Island, Yesong-ri, near

Yesong beach trail, 34°08'30.2"N, 126°33'48.3"E, alt. 2 m, on bark, 06 February 2010, Y.

Joshi & party 100241, 100244, 100247, 100246-2, 100250, 100251 (KoLRI).

Lichens of South Korea... 65

BRIEF DESCRIPTION — ‘Thallus corticolous, continuous, cracked-areolate,

smooth, gray to green. Hypothallus + present, brownish. Photobiont

Trentepohlia. Apothecia numerous, scattered, sessile, constricted at base,

0.3-1.2 mm diam., with a smooth to +crenulate margin. Disc plane to slightly

convex, black, with green to ochraceous-yellowish pruina. Hymenium I+

reddish. Paraphysoids with thickened apical cell having dark brown cap formed

in the outer part of cell wall. Spores hyaline, fusiform, 25-35(-38) x 5-7 um,

(1-)5-7 septate, straight to slightly curved.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

ECOLOGY & DISTRIBUTION — ‘The collections were found growing over bark

along the coast at elevations of 2-46 m. Known also from Indo-Malaya, the

Caribbean, Brazil, China, and Japan (Egea & Torrente 1993).

REMARKS — So far C. proximata is the only Cresponea species known from

South Korea. It is often confused with C. premnea (Ach.) Egea & Torrente, which

can be distinguished by smaller spores (20-27 um) and paraphysoids with dark

brown cap formed in the inner part of the cell wall. For further descriptions see

Egea & Torrente (1993).

Diploschistes actinostomus (Pers. ex Ach.) Zahlbr., Hedwigia 31: 34,1892 Fic. 1E

= Verrucaria actinostoma Pers. ex Ach., Lich. univ.: 288, 1810

SPECIMENS EXAMINED — SOUTH KOREA: Jeonnam Prov.: JANGHEUNG Co., Gwansan-

eup, Mt. Cheonkwan, 34°32'47.2""N, 126°55'39.6"E, alt. 300 m, on rock, 06 October 2005,

J.-S. Hur 050648 (KoLRI); Mt. Cheonkwan, near Gwansan, 34°32'50.6"N, 126°55'43.3"E,

alt. 200 m, on rock, 06 October 2005, L. L6kés (BP).

BRIEF DESCRIPTION — ‘Thallus saxicolous, thick, glossy, smooth, rimose-

areolate, whitish-gray, epruinose. Photobiont trebouxioid. Apothecia numerous,

conspicuous, solitary, perithecioid. Pores small, rounded, black. Spores broadly

ellipsoid, brown, non-amyloid, 16-22 x 10-12 um, with 4-6 transverse septa

and 1-3 longitudinal septa.

CHEMISTRY — Spot test reactions: thallus K-, C+ red, KC-, P-. Secondary

metabolites: lecanoric acid (major) and gyrophoric acid (minor).

ECOLOGY & DISTRIBUTION — The collections were found growing over non-

calcareous rocks at 200-300 m. Cosmopolitan (Rivas Plata et al. 2010).

REMARKS — Diploschistes scruposus, the other species of this genus known

from South Korea and reported here as new record, differs in having urceolate

ascomata, while ascomata in D. actinostomus are perithecioid. Diploschistes

diploschistoides (Vain.) G. Salisb., a related species with perithecioid ascomata,

differs in longer (30-52 um), amyloid spores. For further descriptions see

Mangold et al. (2009).

66 ... Joshi & al.

Diploschistes scruposus (Schreb.) Norman, Nytt Mag. Natur. 7: 232,1853 Fic. 1F

= Lichen scruposus Schreb., Spic. Fl. Lips.: 133, 1771

SPECIMEN EXAMINED — SOUTH KOREA: Jeonnam Prov., GOHEUNG Co., Sorok Island,

alt. 15 m, on rock, 23 March 2003, J.-S. Hur 030065 (KoLRI).

BRIEF DESCRIPTION — ‘Thallus saxicolous, thick, rimose-areolate, greenish-

gray, surface uneven to verrucose, scabrid, epruinose. Photobiont trebouxioid.

Apothecia numerous, conspicuous, solitary, urceolate, 1-1.5 mm diam. Disc

visible from above, blackish-brown, + grayish pruinose. Spores ellipsoid,

brown, non-amyloid, 22-36 x 10-16 um, with 4-7 transverse septa and

1-3 longitudinal septa.

CHEMISTRY — Spot test reactions: thallus K-, C+ red, KC-, P-. Secondary

metabolites: lecanoric acid.

ECOLOGY & DISTRIBUTION — The collection was found growing over non-

calcareous rocks. Cosmopolitan (Rivas Plata et al. 2010).

REMARKS — D. gypsaceus (Ach.) Zahlbr., a related species with urceolate

ascomata, differs in pruinose thallus, 4-spored asci and its typifically calcareous

substratum. D. rampoddensis (Nyl.) Zahlbr. differs in having yellowish-gray

to orange-yellow thallus, narrowly ellipsoid spores (7-12 um wide), and

pantropical distribution. For further descriptions see Mangold et al. (2009).

Fissurina insidiosa C. Knight & Mitt., Trans. Linn. Soc. London 23: 102, 1860

Fig. 1G

SPECIMEN EXAMINED - SOUTH KOREA: Jeju Island, Mt. Halla, 33°22'77.5"N,

126°33'74.9"E, alt. 1000 m, on bark, 21 April 2009, J.-S. Hur 090100 (KoLRI).

BRIEF DESCRIPTION — ‘Thallus corticolous, thin, cracked and fissured, glossy,

greenish-gray. Apothecia (lirellae) inconspicuous, immersed, indicated by

a thin line between two lips, sometimes slightly open, somewhat raised and

paler than the thallus, straight, curved or sinuous, 1-3 mm long. Thalline

exciple sometimes raised around the lirellae or sometimes flush. Apothecia

subcontexta-type (see Staiger 2002). Spores 4-locular, 18-20 x 8-10 um.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

ECOLOGY & DISTRIBUTION— ‘The collection was found growing over bark at

1000 m. Known also from New Zealand (Hayward 1977).

REMARKS — So far F. insidiosa is the Fissurina species known from South Korea.

In external morphology it is close to F dumastii Fée, which differs in having

dumastii-type apothecia. Dumastii-type mature apothecia are characterized by

erumpent lirellae with thin labia and a partly exposed disc, while subcontexta-

types have prominent lirellae with thick labia and hidden discs. For further

descriptions see Staiger (2002).

Lichens of South Korea ... 67

Fic. 1. New records examined in the present study. A. Bacidia arceutina, 080229 (KoLRI);

B. B. schweinitzii, 080329 (KoLRI); C. B. subincompta, 080404 (KoLRI); D. Cresponea

proximata, 070131 (KoLRD); E. Diploschistes actinostomus, 050648 (KoLRI); F. D. scruposus,

030065 (KoLRI); G. Fissurina insidiosa, 090100 (KoLRD); H. Fuscidea recensa var. arcuatula,

070097 (KoLRI). Scale = 3 mm.

68 ... Joshi & al.

Fuscidea recensa var. arcuatula (Arnold) Fryday, Lichenologist 40: 313, 2008

Fic. 1H

= Biatora arcuatula Arnold, Flora 71: 107, 1888

SPECIMEN EXAMINED — SOUTH KOREA: Jeonnam Prov.: YEOsu ciTy, Geomun Island,

34°00'38.7"N, 127°19'01.2"E, alt. 10 m, on rock, 24 March 2007, J.-S. Hur 070097

(KoLRI).

BRIEF DESCRIPTION — Thallus saxicolous, areolate, areoles usually contiguous,

discrete, rounded and tuberculate, pale gray, growing in zoned patches one

against another. Prothallus dark brownish-black. Soredia absent. Apothecia

numerous, sessile, broadly attached, lead coloured, brownish when wet, 0.4-0.6

mm diam. Disc plane to slightly convex. Spores hyaline, ellipsoid, curved, 9-12

x 3.5-4.5 um.

CHEMISTRY — Spot test reactions: thallus and medulla K-, C-, KC-, P-.

Medulla UV+ white. Secondary metabolites: divaricatic acid.

ECOLOGY & DISTRIBUTION — The collection was found growing over non-

calcareous rocks at 10 m along the coast. Also known from Europe, North

America, and Indonesia (Smith et al. 2009); new to East Asia.

REMARKS — So far F. recensa var. arcuatula is the only Fuscidea representative

known from South Korea. It is often confused with F. recensa (Stirt.) Hertel

et al. var. recensa, which has a sorediate thallus and thick persistent proper

margin. Although Fryday (2008) noted the non-sorediate nature of some

specimens of F recensa var. recensa, their thick white thallus and thin proper

margin separate them from E recensa var. arcuatula. For further descriptions

see Fryday (2008).

Micarea elachista (Kérb.) Coppins & R. Sant., Bull. Br. Mus. nat. Hist. (Bot.)

TEE 131, 1983 Fig. 2A

= Biatora elachista Korb., Parerga lichenol. (Breslau): 159, 1860

SPECIMEN EXAMINED - SOUTH KOREA: Kangwon Prov.: PYEONGCHANG Co., Mt.

Odae, Duro Peak, 37°45'52.7"N, 128°36'35.2"E, alt. 1081 m, on bark, 15 July 2008, J.-S.

Hur 080488 (KoLRI).

BRIEF DESCRIPTION — Thallus corticolous, continuous, areolate, greenish-

gray. Photobiont micareoid. Apothecia numerous, immarginate, convex to

globose, 0.2-0.4 mm diam., brown to brownish-black. Epithecium dark brown.

Hymenium colourless, with yellowish brown vertical streaks. Spores hyaline,

fusiform to oblong-fusiform, 0-1(-3) septate, 11-15 x 2-3 um.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

ECOLOGY & DISTRIBUTION — ‘The collection was found growing over bark

at an elevation of 1081 m. Known also from Europe, Australia, and North

America (Coppins 1983; Smith et al. 2009); new to Asia.

Lichens of South Korea... 69

REMARKS — So far M. elachista is the only Micarea species known from South

Korea. It has often been confused with M. denigrata (Fr.) Hedl., which can be

distinguished by C+ red apothecia in sections (due to presence of gyrophoric

acid). The related species M. rhabdogena (Norman) Hedl. can be differentiated

by an endoxylic thallus and smaller, mostly simple spores. For further

descriptions see Coppins (1983).

Mycoblastus sanguinarius (L.) Norman, Cat. Lich. Univers. 4: 5, 1926 Fic. 2B

= Lichen sanguinarius L., Sp. pl. 2: 1140, 1753

SPECIMENS EXAMINED — SOUTH KOREA: Gyeongnam Prov.: SANCHUNG Co., Jungsan-

ri, Mt. Jiri, 35°20'06.9"N, 127°42'47.1"E, alt. 1530 m, on Acer bark, 16 September

2006, J.-S. Hur 060735 (KoLRI); Jeonnam Prov.: KWANGYANG City, Mt. Baekwoon,

35°37'18.5"N, 127°37'51.1"E, alt. 1160 m, on rock, 17 August 2006, J.-S. Hur 060626

(KoLRI).

BRIEF DESCRIPTION — Thallus corticolous, thick, verrucose or papillose-warted,

continuous to + cracked, gray. Prothallus present, grayish-black. Hypothecium

carmine to blood-red. Apothecia numerous, 0.3-1.2 mm diam., appressed or

sessile leaving a bright carmine-red spot when damaged, convex, black. Asci

1-spored. Spores hyaline, broadly ellipsoid, 70-84 x 30-40 um. Pycnidia not

seen. For further descriptions see Smith et al. (2009) and Kantvilas (2009).

CHEMISTRY — Spot test reactions: thallus K+ yellow, C-, KC-, P+ yellow.

Secondary metabolites: atranorin and caperatic acid.

ECOLOGY & DISTRIBUTION — ‘The collections were found growing over

Acer bark and siliceous rocks at 1160-1530 m. Known also from Asia, Africa,

Europe, Macaronesia, and North America (Smith et al. 2009, Kantvilas 2009).

REMARKS — So far M. sanguinarius is the only Mycoblastus species known from

South Korea. It has often been confused with M. sanguinarioides Kantvilas,

which differs in rather thin, smooth thallus and its distribution in the Southern

Hemisphere (Tasmania, Australia). The related species M. affinis (Schaer.)

T. Schauer differs by lacking red pigment at the apothecial base, smaller spores

(less than 70 um), and the presence of planaic acid.

Phyllopsora corallina (Eschw.) Mill. Arg., Bot. Jb. 20: 264, 1894 Fig. 2C

= Lecidea corallina Eschw., Fl. Bras. Enum. Pl. 1: 256, 1833

SPECIMEN EXAMINED - SOUTH KOREA: Kangwon Prov.: INJAE Co., Baekdam Temple,

38°11'16.4"N, 128°21'42.7"E, alt. 450 m, on rocks, 11 October 2004, J.-S. Hur 041503

(KoLRI).

BRIEF DESCRIPTION — Thallus corticolous, squamulose, squamules round or

elongate, adnate to tascending, scattered when young, later contiguous or

imbricate, tan coloured. Upper surface glabrous. Prothallus well developed,

white. Cortex type 2. Isidia numerous, attached marginally to the squamules,

cylindrical, simple or rarely branched, tips brownish-black coloured. Apothecia

not seen.

70 ... Joshi & al.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

ECOLOGY & DISTRIBUTION — The collection was found growing over non-

calcareous rock at an elevation of 450 m. Known also from Asia, North and

South America, Australia, and Africa (Brako 1991).

REMARKS — So far P. corallina is the only Phyllopsora species known from

South Korea. It is close to P. africana Timdal & Krog, P. kiiensis (Vain.) Gotth.

Schneid., P. martinii Swinscow & Krog, and P. swinscowii Timdal & Krog,

in having marginally attached isidia, but it differs in chemistry. Phyllopsora

africana and P. martinii contain argopsin and chlorophyllopsorin, P. kiiensis

has furfuracein, and P swinscowii contains methyl 2,7-dichloropsoromate and

methyl 2,7-dichloronorpsoromate. For further descriptions see Brako (1991)

and Timdal & Krog (2001).

Sarcogyne privigna (Ach.) A. Massal., Geneac. Lich.: 10, 1854 Fic. 2D

= Lecidea privigna Ach., Method. Lich.: 49, 1803

SPECIMEN EXAMINED — SOUTH KOREA: Jeonnam Prov.: Hwasun Co., Mt. Baega,

35°10'14.1"N, 127°08'44.9"E, alt. 490 m, on rock, 08 October 2005, L. Lék6s 050671

(BP, KoLRI); Kangwon Prov.: OkGYE Co., Gangneung city, Mt. Jabyung, 37°33'16.6"N,

128°58'46.5"E, alt. 360 m, on rock, 19 May 2010, J.-S. Hur s.n. (KoLRI).

BRIEF DESCRIPTION — ‘Thallus saxicolous, endolithic or inconspicuously

developed, with gray ecorticate regions occurring below the apothecia.

Apothecia numerous, sessile, round to +irregular, dispersed or contiguous,

0.5-1.3 mm diam., red to blackish-red. True exciple black, persistent. Hymenium

up to 95 um tall. Hypothecium pale brown. Asci multi spored, c. 200-spored.

Spores hyaline, cylindrical to oblong, 4-5 x 1-2 um.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

ECOLOGY & DISTRIBUTION — The collections were found growing over

siliceous rocks at an elevation of 490 m. Known also from Asia, Europe, North

America, Africa, and Australia (Smith et al. 2009); new to East Asia.

REMARKS — So far S. privigna is the only Sarcogyne species known from

South Korea. In external morphology it is often confused with S. clavus (DC.)

Kremp., which differs in having dark brown hypothecium, a taller hymenium,

a crenulate true exciple, and a more or less coastal distribution. For further

descriptions see Magnusson (1935) Knudsen & Standley (2007).

Scoliciosporum umbrinum (Ach.) Arnold, Flora, Jena 54: 50, 1871 Fic. 2E

= Lecidea umbrina Ach., Lich. univ.: 183, 1810

SPECIMENS EXAMINED — SOUTH KOREA: Jeonnam Prov.: KWANGYANG City, Mt.

Baekwoon, 35°06'55.8"N, 127°36'26.5"E, alt. 875 m, on bark, 27 June 2006, J.-S. Hur

060393 (KoLRI); 35°06'55.3"N, 127°36'31.6"E, alt. 904 m, on bark, 27 June 2006, J.-S.

Lichens of South Korea ... 71

Fic. 2. New records examined in the present study. A. Micarea elachista, 080488 (KoLRI);

B. Mycoblastus sanguinarius, 060735 (KoLRI); C. Phyllopsora corallina, 041503 (KoLRI);

D. Sarcogyne privigna, s.n. (KoLRI); E. Scoliciosporum umbrinum, 060396 (KoLRI);

E. Toninia cinereovirens, s.n. (KoLRI). Scale = 3 mm

Hur 060396 (KoLRI); Kangwon Prov.: Mt. Seokpyeong, 37°34'29.9"N, 128°51'21.8"E,

alt. 686 m, on bark, 24 May 2008, J.-S. Hur 080193 (KoLRI).

BRIEF DESCRIPTION — ‘Thallus corticolous, variable, thin, cracked, to rather

thick, warted to almost granular, continuous, gray to greenish-gray. Apothecia

numerous, plane to convex at maturity, 0.2-0.6 mm diam., brown to black.

Epithecium blue-green (K-, N+ purple) or olive brown (K-, N-). Spores

hyaline, acicular, 3-7 septate, 18-27 x 2-3 um +spirally arranged in the ascus.

Pycnidia not seen.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

72 ... Joshi & al.

ECOLOGY & DISTRIBUTION — The collections were found growing over bark

at 686-904 m. Cosmopolitan (Smith et al. 2009).

REMARKS — So far S. umbrinum is the only Scoliciosporum species known from

South Korea. Its external morphology and anatomical features resemble those

of S. sarothamni (Vain.) Vézda, which differs in having gyrophoric acid. For

further descriptions see Ekman & Tonsberg (2004).

Toninia cinereovirens (Schaer.) A. Massal., Ric. auton. lich. crost. (Verona): 107,

1852 Fic. 2F

= Lecidea cinereovirens Schaer., Lich. helv. spicil. 3: 109, 1828

SPECIMEN EXAMINED — SOUTH KOREA: Jeonnam Prov.: GWANGYANG CITY, Tae in-

dong, 34°56'63.2"N, 127°44'61.8"E, alt. 2 m, on rock, 2009, J.-S. Hur GW1028 (KoLRI).

BRIEF DESCRIPTION — ‘Thallus saxicolous, squamulose, indeterminate.

Squamules up to 2 mm diam., contiguous to irregularly imbricate with free to

ascending darker gray margin. Upper side brown, epruinose, lacking pores and

pseudocyphellae. Under surface pale brownish. Apothecia up to 0.8 mm diam.,

epruinose. Epithecium olivaceous brown, K-, N-. Spores hyaline, bacilliform,

1—3 septate, 14-26 x 3-4 um.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: None detected.

ECOLOGY & DISTRIBUTION — ‘The collection was found at 2 m along the

coast growing over non-calcareous rocks. Known also from Europe (Timdal

1991); new to East Asia.

REMARKS — So far T’ cinereovirens is the only Toninia species known from South

Korea. It has often been confused with T. squalida (Ach.) A. Massal., which

differs in having acicular, 3-7 septate spores and non-imbricate squamulose

thallus with margins +appressed to the substratum. For further descriptions

see Timdal (1991).

Acknowledgments

This work was supported by a grant from The Korea National Research Resource

Center Program (Grant 2010-0000660) and Korean Forest Service Program (KNA

2010) through Korea National Arboretum. The authors are thankful to Drs. R. Liicking,

FE Crison, and S. Nayaka for reviewing the manuscript and providing valuable

comments.

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tricosa (lichenized Ascomycota, Graphidaceae) in Korea. Mycobiology 37: 152-154. doi:10.4489/

MYCO:2009:37.2,.152

Joshi Y, Wang XY, Lee MY, Byun BK, Koh YJ, Hur JS. 2009b. Notes on some new records of macro-

and micro-lichens from Korea. Mycobiology 37: 197-202. doi:10.4489/MYCO.2009.37.3.197

Joshi Y, Wang XY, Koh YJ, Hur JS. 2009c. Thelotrema subtile and Verrucaria muralis new to South

Korea. Mycobiology 37: 302-304. doi:10.4489/MYCO.2009.37.4.302

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(Physciaceae) in South Korea. Mycobiology 38: 62-64. doi:10.4489/MYCO.2010.38.1.062

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Ascomycetes) from South Korea. Mycobiology 38: 65-69. doi:10.4489/MYCO.2010.38.1.065

Joshi Y, Wang XY, Nguyen TT, Koh YJ, Hur JS. 2010c. Notes on the existence of Leucodecton

desquamescens (thelotremoid Graphidaceae) from South Korea. Mycobiology 38: 149-152.

doi:10.4489/MYCO.2010.38.2.149

Joshi Y, Wang XY, Koh YJ, Hur JS. 2010d. Notes on lichen genus Lepraria Ach. (Stereocaulaceae)

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Joshi Y, Licking R, Wang XY, Yamamto Y, Koh YJ, Hur JS. 2010e. A new species of Graphis

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Joshi Y, Wang XY, Yamamto Y, Koh YJ, Hur JS. 2010h. A first modern contribution to Caloplaca

biodiversity in South Korea; two new species and some new country records. Lichenologist 42:

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7A ... Joshi & al.

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MY COTAXON

Volume 116, pp. 75-120 April-June 2011

DOI: 10.5248/116.75

Glomeromycota: three new genera

and glomoid species reorganized

FRITZ OEHL', GLADSTONE ALVES DA SILVA?,

BRUNO TOMIO GOTO? & EWALD SIEVERDING*

'Federal Research Institute Agroscope Reckenholz-Tanikon ART, Ecological Farming Systems,

Reckenholzstrasse 191, CH-8046 Ziirich, Switzerland

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

Av. Prof. Nelson Chaves s/n, Cidade Universitaria, 50670-420, Recife, PE, Brazil

*Departamento de Botanica, Ecologia e Zoologia, CB, Universidade Federal do Rio Grande

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

‘Institute for Plant Production and Agroecology in the Tropics and Subtropics,

University of Hohenheim, Garbenstrasse 13, D-70599 Stuttgart, Germany

“CORRESPONDENCE TO: fritz.oehl@art.admin.ch & sieverdinge@aol.com

ABSTRACT —Species in the orders Glomerales and Diversisporales (Glomeromycetes) with

glomoid spore formation are reorganized based on combined ribosomal sequence and

morphological analyses. Within the Glomerales two genera in the Glomeraceae (Septoglomus,

Simiglomus) and one genus in the Claroideoglomeraceae (Viscospora) are proposed as new.

Paraglomerales species (thus far monogeneric) also form glomoid spores that may all germinate

directly through the spore wall instead through subtending hyphae as in Glomerales.

Key worps — Glomus group A, Glomus group B, molecular phylogeny, rDNA

Introduction

Molecular biological studies have dramatically improved the understanding

of phylogenetic relationships across a broad swathe of organisms. Arbuscular

mycorrhizal (AM) fungi are no exception, as molecular analyses have expanded

our knowledge of this group (e.g. Simon et al. 1992, Redecker et al. 2000a, b,

Morton & Redecker 2001, Schwarzott et al. 2001, Schiifler et al. 2001, Souza et

al. 2005, Palenzuela et al. 2008, 2010). This has led to taxonomic re-organization

and the description of several new genera and families (e.g. Morton & Redecker

2001, Walker et al. 2007, Oehl et al. 2008). SchiifBler et al. (2001) established

the phylum Glomeromycota with four orders (Archaeosporales, Diversisporales,

Glomerales, Paraglomerales) within one class (Glomeromycetes) described

earlier by Cavalier-Smith (1998).

76 ... Oehl & al.

Within the Glomeromycota, species with glomoid spore formation represent

the largest group. These species had previously been included within the

Glomeraceae (Pirozynski & Dalpé 1989) of the suborder Glomerineae of the

Glomerales (Morton & Benny 1990). However, after this large group of Glomus

species was found to be polyphyletic (e.g. Redecker et al. 2000b, Schwarzott et al.

2001), several glomoidspore-forming species were separated from the Glomerales

sensu Morton & Benny. Based on DNA sequence analyses, Gl. occultum was

declared type species of the new genus Paraglomus (Paraglomeraceae; Morton &

Redecker 2001) in the new order Paraglomerales (Schifsler et al. 2001). Glomus

spurcum was moved as type species to a new genus Diversispora (Walker &

Schiifler 2004) in the Diversisporaceae (Diversisporales) (Schiifler et al. 2001).

More recently Oehl & Sieverding (2004) established Pacispora (Pacisporaceae,

Walker & Schiifler 2004) with Glomus scintillans S.L. Rose & Trappe (Rose

& Trappe 1980) as type species for all glomoid species possessing two spore

walls and specific germination characteristics. Moreover, some glomoid species

were recently transferred into the Ambisporaceae of the Archaeosporales (e.g.

Gl. callosum, Gl. fecundisporum, Gl. leptotichum; Walker et al. 2007, Walker

2008). However, the remaining large group of Glomus spp. is considered

particularly difficult to differentiate morphologically (Morton & Redecker

2001, Schwarzott et al. 2001, Walker & SchiiSler 2004).

Phylogenetic reconstructions within AM fungi over the last 10 years indicate

that this large group of Glomus species likely originated from a single ancestor,

from which (according to Schwarzott et al. 2001) descend the Glomus groups

Aa (related to Gl. mosseae and Gl. geosporum), Ab (related to Gl. intraradices

and Gl. irregulare), Ac (related to Glomus sp. W3347, now Gl. hoi; see Daniell

et al. 2001, Opik et al. 2006), and B (related to Gl. etunicatum). SchiiGler et

al. (2001) also showed that the clades of groups Aa, Ab and B divide further.

Adapting and slightly modifying the numbering system of Schwarzott et al.

(2001), we differentiate the clades into group Aal (lead sp. Gl. mosseae), Aa2

(lead sp. GI. hoi), Aa3 (lead sp. Gl. constrictum), Ab1 (lead spp. Gl. microcarpum,

Gl. macrocarpum), Ab2 (lead spp. Gl. intraradices, Gl. irregulare, Gl. aggregatum),

Ab3 (lead spp. Gl. sinuosum, GI. coremioides), B1 (lead sp. Gl. etunicatum), and B2

(lead sp. GI. viscosum). In the present study, “lead species” are those with typical

morphological characters and not necessarily type species. Under “Results &

discussion” we present the supporting phylogenetic clades and detail the major

morphological spore characteristics for each of our groups — Aal, Aa2, Aa3,

Ab1, Ab2, Ab3, B1, B2. In those cases where morphological and molecular data

are congruent, species of the same group and number can be classified together

at the genus level (e.g., group Aal represents one genus, while Aa2 represents

another genus), while the major groups A and B are suggested to be family

levels. In the case of clades Ab1, Ab2, Ab3, however, the molecular database

Glomoid species reorganized ... 77

remains under-represented with regard to the known species and the results

are contradictory, as some sequences are derived from species that have been

only poorly characterized. Although the morphological data indicate a division

of Ab into at least three subgroups (genera), we currently lack sufficient genetic

data to support new genera. Thus, clades Ab1, Ab2, Ab3 will be considered in

one single genus (group Ab) at this time.

Previous phylogenetic analyses indicated that several Glomus spp. (e.g.

Gl. versiforme) belong to clade ‘Glomus group ‘C’ (e.g. Schwarzott et al.

2001, Schiifler et al. 2001, Redecker et al. 2007), currently Diversispora

(Diversisporaceae). Only six glomoid spore-forming species have so far been

included in this genus (Walker & Schiifler 2004, Gamper et al. 2009, Schiifler

& Walker 2010). The morphology of Diversispora spores differs, however, from

spores of groups A and B. We call this group of species “group Ca” and will

transfer them to the genus Diversispora.

Recently, Redecker et al. (2007) identified another Glomus group (including

Gl. fulvum and Gl. pulvinatum) that was genetically related to group C

but phylogenetically and morphologically clearly distinct from group Ca

(Diversispora). The authors did not separate this group from the other

species of group C (Diversispora), possibly due to lack of recognition of the

morphologically distinct features of spores. Recently, Schiifler & Walker

(2010) erected the genus Redeckera (“group Cb”) with three species (Gl. fulvum,

Gl. megalocarpum, and Gl. pulvinatum) within the Diversisporaceae.

When Schiifsler et al. (2001) proposed a new phylum for the AM fungi,

although they initiated a principal re-organization of species with the four

orders, they did not propose families and genera that were clearly separated

genetically from each other. For example, Diversisporales and Diversisporaceae,

lacking either type genus or type species, were at that time invalidly described,

while the same authors mentioned only that some Glomus groups in the

Glomerales and Glomeraceae were clearly genetically different. This had the

unfortunate result that other taxonomists or ecological mycorrhizologists

continued to refer to Glomus group A, group B, and ‘the like’ as Schiifler et al.

(2001) had called them. Even new taxa were being justified on the basis of small

differences in partial rDNA sequences within such Glomus groups as A, B or

C (e.g. Blaszkowski et al. 2004, 2006, 2008, Oehl et al. 2005). Furthermore, in a

recent study (Gamper et al. 2009) species of Glomus, Diversispora and others,

e.g. Glomus versiforme and D. celata, were shown to be phylogenetically strongly

related, although no attempts were made to reorganize Gl. versiforme and related

‘Glomus’ spp. into Diversispora. In ecological studies, so-called ‘environmental

sequences (DNA sequences from AM species as yet unrecorded in databases,

often directly recovered from mycorrhizal root or soil analyses instead of from

identified spores) are named and referred to as ‘Glomus sp. uncultured’ or

78 ... Oehl & al.

‘Glomus unknown, not particularly helpful to someone who wishes to know

whether the species so named represents Diversispora, Glomus group A or B

sensu Schiffer et al., or a different group that might have completely different

spore formation.

It is clear that an improved phylogeny of Glomus is urgently needed (see

also Kriiger et al. 2009, Stockinger et al. 2009). Currently, new or unknown

species that are related to ‘groups of genetically related fungi’ but belong to

clearly distinct phylogenetic groups continue to be called ‘Glomus’ and thus

remain incorrectly classified. One motivation for this paper was the feeling that

because GI. versiforme has been supported as belonging to Diversispora, it should

officially be included within that genus. From that it followed that if group

A could phylogenetically be separated from groups B and C (Diversispora),

a new organization of the known and described species is both justified and

necessary. A first attempt was tried only recently by Schiifler & Walker (2010);

however, these authors based their conclusions almost exclusively on molecular

analyses and did not correlate their phylogenetic results with the morphological

characters of the different phylogenetic clades that our analyses indicate.

The objectives of the present study were therefore to: 1) identify and describe

morphological differences or similarities of species in the various ‘Glomus

groups, and 2) reorganize the species into existing or new genera as accurately

as possible according to currently available information. Thus, we aim here

at re-organizing the AM fungal species with glomoid spore formation using

existing genetic information combined with existing or new morphological

spore characteristics within the Glomerales and Diversisporales.

Material & methods

Specimen analyses

We analyzed specimens representing 101 of a total 117 described Glomus and

Diversispora spp. (TABLE 1). Holotype, isotype, paratype and ex-type materials were

examined along with representatives from institutional herbaria — OSC, FH, Z+ZT,

URM (Recife, Brazil), Embrapa Agrobiologia (Seropédica, Brazil), DCS-UFLA (Lavras,

Brazil), International Culture Collection of Vesicular-Arbuscular Mycorrhizal Fungi

(INVAM), Swiss collection of Arbuscular Mycorrhizal Fungi (SAF, Agroscope ART

Zurich) — and private AM fungal collections curated by Sieverding, Oehl, Trappe,

Blaszkowski, McGee, and Goto. The Hall & Abbott (1979) photographic slide collection

was also reviewed. For this paper, all original species descriptions and published species

emendations were also considered.

Older specimens (mounted on microscopic slides prior to 1990) were mostly

mounted in lactophenol, while others were fixed with polyvinyl alcohol-lactic acid-

glycerol (PVLG) or in a mixture of PVLG + Melzer’s reagent, which post-1990 are the

principal fixing media (Brundrett et al. 1994). Newly mounted spores and sporocarps

from collections or from cultures were fixed using the latter two fixing media, or

occasionally also in a mixture of 1:1 lactic acid to water, in Melzer’s reagent, and in water.

Glomoid species reorganized ... 79

When available, spores freshly isolated from soils or bait cultures were also mounted

and analyzed. Spore wall terminology follows the nomenclature of Walker (1983) and

Stiirmer & Morton (1997). Analyses of the spore walls, the germination structures and

all other mycorrhizal structures were performed using compound microscopes at 100-

1000x. Most photographs were taken with a digital camera (Olympus model DP70-

CU) mounted on a Zeiss Axioplan compound microscope, although a few were taken at

OSC, URM or INVAM during short-term visits. Legends and scales were inserted with

Adobe Photoshop C82 9.0.

All spore observations and all information on spore characteristics are based on

spores extracted from soil, from trap cultures or from single or multiple spore-derived

pure cultures. No information is provided from in-vitro-cultured materials.

Molecular and phylogenetic analyses

GENES ANALYZED: Partial sequences of 6-tubulin and rRNA (SSU and LSU) genes

were used to reconstruct, by independent analyses, the phylogeny of the Glomeromycota.

Intron sequences from the B-tubulin gene were excluded and only exon regions were

considered within analyses. ITS sequences were also used to clarify relationships among

members of the Diversisporales.

SEQUENCE ALIGNMENT: The sequences (all obtained from the National Center for

Biotechnology Information-NCBIJ) were aligned using the program ClustalX (Larkin et

al. 2007) and edited with the BioEdit program (Hall 1999) to obtain a final alignment.

TABLE 1. Type and non-type specimens from 115 Glomus and 2 Diversispora species

analyzed for reorganization of the Glomerales.

SPECIES NAME

Diversispora spurca

Di. celata

Glomus achrum

Gl. africanum

Gl. aggregatum

Gl. albidum

Gl. ambisporum

Gl. antarcticum

Gl. arborense

Gl. arenarium

Gl. atrouva

Gl. aurantium

Gl. aureum

Gl. australe

Gl. avelingiae

Gl. badium

Gl. bagyarajii

Gl. bistratum

Gl. boreale

Gl. botryoides

Gl. brohultii

Gl. caesaris

TYPE MATERIAL (OBSERVER)

Ex type at INVAM (Oehl)

Ex type, inclusive pure cultures (Oehl)

Type (Oehl), ex type (Goto)

No access

Type OSC #40254 (Oehl)

Holotype OSC #40400 (Oehl)

Holotype OSC #44289 (Oehl)

No access

Type at OSC (Oehl), ex type (Goto)

Ex type (Oehl, Goto)

Type (Oehl)

Holotype Z+ZT, isotypes OSC #; FB;

Pure culture (Oehl)

No access

Holotype Z+ZT, isotypes deposited at

OSC (Sieverding, Oehl)

No access

Type (Oehl), ex type (Goto)

Photographic slide collection of Hall &

Abbott (1979)

Ex type (Sieverding)

Holotype Z+ZT, isotypes OSC #; FB;

Pure culture (Oehl)

NON TYPE MATERIAL (OBSERVER)

Specimen from Bolivia & Oman

(Oehl)

Specimen from Switzerland (Oehl)

OSC #45840 (Oehl)

Specimen from Bolivia, Benin (Oehl)

Specimen from Chile and UAE

(Sieverding)

Specimen from Germany (Oehl)

Specimen & pure cultures from

Central Europe (Oehl)

(Trappe, McGee, Oehl)

Specimen from Central Europe

(Oehl, Sieverding)

Trappe collection (Oehl)

Specimen from Benin (Oehl), and

Costa Rica (Sieverding)

Specimen and pure cultures from

Germany (Oehl)

80 ... Oehl & al.

TABLE 1, continued

GI. caledonium

GI. canadense

Gl. candidum

Gl. canum

Gl. cerebriforme

Gl. citricola

Gl. claroideum

GI. clarum

Gl. clavisporum

Gl. constrictum

GI. convolutum

GI. coremioides

GI. coronatum

GI. corymbiforme

Gl. cuneatum

Gl. delhiense

Gl. diaphanum

GI. dimorphicum

GI. deserticola

Gl. dolichosporum

GI. drummondii

GI. eburneum

GI. epigaeum

GI. etunicatum

GI. fasciculatum

Gl. flavisporum

GI. formosanum

Gl. fragile

Gl. fragilistratum

Gl. fuegianum

GI. fulvum

GI. geosporum

GI. gibbosum

Gl. globiferum

Gl. glomerulatum

GI. goaense

GI. halonatum

Gl. heterosporum

Gl. hoi

GI. hyderabadense

GI. indicum

GI. insculptum

Gl. intraradices

GI. invermaium

GI. iranicum

Gl. irregulare

Type FH (Oehl)

No access

Ex type (Oehl, Goto)

Ex type (Oehl)

No access

Type OSC #40252 (Oehl)

Ex type specimen (Sieverding)

Ex type (Oehl))

Paratypes OSC #38850 (Oehl,

Sieverding)

Paratype OSC #30986, ex isotype (Oehl)

Ex type (Oehl)

Type OSC # 53909 (Oehl)

Ex type (Goto)

No access

Ex type (Oehl)

No access

Holotype 44474, Paratype OSC #47834

(Oehl)

No access

Type (Oehl), ex type (Goto)

Ex type at INVAM (Oehl)

Holotype OSC #39475 (Oehl)

Holotype OSC (Oehl)

Type sensu (Walker & Koske 1987)

Ex type at INVAM (Oehl)

Type OSC #29419 (Oehl)

Type: Blaszkowski online pages

Holotype OSC #46718 (Oehl)

Isotype OSC #46674, ex type

(Sieverding)

No access

Paratype Trappe 3594 (Oehl)

Holotype 44288 (Oehl)

Paratype Trappe 3058 (Oehl)

No access

Type (Oehl)

Isotype OSC #43941 (Oehl)

No access

Type (Oehl)

Specimen and pure cultures from

Central Europe (Oehl)

Thaxter collection, Trappe collection

(Oehl)

Specimen from Brazil, Europe (Oehl),

cultures from Benin (Oehl)

INVAM; specimen from Colombia

(Sieverding)

Specimen from Benin (Oehl)

Specimen and pure cultures from

Central Europe (Oehl)

Specimen from Brasil (Goto); Trappe

collection (Oehl)

Specimen from India, Mexico, Spain

(Oehl)

Specimen from Central Europe,

Bolivia (Oehl)

Colombia (Sieverding)

Specimen from Bolivia & Oman

(Oehl)

Specimen from Benin, Bolivia,

Paraguay, Europe, Mexico (Oehl)

Specimen from Central Europe

(Oehl, Sieverding)

Trappe collection (Oehl)

Specimen from UAE (Sieverding)

Trappe collection (Oehl)

Specimen from Switzerland (Oehl)

Trappe collection (Oehl)

Specimen and pure culture and from

Central Europe (Oehl)

Specimen from UAE (Sieverding)

Specimen at INVAM (Oehl)

Specimen from Brazil (Goto)

Specimen from Germany (Oehl)

Specimen from Germany (Oehl),

cultures from Benin (Oehl, IITA)

Specimen from Benin (Oehl) and

from Colombia (Sieverding)

Specimen and cultures from Europe

&, specimen from INVAM (Oehl)

Specimen from UAE (Sieverding)

Specimen from Switzerland (Oehl)

TABLE 1, concluded.

Gl. kerguelense

Gl. lacteum

Gl. lamellosum

Gl. liquidambaris

Gl. luteum

Gl. macrocarpum

Gl. magnicaule

GI. manihotis

Gl. megalocarpum

Gl. melanosporum

Gl. microaggregatum

Gl. microcarpum

Gl. minutum

Gl. monosporum

Gl. mortonii

Gl. mosseae

Gl. multicaule

GI. multiforum

Gl. nanolumen

Gl. pachycaule

Gl. pallidum

Gl. pansihalos

Gl. pellucidum

Gl. perpusillum

Gl. proliferum

Gl. przelewicense

Gl. pubescens

Gl. pulvinatum

GI. pustulatum

Gl. radiatum

Gl. rubiforme

Gl. segmentatum

Gl. sinuosum

Gl. spinosum

Gl. spinuliferum

Gl. taiwanense

Gl. tenebrosum

Gl. tenerum

Gl. tenue

Gl. tortuosum

Gl. trimurales

Gl. verruculosum

Gl. versiforme

Gl. vesiculiferum

Gl. viscosum

Gl. walkeri

Gl. warcupii

Gl. xanthium

Type OSC (Oehl)

Isotype OSC #50183 (Oehl)

Type OSC, ex type INVAM (Oehl)

Ex type, photographic slide collection

by Hall & Abbott (1979) (Oehl)

Holotype OSC #41498 (Oehl)

Holotype OSC #28739, ex type (Oehl)

Holotype OSC #, ex type (Oehl)

Holotype OSC #46719 (Oehl)

Ex type (Oehl)

Type OSC (Oehl)

Holotype OSC #49460 (Oehl)

Holotype FH (Oehl)

Holotype OSC #35386 (Oehl)

Type (Oehl)

Isotype OSC #49585

Type at PDD (Oehl)

Holotype OSC (Oehl)

Ex type (Oehl, Goto)

Type (Oehl), Ex type ( Goto)

Ex type (Oehl)

No access

Holotype OSC #46721 (Oehl)

Type OSC #30931, paratypes (Oehl)

Type , ex type (Oehl)

Ex type (Sieverding)

Holotype Z+ZT, isotopes at OSC & FB

(Oehl, Sieverding)

Type OSC #40°251 (Oehl)

Holotype OSC #49584 (Oehl)

Ex type, INVAM (Oehl)

Ex type (Oehl)

Type (Oehl), ex type ( Goto)

Ex type (Oehl)

Type (Oehl) ex type (Goto)

Glomoid species reorganized ... 81

Specimen from UAE (Sieverding)

Specimen from Poland, Germany

(Oehl, Sieverding)

Specimen from Brazil (Oehl, Goto)

INVAM; specimen from Central

Europe, Benin, Mexico (Oehl)

Specimen from Mexico (Oehl)

Specimen from Central Europe

(Oehl)

Specimen from Switzerland (Oehl)

Specimen from Germany (Oehl,

Sieverding)

Specimen and pure cultures from

Germany (Oehl)

Specimen from Switzerland &India

(Oehl)

Specimen from India and Benin

(Oehl)

Specimen at INVAM (Oehl)

Specimen and pure pot cultures from

Switzerland (Oehl, Sieverding)

OSC #30990 (Oehl)

Trappe collection (Oehl)

Btaszkowski collection (Oehl)

Trappe collection (Oehl)

Specimen from Switzerland (Oehl)

Specimen and cultures from Central

Europe, India, Oman, Benin, Bolivia

(Oehl, Sieverding)

Specimen from Switzerland (Oehl)

Specimen from Benin (Oehl),

Thailand (Sieverding), Brasil (Goto)

Specimen from Switzerland (Oehl)

Specimen from Australia (Oehl)

Specimen from alpine areas (Oehl)

OSC #32894

Specimen at INVAM; specimen from

Central Europe (Oehl, Sieverding)

OSC #28727 (Oehl)

Specimen at INVAM

Gl. zaozhangianum No access

82 ... Oehl & al.

PHYLOGENETIC ANALYSES: The main tree was constructed by a heuristic search

using the maximum likelihood (ML) method with the Phylogenetic Analysis Using

Parsimony (PAUP) program version 4 (Swofford 2003). Maximum parsimony (MP) and

neighbor joining (NJ) analyses with 1000 bootstrap replications also were performed

by PAUP. Bayesian (two runs over 1 x 10° generations with a burnin value of 2500) and

maximum likelihood (1000 bootstrap) analyses were executed, respectively, in MrBayes

3.1.2 (Ronquist & Huelsenbeck 2003) and PhyML (Guindon & Gascuel 2003), launched

from Topali 2.5. The model of nucleotide substitution was estimated using Topali 2.5

(Milne et al. 2004). Sequences from Neurospora crassa, Boletus edulis, Rhizophydium

sphaerotheca were used as outgroups for Glomeromycota, and Glomus intraradices and

G. xanthium for Diversisporales (ITS analysis).

Boletus edulis DQ534675

Neurospora crassa AY046271

Glomus proliferum AF213462

G. vesiculiferum L20824

G. fasciculatum Y 17640

G. irregulare FJ009618

G intraradices AJ852526 Ab2

G. intraradices AY635831

— G. aggregatum AY285859

73 62

55 76| ¢ G clarum AJ852597

0.99} 1.00

G, manihotis Y17648

54 G. badium AJ871990

mie G, sinuosum AJ133706 Ab3

ss G. coremioides AJ249715

039 G. perpusillum FJ164236

G. iranicum HM153420 Ab2

G. indicum GU059541

oe G aureum AJ871991 Fav

hives 58. Glomus sp. AJ301857

Aa2

vy dnosB snwoj5

1.00 100

100 ;

10oL G hoi AF485889

G. caledonium Y17653

wes G. fragilistratum AJ276085

56)

ee G. geosporum AJ245637

g7 0-83

Oty G. coronatum AJ276086

4 G mosseae 214007

1.00 G. mosseae AY635833

G. africanum HM153416

Aa3

G. verruculosum AJ301858 Aat

io G constrictum AJ534309

Glomus viscosum Y 17652 $ B2

100

100 G. etunicatum Y17639

100

100 4.00] | G etunicatum AJ852598

G. lamellosum AJ276083

G. luteum AJ276089

G. claroideum AJ276075

1.00

q dnoab snwioj5

Glomoid species reorganized ... 83

100

700 Kuklospora colombiana Z14006

1.00 100

100 Acaulospora spinosa 214004

100] 37

1.00] 94 A. laevis Y17633

1.00 A. scrobiculata AJ306442

+ A. longula AJ306439

p Ne A. rugosa Z14005

Glomus fulvum AM418543 $ Cb

Diversispora spurca AJ276077

99 D. celata AM713421 2

100 i 3

1.00 t D. celata AM713422 5

100 Ca lo

ae 400 G. eburneum AM7 13429 3

100 i=

cas 1.00| ' G eburneum AM713431 ae

1.00 G. versiforme X86687

si G. versiforme AJ276088

0.68 tool Pacispora scintillans AJ619944

eo P. scintillans AJ619948

99 Gigaspora gigantea AJ852602

97 >

79 99 G. rosea AJ852606

73} 1.00 64

on se G. albida AJ852599

1.00 | -— Racocetra castanea AF038590

100 Dentiscutata cerradensis AB041345

ie Fuscutata heterogama AJ852609

a F. heterogama AJ306434

99 =o Scutellospora aurigloba AJ276093

100 a S. calospora AJ306445

ici shed Archaeospora trappei AM114274

700 s

86 1.00” 4: trappei Y17634

74 100 ji ‘

7 400 Geosiphon pyriforme AJ276074

1.00] 98 1eo-— G pyriforme X86686

79 ‘

’ oe 400 Ambispora fennica AM268195

“138 _ A. callosa ABO47308

1.00

A. appendicula AB047302

A. appendicula ABO15052

Paraglomus occultum AJ276081

700) | P. occultum DQ322629

ip P. brasilianum AJ301862

P. laccatum AM295493

Fic. 1A—B. Phylogenetic reconstruction of the Glomeromycota obtained from partial SSU rDNA

sequences (~1800 bp). Group C are Diversisporaceae. The neighbor-joining (NJ), maximum

likelihood (ML) and bayesian analyses were performed with GTR+G+I substitution model.

Sequences are labeled with their database accession numbers. Bootstrap values (in %) are from NJ,

maximum parsimony (MP) and ML analyses (1000 bootstraps), respectively. The lasting numbers

below each branch line denotes the credibility value from the bayesian analysis. Only topologies

with bootstrap values of at least 50% are shown (Consistency Index = 0.4701; Retention Index =

0.8241). (Fic. 1A left on p. 82; Fic. 1B above)

0.01

84 ... Oehl & al.

Results & discussion

Relationships between general phylogenetics and morphological features

All four orders of the Glomeromycota have species that form glomoid spores.

Paraglomerales and Archaeosporales have low phylogenetic relationship with

Glomerales and Diversisporales (e.g. Redecker et al. 2007, Palenzuela et al. 2008;

Fics. 1-2, 4). Representatives of Ambispora, Archaeospora, Intraspora, and

Paraglomus form extraradical mycelia and mycorrhizal structures that stain

faintly to not at all in trypan blue (Spain & Miranda 1996, Spain 2003, Spain

et al. 2006, Sieverding & Oehl 2006). (The reasons for this are unclear: vesicle

formation is rarely reported or might have been misinterpreted in Archaeospora,

Intraspora and Paraglomus.) ‘This differs from the current glomeralean and

diversisporalean species, which have fungal structures that stain blue to deep

Boletus edulis AF336240

Neurospora crassa AF286411

Glomus trimurales FJ461851

Diversispora celata DQ350453

100 D. celata AY639233

G eburneum EF067886

1.00 G. eburneum EF067888 Ca

70 G. versiforme AY842573

G. versiforme AM947664

0.65 G. insculptum FJ461838

G. aurantium EF581864

ae G. aurantium EF581861

mien Scutellospora calospora AJ510231

pete Fuscutata heterogarna AY900503

0.87% F. heterogama AY900500

100 Dentiscutata nigra AY900498

9911 Lb. nigra AY900494

5 dnoib snwoj5

4.00 Gigaspora rosea Y12075

84 G. margarita AF396782

66 G. margarita AF396783

G. gigantea AY900506

G. gigantea AY900504

Racocetra gregaria AJ510232

69 R. verrucosa AY900508

72 79 R. castanea Y12076

76 84 | Cetraspora pellucida AY639313

4.00 105° pellucida AF396784

400 fr Pacispora scintillans FM876831

100 1 P scintilians FM876832

1.00

- Kuklospora colombiana FJ461804

100 K. kentinensis FM876822

39 K. kentinensis FM876830

ene Acaulospora longula AJ510228

Site A. paulinae AY639328

62 A. paulinae AY639263

Posh A. laevis AJ510229

A, lacunosa AJ510230

eri A Ian

Glomoid species reorganized ... 85

blue with trypan blue. They are also reported to form vesicular arbuscular

mycorrhiza (Schiifler et al. 2001), except for Gigasporaceae, Scutellosporaceae,

Racocetraceae and Dentiscutataceae, which form arbuscular mycorrhiza but not

intraradical vesicles, or at least vesicles have never been observed (Bentivenga &

Morton 1995, Oehl et al. 2008). To date Paraglomerales species are monogeneric

and form glomoid spores that may all germinate directly through the spore wall

instead of through the subtending hyphae as in Glomus and Diversispora (Spain

and Miranda 1996; Oehl pers. obs. on P. occultum).

97 Glomus diaphanum AJ972461 Ab2

o> +

99 89 G. diaphanum AJ972460

84 1.00 ref G. sinuosum FJ461846 + Ab3

= 1.00 G. intraradices AY842577

4.00 61 ‘ a G intraradices AF396797

88 72 G proliferum FM992402

93 Ab2

4.00 0.96 G. proliferum FM992398

G. manihotis AM158947

96 G. clarum AJ510242

+e G. clarum AJ510243 Q

Glomus coronatum AF 145740 9

1.00 3

G fragilistratum AF145747 =

100 G coronatum AF 145739 io]

100 Ps

400 G. mosseae AF 145735

1.00 G. mosseae AY639273 Aat a

96 G. caledonium AJ510239

56 99 G. caledonium AJ628059

91 92

a G AF 145743

73 Ba 1.00 geosporum

0.98 58 G geosporum AF 145742

0.62 a G deserticola AJ746249

97 G constrictum FJ461827 Aa3

1.00 G. xanthium AJ849467

Glomus luteum FM876809 ©

G. claroideum AF235007 ro

86 G. claroideum AJ271929 3

85) G. lamellosum AY541863 a

i 24 G. etuni AJ623310 S

1.00) 100 , etunicatum 5

99 G. etunicatum AF 145749 cS

“4.00 . B1

G. drummondii AJ972466 w

63 >

64 G. walkeri AJ972468

ey ges Geosiphon pyriforme AM183920

92 1100 Ambispora gerdemannii AJ271712

1.00 90 A. fennica AM268202

1.00 95

100 Paraglomus occultum DQ273827

100 P. laccatum AM295494

0.1 1.00

Fic. 2. Phylogenetic reconstruction of the Glomeromycota obtained from partial LSU rDNA

sequences (~600 bp). The neighbor-joining (NJ), maximum likelihood (ML) and bayesian analyses

were performed with GTR+G+I substitution model. Sequences are labeled with their database

accession numbers. Bootstrap values (in %) are from NJ, maximum parsimony (MP) and ML

analyses (1000 bootstraps), respectively. The lasting numbers below each branch line denotes the

credibility value from the bayesian analysis. Only topologies with bootstrap values of at least 50%

are shown (Consistency Index = 0.4207; Retention Index = 0.7818). (Fic. 2A left on p. 84; Fic. 2B

above.)

86 ... Oehl & al.

Phylogenetic reconstructions for gloomeromycotan species based on analyses of the 18S,

28S, and ITS regions of the ribosomal gene, and of the B-tubulin gene

The major clades for groups Aa, Ab, B and C were monophyletically obtained

in the phylogenetic trees constructed for the genome regions analyzed (Fics.

1-4). Clade group Aa was divided into three subclades with Gl. mosseae, Gl. hoi,

and Gl. constrictum as lead species of Aal, Aa2 and Aa3, respectively. Further

separation of clade Ab is not clear, although former Sclerocystis spp. (group Ab3)

and Gl. badium (morphologically also in group Ab3) are monophyletic (Fic. 1).

Clades Ab1 and Ab2, which differ consistently in glomoid spore morphology

(see below) and contain the largest number of species, were not re-organized

because the genetic database is not yet complete. Of the species analyzed, only

Gl. aureum represents clade Ab1, which should have Gl. macrocarpum as type

species according to Clements & Shear (1931). However, the 18S rDNA region

(Fic. 1) does not provide sufficient support by itself for creating a new genus.

The two lead species of group Ab2 are Gl. intraradices and Gl. irregulare, but

this group also is not yet recognized at the genus level. The group B clade

clearly stands apart from the others and splits into Glomus groups B1 and B2.

Glomus viscosum is hereafter attributed as a single species to group B2, while

Gl. etunicatum was recognized as the lead species for the group B1 (Fic. 1).

The ITS regions from clade groups Ca and Cb were analyzed because other

sequence data were either unavailable for group Cb species or (for the 18S)

available only for the lead species, Gl. fulvum. ITS analyses showed the Ca and

Cb clades well separated from each other (Fic. 3).

Morphology of glomoid, diversisporoid, and pacisporoid spores

All glomoid spores of the present Glomeraceae and Diversisporaceae have

one single spore wall with several layers. In many species the structural wall

layer is the inner layer, although other species possess additional innermost

(semi-)flexible layers. Because these additional layers are generally thin (often

<lum), closely adhere to the structural layer, and can be separated from the

latter only through hard pressure on the cover slides, they are not considered

separate walls. The presence or absence of innermost flexible layers is not

sufficient to distinguish morphologically between Glomus groups since the

majority of groups have some species with and some species without innermost

flexible layers. Such characters as mycelial/ mycorrhizal infection features, spore

size, spore wall color, wall ornamentation, and layer numbers or their staining

features could rarely be attributed to all species within a specific clade.

Spores of groups A and B have a structural layer that forms a continuum

from attached hyphal to spore wall layers (Fics. 5-17). We refer to such spore

types as GLOMOID.

Because Ca and Cb group spores have a structural layer that appears to

be inserted into the wall of the terminal element of the attached hypha, the

Glomoid species reorganized ... 87

Glomus xanthium AJ849467

G. intraradices AM980842

Acaulospora lacunosa AJ891113

A. denticulata AJ239115

A. paulinae AJ891114

A. morrowiae AJ242500

Pee A. morrowiae AM905249

9¢ A. laevis AJ242499

A. colossica AF 133772

A. alpina AJ891105

A. alpina AJ891109

Racocetra persica AJ410740

1.00

oe 5 R. castanea AJ313169

ol R. fulgida AM503938

0.58 Cetraspora pellucida AY035663

100 C. pellucida AJ239121

190 Dentiscutata cerradensis AB048690

ea 89 4.00 D. cerradensis ABO48686

52 ioe es Fuscutata heterogama AY997088

0.62 ved F. heterogama AJ245961

1.00 ... Gigaspora margarita AJ410742

a G gigantea AJ410755

1.00 G rosea AJ410750

G rosea AF004699

+ Pacispora scintillans FM876832

#00 Son P. scintillans FM876831

nen Glomus fulvum AM418547

1.00

G fulvum AM418548

G. fulvum AM418546

G. fulvum AM418545 Cb

toe G. megalocarpum AM418552

00 G megalocarpum AM418551

G. pulvinatum AM418550

G. pulvinatum AM418549

Bb G. aurantium AJ849468

Diversispora celata AM713403

ae D. celata AM713404

ane D. celata AM713402 Ca

G eburneum AM713409

G eburneum AM713408

G eburneum AM713407

G. versiforme AY 842569

G. versiforme AY842567

4 dnoib snwoj5

0.1

Fic. 3. Phylogenetic reconstruction of the Diversisporales obtained from ITS sequences (~500 bp).

The neighbor-joining (NJ), maximum likelihood (ML) and bayesian analyses were performed

with GITR+G+I substitution model. Sequences are labeled with their database accession numbers.

Bootstrap values (in %) are from NJ, maximum parsimony (MP) and ML analyses (1000 bootstraps),

respectively. The lasting numbers below each branch line denotes the credibility value from the

bayesian analysis. Only topologies with bootstrap values of at least 50% are shown (Consistency

Index = 0.5539; Retention Index = 0.8092).

structural spore wall layers may not form a continuum with the hyphal wall

layers (Fics. 18-21). The inner wall layer appears to differentiate de novo,

similar (but not equal) to inner walls in Acaulosporaceae or Entrophosporaceae

88 ... Oehl & al.

Neurospora crassa M13630

Rhizophydium sphaerotheca AY944848

68 Archaeospora trappei FJ174311

64 Ambispora leptoticha FJ174312

992» Os Glomus deserticola FJ174295 Aa3

G. diaphanum FJ174300 Ab2

50 78 G. sinuosum Fj174298 Ab3

80 100 ‘4 G. clarum FJ174296

1.00 1.0019 G manihotis FJ174297 Ab2

G. intraradices FJ174299

G proliferum AJ717321

ae s2[~ G mosseae FJ174292

72 99 G. coronatum FJ174290

0.99 4.08% G geosporum FJ174291 Aat

60 G fragilistratum FJ174293

57 G geosporum AJ717323

109 Glomus luteum Fj174282

toor” G etunicatum FJ174280

0.74 1.00 ]}- G claroideum FJ174283

G etunicatum FJ174279

Fuscutata heterogama FJ174315

Gigaspora rosea FJ174274

G albida FJ174275

100 G. gigantea FJ174276

991.09 G decipiens FJ174277

94 54 G. margarita FJ174278

il 0.96 53 Scutellospora calospora FJ174268

Racocetra persica FJ174272

ee R. verrucosa FJ174270

86 R. gregaria FJ174271

100 93 Cetraspora pellucida FJ174269

R. fulgida FJ174273

1.00 R. castanea AJ717326

100 —— Glomus versiforme FJ174285

50 G. trimurales FJ174289

73] 1:00 Ip Diversispora spurca FJ174287

G. eburneum FJ174288

4.00 - Kuklospora colombiana FJ174310

97 Acaulospora koskei FJ174309

rhe A. morrowiae FJ174302

= mm A. dilatata FJ174301

ry A. mellea FJ174303

0.66 K. kentinensis FJ174306

A. foveata FJ174304

A. lacunosa FJ174305

A. scrobiculata FJ174307

A. laevis FJ174308

100 Paraglomus occultum FJ174313

96 L. P pbrasilianum FJ174314

vy dnoi6 snwoj5

q dnosb snuoj5

9 dnos6 snwoj5

0.1

Fic. 4. Phylogenetic reconstruction of the Glomeromycota obtained from partial B-tubulin

sequences (~600 bp). The neighbor-joining (NJ), maximum likelihood (ML) and bayesian analyses

were performed with GTR+G+I substitution model. Sequences are labeled with their database

accession numbers. Bootstrap values (in %) are from NJ, maximum parsimony (MP) and ML

analyses (1000 bootstraps), respectively. The lasting numbers below each branch line denotes the

credibility value from the bayesian analysis. Only topologies with bootstrap values of at least 50%

are shown. (Consistency Index = 0.3709; Retention Index = 0.6969).

(Oehl et al. 2006, Sieverding & Oehl 2006). In Ca the structural and (if present)

inner flexible spore wall layers form a septum at the base or at a short distance

from the base of the spore within the subtending hypha. In Cb the inner lamina

of the structural wall layer always forms a septum at the spore base and the

outer lamina may continue for 1-10 um into the subtending hypha. We refer to

this spore type as DIVERSISPOROID.

Glomoid species reorganized ... 89

Pacispora spores superficially resemble glomoid spores, as the spore wall

structural layer forms a continuum with the hyphal wall layers. However,

Pacispora spores strongly differ from glomoid spores: i) in possessing an

additional thick inner germinal wall that forms de novo and is clearly separate

from the outer wall; ii) by species-specific germination structures resembling

a germ shield; and iii) by germ tubes that arise from the ‘shields’ on the inner

wall and penetrate directly the adjacent outer wall. We refer to this spore type

as PACISPOROID.

Morphology of the subtending hypha, spore base, and spore pore closure

A combination of two major morphological features and one secondary

feature are congruent with the major molecular phylogenetic groups of the

glomoid, diversisporoid, and pacisporoid species (TABLE 2): i) the morphology

of the spore base including the pore closure and position of the septum

(Fics. 5-23, 24-114); ii) the morphology of the subtending hypha (su)

(Fics. 5-23, 24-114); and iii) secondarily, the prevailing formation and spore

organization level within either sporocarps or spore clusters or singly in soil.

Spores of Glomus groups Aa(1,2,3) and Ab(1,2,3) all have su that are

concolorous with (or slightly lighter in color than) the structural spore wall,

and the spore wall differentiation extends completely into the sH (TABLE 2,

FIGs. 5-14, 24-89). In group Bl and B2 spores the structural spore wall

differentiation also extends into the su, but the su colour changes abruptly to

hyaline or white at the spore base, even when the spore wall layers are distinctly

pigmented (TABLE 2, FIGS. 15-17, 90-98).

TABLE 2. Morphology of subtending hyphae (sH) and spore formation in glomoid

spored species in Glomerales and Diversisporales

GROUP AAI AA2 AA3 ABI AB2

LEAD Gl. mosseae Gl. hoi Gl. constrictum GI. macrocarpum _ Gl. intraradices

SPECIES GI. microcarpum Gl. irregulare

RECENT OR Funneli- Simiglomus Septoglomus Glomus Glomus

NEW GENUS _formis

SH CHARACTERS

Color Same as spore Sameas Same as Same as Same as

wall spore wall spore wall spore wall spore wall

Shape Funnel-shape — Cylindric/sl. Cylindric to Cylindric to Cylindric,

to cylindric, funnel- constricted, slightly funnel-

& shape, rarely sl. shape,

funnel-shape,

thickness thick walled thick walled +/- thick walled thick walled thick walled

Pore closure | Conspicuous Sev. conspic. Septum under Introverted sw Open or septum

septum under septadistant —_/ at spore base thickening; +/- distant to spore

/ at spore to spore bridging septum _base; introverted

base base sw thickening

rudimentary

SPORE Singly or in Singly or Singly or open Compact, eee d

FORMATION — few-spored open spore spore clusters unorganized Pet

: unorganized

sporocarps clusters multi-spored i Tasteeonlen

sporocarps, : i

singly

clusters or singly

90 ... Oehl & al.

TABLE 2, concluded.

GROUP AB3 Bi B2 Ca CB

ae Gl. coremioides Gl. etunicatum Gl.viscosum Gl. versiforme Gl. fulvum

RECENT OR Glomus patie Viscospora Diversispora Redeckera

NEW GENUS teria t2

SH CHARACTERS

Color Same as spore White to hyaline, | White to Hyaline distant Hyaline distant

wall abrupt color hyaline to septum that to septum that

change at spore is concolorous is concolorous

base if spore wall with structural with structural

pigmented SWL SWL

Shape; Cylindric, Conspicuously Cylindric; Generally fragile, | Generally

funnel-, or bill-shaped; cylindric distant fragile and

& bill-shaped,; to septum, or inflated ;

constricted;

Thickness thick-walled +/- thick-walled thick-walled thin-walled thin-walled

Pore closure Introvertedsw Septum at spore Constricted | Septum at or Broad septum

thickening base wall or beneath spore at spore base

open base, or open

SPORE In compact, Singly Singly or In large, compact In large,

FORMATION _ organized in loose compact

sporocarps,

sporocarps, clusters eee Nee sporocarps/

around hyphal clusters, open

Roe aggregates,

center; initially ; aggregates,

: or singly :

also singly or singly

In contrast, the spore wall differentiation generally does not extend into the

SH in group Ca and Cb spores (TABLE 2, Fics. 14-19, 99-114), as lamination

and differentiation of the structural spore wall layer either does not extend into

the sH or extends only a short distance from the base. Within these Ca and Cb,

therefore, the sH are usually quite fragile (Figs. 99-107) and hyaline to white,

distal to the pore closure at the spore base (GI. spurcum = Di. spurca) or in the

sH (GI. eburneum). The hyaline to white color of the sH behind the septum

changes conspicuously when spores are pigmented. Cb species have broad

sH and broad septa at the spore bases (Fics. 108-114). Their su are inflated

(GI. pulvinatum) or (sometimes) very short within compact sporocarps (e.g.

GI. fulvum and Gl. canadense).

Type of spore formation

Glomus group spore formation is intercalary or terminal on su, both for

spores formed either singly in soil (e.g. Gl. arborense, Gl. etunicatum) or in

loose/dense spore clusters or sporocarps (e.g. Gl. fasciculatum, Gl. pallidum,

Gl. glomerulatum).

Fics 5-17. Examples of spore bases (with 1-3 spore wall layers; swL1-3) and subtending hyphae

(sH) of species assigned to phylogenetic clades Aal, Aa2, Aa3, Ab1, Ab2, Ab3, B1 and B2; structural

spore wall layers are continuous with the su wall. 5-14. su of A clade spores are concolorous

with or slightly lighter than the spore wall. 5-8. Clade Aa species never have an introverted wall

thickening at the base; spore pores are generally closed by a conspicuous septum (sp) arising

Glomoid species reorganized ... 91

5 6 SWL3 7

sp SWL2 8

SWL2 wg sp SWL2

: Z

a A\ de |

SWL1

: ak

9 swi2 10 sp SWL2 11 sp gle

p 2A ( |

SWL1 SWL1

Sp SWL2

12 i se. “SRS aid SWL2

SWL2 | |

m4 at

1 Sp 6 17 swi3

sp Sp

SWL2 SWL2 / SWL2

SWL1 SWL1

from the structural layer and/or the (semi-)flexible layer beneath. 5-6. Aal. 5. Lead species

Glomus mosseae. 6. Gl. geosporum. 7. Aa3: lead species Gl. constrictum. 8. Aa2: so far single lead

species Gl. hoi. 9. Ab2: without introverted wall thickening; spore base pore regularly open; lead

species Gl. intraradices. 10-13. Ab1: typical Glomus species with an introverted wall thickening at the

spore base; sometimes a short bridging septum arising from the structural wall layer or innermost

flexible lamina closes the spore base pore. 10. Lead species Gl. microcarpum. 11. Gl. aureum.

12. Lead species Gl. macrocarpum. 13. Gl. pellucidum. 14. Ab3; also with a slight introverted

wall thickening but pores generally closed by a short septum arising from the structural layer

and/or the (semi-)flexible layers beneath. In some clade Ab3 species the spore head sometimes

is also thickened towards the sporocarp surface; lead species Gl. coremioides and Gl. sinuosum.

15-17. Bl: spore su bill-shaped and hyaline to white, even when the spore wall is pigmented.

15-16. lead species Gl. etunicatum. 17. Gl. claroideum.

92 ... Oehl & al.

18 SWL2

= 19 swi2

abs a 22

SWL2 “

SWL1

Fics 18-23. Clades Ca (Diversispora) and Cb (Redeckera) spore bases (with 1-3 spore wall layers;

swL 1-3) and subtending hyphae (sx): structural spore wall layer seemingly extends only a very

short distance into the subtending hyphae, soon becoming hyaline to white and undetectable; often

only the adherent outer spore wall layer(s) appears continuous with the mycelial hyphal wall to

connect the two neighboring spores in compact sporocarps. ‘The structural layer often appears

inserted into the ‘swollen hyphal tips. 18-22. Ca: spore sH generally white and cylindrical at the

spore base, soon fragile. Septum (sp) arises from the structural spore wall layer and either closes

the pore immediately or at some distance from the spore base (18-21) or the spore pore appears

open (22). 23. Cb: spore sH generally inflated a very short distance from the spore base with the

structural layer extending an equally short distance in the subtending hypha. At base, spores are

‘endospore-like, with a broad, thick septum formed by the structural layer.

Aal species generally form spores singly (e.g. Gl. geosporum, Gl. caledonium)

or occasionally in sporocarps containing just a few (2-20) spores (e.g. Gl.

mosseae, Gl. monosporum, Gl. coronatum). Aa2 and Aa3 species form spores

preferably singly in soil or roots, or in very loose spore clusters (Gl. hoi,

Gl. constrictum, Gl. deserticola, Gl. xanthium). Ab1 and Ab2 species generally

form spores in multi-spored loose/dense variably sized spore clusters. During

initial spore formation they may also form spores singly, although some isolates

appear never to form clusters in single species cultures on certain hosts. Ab3

species form spores strictly in sporocarps but may initially form single spores

on their mycelia (GI. sinuosum).

Glomoid species reorganized ... 93

B1 species generally form spores either singly in soil (e.g. Gl. etunicatum,

Gl. claroideum, Gl. walkeri) or in variably sized loose spore clusters (Gl. viscosum).

Sporocarp formation is extremely rare, if ever. The solitary B2 species —

Gl. viscosum — forms spores in clusters of highly variable size. Ca species form

spores singly in soils (e.g. Gl. spurcum = Di. spurca; Gl. eburneum) or in small

to large and irregular shaped compacted clusters with numerous spores (e.g.

Gl. versiforme, Gl. epigaeum) depending on culture and site conditions. Finally,

Cb species generally form spores in large, compact sporocarps (e.g. Gl. fulvum,

Gl. fragile) that are among the largest known within the Glomeromycota

(especially Gl. megalocarpum). As the formation of a peridium around the

sporocarps or single spores does occur in groups Aal, Abl, Ab3, and Cb, it is

not a consistent morphological indicator at the family or genus level. Peridium

formation has not yet been observed for B1, B2, and Ca species.

Organization level of sporocarps

Except for group Ab3, spores of other Glomus groups have (with some

exceptions) limited to no organization in sporocarps or compacted spore

clusters. As spores in sporocarps appear to be randomly distributed, su are

variably long (e.g. Gl. mosseae, Gl. coronatum, GI. macrocarpum, Gl. ambisporum,

Gl. versiforme, Gl. fulvum, Gl. pulvinatum). In contrast, Ab3 sporocarps are

highly organized with spores forming around a central hyphal plexus. Variation

in sH length is much lower (e.g. Gl. sinuosum, Gl. rubiforme). Sporocarp size

variability of Ab3 species is low, largely dependent on the diameter of the central

hyphal plexus and spore size in individual species. In unorganized sporocarps

of the other “Glomus groups’ sporocarp size is much more variable due to a lack

of symmetry, greater su length variability, and greater variability of numbers

of spores formed per sporocarp. According to our analyses, species that

form dense, highly compact sporocarps (e.g., Gl. radiatum, Gl. vesiculiferum,

Gl. segmentatum, GI. convolutum, Gl. fuegianum) do not form spores around a

central hyphal plexus arising from a single hypha but sporocarps appear to have

several hyphal attachments. They thus probably belong to group Ab1 and so are

not attributed to group Ab3.

Taxonomic revision

Glomeraceae Piroz. & Dalpé emend. Oehl, G.A. Silva & Sieverd. (Glomus group A)

EMENDED DESCRIPTION: Spores formed terminally on or intercalary in hyphae,

in soil and sometimes in roots, either singly, in spore clusters, or in multiple-

spored loose to compact sporocarps when compact spores are randomly

distributed or organized around a central plexus of hyphae. Compact sporocarps

with or without peridium. Spores with one mono-to-multiple layered wall.

Wall of the sH conspicuously continuous and concolorous with the spore wall,

94 ... Oehl & al.

or slightly lighter in color than the spore wall; su funnel-shaped, cylindrical or

constricted; forming typical vesicular-arbuscular mycorrhiza, with mycorrhizal

structures that stain blue to dark blue in trypan blue.

TYPE GENUS: Glomus Tul. & C. Tul.

OTHER GENERA: Funneliformis C. Walker & A. Schiissler, Septoglomus Sieverd.

et al., Simiglomus Sieverd. et al.

Glomus Tul. & C. Tul. emend. Oehl, G.A. Silva & Sieverd. Figs. 25-74

(Glomus groups Ab1, Ab2 & Ab3)

EMENDED DESCRIPTION: Spores formed singly within soil or sometimes roots,

in disorganized, multiple-spored loose spore clusters or in compact sporocarps;

compact sporocarps without or with peridium, spores are either not organized

in sporocarp, or organized around a central hyphal plexus. Spores with a

mono-to-multiple layered wall. Wall of the sH conspicuously continuous and

concolorous with the spore wall, or slightly lighter in color than the spore wall.

Spore pore closure often by introverted wall thickening, sometimes supported

by a short bridging septum, rarely open. Forming typical vesicular-arbuscular

mycorrhiza, with mycorrhizal structures that stain blue to dark blue in trypan

blue.

TYPE SPECIES: Glomus macrocarpum Tul. & C. Tul.

Glomus achrum Blaszk., D. Redecker, Koegel, Schiitzek, Oehl & Kovacs, Botany 87:

262. 2009.

Glomus aggregatum N.C. Schenck & G.S. Sm., Mycologia 74: 80. 1982.

Glomus ambisporum G.S. Sm. & N.C. Schenck, Mycologia 77: 566. 1985.

Glomus antarcticum Cabello, Mycotaxon 51: 124. 1994

Glomus arborense McGee, Trans. Br. Mycol. Soc. 87: 123. 1986.

Glomus atrouva McGee & Pattinson, Austral. Syst. Bot. 15: 115. 2002.

Glomus aureum Oehl & Sieverd., J. Appl. Bot. 77: 111. 2003.

Glomus australe (Berk.) S.M. Berch, Can. J. Bot. 61: 2611. 1983.

= Endogone australis Berk., Fungi Flora Tasman. 2: 282. 1859.

Glomus badium Oehl, D. Redecker & Sieverd., J. Appl. Bot. Food Qual. 79: 39. 2005.

= Funneliformis badius (Oehl, D. Redecker & Sieverd.) C. Walker & A. Schiissler,

The Glomeromycota - a species list: 13. 2010.

Glomus bagyarajii V.S. Mehrotra, Philipp. J. Sci. 126: 235. 1997.

Glomus bistratum Blaszk., D. Redecker, Koegel, Symanczik, Oehl & Kovacs, Botany

87: 267. 2009.

Glomus boreale (Thaxt.) Trappe & Gerd., Mycol. Mem. 5: 58. 1974.

= Endogone borealis Thaxt., Proc. Am. Acad. Arts Sci. 57: 318. 1922

Glomoid species reorganized ... 95

Glomus botryoides EM. Rothwell & Victor, Mycotaxon 20: 163. 1984.

Glomus brohultii Sieverd. & R.A. Herrera, J. Appl. Bot. 77: 37. 2003

Glomus canum McGee, Austral. Syst. Bot. 15: 116. 2002.

Glomus cerebriforme McGee, Trans. Br. Mycol. Soc. 87: 123. 1986.

Glomus citricola D.Z. Tang & M. Zang, Acta Bot. Yunn. 6: 301. 1984.

Glomus clarum T.H. Nicolson & N.C. Schenck, Mycologia 71: 182. 1979.

= Rhizophagus clarus (T.H. Nicolson & N.C. Schenck) C. Walker & A.

Schiissler, The Glomeromycota - a species list: 19. 2010.

Glomus clavisporum (Trappe) R.T. Almeida & N.C. Schenck, Mycologia 82: 710.

1990.

= Sclerocystis clavispora Trappe, Mycotaxon 6: 359. 1977.

= Sclerocystis microcarpus S.H. Iqbal & Perveen, Trans. Mycol. Soc. Japan 21: 58. 1980.

Glomus convolutum Gerd. & Trappe, Mycol. Mem. 5: 42. 1974.

Glomus coremioides (Berk. & Broome) D. Redecker & J.B. Morton, Mycologia 92:

284. 2000.

= Sclerocystis coremioides Berk. & Broome, J. Linn. Soc., Bot. 14: 137. 1873.

= Xenomyces ochraceus Ces., Atti Accad. Sci. Fis. 8: 26. 1879.

= Ackermannia coccogena Pat., Bull. Soc. Mycol. Fr. 18: 182. 1902.

= Sphaerocreas coccogena (Pat.) Hohn., Sitzungsber. K. Akad. Wiss.

Wien, Math.-Naturwiss. K]., Abt. 1, 118: 401. 1909.

= Sclerocystis coccogena (Pat.) Hohn., Sitzungsber. K. Akad. Wiss.

Wien, Math.-Naturwiss. K]., Abt. 1, 119: 399. 1910.

= Ackermannia dussii Pat., Bull. Soc. Mycol. Fr. 18: 181. 1902.

= Sphaerocreas dussii (Pat.) Hohn., Sitzungsber. K. Akad. Wiss.

Wien, Math.-Naturwiss. K]., Abt. 1, 118: 401. 1909.

= Sclerocystis dussii (Pat.) Hohn., Sitzungsber. K. Akad. Wiss.

Wien, Math.-Naturwiss. Kl., Abt. 1, 119: 399. 1910.

Glomus corymbiforme Biaszk., Mycologia. 87: 732. 1995.

Glomus cuneatum McGee & A. Cooper, Austral. Syst. Bot. 15: 117. 2002.

Glomus custos C. Cano & Dalpé, Mycotaxon 109: 502. 2009.

= Rhizophagus custos (C.Cano & Dalpé) C. Walker & A. Schiissler,

The Glomeromycota — a species list: 19. 2010.

Glomus delhiense Mukerji, Bhattacharjee & J.P. Tewari, Trans. Br. Mycol. Soc 81: 643.

1983.

Glomus diaphanum J.B. Morton & C. Walker, Mycotaxon 21: 433. 1984.

= Rhizophagus diaphanus (J.B. Morton & C. Walker) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 19. 2010.

Glomus dolichosporum M.Q. Zhang & You S. Wang, Mycosystema 16: 241. 1997.

96 ... Oehl & al.

Glomus fasciculatum (Thaxt.) Gerd. & Trappe, Mycol. Mem. 5: 51. 1974.

= Endogone fasciculata Thaxt., Proc. Am. Acad. Arts & Sci. 57: 308. 1922.

= Rhizophagus fasciculatus (‘Thaxt.) C. Walker & A. Schiissler,

The Glomeromycota - a species list: 19. 2010.

= Endogone arenacea Thaxt., Proc. Am. Acad. Arts & Sci. 57: 317. 1922

= Rhizophagites butleri Rosend., Bull. Torrey Bot. Club 70: 131. 1943.

Glomus flavisporum (M. Lange & E.M. Lund) Trappe & Gerd., Mycol. Mem. 5: 58.

1974.

= Endogone flavispora M. Lange & E.M. Lund, Friesia 5: 93. 1954.

Glomus formosanum C.G. Wu & Z.C. Chen, Taiwania 31: 71. 1986.

Glomus fuegianum (Speg.) Trappe & Gerd., Mycol. Mem. 5: 58. 1974.

= Endogone fuegiana Speg., Anal. Soc. Cient. Argent. 24:125. 1887.

Glomus globiferum Koske & C. Walker, Mycotaxon 26: 133. 1986.

Glomus glomerulatum Sieverd., Mycotaxon 29: 74. 1987.

Glomus goaense Khade, Mycorrhiza News 20(4): 21. 2010.

Glomus heterosporum G.S. Sm. & N.C. Schenck, Mycologia 77: 567. 1985.

Glomus hyderabadense Swarupa, Kunwar, G.S. Prasad & Manohar., Mycotaxon 89:

247. 2004.

Glomus indicum Blaszk., Wubet & Harikumar, Botany 88: 134. 2010.

Glomus intraradices N.C. Schenck & G.S. Sm., Mycologia 74: 78. 1982.

= Rhizophagus intraradices (N.C. Schenck & G.S. Sm.) C. Walker &

A. Schiissler, The Glomeromycota - a species list: 19. 2010.

Glomus invermaium LR. Hall, Trans. Br. Mycol. Soc. 68: 345. 1977.

Fics 24-46. Glomus clade Ab1 forms spores in unorganized clusters/sporocarps or (sometimes)

singly in soil; subtending hyphae (su) are concolorous with (or slightly lighter than) the spore

wall, normally with introverted, species-specific basal wall thickenings; sH often cylindrical

(very short if formed in compact sporocarps). Species-specific sH wall thickening may continue

over rather long distances (< 200-500 um) even in sporocarps. Spores with 1-4 wall layers

(SwL1-swL4); pore at spore base pores often closed by introverted wall thickenings or with an

additional septum (sp) that arises from the structural layer, from an adherent innermost (semi-

)flexible layer, or from both layers. Individual layers might be species-specifically ornamented

(ORN). 24-25. Gl. microcarpum. 26. Gl. macrocarpum. 27. Gl. spinuliferum. 28. Gl. magnicaule.

29. Gl. multicaule. 30. Gl. heterosporum. 31. Gl. aureum. 32. Gl. invermaium. 33. Gl. glomerulatum.

34. Gl. ambisporum. 35. Gl. warcupii. 36. Gl. melanosporum. 37. Gl. atrouva. 38. Gl. botryoides.

39. Gl. spinosum. 40. Gl. mortonii. 41. Gl. australe. 42. Gl. fuegianum. 43. Gl. tenerum.

44. Gl. pellucidum. 45. Gl. segmentatum. 46. Gl. convolutum: spore peridium was removed by strong

pressure on the cover slide.

Glomoid species reorganized ... 97

98 ... Oehl & al.

Fics 47-58. Glomus clade Ab2 form spores in disorganized loose/dense clusters/sporocarps

(sometimes singly in soil); subtending hyphae (sH) concolorous with (or slightly lighter than) the

spore wall. sH generally openly cylindric; introverted wall thickenings rudimentary, but usually not

existent; spore base pore diagnostic: open or (rather rarely) with a septum arising from innermost

adherent (semi-)flexible layer. Spores with 1-4 wall layers (swLl-sw14). 47. Gl. intraradices.

48. Gl. aggregatum. 49. Gl. vesiculiferum. 50. Gl. globiferum. 51. Gl. tortuosum. 52. Gl. corymbiforme.

53. Gl. microaggregatum. 54. Gl. clarum. 55. Gl. manihotis. 56. Gl. fasciculatum. 57. Gl. diaphanum.

58. Gl. pallidum.

Glomus iranicum Blaszk., Kovacs & Balazs, Mycologia 102: 1457. 2010.

= Rhizophagus iranicus (Blaszk., Kovacs & Balazs) C. Walker & A.

Schiissler, The Glomeromycota — a species list: 19. 2010.

Glomus irregulare Btaszk., Wubet, Renker & Buscot, Mycotaxon 106: 252. 2008.

= Rhizophagus irregularis (Blaszk., Wubet, Renker & Buscot) C. Walker

& A. Schiissler, The Glomeromycota — species list: 19. 2010.

Glomus liquidambaris (C.G. Wu & Z.C. Chen) YJ. Yao, Kew Bull. 50: 306. 1995.

= Sclerocystis liquidambaris C.G. Wu & Z.C. Chen, Trans.

Mycol. Soc. Rep. China, 2: 74. 1987.

= Sclerocystis cunninghamia H.T. Hu, Quart. J. Chinese For. 21: 52. 1988.

Glomus macrocarpum Tul & C. Tul., Giorn. Bot. Ital., Anno 1, 2(7-8): 63. 1845.

= Endogone macrocarpa (Tul. & C. Tul.) Tul. & C. Tul., Fungi Hypog.: 182. 1851.

Glomoid species reorganized ... 99

= Endogone guttulata E. Fisch., Ber. Schweiz. Bot. Ges. 32: 13. 1923.

= Endogone nuda Petch., Ann. R. Bot. Gdns Peradeniya 9: 322. 1925.

= Endogone pampaloniana Bacc., Nuovo Giorn. Bot. Ital., n.s. 10: 90. 1903.

= Paurocotylis fulva var. zelandica Cooke, Grevillea 8: 59. 1879.

Glomus magnicaule LR. Hall, Trans. Br. Mycol. Soc. 68: 345. 1977.

Glomus manihotis R.H. Howeler, Sieverd. & N.C. Schenck, Mycologia 76: 695. 1984.

= Rhizophagus manihotis (R.H. Howeler, Sieverd. & N.C. Schenck) C. Walker

& A. Schiissler, The Glomeromycota - a species list: 19. 2010.

Glomus melanosporum Gerd. & Trappe, Mycol. Mem. 5: 46. 1974.

Glomus microaggregatum Koske, Gemma & P.D. Olexia, Mycotaxon 26: 125. 1986.

Glomus microcarpum Tul. & C. Tul., Giorn. Bot. Ital., Anno 1, 2(7-8): 63. 1845.

= Endogone microcarpa (Tul. & C. Tul.) Tul. & C. Tul., Fungi Hypog.: 182. 1851.

= Endogone neglecta Rodway, Proc. Roy. Soc. Tasmania 1917: 107. 1918.

Glomus minutum Blaszk., Tadych & Madej, Mycologia 76: 189. 2000.

Glomus mortonii Bentiv. & Hetrick, Mycotaxon 42: 10. 1991.

Glomus multicaule Gerd. & B.K. Bakshi, Trans. Br. Mycol. Soc. 66: 340. 1976.

Glomus nanolumen Koske & Gemma, Mycologia 81: 935. 1990.

Glomus pachycaule (C.G. Wu & Z.C. Chen) Sieverd. & Oehl, comb. nov.

MycoBank MB 519622

= Sclerocystis pachycaulis C.G. Wu & Z.C. Chen, Taiwania 31: 74. 1986.

Glomus pallidum LR. Hall, Trans. Br. Mycol. Soc. 68: 343. 1977.

Glomus pansihalos S.M. Berch & Koske, Mycologia 78: 832. 1986.

Glomus pellucidum McGee & Pattinson, Austral. Syst. Bot. 15: 120. 2002.

Glomus perpusillum Blaszk. & Kovacs, Mycologia 101: 249. 2009.

Glomus proliferum Dalpé & Declerck, Mycologia 92: 1180. 2000.

= Rhizophagus prolifer (Dalpé & Declerck) C. Walker & A.

Schiissler, The Glomeromycota — a species list: 19. 2010.

Glomus pubescens (Sacc. & Ellis) Trappe & Gerd., Mycol. Mem. 5: 57. 1974.

= Sphaerocreas pubescens Sacc. & Ellis, Michelia 2: 582. 1882.

= Stigmatella pubescens (Sacc. & Ellis) Sacc., Syll. Fung. 4: 680. 1886.

= Sclerocystis pubescens (Sacc. & Ellis) Hohn., Sitzungsber. K. Akad.

Wiss. Wien, Math.-Naturwiss. K1., Abt. 1, 119: 399. 1910.

= Endogone pubescens (Sacc. & Ellis) Zycha, Krypt.-Fl. Brandenburg 6a: 214. 1935.

Glomus radiatum (Thaxt.) Trappe & Gerd., Mycol. Mem. 5: 46. 1974.

= Endogone radiata Thaxt., Proc. Am. Acad. Arts & Sci. 57: 316. 1922.

100 ... Oehl & al.

Fics 59-74. Glomus clade Ab3, with species virtually all belonging to the former genus Sclerocystis;

form spores in highly organized, compact sporocarps, generally with > 50 spores per sporocarp;

subtending hyphae (sh) generally short, bill-shaped or cylindrical, and concolorous with (or slightly

lighter than) the spore wall. Spores species-specifically with (1-)2 wall layers (SWL1-SWL2) and

wall thickening at spore base, but pore generally closed by a septum that regularly arises from

the structural wall layer. 59-60. Gl. coremioides. 61-63. Gl. clavisporum. 64-65. Gl. badium.

66-67. GI. taiwanense. 68-70. GI. sinuosum. 71-72. Gl. pachycaule. 73-74. Gl. rubiforme.

Glomus rubiforme (Gerd. & Trappe) R.T. Almeida & N.C. Schenck, Mycologia 82:

709. 1990.

= Sclerocystis rubiformis Gerd. & Trappe, Mycol. Mem. 5: 60. 1974.

= Sclerocystis indica Bhattacharjee & Mukerji, Acta Bot. Indica 8: 99. 1980.

Glomus segmentatum Trappe, Spooner & Ivory, Trans. Br. Mycol. Soc. 73: 362. 1979.

Glomoid species reorganized ... 101

Glomus sinuosum (Gerd. & B.K. Bakshi) R.T. Almeida & N.C. Schenck, Mycologia

82: 710. 1990

= Sclerocystis sinuosa Gerd. & B.K. Bakshi, Trans. Br. Mycol. Soc. 66: 343. 1976.

= Sclerocystis pakistanica S.H. Iqbal & Perveen, Trans. Mycol. Soc. Japan 21: 59. 1980.

Glomus spinosum H.T. Hu, Mycotaxon 83: 160. 2002.

Glomus spinuliferum Sieverd. & Oehl, Mycotaxon 86: 158. 2003.

Glomus taiwanense (C.G. Wu & Z.C. Chen) R.T. Almeida & N.C. Schenck

ex Y.J. Yao, Kew Bull. 50: 306. 1995.

= Sclerocystis taiwanensis C.G. Wu & Z.C. Chen, Trans.

Mycol. Soc. Rep. China 2: 78. 1987.

Glomus tenebrosum (Thaxt.) S.M. Berch. Can. J. Bot. 60: 2615. 1983.

= Endogone tenebrosa Thaxt., Proc. Am. Acad. Arts Sci. 57: 314. 1922.

Glomus tenue (Greenall) I.R. Hall, Trans. Br. Mycol. Soc. 68: 350. 1977.

= Rhizophagus tenuis Greenall, New Zealand J. Bot. 1: 398. 1963.

Glomus tortuosum N.C. Schenck & G.S. Sm., Mycologia 74: 83. 1982.

Glomus vesiculiferum (Thaxt.) Gerd. & Trappe, Mycol. Mem. 5: 49. 1974.

= Endogone vesiculifera Thaxt., Proc. Am. Acad. Arts Sci. 57: 309. 1922.

= Funneliformis vesiculifer (Thaxt.) C. Walker & A. Schiissler,

The Glomeromycota - a species list: 14. 2010.

Glomus warcupii McGee, Trans. Br. Mycol. Soc. 87: 125. 1986.

Glomus zaozhuangianum FY. Wang & RJ. Liu, Mycosystema 21: 522. 2002.

Funneliformis C. Walker & A. Schiissler, emend. Oehl, G.A. Silva & Sieverd.

(Glomus group Aal) FIGs. 75-84

EMENDED DESCRIPTION: Spores formed within soil or rarely roots, singly or

sometimes in sporocarps with a few to several spores per sporocarp only; the

conspicuous sH is concolorous with spore wall color (or slightly lighter in

color), SH is species-specific and generally funnel-shaped to cylindrical. Wall

differentiation and pigmentation may continue over long distances from the

spore base (often > 50-250 um), then mycelium may become hyaline. Pore

regularly closed by a conspicuous septum that species-specifically arises from

the structural wall layer, from an additional adherent innermost, (semi-)flexible

lamina, or from both but not by introverted wall thickening, which is lacking.

Forming typical vesicular-arbuscular mycorrhiza, with mycorrhizal structures

that stain blue to dark blue in trypan blue.

GENDER: Funneliformis is a compound Latin adjective with a masculine/

feminine termination. Although under ICBN [Vienna Code] Article 62.3, the

authors had the prerogative to choose either of these genders, their actual choice

102 ... Oehl & al.

of neuter gender was not an available option. We therefore choose to treat this

genus as masculine (Article 62.3) and have made the appropriate corrections to

all adjectival epithets.

TYPE SPECIES: Funneliformis mosseae (T.H. Nicolson & Gerd.) C. Walker & A.

Schiissler

Funneliformis caesaris (Sieverd. & Oehl) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518445

= Glomus caesaris Sieverd. & Oehl, Mycotaxon 84: 381. 2002.

Funneliformis caledonius (T.H. Nicolson & Gerd.) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 13. 2010.

= Endogone macrocarpa var. caledonia T.H. Nicolson & Gerd., Mycologia 60: 318. 1968.

= Glomus caledonium (T.H. Nicolson & Gerd.) Trappe & Gerd., Mycol. Mem. 5: 56. 1974.

Funneliformis coronatus (Giovann.) C. Walker & A. Schiissler, The Glomeromycota

— a species list: 13. 2010.

= Glomus coronatum Giovann., Can. J. Bot. 69: 162. 1990.

Funneliformis dimorphicus (Boyetchko & J.P. Tewari) Oehl, G.A. Silva & Sieverd.,

comb. nov.

MycoBank MB 518447

= Glomus dimorphicum Boyetchko & J.P. Tewari, Can J. Bot. 64: 90. 1986.

Funneliformis fragilistratus (Skou & I. Jakobsen) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 13. 2010.

= Glomus fragilistratum Skou & I. Jakobsen, Mycotaxon 36: 276. 1989.

Funneliformis geosporus (T.H. Nicolson & Gerd.) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 14. 2010.

= Endogone macrocarpa var. geospora T.H. Nicolson & Gerd. Mycologia 60: 318. 1968.

= Glomus macrocarpum var. geosporum (T.H. Nicolson &

Gerd.) Gerd. & Trappe, Mycol. Mem. 5: 55. 1974.

= Glomus geosporum (T.H. Nicolson & Gerd.) C. Walker, Mycotaxon 15: 56. 1982.

Funneliformis halonatus (S.L. Rose & Trappe) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518453

= Glomus halonatum S.L. Rose & Trappe, Mycotaxon 10: 413. 1980.

Funneliformis kerguelensis (Dalpé & Strullu) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518454

= Glomus kerguelense Dalpé & Strullu, Mycotaxon 84: 53. 2002.

Funneliformis monosporus (Gerd. & Trappe) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518456

= Glomus monosporum Gerd. & Trappe, Mycol. Mem. 5: 41. 1974.

Funneliformis mosseae (T.H. Nicolson & Gerd.) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 13. 2010.

= Endogone mosseae T.H. Nicolson & Gerd., Mycologia 60: 314. 1968.

= Glomus mosseae (‘T.H. Nicolson & Gerd.) Gerd. & Trappe, Mycol. Mem. 5: 40. 1974.

Glomoid species reorganized ... 103

Woks

WV SWL2 omy

| r ae

50 um swt ~9oum ye

Fics 75-84. Funneliformis species (clade Aa1) generally form big (150-400 um) spores, singly and

(species-specifically) also in ‘small’ sporocarps with only a few to several spores per sporocarp;

subtending hyphae (sH) are concolorous with (or slightly lighter than) the spore wall without

introverted wall thickening at the spore base and are species-specifically cylindrical, funnel-

shaped, or (rarely) slightly constricted. sH wall thickening may extend over long distances (>

100-300 um). Spores with 1-5 wall layers (swL1-sw15); pore regularly closed by a conspicuous

septum (sp) that species-specifically arises from the structural layer, from an adherent innermost,

(semi-) flexible layer, or from both layers. 75. Fu. mosseae. 76. Fu. monosporus. 77. Fu. fragilistratus.

78. Fu. coronatus. 79. Fu. multiforus. 80. Fu. geosporus. 81. Fu. verruculosus. 82. Fu. kerguelensis.

83. Fu. caledonius. 84. Fu. caesaris.

Funneliformis multiforus (Tadych & Btaszk.) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518459

= Glomus multiforum Tadych & Blaszk., Mycologia 89: 805. 1997.

104 ... Oehl & al.

Funneliformis verruculosus (Btaszk.) C. Walker & A. Schiissler, The Glomeromycota

—a species list: 14. 2010.

= Glomus verruculosum Blaszk., Mycologia 89: 809. 1997.

Simiglomus Sieverd., G.A. Silva & Oehl, gen. nov. Fics. 85-86

MycoBAnk MB 518435 (Glomus group Aa2)

Sporae singulariter vel in congregatos apertos cum sporis pauciaribus efformatae; tunica

sporarum cum tunica hypharum coniuncta; porum sporarum apertum; hyphae cylindricae

vel infundiles pori sporarum aperti, pauciores septa in distanica ex tunicae hyphalis in

hypha; mycorrhizas vesicular-arbusculares formans caeruleas colorantes cum ‘trypan

blue’.

ErymMo.oey: from the Latin: simi(laris) = similar; glomus = cluster; referring to the

relation with Glomus, to which species of the new genus previously belonged.

KEY CHARACTERS: Spores formed singly or in very loose, small clusters. Spores

with a mono-to-multiple layered spore wall. Wall of the sH conspicuously

continuous and concolorous with the spore wall, or slightly lighter in color than

the spore wall. sH are funnel-shaped to cylindrical. Wall at spore attachment

not with introverted wall thickening. Pore at spore base open but several septa

in hyphae in some distance from spore base can separate spore contents from

mycelia contents. Walls of su thickened over very long distances from the spore

base (up to > 1000 um). Forming typical vesicular-arbuscular mycorrhiza, with

mycorrhizal structures that stain blue to dark blue in trypan blue.

TYPE SPECIES: Simiglomus hoi (S.M. Berch & Trappe) G.A. Silva et al.

Simiglomus hoi (S.M. Berch & Trappe) G.A. Silva, Oehl & Sieverd., comb. nov.

MycoBank MB 518461

= Glomus hoi S.M. Berch & Trappe, Mycologia 77: 654. 1985.

\\—NA& ae \

Fics 85-86. Simiglomus hoi (clade Aa2). Spores formed singly or in very loose small clusters; the

subtending hyphae (sH) are concolorous with (or slightly lighter than) the spore wall without

introverted wall thickening at the spore base and generally cylindrical to slightly funnel-shaped

or slightly constricted. sH wall thickening may extend over very long distances. The spore pore is

generally closed by conspicuously recurved septa at varying distances from the spore base. Often

several recurved septa (sp) visible that arise from the structural layer in the subtending hyphae.

Glomoid species reorganized ... 105

Fics 87-89. Septoglomus (clade Aa3) species form spores singly in soil or in very loose small

clusters; subtending hyphae (sH) are concolorous with (or slightly lighter than) the spore wall

without introverted wall thickening at the spore base and generally cylindrical to constricted.

sH wall thickening may extend over long distances. Spores with 1-3 wall layers (swL1-sw13); if

closed, pore often with one to several septa (sp) that species-specifically arise from the structural

layer in the subtending hyphae. 87. Se. constrictum. 88. Se. deserticola. 89. Se. xanthium.

Septoglomus Sieverd., G.A. Silva & Oehl, gen. nov. Fics. 87-89

MycoBank MB 518436 (Glomus group Aa3)

Sporae singulariter vel in congregatos apertos efformatae; tunica sporarum cum tunica

hypharum coniuncta; hyphae cylindricae vel infundiles pori sporarum occlusi septo.

Mycorrhizas vesicular-arbusculares formans caeruleas colorantes cum ‘trypan blue’.

EryMo_oey: from the Latin: septum = septum; glomus = cluster; referring to the relation

with Glomus, to which species of the new genus previously belonged.

KEY CHARACTERS: Spores formed singly or in very loose, small clusters. Spores

with a mono-to-multiple layered spore wall. Wall of the sH conspicuously

continuous and concolorous with the spore wall, or slightly lighter in color

than the spore wall. sH are cylindrical to constricted or slightly funnel shaped

at spore base. Pore at spore base or in some distance from spore based closed by

a septum. Forming typical vesicular-arbuscular mycorrhiza, with mycorrhizal

structures that stain blue to dark blue in trypan blue.

TYPE SPECIES: Septoglomus constrictum (Trappe) Sieverd. et al.

Septoglomus africanum (Btaszk. & Kovacs) Sieverd., G.A. Silva & Oehl, comb. nov.

MycoBank MB 519466

= Glomus africanum Blaszk. & Kovacs, Mycologia 102: 1452. 2010.

= Funneliformis africanus (Blaszk. & Kovacs) C. Walker & A.

Schiissler, The Glomeromycota - a species list: 13. 2010.

Septoglomus constrictum (Trappe) Sieverd., G.A. Silva & Oehl, comb. nov.

MycoBank MB 518462

= Glomus constrictum Trappe, Mycotaxon 6: 361. 1977.

= Funneliformis constrictus (Trappe) C. Walker & A. Schiissler,

The Glomeromycota — a species list: 14. 2010.

106 ... Oehl & al.

Septoglomus deserticola (Trappe, Bloss & J.A. Menge) G.A. Silva, Oehl & Sieverd.,

comb. nov.

MycoBank MB 518463

= Glomus deserticola Trappe, Bloss & J.A. Menge, Mycotaxon 20: 123. 1984.

Septoglomus xanthium (Blaszk., Blanke, Renker & Buscot) G.A. Silva, Oehl &

Sieverd., comb. nov.

MycoBank MB 518464

= Glomus xanthium Blaszk., Blanke, Renker & Buscot, Mycotaxon 90: 459. 2004.

= Funneliformis xanthius (Blaszk.) C. Walker & A. Schiissler,

The Glomeromycota - a species list: 14. 2010.

Claroideoglomeraceae C. Walker & A. Schiissler, emend. Oehl, G.A. Silva & Sieverd.

(Glomus group B)

EMENDED DESCRIPTION: Spores formed singly or in clusters with low spore

numbers, extremely rarely in sporocarps; in soils or rarely in roots; sH hyaline

to white, rarely subhyaline, often conspicuously bill-shaped. Spores with 1-4

wall layers (swL1-sw14); pore closure at spore base often with a septum that

species-specifically may arise from the structural layer, from an adherent inner,

(semi-)flexible layer, or from both layers.

TYPE GENUS: Claroideoglomus C. Walker & A. Schiissler

OTHER GENUS: Viscospora Sieverd. et al.

Claroideoglomus C. Walker & A. Schiissler, emend. Oehl, G.A. Silva & Sieverd.

(Glomus group B1) FIGs. 90-98

EMENDED DESCRIPTION: Spores generally formed singly in soil or rarely in

roots; sH hyaline to white, rarely subhyaline, conspicuously bill-shaped. Spores

with 1-4 wall layers (swLl-sw14); pore closure at spore base often with a

septum that species-specifically may arise from the structural layer, from an

adherent innermost, (semi-)flexible layer, or from both layers.

TYPE SPECIES: Claroideoglomus claroideum (N.C. Schenck & G.S. Sm.)

C. Walker & A. Schiissler

Claroideoglomus candidum (Furrazola, Kaonongbua & Bever), Oehl, G.A. Silva &

Sieverd., comb. nov.

MycoBANnkK MB 519459

= Glomus candidum Furrazola, Kaonongbua & Bever, Mycotaxon 113: 103. 2010.

Claroideoglomus claroideum (N.C. Schenck & G.S. Sm.) C. Walker & A. Schiissler,

The Glomeromycota - a species list: 21. 2010.

= Glomus claroideum N.C. Schenck & G.S. Sm., Mycologia 74:

84. 1982 (emend. Walker & Vestberg 1998).

Claroideoglomus drummondii (Blaszk. & Renker) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 22. 2010.

= Glomus drummondii Blaszk. & Renker, Mycol. Res. 110: 559. 2006.

Glomoid species reorganized ... 107

Fics 90-98. Claroideoglomus (clade B1) species generally form spores singly in soils or (rarely) in

roots; rarely also in very loose spore clusters; subtending hyphae (sH) conspicuously bill-shaped but

hyaline to white, rarely subhyaline. Spores with 1-4 wall layers (swL1-sw14); pore closure at spore

base generally with a septum (sp) that species-specifically may arise from structural wall layer,

an adherent inner (semi-)flexible layer, or both layers. 90-91. Cl. etunicatum. 92. Cl. lamellosum.

93-94. CI. claroideum. 95-96. Cl. luteum. 97. Cl. drummondii. 98. Cl. walkeri.

Claroideoglomus etunicatum (W.N. Becker & Gerd.) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 22. 2010.

= Glomus etunicatum W.N. Becker & Gerd., Mycotaxon 6: 29. 1977.

Claroideoglomus lamellosum (Dalpé, Koske & Tews) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 22. 2010.

= Glomus lamellosum Dalpé, Koske & Tews, Mycotaxon 43: 289. 1992.

Claroideoglomus luteum (L.J. Kenn., J.C. Stutz & J.B. Morton) C. Walker & A.

Schiissler, The Glomeromycota — a species list: 22. 2010.

= Glomus luteum L.J. Kenn., J.C. Stutz & J.B. Morton, Mycologia 91: 1090. 1999.

Claroideoglomus walkeri (Btaszk. & Renker) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 22. 2010.

= Glomus walkeri Blaszk. & Renker. Mycol. Res. 110: 563. 2006.

108 ... Oehl & al.

Viscospora Sieverd., Oehl & G.A. Silva, gen. nov. (Glomus group B2)

MycoBank MB 518439

Sporae in congregatos apertos efformatae; tunica sporarum cum tunica hypharum

coniuncta; hyphae hyalinae vel albae; stratum exterior hyphae et sporarum cum materia

mucilagina; porum sporarum non occlusum vel occlusum hyphae crassae. Mycorrhizas

vesicular-arbusculares formans caeruleas colorantes cum ‘trypan blue’.

Erymo.oey: from the Latin: viscosus = sticky; spora = spore; referring to the adhesive

nature of the spore surface of the type species of the genus.

KEY CHARACTERS: Spores generally formed in loose clusters; sH hyaline

to white, rarely subhyaline, often thick-walled. Spores with 1-4 wall layers

(SwL1-swL4); outer wall layer exuding a mucigel-like substance. Pore closure

at spore base often open, or semi-closed by wall thickening.

TYPE SPECIES: Viscospora viscosa (‘T.H Nicolson) Sieverd. et al.

Viscospora viscosa (T.H Nicolson) Sieverd., Oehl & G.A. Silva, comb. nov.

MycoBank MB 518471

= Glomus viscosum T.H. Nicolson, Mycol. Res. 99: 1502. 1995.

Diversisporaceae C. Walker & A. Schiissler, emend. Oehl, G.A. Silva & Sieverd.

(Glomus group C)

EMENDED DESCRIPTION: Spore formation either diversisporoid or otosporoid.

Diversisporoid spores either formed singularly, in clusters or in large

sporocarps with high spores numbers, not organized; in pigmented spores sH

conspicuously change color, becoming hyaline to white behind the septum,

i.e. immediately or in a minimal distance to this septum; sH generally straight,

cylindrical, sometimes species-specifically constricted or inflated. Spores

with 1-3 wall layers (swL1l-sw13); pore closure often with a septum that may

species-specifically arise from innermost wall lamina or overlaying laminate

layer, or from both; pore of hyphal attachment rarely (species-specifically)

open. Otosporoid spores with two multiple-layered walls, spore attachment

generally closed by a septum.

TYPE GENUS: Diversispora C. Walker & A. Schiissler

OTHER GENERA: Ofospora Oehl et al., Redeckera C. Walker & A. Schiissler

Diversispora C. Walker & A. Schiissler, emend. G.A. Silva, Oehl & Sieverd.

(Glomus group Ca) FIGS. 99-107

EMENDED DESCRIPTION: Spores either formed singularly, in small open clusters

or in large multi-spored clusters or sporocarps where spores are not organized.

In pigmented spores sH conspicuously change color, becoming hyaline to

white behind the septum; sH generally straight, cylindrical, sometimes species-

specifically constricted, often hyphal attachment looks like inserted in spore

wall. Spores with 1-3 wall layers (swLl-sw13); pore closure often with a

Glomoid species reorganized ... 109

Fics 99-107. Diversispora (clade Ca) species form spores singly or in unorganized clusters or

sporocarps; in pigmented spores subtending hyphae (sH) with a conspicuous color change

distant to the first septum at or near the spore base; sH generally cylindrical, species-specifically

sometimes constricted. Spores species-specifically with 1-3 wall layers (swL1-sw13); pore closure

at spore base often with a septum (sp) that species-specifically may arise from a structural wall

layer, an adherent inner (semi-) flexible layer, or both layers; rarely (species-specifically) pore open.

99. Di. spurca. 100. Di. eburnea. 101. Di. insculpta. 102. Di. arenaria. 103-104. Di. aurantia.

105. Di. epigaea. 106. Di. tenera. 107. Di. pustulata.

septum that may species-specifically arise from the innermost wall layer; rarely

(species-specifically) pore of attachment open.

TYPE SPECIES: Diversispora spurca (C.M. Peiff. et al.) C. Walker & A. Schiissler

Diversispora arenaria (Blaszk., Tadych & Madej) Oehl, G.A. Silva & Sieverd.,

comb. nov.

MycoBank MB 518472

= Glomus arenarium Bliaszk., Tadych & Madej, Acta Soc. Bot. Pol. 70: 97. 2001.

Diversispora aurantia (Blaszk., Blanke, Renker & Buscot) C. Walker & A. Schiissler,

—a species list: 43. 2010.

= Glomus aurantium Blaszk., Blanke, Renker & Buscot, Mycotaxon 90: 450. 2004.

110... Oehl & al.

Diversispora celata C. Walker, Gamper & A. Schuessler, New Phytol. 182: 497. 2009.

Diversispora eburnea (L.J. Kenn., J.C. Stutz & J.B. Morton) C. Walker & A. Schiissler,

The Glomeromycota - a species list: 43. 2010.

= Glomus eburneum L.]J. Kenn., J.C. Stutz & J.B. Morton, Mycologia 91: 1084. 1999.

Diversispora epigaea (B.A. Daniels & Trappe) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 43. 2010.

= Glomus epigaeum B.A. Daniels & Trappe, Can. J. Bot. 57: 540. 1979.

Diversispora gibbosa (Btaszk.) Btaszk. & Kovacs, comb. nov.

MycoBank MB 518475

= Glomus gibbosum Btaszk., Mycologia 89: 339. 1997.

Diversispora insculpta (Btaszk.) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 519546

= Glomus insculptum Blaszk., Mycotaxon 89: 227. 2004.

Diversispora przelewicensis (Btaszk.) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBANK MB 518477

= Glomus przelewicense Blaszk., Bull. Pol. Acad. Sci., Biol. Sci. 36: 272. 1988.

Diversispora pustulata (Koske, Friese, C. Walker & Dalpé) Oehl, G.A. Silva &

Sieverd., comb. nov.

MycoBank MB 518478

= Glomus pustulatum Koske, Friese, C. Walker & Dalpé, Mycotaxon 26: 143. 1986.

Diversispora spurca (C.M. Pfeiff., C. Walker & Bloss) C. Walker & A. Schiissler,

Mycol. Res. 108: 982. 2004.

= Glomus spurcum C.M. Pfeiff., C. Walker & Bloss, Mycotaxon 59: 374. 1996.

Diversispora tenera (P.A. Tandy) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518479

= Glomus tenerum P.A. Tandy, Austral. J. Bot. 23: 864. 1975.

Diversispora trimurales (Koske & Halvorson) C. Walker & A. Schiissler, The

Glomeromycota — a species list: 43. 2010.

= Glomus trimurales Koske & Halvorson, Mycologia 81: 930. 1990.

Diversispora versiformis (P. Karst.) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518481

= Endogone versiformis P. Karst., Hedwigia 23: 39. 1884.

= Glomus versiforme (P. Karst.) S.M. Berch, Can. J. Bot. 61: 2614. 1983.

Redeckera C. Walker & A. Schiissler, emend. Oehl, G.A. Silva & Sieverd.

(Glomus group Cb) Fics. 108-114

EMENDED DESCRIPTION: Spore formation disorganized in large and compact

sporocarps, containing hundreds to thousands of spores per sporocarp; spores

Glomoid species reorganized ... 111

Fics 108-114. Redeckera (clade Cb) with spore formation in unorganized but large and compact

sporocarps (usually with hundreds to thousands spores per sporocarp); spores species-specifically

with 2(-3) wall layers (swL1-sw13); subtending hyphae (sH) generally broad at spore base and with

a conspicuous thick, broad septum (sp) arising from the inner lamina of the generally bi-laminated

spore wall layer (sw12); structural swL2 generally extend over very short (2-10 um) distances

into sH; SWL1 fragile and usually inflating where sw12 becomes invisible in the su. 108. Re. fulva.

109. Re. megalocarpa. 110. Re. pulvinata. 111-112. Re. canadensis. 113-114. Re. fragilis.

with 2 to rarely 3 wall layers (SwL1-sw13); sH generally broad at spore base and

with a conspicuous, thick and broad septum that arises from the inner lamina

(sw12) of the generally bi-laminated, structural wall layer; structural swL2

generally continue over very short distances (2-10 um) into sH; sw1l fragile,

usually inflating in a short distance to the spore base where swL2 becomes

invisible in the su.

GENDER: Redeckera is a Latin feminine noun, formed according to ICBN

[Vienna Code] Recommendation 60B.1(b). Because the feminine gender

is clearly apparent (and reinforced by the implication of Recommendation

20A.1(i)), Article 62.3 does not apply, and the authors did not have the

prerogative to choose any other gender. We therefore correct all adjectival

epithets to feminine.

TYPE SPECIES: Redeckera megalocarpa (D. Redecker) C. Walker & A. Schiissler

Redeckera avelingiae (R.C. Sinclair) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 518482

= Glomus avelingiae R.C. Sinclair, Mycotaxon 74: 338. 2000.

Redeckera canadensis (Thaxt.) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBAnk MB 518483

= Endogone canadensis Thaxt., Proc. Am. Acad. Arts Sci. 57: 317. 1922.

= Glomus canadense (Thaxt.) Trappe & Gerd., Mycol. Mem. 5: 59. 1974.

112 ... Oehl & al.

Redeckera fragilis (Berk. & Broome) Oehl, G.A. Silva & Sieverd., comb. nov.

MycoBaAnk MB 518484

= Paurocotylis fragilis Berk. & Broome, J. Linn. Soc. Bot. 14: 137. 1873.

= Glomus fragile (Berk. & Broome) Trappe & Gerd., Mycol. Mem. 5: 59. 1974.

Redeckera fulva (Berk. & Broome) C. Walker & A. Schiissler, The Glomeromycota -

a species list: 44. 2010.

= Paurocotylis fulva Berk. & Broome, J. Linn. Soc. Bot. 14: 137. 1873.

= Glomus fulvum (Berk. & Broome) Trappe & Gerd., Mycol. Mem. 5: 59. 1974.

Redeckera megalocarpa (D. Redecker) C. Walker & A. Schiissler, The

Glomeromycota: 44. 2010.

= Glomus megalocarpum D. Redecker, Mycol. Progress 6: 38. 2007.

Redeckera pulvinata (Henn.) C. Walker & A. Schiissler, The Glomeromycota: 44. 2010.

= Endogone pulvinata Henn., Hedwigia 36: 212. 1897.

= Glomus pulvinatum (Henn.) Trappe & Gerd., Mycol. Mem. 5: 59. 1974.

Otospora Oehl, Palenz. & N. Ferrol

TYPE SPECIES: Otospora bareae Palenz. et al.

Otospora bareae Palenz., N. Ferrol & Oehl, Mycologia 100: 298. 2008.

Paraglomeraceae J.B. Morton & D. Redecker

Paraglomus J.B. Morton & D. Redecker

TYPE SPECIES: Paraglomus occultum (C. Walker) J.B. Morton & D. Redecker

Paraglomus albidum (C. Walker & L.H. Rhodes) Oehl, G.A. Silva & Sieverd.,

comb. nov.

MycoBank MB 518488

= Glomus albidum C. Walker & L.H. Rhodes, Mycotaxon 12: 509. 1981.

Paraglomus brasilianum (Spain & J. Miranda) J.B. Morton & D. Redecker,

Mycologia 93: 190. 2001.

= Glomus brasilianum Spain & J. Miranda, Mycotaxon 60: 139. 1996.

Paraglomus laccatum (Btaszk.) Renker, Blaszk. & Buscot, Nova Hedwigia 84: 400.

2007.

= Glomus laccatum Btaszk., Bull. Pol. Acad. Sci., Biol. Sci. 36: 271. 1988.

Paraglomus lacteum (S.L. Rose & Trappe) Oehl, G.A. Silva & Sieverd. comb. nov.

MycoBank MB 518489

= Glomus lacteum S.L. Rose & Trappe, Mycotaxon 10: 415. 1980.

Paraglomus occultum (C. Walker) J.B. Morton & D. Redecker, Mycol. Res. 93:190.

2001.

= Glomus occultum C. Walker, Mycotaxon 15: 50. 1982.

Glomoid species reorganized ... 113

Taxonomic problems

We were unsure whether we had attributed Gl. dolichosporum and

Gl. citricola to the correct genus, as the available descriptions were either short

or in Chinese, images were missing or relatively poor, or the type specimen

was not accessible. Moreover, in several species descriptions (some even very

recent), the focus on the spore base and su features was minimal to absent. For

such cases, we decided to retain these species within Glomus, pending future

revision. Placement of Gl. pubescens was similarly difficult: Already transferred

to four different genera other than its basionym genus, this particular species

may not even represent an AM fungus (e.g. Trappe, pers. comm.). Of all clades

discussed here and fungal genera known, this species appears to fit best in

Glomus group Abl.

During our spore analyses, it was apparent that several genetic groups

(clades) possess species with spores so similar that either the species are

difficult to differentiate morphologically or may be conspecific. Examples

include Aa3—Se. deserticola, Se. xanthium; Ab2—Gl. clarum, Gl. manihotis,

Gl. zaozhuangianum; and Ca—Di. versiformis, Di. przelewicensis, Di. epigaea.

As it was beyond the scope of this study to clarify all vagaries and possible

taxonomic uncertainties at the species level, we have placed them into genera

based on morphological characteristics, especially as many type specimens

were unavailable (see TABLE 1). In some cases confusion may have resulted

from misidentification or incorrect re-naming prior to genetic analysis. For

example, the GI. clarum isolate BR147B (supplied by Bioplanta) that grouped

with Gl. manihotis in the initial phylogenetic study by Schifler et al. (2001)

was an ex-type of Gl. manihotis when sent in 1987 to Bioplanta (Sieverding

pers. com.) but which some time later was relabeled as Gl. clarum by someone

else. Not surprisingly, this particular ‘Gl. clarum’ isolate grouped with other

Gl. manihotis ex-type material in the study by Schii®ler et al. (2001).

Consequently, genetic information provided for some species as well as the

conclusion that other species are conspecific may be based on erroneous

information.

Until recently, both germination and germination structures were regarded

as important features to distinguish between AM fungal genera and families

(e.g., Pacisporaceae vs. Glomeraceae by Oehl & Sieverding 2004, Walker &

Schiifler 2004; among Gigasporaceae, Scutellosporaceae, Racocetraceae, and

Dentiscutataceae by Oehl et al. 2008, 2011a; and Archaeospora vs. Ambispora by

Spain et al. 2006). There are clear indications that Paraglomus species germinate

directly through the spore wall as observed for Pa. occultum, Pa. brasilianum

and Pa. lacteum (Spain & Miranda 1996, Rose & Trappe 1980, Oehl pers. obs.)

in contrast to all other species forming single-walled glomoid or diversisporoid

spores where a germ tube emerges apparently through the su (e.g. Glomus,

114... Oehl & al.

Diversispora). There are uncertainties surrounding germ tube formation in some

Glomus spp., such as Gl. pallidum, which Hall (1977) reported as germinating

directly through the spore wall, although his description and photos suggest

instead either intercalary spore formation or two hyphal attachments on the

same spore. Intercalary spore formation (= two attachments) is a frequent

feature for many Glomus sporocarpic (Ab) species.

Molecular difficulties

The major problem encountered during the current study revolved around

the limited molecular database, especially for species of the former Glomus

groups Ab1, Ab3, and Cb. Moreover, only a limited number of Glomeromycota

species in the public databases have been sequenced for all currently available

markers (LSU rDNA, SSU rDNA, ITS region, B-tubulin). Some individual

clades were supported in just one dataset. Another major difficulty was that

either many cultures of glomoid or diversisporoid spore-forming species are

lost or yet to be cultured successfully in single species cultures or the species

were not culturable. This renders analyses difficult or impossible. During the

study, DNA extraction was attempted from several species using field spores or

sporocarps but they largely failed (e.g., Gl. invermaium, Gl. taiwanense). These

obstacles have left the database incomplete and slowed progress in complete

molecular AM fungal identification.

Discussion

In this paper, we have revised the taxonomic organization of AM fungal

species with glomoid spore formation, based largely on newly available molecular

data retrieved from the literature and public databases. This organization

is also based upon morphological spore characteristics, which were either

already well established or newly established during this study. Phylogenetic

trees support all genera proposed in this paper and some genera proposed by

Walker & Schiifer (2010), with bootstrap support > 80% (except Septoglomus

in the beta-tubulin phylogeny) in at least one analysis (NJ, ML, Bayesian or

MP). However, Rhizophagus (Ab2) and Sclerocystis (Ab3) were not supported

by our phylogenetic analyses and thus, the resurrection of these genera Walker

& SchiifSer (2010) cannot be accepted. The genera proposed or confirmed in

the present study are shown as clusters in the trees: Glomus (Ab1, Ab2, Ab3),

Funneliformis (Aa1), Simiglomus (Aa2), Septoglomus (Aa3), Claroideoglomus

(B1), Viscospora (B2), Diversispora (Ca) and Redeckera (Cb). In TABLE 3, we

summarize the new taxonomic organization of AM fungal species within the

Glomeromycota.

There is no doubt that glomoid and diversisporoid spores in Glomeraceae,

Claroideoglomeraceae and Diversisporaceae can clearly be differentiated by

molecular phylogeny (Fics. 1-4), as shown earlier (Schiifler et al. 2001, Silva

Glomoid species reorganized ...

TABLE 3. New classification of the Glomeromycota

CLASS ORDER : FAMILY : GENUS

Glomeromycetes Glomerales Glomeraceae Glomus

Funneliformis

Simiglomus

Septoglomus

Claroideoglomeraceae Claroideoglomus

i Viscospora

Diversisporales Diversisporaceae Diversispora

i : Redeckera

: Otospora

Entrophosporaceae Entrophospora

Acaulosporaceae Acaulospora

Kuklospora

Pacisporaceae Pacispora

Gigasporales Gigasporaceae Gigaspora

Scutellosporaceae Scutellospora

: Orbispora

Racocetraceae Racocetra

Cetraspora

Dentiscutataceae Dentiscutata

Fuscutata

Quatunica

Archaeosporomycetes Archaeosporales Archaeosporaceae Archaeospora

i i Intraspora

Ambisporaceae Ambispora

Geosiphonaceae Geosiphon

Paraglomeromycetes Paraglomerales Paraglomeraceae Paraglomus

LA'S

The taxa Archaeosporomycetes, Paraglomeromycetes, Gigasporales and Orbispora are presented in this

volume (Oehl et al. 2011a, 2011b).

et al. 2006, Stockinger et al. 2010). Glomeraceae and Claroideoglomeraceae

differ morphologically in the hyphal attachments at the spore base and in

spore clustering/aggregation or sporococarp formation. The phylogenetic trees

also show Glomeraceae and Claroideoglomeraceae as separate clades with high

bootstrap values in SSU, LSU, and beta-tubulin analyses.

Differences in hyphal connections at the spore base are also basic

morphological indicators for other related but clearly genetically separate

families of the Glomeromycota (e.g., Acaulosporaceae, Pacisporaceae,

Scutellosporaceae). It has been argued earlier that unequivocal assignment

of glomoid and diversisporoid spore-forming species to the correct major

phylogenetic clade through morphological spore analyses alone would be

116... Oehl & al.

impossible (e.g. Schiifler et al. 2001, Morton & Redecker 2001, Walker et

al. 2007), primarily because glomoid and diversisporoid spores have but few

differentiating morphological characters (which may yet hold true). Features

other than hyphal connections (such as spore size and color) overlap between

many glomoid and diversisporoid species, and intraspecific variation might be

considerable. Spore aggregation and cluster/sporocarp formation in glomoid

and diversisporoid species are not well investigated and have thus far received

only limited attention. However, sporocarp formation and organization may

be stable characteristics, as found in the former genus Sclerocycstis, which at

some point may need to be raised again to genus level within the Glomeraceae.

Other species of Glomeraceae are also always formed in sporocarps (e.g.

Gl. glomerulatum, Gl. formosanum) but as yet there are insufficient molecular

data to support their placement within their own genus. Other glomoid spores

form in clusters or sporocarps occurring in relatively small to sometimes huge

spore numbers (Gl. macrocarpum, Gl. microcarpum, Gl. aureum) as well as

form singly in soil or in roots, so that spore aggregation alone cannot be used

to diagnose the taxonomic order. We acknowledge that the large genus Glomus

currently has morphologically heterogeneous spores, and recent findings

by Blaszkowski et al. (2009a, b) show that several clades with small-spored

species also belong to this group. To date, there are insufficient sequence sets

available for more detailed analyses, although new genera could result from

such analyses. This applies also for Gl. intraradices versus Gl. macrocarpum

and Gl. microcarpum and Glomus Ab1 species in general. Recent research

has also shown that genetic information generated years ago may actually be

derived from another species, Gl. irregulare (Stockinger et al. 2009). Based on

morphology alone, species such as Gl. intraradices should be excluded from

Glomus.

Within Diversispora (respective former Glomus group C, our clade Ca)

and Redeckera (our clade Cb), there has been no previous attempt to explain

how these species differ morphologically from the former Glomeraceae

groups A and B (Schiifler et al. 2001, Walker & Schiifler 2004). While

morphological differentiation between Diversispora and Redeckera is possible,

the morphology of Diversispora spores that differentiates them from spores of

some Claroideoglomus spp. may be difficult to recognize, even for taxonomic

specialists.

To conclude, our paper reports on the congruence of morphological groups

(genera and family) with molecular phylogeny that has shown the need for a

substantial reorganization of taxa in Glomeromycota.

Unsolved obstacles in the Glomeraceae

With approximately 70 species, the revised Glomus remains morphologically

heterogeneous and the largest genus in the Glomeromycota. Species such as

Glomoid species reorganized ... 117

Gl. intraradices, Gl. aggregatum, and Gl. proliferum are atypical in the genus

regarding spore base closure by wall thickening and/or septum. These are

normally quite stable features and not highly variable within species: a septum

not readily seen at the spore base is either species specific or indicates the spore

development stage. Mature spores have septa more frequently formed by the

innermost lamina of the inner spore wall layer. Morphologically speaking,

species like Gl. intraradices, Gl. aggregatum, and Gl. proliferum should be

included in a separate genus, but the absence of sufficient genetic information

(in particular of the other Glomus spp.) does not support this hypothesis.

Additionally, we include several small-spored, cluster-forming species within

the revised Glomus (e.g. Gl. perpusillum, Gl. indicum, G. iranicum) whose spores

quickly decompose, suggesting that some earlier may have erroneously been

designated ‘non-sporulating AM fungi’ (Blaszkowski et al. 2009b). Verification

is still needed to establish whether there are indeed species of ‘non-sporulating’

fungi within the Glomus clade.

Acknowledgments

We acknowledge the help of Joey Spatafora and Richard Halse for giving the

possibility to visit the OSU herbarium in Corvallis (Oregon) in 2002 and 2005. We

are also thankful to OSC (Oregon State University herbarium) and to FH (Farlow

Herbarium, at Harvard University) for providing us with type and non-type material

of several Glomus species. We especially thank Joseph B. Morton and William Wheeler

(West Virginia State University; Morgantown, USA) for giving F. Oehl the opportunity

to investigate, during his visit in 2002, many Glomus isolates maintained at INVAM.

James M. Trappe (USDA-Corvallis, Oregon, USA), Janusz Blaszkowski (Department of

Plant Pathology, Academy of Agriculture, Szczecin, Poland), Peter McGee (University

of Sydney, Australia), Manuela Giovannetti (Universita di Pisa, Italy), Claudia Castillo

(Universidad Catélica de Temuco, Chile), provided type or non type specimen of several

Glomus species. Fabien Hountondji, Atti Tchabi and Louis Lawouin (International

Institute of Tropical Agriculture, Benin) provided soil samples and specimen from

their country. EA. Souza’s great help in discussing specific parts of the paper, and in

calculating clades is highly appreciated. We acknowledge the valuable comments and

revisions of several experts on the manuscript. We appreciate the corrections by Shaun

Pennycook, Nomenclatural Editor, and suggestions by Lorelei L. Norvell, Editor-in-

Chief. This study was supported by the Swiss National Science Foundation (SNSF, Project

315230_130764/1) and by the Fundagao de Amparo a Ciéncia e Tecnologia do Estado

de Pernambuco (FACEPE) which provided a grant to F. Oehl as ‘visiting researcher.

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MY COTAXON

Volume 116, pp. 121-124 April-June 2011

DOI: 10.5248/116.121

Ileodictyon gracile, new to Italy

ALESSANDRO SAITTA, RICCARDO COMPAGNO, ALFONSO LA ROSA,

DANILO LOPEZ & GIUSEPPE VENTURELLA*

Dipartimento di Biologia vegetale e Biodiversita, Universita di Palermo,

Via Archirafi 38, I-90123 Palermo, Italy

* CORRESPONDENCE TO: gvent@unipa.it

ABSTRACT — The first record of Ileodictyon gracile from Italy is reported, accompanied by

notes on the taxonomy, ecology, and distribution of this rare gasteromycete.

Key worps — Phallaceae, rare Mediterranean fungi, gasteromycetoid fungi

Introduction

Gasteromycetoid fungi encompass terrestrial, epigeous and hypogeous and

saprobic and ectomycorrhizal species. They are cosmopolitan but are more

frequently found in warm, dry habitats (Kirk et al. 2008). Modern accounts of

this interesting but taxonomically complex group is particularly limited in Italy

where after the monograph by Petri (1909), only Sarasini (2005) has written on

epigeous gasteromycetes in the country. In the absence of funding for studies

of gasteromycetoid fungi, reports of undescribed species or new records from

Italy are usually restricted to an occasional observation and/or generalized

papers devoted to fungal diversity. This is the first record of Ileodictyon gracile

in Italy, recently collected by us on the island of Pantelleria, Egadi Archipelago

in Trapani province in south-eastern Sicily.

Materials & methods

Basidiomata were identified while fresh and microscopic features were observed

in H,O using a Leica microscope DMLB; spore measurements were based on 50

observations. Nomenclature follows Index Fungorum (http://www.indexfungorum. org/

Names/Names.asp). The description is based both on personal observation, augmented

by characters cited in Sarasini (2005) and Bougher & Syme (1998). The collection is

curated in the fungal dried reference collection of the Herbarium Mediterraneum

Panormitanum, Palermo (PAL).

122 ... Saitta & al.

Taxonomy

Ileodictyon gracile Berk., London J. Bot. 4: 69. 1845. FIG. 1

Unripe basidiomata, sessile, irregularly rupturing at apex, egg-shaped, globose,

subglobose or sphaerical, consistency jelly-like, 1.5 cm in diam. Peridium white,

with long white unbranched rhizomorphs emanating from various points.

Ripe basidiomata, sessile, with receptacle (lattice) up to 80-200 mm broad,

white or cream, sphaerical or broadly ellipsoidal, sessile in a ruptured, white,

membranous volva. Lattice of isodiametric-polygonal interspaces and flattened,

firm struts that are grooved but not broadened at junctions and have glebal

slime adhering to inside surface. Peridium in two layers with hyaline hyphae,

3.5-6.5 wide: exoperidium, white, mat, thin and membranous; endoperidium

thick and gelatinous. Gleba olive-brown. Basal mycelium of white mycelia

strands. Odour foetid or like sour milk. Spores hyaline, (4—)4.5-5.5(-6) x 1.8-

2.4 um, narrowly ellipsoidal, smooth, thin-walled. Basidia clavate, 15-25 x 4-6

um. Cystidia absent.

SPECIMEN EXAMINED: ITALY. SIciLy: province of Trapani, Egadi archipelago, Island of

Pantelleria, Bugeber, 200 m, wood of Quercus ilex L. (Fagaceae) with shrubs of Cistus

salviifolius L. (Cistaceae) in the ground cover, 2 Jan 2009, coll. R. Compagno, A. La Rosa

& D. Lopez (PAL 00001/2011).

COMMENTS — Hosaka et al. (2006) recently reclassified gomphoid-phalloid

fungi based on molecular phylogenetic characters, and Kirk et al. (2008)

included the genus Ileodictyon Tul. & C. Tul. (Phallaceae Corda) in the Phallales

E. Fisch. Berkeley (1845) separated the genus Ileodictyon from Clathrus

P. Micheli ex L. principally based on possession of tubular (not cellular) ribs

and a different volval morphology. Dring & Rose (1977) found that the virtually

sphaeroidal symmetry further differentiated I. gracile and I. cibarium Tul. &

C. Tul. from Clathrus s. str. Ileodictyon gracile is separated from I. cibarium

based on morphological differences of the receptacle, intersections, and creases

(Dring 1980).

Discussion

The so-called lattice or basket fungus is a saprotrophic gasteromycete usually

collected on ground, commonly on the edge of tracks in forests and urban areas

(Bougher & Syme 1998). Ileodictyon gracile has been frequently reported from

Australia (Berkeley & Broome 1879, Cunningham 1944, Bougher & Syme

1998); May et al. 2003) also provide color and black and white illustrations

of basidiomata and microscopic characters. The species also occurs in New

Zeland, Samoa, Japan, Korea, and Africa (Calonge & Romero Zarco 1989, Dring

& Rose 1977, Ka et al. 2004, Malencon & Bertault 1970, Pennycook & Galloway

2004). European distribution is restricted to Spain (Calonge & Romero Zarco

Ileodictyon gracile, new to Italy ... 123

Fic. 1. Basidiomata of Ileodictyon gracile.

1989), Canary Islands (Chavez Barreto & Escobio Garcia 2009), and Portugal

(Pinto-Lopes 1942, 1944; Louro et al. 2009).

We found that Ileodictyon gracile shares similar habitats and phenology

(winter) in Portugal, Spain, and Sicily. In Portugal it was collected under Cistus

ladanifer L. in a Quercus rotundifolia Lam. forest, in Spain it was recorded

in Pinus pinea L. woods with Cistus salviifolius as ground cover over sandy

soil (Calonge & Romero Zarco 1989), and in Sicily in a Q. ilex wood with

C. salviifolius shrubs. Other reported habitats include Eucalyptus reforestations

(Chavez Barreto & Escobio Garcia 2009), open ground near forest trails and

urban areas (Bougher & Syme 1998), and on sandy soil in glades, woods, parks,

and gardens (Sarasini 2005).

Our report represents the southernmost limit of the European distribution

of I. gracile.

Acknowledgements

The authors wish to thank Dr Georgios I. Zervakis (Greece) and Dr Alfredo Vizzini

(Italy) for critically reviewing the manuscript.

124 ... Saitta & al.

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catalogo micoldgico espanol. Boletin Sociedad Micoldgica de Madrid 13: 171-174.

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Fondazione Centro Studi Micologici dell’AMB. 406 p.

MY COTAXON

Volume 116, pp. 125-131 April-June 2011

DOI: 10.5248/116.125

Observations on two rarely collected species of Russula

P. MANIMOHAN & K. P. DEEPNA LATHA

Department of Botany, University of Calicut, Kerala, 673 635, India

CORRESPONDENCE TO: pmanimohan@gmail.com & deepnalathakp@gmail.com

ABSTRACT — Russula periglypta and R. purpureonigra, originally reported from Sri

Lanka several decades back, were collected from Kerala State, India. Full descriptions and

photo-illustrations of these rediscovered species are provided along with some taxonomic

observations.

Key worps — Basidiomycota, Russulales, Russulaceae, taxonomy, floristics

Introduction

During our investigations on the agaric mycota of Kerala State, India, we

came across two rarely collected species of the genus Russula (Basidiomycota,

Russulales, Russulaceae), viz. R. periglypta and R. purpureonigra, originally

reported from Sri Lanka almost a century or more back. As we had the

opportunity to collect and study them on several occasions, we thought it

prudent to provide full descriptions and photo-illustrations of these rediscovered

species along with our taxonomic observations.

Materials & methods

Conventional morphology-based taxonomic methods were employed for this study.

Chemical spot tests were made on stipe surface with ferrous sulfate, aniline and aqueous

phenol. Microscopic observations were made on material stained with 1% aqueous

solutions of phloxine and Congo red and mounted in 3% aqueous KOH. Melzer’s

reagent was used to observe whether the spores and tissues were amyloid. Reactions

of the macrocystidial contents to sulfovanillin were noted. For evaluation of the range

of spore-size, 20 basidiospores each from one specimen of each collection cited were

measured. Colour codes used in the descriptions are from Kornerup & Wanscher

(1978). The examined collections cited are deposited at the Kew (Mycology) Herbarium

and the Kew accession numbers (e.g., K(M) 165811) are indicated. The infrageneric

classification followed is that of Sarnari (1998).

126 ... Manimohan & Deepna Latha

Taxonomy

Russula periglypta Berk. & Broome, Journ. Linn. Soc., Bot. 11: 566 (1871). -Fic.1

ORIGINAL DIAGNOsIS: “Pileus 2% inches across, hemispherical, viscid, margin regularly

and strongly sulcate; stem nearly 3 inches high, % thick in the middle, attenuated at the

base, somewhat swollen in the centre, solid; gills regular, arched, %4 inch wide, acute

behind, reaching to the top of stem; interstices reticulate. Very regular in form. Spores

globose, echinulate, .00025 in diameter.”

BASIDIOMATA medium-sized to somewhat large. PILEUS 25-90 mm diam.,

initially convexo-hemispherical with a slightly depressed centre, then applanate

with a depressed centre and finally almost infundibuliform; surface initially

pale yellow (4A2, 4A3), later greyish brown (5B2, 5D2, 5E2), slightly viscid,

initially smooth and pellucid-striate towards margin, becoming tuberculate-

striate and sulcate up to half the way from margin with age; margin initially

slightly incurved and entire, becoming straight or almost upturned and fissile.

LAMELLAE adnexed to adnate, whitish or marble white (5A2), close to almost

crowded, up to 10 mm broad, rarely furcate, with occasional lamellulae, with

abundant interveining; edge entire, concolorous with the sides. STIPE 25-90 x

6-22 mm, central, terete, mostly tapering towards base, occasionally tapering

towards both ends, stuffed or hollow; surface whitish or marble white (5A2),

glabrous. OpouR mild, spicy. SPORE-PRINT whitish. CHEMICAL SPOT TESTS

on stipe surface: FeSO,: no reaction in the beginning, slowly turning pinkish

brown; aniline: no reaction; aqueous phenol: chocolate brown.

BASIDIOSPORES 5-8 x 4-7.5 um, Q = 1-1.5, QM = 1.2, globose, subglobose

or ovo-ellipsoid, thin-walled, hyaline, strongly amyloid, with 1-3 um long

spinose projections and connectives forming a partial reticulum. BAsIpIA

22-60 x 4-16 um, clavate, thin-walled, hyaline, 4-spored; sterigmata up to

7.5 um long. LAMELLA-EDGE heteromorphous, with scattered macrocystidia.

MAacrocysTIpDIA 22-70 x 3-14 um, sinuoso-fusoid, with an apex that is either

mucronate or capitate, or with a moniliform appendage, hyaline, thin-walled,

projecting up to 22 um beyond the level of tips of basidia, with amorphous

contents, not staining with sulfovanillin. LAMELLAR TRAMA irregular; hyphae

2-12.5 um wide, hyaline, thin-walled, intermixed with groups of sphaerocytes,

8-35 x 7.5-32 um, inamyloid. SUBHYMENIUM poorly developed. PILEUS

TRAMA interwoven; hyphae 2-8 um wide, hyaline, thin-walled, intermixed

with numerous sphaerocytes, 11-40 x 11-40 um, inamyloid. PILEIPELLIs an

ixocutis tending to form an ixotrichodermium at places; hyphae 2-8 um wide,

loosely arranged, suspended in a gelatinized matrix, devoid of encrustations;

pileocystidia scattered or not seen in some collections, 30-70 x 2-6 um,

sinuoso-fusoid, with an apex that is either mucronate or capitate, or with a

moniliform appendage, hyaline, thin-walled, not staining with sulfovanillin.

STIPITIPELLIS a cutis; hyphae 2-6.5 um wide, hyaline, thin- walled, inamyloid,

Russula periglypta and R purpureonigra (India) ... 127

Figure 1: Russula periglypta. A, basidiomata; B, basidiospores; C, basidium; D, macrocystidium; E,

pileipellis. Scale bars, 10 mm for basidiomata; 10 um for microstructures.

128 ... Manimohan & Deepna Latha

devoid of encrustations; caulocystidia absent. CLAMP-CONNECTIONS not

observed on any hyphae.

Hasitat: On the ground, scattered in the proximity of a Vatica chinensis

(Dipterocarpaceae) tree.

SPECIMENS EXAMINED: INDIA. KERALA STATE, Malappuram District, Calicut University

Campus, Botanical Garden: 29 June 1994, P. Manimohan M599a (K(M)165817); 5

July 1994, P. Manimohan M599b (K(M)165818); 12 July 1995, P. Manimohan M599c

(K(M)165819); 30 June 1997, P. Manimohan M685a (K(M)165815); 01 July 1997, P.

Manimohan M685b (K(M) 165816). SRILANKA. CENTRAL PROVINCE, Kandy District,

Peradeniya, Nov. 1868, Thwaites 800 cum icon (K(M)165414, holotype).

Discussion: Russula periglypta was first described by Berkeley & Broome (1871)

with a very scanty description based on material (Thwaites 800, K) collected in

Peradeniya in 1868. Petch (1910) gave a better macroscopic description of the

species, presumably based on his own field observations. Patouillard (1913)

recorded it from Vietnam. Pegler (1986) provided a modern description of the

species based on his examination of the type material now preserved in Kew

Herbarium.

Our examination of the Kerala collections and the type material revealed

that they were conspecific. The pale yellow, slightly viscid pileus that becomes

tuberculate-striate and sulcate up to half the way from margin with age is a

diagnostic field character of this species. The shape of the basidiospores and

the length of the spinose projections showed considerable variations amongst

the five Kerala collections studied. The range of Q value was 1-1.5 and the

range of length of the spinose projections was 1-3 tm. Similarly, the presence

of pileocystidia is another variable character, as we could not see them in some

collections. We are of the opinion that this variability in the basidiospore

morphology and distribution of pileocystidia should be considered during

microscopic identification of this species. Also, unlike Pegler (1986), we

observed scattered macrocystidia on both edges and sides of lamellae, and they

projected up to 22 um beyond the level of tips of basidia.

Russula periglypta belongs to subgen. Ingratula Romagn. sect. Ingratae

(Quel.) Maire subsect. Foetentinae (Melzer & Zvara) Singer.

Russula purpureonigra Petch, Ann. Roy. Bot. Gard., Peradeniya 6: 200 (1917) Fic. 2

ORIGINAL DIAGNOsIs: “Whole fungus at first white, then blackish-gray, finally purple-

black. Pileus up to 15 cm. diameter, infundibuliform, glabrous, with a viscid separable

cuticle. Flesh thick, at first white, becoming black when cut. Stalk up to 6 cm. high,

2.5 cm. diameter, minutely pruinose, equal, solid. Gills crowded, narrow, attenuated

outwards, adnate or adnato-decurrent. Spores white, globose, nodular, 5-7 1 diameter.”

BASIDIOMATA medium-sized to large, fleshy, brittle, all parts blackening

on bruising or on drying. Prteus 30-120 mm diam., initially convex with a

slightly depressed centre, becoming deeply infundibuliform; surface greyish

Russula periglypta and R purpureonigra (India) ... 129

FIGURE 2: Russula purpureonigra. A, basidioma; B, basidiospores; C, basidium; D, macrocystidium;

E, pileipellis. Scale bars, 10 mm for basidioma; 10 um for microstructures.

130 ... Manimohan & Deepna Latha

with pale pinkish or pale brown tints (5B2, 5C2, 6B2, 6C2), slightly sticky or

somewhat viscid, not striate, smooth and glabrous; margin initially distinctly

incurved to slightly inrolled, becoming straight and finally upturned, initially

entire, becoming lobate and fissile. LAMELLAE subdecurrent to decurrent, off-

white to cream-coloured (4A2), crowded, with lamellulae of several lengths,

up to 4 mm wide; edge entire, concolorous with the sides. Stipe 20-50 x

7-20 mm, central, terete, tapering towards base, stuffed; surface off-white or

cream-coloured (4A2), glabrous. ODouR and taste not distinctive. SPORE-

PRINT white. MACROCHEMICAL SPOT TESTS on stipe surface: FeSO,: bluish

green; aniline: chocolate brown; aqueous phenol: chocolate brown.

BASIDIOSPORES 4.5-7 x 4.5-6.5 um, Q = 1-1.33, QM = 1.1, globose to

subglobose or rarely ovo-ellipsoid, thin-walled, hyaline, with amyloid, spinose

projections up to 0.25 um long and connectives forming a partial reticulum.

Basip1a 22-42.5 x 5-11.5 um, clavate, thin-walled, hyaline, 4-spored;

sterigmata up to 6.5 um long. Lamella-edge heteromorphous, with crowded

macrocystidia. Macrocystip1a 40-85 x 3.5-9 um, sinuoso-cylindric or

clavato-mucronate, thin-walled, with brownish contents, not reacting with

sulfovanillin, present both on the edge and sides of lamellae. LAMELLAR

TRAMA interwoven; hyphae 2-6.5 um wide, thin-walled, hyaline, inamyloid,

intermingled with clusters of sphaerocytes, 16-56 x 11-39 um, thin-walled,

hyaline, inamyloid. PILEUs TRAMA interwoven; hyphae 3-7 um broad, thin-

walled, hyaline, inamyloid, intermingled with clusters of sphaerocytes, 11-57 x

10-49 um, thin-walled, hyaline, inamyloid. PILEIPELLIs an ixocutis composed

of gelatinised hyphae; hyphae 2-10 um broad, thin-walled, hyaline, devoid of

encrustations; pileocystidia absent. STIPITIPELLIS a cutis; hyphae 1.5-6.5 um

broad, thin-walled, hyaline, devoid of encrustations; caulocystidia absent.

CLAMP-CONNECTIONS absent.

HasitTat: On the ground, scattered in the proximity of a Vatica chinensis

LEeG,

SPECIMENS EXAMINED: INDIA. KERALA STATE, Malappuram District, Calicut University

Campus, Botanical Garden: 25 August 1993, P. Manimohan M572b (K(M)165811); 22

July 1994, P. Manimohan M572c (K(M)165812); 24 June 1994, P. Manimohan M572d

(K(M)165813); 25 June 1997, PB.) Manimohan M572e (K(M)165814). SRI LANKA.

CENTRAL PROVINCE, Kandy District, Peradeniya, July 1912, Petch 3498 (K(M) 165416);

15 Oct. 1914, Petch 4493 (K(M) 165417).

Discussion: Russula purpureonigra was originally published without any

microscopic characters except those of the basidiospores (Petch 1917) and

the type collection of the species (Petch 4175) remains untraceable. A modern

description of this species provided by Pegler (1986) is based on two authentic

Kew collections (Petch 3498 (KM 165416) and Petch 4493 (KM 165417)). We

examined both collections and found them to be heavily mold-infested but

still retaining all microscopic features including basidiospores. We agree with

Russula periglypta and R purpureonigra (India) ... 131

Pegler’s (1986) observations on most characters. However, according to Pegler

(1986), the basidiospores of the Sri Lankan material measured 7-10 x 6-7.5

uum. We found the basidiospores to be smaller in them (5.5-8 x 4.5-6.5 um

in Petch 3498 and 4.5-6 x 4-5.5 um in Petch 4493). Remarkably, the present

Indian collections also have spores with a similar size range (4.5-7 x 4.5-6.5

um). Petch (1917) in his original description of the species also recorded that

the spores were 5-7 um in diam. Singer (1955) also measured the spores of the

Kew material as 7 x 5.8 um. While Pegler (1986) observed macrocystidia only

on the edges of the lamellae, we found them scattered on both lamellar edges

and sides.

As noted by both Singer (1955) and Pegler (1986), R. purpureonigra is a good

representative of the section Compactae Fr. (= Nigricantes Maire, Nigricantinae

Bataille) of subgenus Compacta (Fr.) Bon owing to its smooth, white pileus,

white spore-print, non-separable pileipellis, somewhat decurrent lamellae, and

basidiomata that blacken on bruising, ageing or drying. The dried basidiomata

of this species have a very characteristic charred appearance. In the field,

basidiomata growing amongst litter in wooded areas, although large, are rather

difficult to spot owing to the camouflaging effect caused by the blackened

parts.

Acknowledgments

We are thankful to Dr Bart Buyck and Dr Machiel Noordeloos for pre-submission

reviews of our manuscript. The staff of the Kew (Mycology) Herbarium are thanked for

arranging a loan of the type and authentic collections of the Russula species discussed

in this paper.

Literature cited

Berkeley MJ, Broome CE. 1871. The fungi of Ceylon (Hymenomycetes, from Agaricus to

Cantharellus). Journal of the Linnean Society, Botany 11: 494-567.

Kornerup K, Wanscher A. 1978. Methuen handbook of colour, 3rd edn., London, Methuen.

Patouillard N. 1913. Quelques champignons du Tonkin. Bulletin trimestriel de la Société

mycologique de France 29: 206-228.

Pegler DN. 1986. Agaric flora of Sri Lanka. Kew Bulletin Additional Series 12: 1-519.

Petch T. 1910. Revisions of Ceylon fungi II. Annals of the Royal Botanic Garden, Peradeniya 4(6):

373-444,

Petch T. 1917. Additions to Ceylon fungi. Annals of the Royal Botanic Garden, Peradeniya 6(3):

195-256.

Sarnari M. 1998. Monografia illustrata del genere Russula in Europa. Vicenza, Fondazione Centro

Studi Micologici.

Singer R. 1955. Type studies on Basidiomycetes VII. Sydowia 9: 367-431.

MY COTAXON

Volume 116, pp. 133-136 April-June 2011

DOT: 10.5248/116.133

Syncephalis clavata (Zoopagales, Zygomycetes),

a first record from the neotropics

ROGER FAGNER RIBEIRO MELO, ANDRE Luiz C. M. DE A. SANTIAGO

& MARIA AUXILIADORA DE Q. CAVALCANTI

Universidade Federal de Pernambuco Av. Prof. Nelson Chaves, s/n,

5670-420, Recife, Pernambuco, Brasil

CORRESPONDENCE TO: rogerfrmelo@gmail.com, andrelcabral@msn.com, xiliamac@terra.com.br

ABSTRACT — During a study on the coprophilous fungi from Brazil, the mycoparasitic fungus

Syncephalis clavata was isolated from lowland paca dung collected at the Reserva Ecoldgica

de Dois Irmaos, located in Recife, State of Pernambuco, Northeastern Brazil. This is the first

record of this species in the neotropics and the second occurrence worldwide. Aspects of the

morphology, biology and ecology of this species are discussed.

Key worps — dung, Zoopagomycotina, taxonomy, herbivorous

Introduction

Syncephalis belongs to the subphylum Zoopagomycotina, order Zoopagales

(Hibbett et al. 2007), family Piptocephalidaceae (Kirk et al. 2008) and has been

described as a haustorial mycoparasite of Mortierellales and Mucorales, or

less commonly of Ascomycetes (Benny 2005). This genus was first described

by van Tieghem & Le Monnier in 1873, and 60 names have been published

in Syncephalis (Ho & Benny 2008), representing about 45 species (Kirk et al.

2008). Species in this genus form thin coenocytic hyphae that may grow over

the host; the fungi have a well developed rhizoidal system or digitate appressoria

that bear simple sporangiophores with an apical fertile swollen head or vesicle

giving rise to one to many spored usually cylindrical merosporangia that form

directly on the vesicle apex or sporiferous structures (like basal cells) that arise

from it, variously shaped smooth or ornamented merospores, and warty, more

or less globose zygospores borne on tongue-like suspensors, (Benjamin 1959).

Syncephalis can be isolated from any habitat that also supports the growth of

a suitable host, often being found parasitizing mucoraceous fungi growing in

animal dung or soil samples (Indoh 1962).

134 ... Melo, Santiago & Cavalcanti

Syncephalis clavata, characterized by its club-shaped vesicle, doliiform

merospores produced in a branched merosporangium and cordate basal cells

that usually detach from the vesicle at maturity, was first isolated and described

by Ho & Benny (2007) in Taiwan as parasitizing Thamnostylum piriforme

(Bainier) Arx & H.P. Upadhyay growing in mouse dung.

The material of S. clavata described below was isolated from lowland paca

dung in Brazil.

Materials & methods

Samples of lowland paca (Cuniculus paca) dung were collected in the Zoological

Park at Reserva Ecolégica de Dois Irmaos (8°7°30”S and 34°52’30”W), located in

Recife, State of Pernambuco, Northeast of Brazil. The area comprises an Atlantic Forest

Ecological Reserve. The samples were collected with a sterilized spatula, transferred

to clean plastic bags, taken to the laboratory and incubated in a moist chamber in

9-cm petri dishes at 28 + 2°C for 7 days under alternating light and dark periods. The

specimen was studied directly from the substrata under a stereomicroscope and a light

microscope and described according to Ho & Benny (2007). A slide is deposited in the

Pe. Camille Torrend (URM) Herbarium of the Universidade Federal de Pernambuco,

Recife, Brazil.

Taxonomy

Syncephalis clavata H.M. Ho & Benny, Bot. Studies 48: 319. 2007. PLATE 1

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO, Recife, Reserva Ecoldgica de Dois

Irmaos, 1.2010, R-LER. Melo s.n. (URM 82246); idem, IV.2010, R.ER. Melo s.n. (URM

82247).

MyceELiIum composed of thin vegetative hyphae, 1-2 um wide, hyaline.

SPORANGIOPHORES erect, simple, (105—)175-195(-197.5) um in length, 75-10

um wide at the broadest portion near the base, tapering gradually upwards.

RHIzoIDs stout, short. VESICLE indistinct, club-shaped, 7.5-11 um diam,

slightly rounded at the apex. MEROSPORANGIA cylindrical, dichotomously

branched once at the base, each containing 3 merospores with cordate basal

cells measuring 7.5-10 x 3.5-5 um, usually detached at maturity. MEROSPORES

doliiform, 7.5-12.5 x 3.5-5 um. ZYGOSPORES not observed.

Hasirat: Parasitizing Thamnostylum piriforme growing on lowland paca

dung.

DISTRIBUTION: Brazil and Taiwan. This is the first record from Brazil.

Notes: The characteristics of Syncephalis clavata reported here show a close

similarity with the descriptions of Ho & Benny (2007). However, we found

taller (175-195 um) and thinner (7.5-10 um) sporangiophores, smaller

vesicles (7.5-11 um), and longer and shorter merospores (7.5-12.5 um) than

described by Ho & Benny (2007). Despite these minor discrepancies, we do

not consider them sufficient to propose a new species. Syncephalis clavata can

Syncephalis clavata, new to the neotropics ... 135

PiatE 1. Syncephalis clavata. A: Sporangiophore with mature merosporangia. A sporangia

of Thamnostylum piriforme is also visible (arrow). B-B1: Sporangiophore bearing cordate

basal cells (arrow) attached to the vesicle. C-C1: Rhizoids. D: Merospores. Scale bars: 10

be distinguished from other species of the genus by its club-shaped vesicle,

doliiform merospores, by having 3 merospores per merosporangium, and by

spore size.

136 ... Melo, Santiago & Cavalcanti

According to Ho & Benny (2007), the clavate sporangiophore of S. clavata

closely resembles S. pygmae, described by Patil & Patil (1994), but the later differs

by the size of the sporangiophores (25-30 x 7-8 um), number of merospores

per merosporangium (4-6) and size of merospores (4-6 x 1-2 um).

Like the Taiwanese isolate, a Brazilian isolate of Syncephalis clavata was also

found parasitizing Thamnostylum piriforme growing on rodent dung. In Brazil,

Trufem (1984) isolated Syncephalis cornu and S. tengi parasitic on Mucor mucedo

and Viriato & Trufem (1985) isolated S. asymmetrica, S. cornu, S. penicillata,

S. sphaerica, and S. tengi from herbivore dung. This report describes the first

occurrence of Syncephalis clavata in Brazil, contributing to the knowledge of

the geographical distribution of Syncephalis.

Acknowledgements

The authors thank the Conselho Nacional de Desenvolvimento Cientifico e

Tecnolégico (CNPq) for providing a MSc Scholarship and a research grant to the first

and third authors, respectively, and the Fundagao de Amparo a Ciéncia e Tecnologia

do Estado de Pernambuco (FACEPE) for providing a Post-doc fellowship to the second

author. Drs. José Luiz Bezerra and Merlin White are gratefully acknowledged for helpful

presubmission reviews.

References

Benjamin RK. 1959. The merosporangiferous Mucorales. Aliso 4: 321-433.

Benny GL. 2005. Zygomycetes. Published on the Internet at http://www.zygomycetes.org.

Hibbett DS, Binder M, Bischoff JE, Blackwell M, Cannon PF, Eriksson OE, Huhndorf S, James

T, Kirk PM, Licking R, Lumbsch HT, Lutzoni F, Matheny PB, McLaughlin DJ, Powell MyJ,

Redhead S, Schoch CL, Spatafora JW, Stalpers JA, Vilgalys R, Aime MC, Aptroot A, Bauer

R, Begerow D, Benny GL, Castlebury LA, Crous PW, Dai YC, Gams W, Geiser DM, Griffith

GW, Gueidan C, Hawksworth DL, Hestmark G, Hosaka K, Humber RA, Hyde KD, Ironside

JE, Kéljalg U, Kurtzman CP, Larsson KH, Lichtwardt R, Longcore J, Miadlikowska J, Miller A,

Moncalvo JM, Mozley-Standridge S, Oberwinkler F, Parmasto E, Reeb V, Rogers JD, Roux C,

Ryvarden L, Sampaio JP, Schiifler A, Sugiyama J, Thorn RG, Tibell L, Untereiner WA, Walker

C, Wang Z, Weir A, Weiss M, White MM, Winka K, Yao YJ, Zhang N. 2007. A higher-level

phylogenetic classification of the Fungi. Mycological Research 111: 509-547. doi: 10.1016/

j-mycres.2007.03.004

Ho HM, Benny GL. 2007. Two new species of Syncephalis from Taiwan, with a key to the Syncephalis

species found in Taiwan. Botanical Studies 48:319-324.

Ho HM, Benny GL. 2008. A new species of Syncephalis from Taiwan. Botanical Studies 49:45-48.

Indoh H. 1962. Studies on Japanese Mucorales. I. On the genus Syncephalis. Sci. Rep. Tokyo Bunrika

Daigaku, Sect. B 11: 1-26.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the Fungi. CABI: Wallingford

(United Kingdom).

Patil MS, Patil BJ. 1994. Studies in Mucorales: Piptocephalidaceae. Indian Phytopathol. 47:

DIF =225,

Trufem SFB. 1984. Mucorales do estado de Sao Paulo. 4. Espécies coprofilas. Rickia 11: 53-64.

Viriato A, Trufem SFB. 1985. Mucorales do estado de Sao Paulo. 7. Espécies merosporangiadas.

Rickia 12:147-154.

MY COTAXON

Volume 116, pp. 137-142 April-June 2011

DOI: 10.5248/116.137

Jahnula aquatica and its anamorph Xylomyces chlamydosporus

on submerged wood in Thailand

SOMSAK SIVICHAI, VEERA SRI-INDRASUTDHI & E.B. GARETH JONES

BIOTEC-Mycology Laboratory, Bioresources Technology Unit, National Center

for Genetic Engineering and Biotechnology, 113 Thailand Science Park,

Paholyothin Rd., Khlong 1, Klong Luang, Pathum Thani 12120, Thailand

CORRESPONDENCE TO: sivichai@gmail.com, veera@biotec.or.th & remispora@gmail.com

ABSTRACT — Single ascospore isolates of Jahnula aquatica germinated to give rise to the

anamorphic fungus Xylomyces chlamydosporus. This is the first confirmed connection

between a Jahnula species and the anamorphic genus Xylomyces. Colony morphology and

sporulation of Xylomyces in culture are reported, with also a fungal list of Jahnula and related

genera found in Thailand.

KEY worpDs — anamorph-teleomorph connection, freshwater Ascomycota, lignicolous

Introduction

In an ongoing study of freshwater lignicolous fungi in Thailand, natural

wood from various locations was collected and yielded a number of Jahnula

spp. Of these, a strain of Jahnula aquatica (BCC21544) contaminated by

bacteria on CMA gave rise to chlamydospores and was identified as Xylomyces

chlamydosporus. The order Jahnulales (Pang et al. 2002) comprises five

teleomorphic genera and three anamorphic genera (Brachiosphaera, Speiropsis,

Xylomyces), but no specific anamorph-teleomorph connections have been made

(Campbell et al. 2007). Eight Xylomyces taxa have been described, all from

freshwater except for X. rhizophorae (Kohlmeyer & Volkmann-Kohlmeyer

1998), which occurs on senescent mangrove wood of Rhizophora. This is the

first confirmed account of the connection of a Jahnula species to its anamorph

and is described and illustrated herein.

Materials & methods

Dead, decorticated, and barked twigs and wood were collected at a stream in Mu Ko

Chang National Park, Trat province, Thailand. On return to the laboratory samples were

washed, incubated in sterile plastic boxes on a layer of moist, sterile, tissue paper, and

periodically examined for fungi using a stereomicroscope. After the fungi were isolated

138 ... Sivichai, Sri-Indrasutdhi & Jones

and identified, preparations were mounted in lactophenol-cotton blue and sealed with

polyvinyl] alcohol.

Single-ascospore isolates of Jahnula aquatica were prepared by spreading ascospores

over Corn Meal Agar (CMA) surface with a flame-sterilized inoculation loop dipped in

0.05% (w/v) Triton X-100. Plates were incubated at 20°C in a cabinet with cool white

fluorescent light and daily examined with a microscope for signs of germination. Six

to eight germinated ascospores were transferred to new plates and incubated in the

same cabinet. After isolation, specimens were dried down and deposited in the BIOTEC

Bangkok Herbarium (BBH); live cultures are maintained in the BIOTEC Culture

Collection (BCC).

Taxonomy

Jahnula aquatica (Kirschst.) Kirschst., Ann. Mycol. 34: 196. 1936. Fics. 1-6

MycoBank MB 257114

= Amphisphaeria aquatica Plottn. & Kirschst., Verh. Bot. Ver. Prov. Brandenb.

48: 52. 1906, nom. illegit., non (Ellis & Everh.) Berl. & Voglino 1886.

= Melanopsamma aquatica Kirschst., Krypt. Fl. Brandenb. 7(2): 226. 1911.

Anam.: Xylomyces chlamydosporus Goos, R.D. Brooks & Lamore, Mycologia 69: 282. 1977.

Ascomata 345-380 x 265-330 um, hyaline when young, becoming translucent

black with age, membranous, globose to subglobose, ostiolate, papillate,

superficial to partially immersed in wood, scattered, attached to adjacent

ascomata by broad, brown, superficial, stoloniferous hyphae. Neck 60-95

x 74-80 um, periphysate; periphyses short, hyaline (Fics. 1). Peridium

20-30 mm wide, of textura angularis in surface view, in longitudinal section

peridial wall 3-4 cell layers wide, composed of an outermost layer of large,

moderately thick-walled brown cells, and an inner layer of elongated, flattened,

thin-walled hyaline cells. Pseudoparaphyses septate, narrow, 1.5-2 um wide,

hyaline, filiform, branched and anastomosing above the asci. Asci 170-200 x

15-18 um, fissitunicate, cylindrical, pedicellate, with an apical chamber and

eight, overlapping uniseriate ascospores. Ascospores (28-)33-40 x (8-)11-12

um, brown to dark brown, ellipsoidal-fusiform, 1-septate, constricted at the

septum, upper cell broader than basal cell, multiguttulate, without a gelatinous

sheath or appendages (Fics. 2-6).

ANAMORPH: The anamorph is identified by the lack of conidiophores and the

production of abundant chlamydospores. Colonies on CMA slow growing, dark

brown to black, effuse; hyphae thick-walled, septate, constricted at the septa;

hyphal cells cylindrical to subglobose; after a month bacteria-contaminated

colonies developed chlamydospores that were fusiform, intercalary, straight or

curved, solitary or in chains (95-420 x 26-42 um), occasionally branched,

with thickened septa, constricted at the septa, dark brown to blackish, end cells

paler (Fics. 7-10).

Hasirat: Saprobic on submerged natural wood in a stream.

Jahnula aquatica—Xylomyces chlamydosporus connection ... 139

Fics. 1-6. Jahnula aquatica. (BCC 21544). 1. Ascospores mass at the ostiole. 2. Ascospores.

3-6. Ellipsoidal-fusiform ascospores. Scale bars = 10 um.

SPECIMEN EXAMINED: THAILAND. Trat: Mu Ko Chang National Park, Khlong Phlu

Waterfall on submerged natural wood, 3 April 2006, Somsak Sivichai & Veera Sri-

Indrasutdhi, BIOTEC SS3895.

GEOGRAPHICAL DISTRIBUTION: Germany, Hong Kong, South Africa, Thailand, USA (IL,

TN).

ComMENnTs: The collection of Jahnula aquatica is in accord with other

descriptions of this species (Hawksworth 1984; Hyde & Wong 1999; Raja &

Shearer 2006), except no anamorph was reported previously. Unlike some

other members of the genus, the ascomata are immersed in the substratum or

superficial (lacking long stalks), and the ascospores lack a gelatinous sheath or

appendages (Fics. 2-6) (Raja & Shearer 2006).

140 ... Sivichai, Sri-Indrasutdhi & Jones

10. Xylomyces chlamydosporus produced from Jahnula aquatica ascospores.

7. Chlamydospores produced on CMA contaminated by bacteria. 8-10. Chlamydospores

produced on CMA. Scale bars = 10 pm.

Fics. 7-

Jahnula aquatica—Xylomyces chlamydosporus connection ... 141

Xylomyces chlamydosporus was described by Goos et al. (1977) from

submerged wood in Rhode Island, U.S.A.; Goh et al. (1997) reported four

different Xylomyces species also on submerged wood in freshwater streams. No

teleomorph has been described for any of these. Xylomyces was referred to the

Jahnulales based on the wide mycelium and molecular evidence (Campbell et

al., 2007), but no connection has previously been made to a specific Jahnula

species. Our study confirms that X. chlamydosporus has been correctly referred

to the Jahnulales, with J. aquatica supported as its teleomorph.

The taxa in the Jahnulales that have been collected in tropical streams of

Thailand are listed in TABLE 1.

TABLE 1. List of species of Jahnula and related genera collected in Thailand

Are ASCOSPORES REFERENCES

(sheath or polar pad)

Aliquandostipite khaoyaiensis Thick sheath Inderbitzin et al. 2001

Inderb.

A. siamensiae (Sivichai &

E.B.G. Jones) J. Campb. et al.

Jahnula appendiculata

= Campbell et al. 2007

Mette oe Thick sheath & appendages Pinruan et al. 2002

i eS Kirschst. - Sess

J. ae Ls Current study

J. es de) KD. Hyde Polar pad Current study

‘ Sa & S.W. Wong ae eae Tk

J. seychellensis Polar pad Current study

K.D. Hyde & S.W. Wong

J. morakotii 5 eS

SivichaiBe Boonyien - Sivichai & Boonyuen 2010

Megalohypha aqua-dulces - Ferrer et al. 2007

A. Ferrer & Shearer

Acknowledgements

We would like to thank Dr. Pang Ka-Lai and Dr. Mohamed A. Abdel-Wahab, for

their comments and suggestions to correct the manuscript. This work was supported

by the TRF/BIOTEC special Programme for Biodiversity Research and Training grant

BRT R_647001 We would like to thank Prof. Morakot Tanticharoen and Dr. Kanyawim

Kirtikara at BIOTEC for their constant interest and support in our study.

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Hawksworth DL. 1984. Observations on Jahnula Kirschst., a remarkable aquatic pyrenomycete.

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Hyde KD, Wong SW. 1999. Tropical Australian freshwater fungi. XV. The ascomycete genus Jahnula,

with five new species and one new combination. Nova Hedwigia 68: 489-509.

142 ... Sivichai, Sri-Indrasutdhi & Jones

Ferrer A, Sivichai S, Shearer CA. 2007. Megalohypha, a new genus in the Jahnulales from aquatic

habitats in the tropics. Mycologia 99: 456-460.doi:10.3852/mycologia.99.3.456

Goh TK, Ho WH, Hyde KD, Tsui CKM. 1997. Four new species of Xylomyces from submerged

wood. Mycological Research 101: 1323-1328. doi:10.1017/S0953756297004164

Goos RD, Brooks RD, Lamore BJ. 1977. An undescribed hyphomycete from wood submerged in a

Rhode Island stream. Mycologia 69: 280-286. doi:10.2307/3758653

Inderbitzin P, Landvik S, Abdel-Wahab MA, Berbee ML. 2001. Aliquandostipitaceae, a new family

for two new tropical ascomycetes with unusually wide hyphae and dimorphic ascomata.

American Journal of Botany 88: 52-61. doi:10.2307/2657126

Kohlmeyer J, Volkmann-Kohlmeyer B. 1998. A new marine Xylomyces on Rhizophora from the

Caribbean and Hawaii. Fungal Diversity 1: 159-164.

Pang KL, Abdel-Wahab MA, Sivichai S, Jones EBG. 2002. Jahnulales (Dothideomycetes,

Ascomycota): a new order of lignicolous freshwater ascomycetes. Mycological Research 106:

1031-1042. doi:10.1017/S095375620200638X

Pinruan U., Jones EBG., Hyde KD. 2002. Aquatic fungi from peat swamp palms: Jahnula

appendiculata sp. nov. Sydowia 54: 242-247.

Raja H, Shearer CA. 2006. Jahnula species from North and Central America, including three new

species. Mycologia 98: 319-332. doi:10.3852/mycologia.98.2.319

Sivichai S$, Boonyuen N. 2010. Jahnula morakotii, sp. nov. and J. appendiculata from a peat swamp

in Thailand. Mycotaxon 112: 475-481. doi:10.5248/112.475

MY COTAXON

Volume 116, pp. 143-149 April-June 2011

DOT: 10.5248/116.143

Glomus crenatum (Glomeromycetes), anew ornamented species

from Cuba

EDUARDO FURRAZOLA!, YAMIR TORRES-ARIAS', ROBERTO L. FERRER",

RICARDO A. HERRERA", RICARDO L.L. BERBARA*& BRUNO TOMIO GOTO?

'Instututo de Ecologia e Sistematica, IES-CITMA, A.P. 8029, de la Habana, Cuba

*Departamento de Ciéncias do Solo, Universidade Federal Rural do Rio de Janeiro,

Seropédica, Rio de Janeiro, Brazil

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

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

*CORRESPONDENCE TO: eduardofurrazola@ama.cu

ABSTRACT — A new ornamented species of the Glomeromycetes found in Pinar del Rio and

Moa, western and east Cuba, respectively, is proposed here as Glomus crenatum. The fungus

differs from previously described species by the darkly pigmented spore walls that possess a

hemispherical dome-like surface ornamentation.

Key worps — Caribbean, Glomeromycota, Glomerales, tropical forest

Introduction

Among several recently described species of the genus Glomus (Blaszkowski

et al. 2009a,b, 2010a,b; Cano et al. 2009; Furrazola et al. 2010) only few produce

ornamented spores (Blaszkowski et al. 2004, Hu 2002, Oehl et al. 2003). In

South and Central America few species with glomoid glomerospores have

been described, such as Glomus brohultii Sieverd. & R.A. Herrera 2003,

G. patagonicum [= Pacispora patagonica (Novas & Francchia) C. Walker et al.

2007], P. boliviana Sieverd. & Oehl 2004, G. megalocarpum D. Redecker 2007,

and a glomoid synanamorph of Ambispora brasiliensis B.T. Goto et al. 2008

(Herrera-Peraza et al. 2003, Oehl & Sieverding 2004, Novas et al. 2005, Walker

et al. 2007, Redecker et al. 2007, Goto et al. 2008). Only Pacispora patagonica and

P. boliviana have an ornamented outer spore wall but these species differentiate

anew an inner wall during spore development and a germinal orb specialized to

produce germinal tube initiation (Oehl & Sieverding 2004, Walker et al. 2004).

144 ... Furrazola & al.

Cuba has a high diversity of soils, many endemic plants, and different

ecosystems that probably harbour many undescribed microorganisms, including

arbuscular mycorrhizal fungi (AMF). The native soils of the Cuban biosphere

reserve “Sierra del Rosario” have been surveyed for arbuscular mycorrhizal

(AM) fungi within a long-term ecological and biodiversity study (Herrera et

al. 1988, Herrera-Peraza et al. 1994, Ferrer & Herrera 1988). One glomerospore

found during this work is described herein as Glomus crenatum.

Material & methods

Site ecology

The UNESCO Biosphere Reserve “Sierra del Rosario” is located at the eastern section of

the “Sierra del Rosario” mountains (22°45' to 23°00'N, 82°50' to 83°10'W) extending over

both provinces of Pinar del Rio and La Habana. The mountain range reaches altitudes up

to 565 m. The 20-year annual temperature and rainfall averages at the reserve are 24.4°C

and 2014 mm (Herrera et al. 1988). Ten soil samples were collected in native evergreen

mesophyllous forests or secondary forests dominated by Syzygium jambos (L.) Alston

(locally called “Pomarrozal”), an exotic, invasive tree species. The native evergreen

mesophyllous forest community is commonly dominated by Pseudolmedia spuria (Sw.)

Griseb., Oxandra lanceolata (Sw.) Baill., Trophis racemosa (L.) Urb., Matayba apetala

(Macfad.) Radlk., Dendropanax arboreus (L.) Decne. & Planch. and Calophyllum

antillanum Britton, although the composition varies with topography. Trees heights are

commonly 10-15 m on exposed, sunny hilltops, 20-25 m on lower slopes, and < 35-40

m in deep protected “V” valleys where nutrient and water usually is available all year.

Syzygium jambos, an opportunistic invasive species that quickly colonizes cut areas,

can also colonize pure stands, being then very difficult to eradicate. Another sample

was collected in Moa, Northeastern Cuba, from a plantation of Pinus cubensis Griseb.

saplings shorter than 2 m, where AM hosts, such as Cecropia spp. and other indigenous

plants, were also present.

Morphological analyses

Spores were separated from soil samples by wet-sieving and decanting (Gerdemann

& Nicolson 1963) followed by gradient centrifugation in 1M sucrose (Sieverding 1991).

Glomerospores were then placed in dishes with water and separated under a dissecting

microscope. Spores were mounted on microscope slides either in water (to check unmodified

spore wall components; Spain 1990) or permanently in polyvinyl-lacto-glycerol (PVLG;

Omar et al. 1979) or PVLG with Melzer’s reagent. Permanent slides were also prepared

using Farrant’s (Arabic gum, 40 g; water, 40 ml; glycerol, 20 ml; phenol, 20 mg) and Hoyer’s

(arabic gum, 20 g; water, 25 ml; glycerol, 10 ml; chloral hydrate, 100 g) mounting fluids.

Spores were also examined in PVLG-Cotton Blue 0.05 % to assess the reaction of the spore

wall components.

Terminology for the species description was adopted from Oehl et al. (2003) and

Goto & Maia (2006). Zeiss Axioskop compound microscopes with or without Nomarski

differential interference contrast (DIC) were used for observations, and digital images

were taken with Axiocam and AxioVision (v. 3.1 software at 1300 x 1030 dpi) or Canon

digital cameras.

Glomus crenatum sp. nov. (Cuba) ... 145

Arbuscular mycorrhizal cultures

Bait cultures with soils from primary evergreen forest (Sierra del Rosario) were

established in greenhouse at Ecology and Systematics Institute on Sorghum bicolor

(L.) Moench and Plantago major L. as host plants (Sieverding 1991) in 1.0 L pots. An

approximately 200 g field soil sample previously sieved by 2 mm mesh and including

rootlets of native plants formed a sparse layer in the 1L plastic pot, which was previously

filled to the midpoint with autoclaved primary evergreen forest soil: quartz sand mixture

3:1 (v:v). The pot was refilled with sterilized mixture and sown with seeds of the selected

plants.

Voucher specimens were deposited in the herbaria of Instituto de Ecologia y

Sistematica, la Habana, Cuba (HAC) and Universidade Federal de Pernambuco, Recife,

Brazil (URM).

Pure cultures were made by inoculating Sorghum bicolor and Plantago major seedlings

germinated in autoclaved quartz sand with 15-20 spores of G. crenatum isolated from

forest soils. 1L pots were filled with autoclaved primary evergreen forest soil: quartz

sand mixture 3:1 (v:v) and watered to field capacity. A small hole was punched in the

pot center using a glass shaker and the spores were discharged in the hole using Pasteur

pipettes. Thereafter plant seeds were sown and the plantlets grew for 3-4 months in 10

pots.

Taxonomy

Glomus crenatum Furrazola, R.L. Ferrer, R.A. Herrera & B.T. Goto, sp. nov.

MycoBAnk MB 518874 FIGs 1-9

Sporocarpia ignota. Sporae singulatim in solo vel rarum in radicibus efformatae, brunneae

vel luteo-brunneae, globosae vel subglobosae, 100-205 um diam. Tunica sporae stratis

duobus: stratum exterior hyalinum, 0.5-2.0 um crassum, secundum stratum, laminatum,

4.5-11.5 um crassum, superficie exterioris colliculare, colliculis 5.5-18 um altis, 7.5-28

um diam ad basem. Strata sporarum continuantes stratis hypharum subtendarum; strata

secundum hypharum subtendarum concolorata et continuantes cum stratae sporarum,

hypha subtenda lobulata vel irregulariter ramosa, valde recta, acute recurvata, ad basem

sporae 11-30 umcrassa, ad basem sporarum; hyphae 4.0-15 um crassae pariete hypharum

hyalinohyalina.

Type: Cuba, Pinar del Rio province, Reserva de la Biosfera “Sierra del Rosario’, Loma

El Salon, Vallecito Izquierda, primary evergreen forest on a convex hillside with N-NE

exposition (400 to 425 m.a.s.l), 30 Nov.1994, R.A. Herrera, permanent slide mounted in

PVLG (Holotype: HAC, Isotype: URM82278).

ETyMOLoGy: crenatum (Latin), referring to the distinctive ornamentation on the outer

surface of the glomerospore.

GLOMEROSPORES formed singly in soil and terminally on hypha (Fic. 1);

yellow brown to orange brown when young (Fics 1-4), darkening slightly to

dark brown when matured in soil (Fics 5-6); globose (100-208 um diam.) to

subglobose to elliptical (82-110 x 140-210 um).

SPORE WALL 5.0-13.0 um thick in total, consisting of two layers (SWLI,

SWL2; Fics 2-3) that do not react in Melzer’s reagent. Outer layer (SWL1) is

146 ... Furrazola & al.

hyaline, semi-persistent, 0.5-2.0 um thick, smooth in young spores (Fics 2-3),

with granulations on the upper surface in mature spores. Inner layer (SWL2) is

yellow brown to dark reddish brown, 4.5-11.5 um thick (Fics 2-3), laminate,

with hemispherical dome-shaped ornamentation, 7.5-28 um diam., 5.5-18

um tall (Fics 4-6). Distance between single projections are (5—)8-20(—40) um.

Ornamentation in planar view is circular to ellipsoid (Fics 5-6). Spore wall

layers are continuous with the layers of the subtending hypha (Fie. 8).

SUBTENDING HYPHA (sh) often detached at spore base; when present,

single or occasionally double, straight to sharply curved, sinuous (Fics 7-9)

or occasionally straight and parallel-sided, concolorous with wall SWL2,

11-30 um wide (mean = 16.7 um) at the point of attachment tapering to approx

2-6 um at distance of 9 um from the spore base; wall 4.0-15 um thick (mean

= 7.2 um) near the spores base, tapering to approx. 1 um distally; occlusion

introverted spore wall thickening and by septum arising from SWL2 (Fic. 9).

Pore at the subtending hypha sometimes partially open (Fic. 8). Subtending

hypha convolutions often clump a lot of organic and soil particles or small

pieces of roots. In PVLG-Cotton Blue, only SWL2 show a slight cyanophilous

reaction in young hyphae.

ADDITIONAL SPECIMENS EXAMINED: CUBA. HOLGUIN PROVINCE, Moa-Sagua-Baracoa,

from plantation of Pinus cubensis Griseb.; AMF hosts (e.g., Cecropia spp.) and other

indigenous plants also present.

GERMINATION is directly by regrowth of the subtending hypha.

ARBUSCULAR MYCORRHIZA FORMATION is unknown to date.

DISTRIBUTION — So far, the new fungus has been detected only in Cuba.

Known in soil and litter of evergreen forests consisting mainly of Pseudolmedia

spuria, Oxandra lanceolata, Trophis racemosa, Matayba apetala, Dendropanax

arboreus, and Calophyllum antillanum in native, ecologically stable (climax)

tropical evergreen forest ecosystems in Sierra del Rosario, Pinar del Rio (Cuba

occidental).

Discussion

Glomus crenatum is readily distinguished from previously described Glomus

species by its ornamented spores covered with collicular outgrowths. Glomus

pustulatum Koske et al. 1986 also produces spores with collicular ornamentation,

but its spores are paler, smaller (40-140 x 60-140 um), and with a three-layered

spore wall (vs. two layers in G. crenatum) (Koske et al 1986). Additionally,

the ornamentation in G. pustulatum is a part of the outermost spore wall

component (SWL1), whereas in G. crenatum the collicular outgrowths cover

the inner structural laminate spore wall layer (SWL2).

Glomus multicaule Gerd. & B.K. Bakshi 1976, which also forms ornamented,

darkly pigmented glomerospores with projections on the laminate layer SWL2,

Glomus crenatum sp. nov. (Cuba) ... 147

aa SMe od

Fics. 1-9. Glomus crenatum. 1. General aspect of glomerospores in PVLG; note the colliculate

ornamentation (Orn) on the spore wall (SW). 2-3. Spore wall layers (SWL1 and SWL2);

micrographs taken using Nomarski interference. 4-6. Dome shaped, hemispheric, ornamentation

(Orn) on spore wall. 7-8. Irregularly branched and/or convoluted subtending hyphae (sh); note the

open pore in the subtending hypha. 9. Septum formed by SW12.

differs from G. crenatum by having two or more attachment hyphae, very

irregular spores (149-249 x 124-162 um), and small projections that are only

1.2-3.7 um high (Gerdemann & Bakshi 1976).

The irregularly convoluted subtending hyphae found in G. crenatum (Fics

7-9) resemble those described for G. fuegianum (Speg.) Trappe & Gerd. 1974

(Gerdemann & Trappe 1974) and G. badium Oehl et al. 2005 (Oehl et al. 2005),

both of which are known to form dense sporocarps in soil. Such structures are

generally difficult to observe due to adherent soil and organic matter particles

(Fic. 9) or because they become detached close to the spore base. However,

such structures suggest that G. crenatum might also form compact sporocarps

in soil. However, spore aggregations have not yet been observed in the Cuban

tropical forest soils investigated.

148 ... Furrazola & al.

Acknowledgements

The authors acknowledge Dr. Janusz Blaszkowski (Department of Plant Protection,

West Pomeranian University of Technology, Szczecin, Poland) and Dr. Fritz Oehl

(Agroscope Reckenholz-Tanikon ART, Zurich, Switzerland) for reviewing the

manuscript and making helpful comments and suggestions. The first author thanks

Dr. Chris Walker for valuable comments regarding the species name. This work was

supported by the Conselho Nacional de Desenvolvimento Cientifico e Tecnoldégico

(CNPq) and by a grant from the Inter-American Institute for Global Change Research

(IAI) CRN 2014 which is supported by the US National Science Foundation (Grant

GEO-04523250). Thanks to Dr. Malcolm Hadley for the permanent support to improve

the ecological research in Cuba. We also acknowledge the careful and accurate revision

by Dr. Pedro Herrera (Latin diagnosis and original languages).

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MY COTAXON

Volume 116, pp. 151-155 April-June 2011

DOI: 10.5248/116.151

A new species of Xylaria from China

HalI-x1A Ma’, LARISSA VASILYEVA” & YU LI’*

"Institute of Mycology, Jilin Agricultural University, Changchun 130118, China

?Institute of Biology & Soil Science, Far East Branch of the Russian Academy

of Sciences, Vladivostok 690022, Russia

CORRESPONDENCE TO *: *'yuli966@126.com & ’vasilyeva@biosoil.ru

AsstTRacT—Xylaria ficicola, a new species of Xylaria (Xylariales, Xylariaceae), is described

from China. It is characterized by its stroma with a tiny subglobose fertile part and a relatively

long slender stipe, as well as ascospores with an appendage on each end.

Key worps—Ascomycota, host-specificity, taxonomy

Introduction

Xylaria Hill ex Schrank is a cosmopolitan genus, widespread throughout

tropical, subtropical and temperate regions (Dennis 1956, 1957, 1958; Martin

1970; Rogers 1983, 1984a,b, 1986; Rogers & Callan 1986; Rogers et al. 1988,

1997; Callan & Rogers 1990, 1993; Lzess@e 1987, 1992, 1993, 1999; Ju & Hsieh

2007; Ju et al. 2009; Trierveiler-Pereira et al. 2009). Chou (1935) was probably

the first Chinese mycologist to report Xylaria in China; he described one species

from Kweichow, China. Tai (1979) listed 53 taxa of Xylaria which were mostly

recorded in the southern provinces of China. Abe & Liu (1995) found a species

of Xylaria in Zhejiang province. The species diversity of Xylaria in China is still

poorly known and needs further investigations.

A number of Xylaria species are host-specific, mainly described on seeds

and fruits (Rogers 1979; Rogers et al. 1992, 2002; Leessoe & Lodge 1994; Xu

1999; San Martin et al. 2001). The species recently found on dried fallen leaves

and petioles of Ficus auriculata in China seems to belong to this group.

Materials & methods

The fungal material was collected in the evergreen forest of Xishuangbanna Tropical

Botanic Garden, China. The methods of collecting, preservation, and identification of

the specimens follow those of Ju and Rogers (1999).

152 ... Ma, Vasilyeva & Li

Taxonomy

Xylaria ficicola H.X. Ma, Lar.N. Vassiljeva & Yu Li, sp. nov. FIGS 1-5

MycoBANnk 519375

Stromata erecta vel prostrata, plerumque solitaria, non ramosa vel interdum ramosa,

capitulum conicum vel subglobosum, 1-2.5 mm diam., 1-2 mm crassum plus minusve

mollium, intus album, extrinsecus nigrum et aliquanto verrucosum; perithecia prominentia

vel inclusa, 0.3-0.5 mm diam.; ostiola leniter papillata vel inconspicua. Stipitis tenuis,

glabris, usque ad 6 cm longis. Asci octospori, cylindrici, longe sipitati, 190-220x8-10 um,

partibus sporiferis 110-132 um, cum annulo apicale in liquore iodato Melzeri cyanescente,

urniformi, 5-6.5(-7.5)x3-3.5 jm. Ascosporae brunneae vel fuscae, unicellulares,

ellipsoideo-inequilaterales (16-)17.5-21(-22.7) x 6.5-85 wm, plerumque sine rima

germinativa, cum extremis rotundatis vel late rotundatis, quisque cum apendicula hyalina

noncellulare, usque ad globosa.

Type — Haixia Ma, HMJAU 22818 (holotype) on dried fallen leaves and petioles of

Ficus auriculata Lour. (Moraceae); Xishuangbanna Tropical Botanical Garden, China,

6. VIII.2010.

ErymMo.ocy - The species is named after the substrate that the fungus inhabits.

STROMATA upright or prostrate, usually solitary, unbranched or sometimes

branched, the fertile head conical to subglobose, 1-2.5 mm diam.,

1-2 mm thick, texture soft, internally white, externally black and smooth, but

sometimes verrucose because of the perithecial mounds; perithecia prominent

or embedded, 0.3-0.5 mm diam.; ostioles slightly papillate or inconspicuous.

Stipes thin, glabrous, up to 6 cm long. Asci eight-spored, cylindrical, long-

stipitate, 190-220 x 8-10 um, the spore-bearing part 110-132 um long, with

apical ring bluing in Melzer’s iodine reagent, hat-shaped, 5-6.5(-—7.5) x 3-3.5

um. Ascospores brown to dark brown, unicellular, ellipsoid-inequilateral,

(16-)17.5-21(-22.7) x 6.5-8.5 um, smooth, usually without an obvious germ

slit, with broadly or narrowly rounded ends, each bearing a round hyaline

noncellular appendage up to 5 x 5 um.

ComMENtTs — Xylaria ficicola is similar to X. guazumae F. San Martin &

J.D. Rogers (San Martin & Rogers 1989) in stromatal morphology, but the

latter grows on the fallen fruits of Guazuma ulmifolia Lam. (Sterculiaceae) and

it has cespitose stromata and relatively smaller ascospores (14—)15-18(-19) x

5.5-6 um. In addition, the apical ring in asci is different: Xylaria guazumae has

rectangular apical ring, 2.8-3.2(-3.8) x 2-2.5(-3) um, but X. ficicola has reversed

hat-shaped apical ring, 5-6.5(-—7.5) x 3-3.5 um. Furthermore, X. ficicola could

be separated from X. guazumae by the ascospores with inconspicuous germ

slits. We did not observe a germ slit on the ascospores, they may be lacking or

extremely faint.

Xylaria filiformoidea Hladki & A.I. Romero (Hladki & Romero 2010) from

Argentina has similar stromata with subglobose to cylindrical fertile part and

long slender stipe, as well as ascospores with appendages, but its ascospores are

very small (8-9 x 4-5 um).

Xylaria ficicola sp. nov. (China) ... 153

Fic. 1-5. Xylaria ficicola.

1: Stromata. 2: Ascal apical ring. 3: Ascospore. 4: Fertile head of a stroma. 5: Asci.

Scale bars: 1 = 6.5 mm, 2 = 6 um, 3 = 5 um, 4= 0.6 mm, 5 = 49 um

154 ... Ma, Vasilyeva & Li

Acknowledgments

This study was supported by the National Natural Science Foundation of China

(grant no. 30770005). We are also grateful to the Ministry of Agriculture of China for

funding the field study in the project entitled “The Project of System Construction of

Modern Agricultural Technology”. We greatly appreciate Dr. J. D. Rogers of Washington

State University and Dr. Guo-zhong Liti of College of Life Science, Dalian Nationalities

University for reviewing presubmitted manuscript and giving helpful comments

and suggestions. We thank Mr. Jian-chun Zhang, Jilin Agricultural University, who

accompanied the field trip and provided general help. We thank Dr. Pu Liu, Jilin

Agricultural University for correcting the manuscript and reviewing Latin description.

Literature cited

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Mt. Fengyangshan and Mt. Baishanzu, Zhejiang Prov. in East China. Bulletin of the National

Science Museum, Tokyo, Series B, 21: 75-86.

Callan BE, Rogers JD. 1990. Teleomorph-anamorph connections and correlations in some Xylaria

species. Mycotaxon 36: 343-369.

Callan BE, Rogers JD. 1993. A synoptic key to Xylaria species from continental United States and

Canada based on cultural and anamorphic features. Mycotaxon 46: 141-154.

Chou ZH. 1935. Notes on some fungi from Kweichow. Bulletin of the Fan Memorial Institute of

Biology (Botany) 6: 161-166.

Dennis RWG. 1956. Some Xylarias of tropical America. Kew Bulletin: 401-444.

Dennis RWG. 1957. Further notes on tropical American Xylariaceae. Kew Bulletin: 297-232.

Dennis RWG. 1958. Some xylosphaeras of tropical Africa. Revista de Biologia 1: 175-208.

Hladki AI, Romero AI. 2010. A preliminary account of Xylaria in the Tucuman Province, Argentina,

with a key to the known species from the Northern Provinces. Fungal Diversity 42: 79-96.

doi:10.1007/s13225-009-0008-6

Ju YM, Hsieh HM. 2007. Xylaria species associated with nests of Odontotermes formosanus in

Taiwan. Mycologia 99: 936-957. doi:10.3852/mycologia.99.6.936

Ju YM. Hsieh HM, Vasilyeva L, Akulov A. 2009. Three new Xylaria species from Russian Far East.

Mycologia 101: 548-553. doi: 10.3852/08-188

Leessoe T. 1987. Xylaria corniformis reconsidered. Mycotaxon 30: 81-85.

Leessoe T. 1992. Xylaria digitata and its allies - delimitation and typification - I. Persoonia 14:

603-613.

Leessoe T. 1993. Xylaria digitata and its allies - delimitation and typification - II. Persoonia 15:

149-153.

Leessoe T. 1999. The Xylaria comosa complex. Kew Bulletin 54: 605-619.

Leessoe T, Lodge DJ. 1994. Three host-specific Xylaria species. Mycologia 86: 436-446.

Martin P. 1970. Studies in the Xylariaceae: VUI. Xylaria and its allies. South African Journal of

Botany 36: 71-83.

Rogers JD. 1979. Xylaria magnoliae sp. nov. and comments on several other fruit-inhabiting species.

Canadian Journal of Botany 57: 941-945. doi: 10.1139/b79-115

Rogers JD. 1983. Xylaria bulbosa, Xylaria curta and Xylaria longipes in continental United States.

Mycologia 75: 457-467. doi:10.2307/3792687

Rogers JD. 1984a. Xylaria acuta, Xylaria cornu-damae, and Xylaria mali in continental United

States. Mycologia 76: 23-33. doi: 10.2307/3792832

Xylaria ficicola sp. nov. (China) ... 155

Rogers JD. 1984b. Xylaria cubensis and its anamorph Xylocoremium flabelliforme, Xylaria allantoidea,

and Xylaria poitei in continental United States. Mycologia 76: 912-923. doi:10.2307/3793147

Rogers JD. 1986. Provisional keys to Xylaria species in continental United States. Mycotaxon 26:

85-97.

Rogers JD, Callan BE. 1986. Xylaria polymorpha and its allies in continental United States. Mycologia

78: 391-400. doi:10.2307/3793042

Rogers JD, Callan BE, Rossman AY, Samuels GJ. 1988. Xylaria (Sphaeriales, Xylariaceae) from

Cerro de la Neblina, Venezuela. Mycotaxon 31: 103-153.

Rogers JD, Ju YM, Hemmes DE. 1992. Hypoxylon rectangulosporum sp. nov., Xylaria psidii sp.

nov., and comments on taxa of Podosordaria and Stromatoneurospora. Mycologia 84: 166-172.

doi:10.2307/3760247

Rogers JD, Ju YM, Hemmes DE. 1997. Xylaria moelleroclavus sp. nov. and its Moelleroclavus

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MY COTAXON

Volume 116, pp. 157-160 April-June 2011

DOT: 10.5248/116.157

Dendrophlebia (Agaricomycetes), a new corticioid genus

from India

G.S. DHINGRA* & PRIYANKA

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

* CORRESPONDENCE TO: dhingragurpaul@gmail.com

ABSTRACT - A new corticioid genus, Dendrophlebia, is described from West Kameng in

Arunachal Pradesh.

Key worps - Eastern Himalaya, Bomdila, angiosperm host

While conducting a fungal foray in Bomdila in West Kameng of Arunachal

Pradesh, India, Dhingra made a collection from a decaying angiospermous

stump. On the basis of macroscopic and microscopic characters it was compared

to similar genera within Corticiaceae s.]. (Rattan, 1977, Eriksson et al. 1981,

Hjortstam et al. 1987, Dhingra 2005) but could not be assigned to any already

known, hence the description of a new genus.

Dendrophlebia Dhingra & Priyanka, gen. nov.

MycoBank MB 517870

Basidiocarpum resupinatum, adnatum, effusum, ceraceum; hymenium laevigatum,

continuum, flavum ad subfuscum, vinaceum in 3% KOH; systema hyphale monomiticum;

hyphae generatoriae, fibulatae; dendrophysis adsunt; cystidia hyphoidae, tenuitunicata;

basidia clavata, 4-sterigmatibus; basidiosporae ellipsoides, leves, crassitunicatae,

inamyloidee, acyanophilee.

Type SpEciEs: Dendrophlebia crassispora Dhingra & Priyanka

Erymo.oey: The name of the genus is based on the presence of dendrohyphidia in

addition to the characters of the genus Phlebia.

Basidiocarp resupinate, closely adnate, effused, ceraceous; hymenial surface

smooth, continuous, yellowish to brownish, turning dark-ruby on putting a drop

of 3% KOH solution; margins not differentiated. Hyphal system monomitic;

generative hyphae branched, septate, clamped; hyphae often agglutinated,

penetrating deep into the substratum, covered by some yellowish-brown

158 ... Dhingra & Priyanka

0 YO@O

: 25 nm

: N 4 (

ACASANG

SEE SS

Fics 1-6. Dendrophlebia crassispora: microscopic structures.

1. basidiospores; 2. basidia; 3. cystidia; 4. dendrohyphidia; 5. generative hyphae;

6. vertical section through basidiocarp.

crystalline matter. Dendrohyphidia present. Cystidia thin-walled, hyphoid.

Basidia clavate to subclavate, 4-sterigmate, with a basal clamp. Basidiospores

ellipsoid, smooth, with thickened walls, inamyloid, acyanophilous.

REMARKS—Dendrophlebia resembles genus Phlebia in having a compact

texture, clavate basidia, clamped generative hyphae, and 4-sterigmate basidia.

Dendrophlebia gen. nov. (India) ... 159

However, the presence of dendrohyphidia and basidiospores with distinctly

thickened walls distinguish this genus from Phlebia. The colour change to red

after addition of KOH is also found in some Phlebia species, as well as among

representatives of other genera. A sample was sent to Prof. Nils Hallenberg,

who also supported the concept of the new genus.

Fic. 7. Dendrophlebia crassispora:

basidiocarp showing hymenial surface.

Dendrophlebia crassispora Dhingra & Priyanka, sp. nov. Fics 1-7

MycoBank MB 517871

Basidiocarpum resupinatum, adnatum, effusum, ceraceum, ad 0.7 mm crassum; hymenium

laevigatum, continuum, maydi-flavum ad ranunculinum, in sicco subfuscum vinaceum

ad purpureum in 3% KOH; margine indeterminato;systema hyphale monomiticum;

hyphae generatoriae ad 2.5 um latae, fibulatae; dendrophysis irregulariter ramosae,

tenuitunicatae, in contexto et in hymenio adsunt; cystidia 22.5-35 x 3-4 um, hyphoidae,

sinuosa, tenuitunicata; basidia 20-32.5 x 5-6.5 um, clavata ad subclavata, vel sinuosa,

4-sterigmatibus; basidiosporae 4.5-5.5 x 3-4 um, ellipsoides ad late ellipsoides vel ovatae,

leves, crassitunicatae, inamyloides, acyanophiles, multiguttatae.

Tye: India, Arunachal Pradesh: West Kameng, Bombila, on a decaying angiospermous

stump, Dhingra 19726 (PAN, holotype), August 26, 1981.

Erymo.oey: The epithet refers to the thick-walled spores.

Basidiocarp resupinate, closely adnate, effused, up to 0.7 mm thick in section,

ceraceous; hymenial surface smooth, continuous, butter-yellow to corn-

yellow or buttercup yellow, somewhat darkening on drying, turning dark-ruby

on putting a drop of 3% KOH solution; margins not differentiated. Hyphal

system monomitic; generative hyphae up to 2.5 um wide, branched, septate,

clamped; basal zone and subhymenium not differentiated from each other

composed of compactly packed to agglutinated hyphae and dendrohyphidia,

covered by yellowish-brown crystalline matter, hyphae penetrate deep into

the substrate. Dendrohyphidia richly and irregularly branched, thin-walled,

present both in context and hymenium. Cystidia 22.5-35 x 3-4 um, hyphoid

160 ... Dhingra & Priyanka

to somewhat subulate, often sinuous, thin-walled, with a basal clamp, negative

to sulphovanillin. Basidia 20-32.5 x 5-6.5 um, clavate to subclavate, often

constricted or somewhat sinuous, 4-sterigmate, with a basal clamp; sterigmata

up to 10 um long. Basidiospores 4.5-5.5 x 3-4 um, ellipsoid to broadly ellipsoid

or ovate, tapering towards the base, smooth, somewhat thick-walled, inamyloid,

acyanophilous, with one to many oil drops.

Acknowledgements

The authors thank Prof. Nils Hallenberg (Gothenburg, Sweden) for valuable

suggestions and peer review; Prof. B.M. Sharma, Department of Plant Pathology, COA,

CSKHPAU, Palampur, H.P., India for peer review; and the Head, Department of Botany,

Punjabi University, Patiala, for providing research facilities.

Literature cited

Dhingra GS. 2005. Genus Phlebia Fr. in the Eastern Himalaya. J. Ind. Bot. Soc. 84: 111-117.

Eriksson J, Hjortstam K, Ryvarden L. 1981. The Corticiaceae of North Europe - VI. Fungiflora,

Oslo. pp. 1051-1276.

Hjortstam K, Larsson KH, Ryvarden L. 1987. The Corticiaceae of North Europe - I. Fungiflora,

Oslo. pp. 1-59.

Rattan SS. 1977. The resupinate Aphyllophorales of the North Western Himalayas. Bibliotheca

Mycologica 60: 1-427.

MY COTAXON

Volume 116, pp. 161-169 April-June 2011

DOI: 10.5248/116.161

Orbispora gen. nov., ancestral in the

Scutellosporaceae (Glomeromycetes)

FRITZ OEHL’, DANIELLE KARLA ALVES DA SILVA’,

LEONOR CosTA MAIA?, NATALIA MIRELLY FERREIRA DE SOUSA’,

HELDER ELisIo EVANGELISTA VIEIRA” & GLADSTONE ALVES DA SILVA?

'Federal Research Institute Agroscope Reckenholz-Tanikon ART,

Ecological Farming Systems, Reckenholzstrasse 191, CH-8046 Ziirich, Switzerland

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

Av. Prof. Nelson Chaves s/n, Cidade Universitaria, 50670-420, Recife, PE, Brazil

* CORRESPONDENCE TO: fritz.oehl@art.admin.ch

AxsstRact — Scutellospora pernambucana and S. projecturata are transferred into

Orbispora, our proposed new genus in the arbuscular mycorrhiza-forming Scutellosporaceae

(Glomeromycota). Orbispora is recognized by spores formed terminally on sporogenous

cells and three spore walls with single mono-lobed hyaline to subhyaline germination orbs

on the inner ‘germinal’ wall that appear identical to the orbs in Kuklospora colombiana,

K. kentinensis, and a few Acaulospora species. DNA sequence analyses show the two Orbispora

species as ancestral within the sporogenous cell-forming Glomeromycetes. An updated key

summarizes the morphological differences among species in the Scutellosporaceae.

Key worps — evolution, Gigasporineae, Gigasporaceae, molecular phylogeny, rDNA

Introduction

Recently, the genus Scutellospora C. Walker & FE. Sanders 1986 was

revised, and Gigasporaceae sensu lato was divided into four families based on

phylogenetic data congruent with spore wall morphology (especially that of

germination shields) and spore germination: Gigasporaceae, Scutellosporaceae,

Racocetraceae and Dentiscutataceae (Oehl et al. 2008, Goto et al. 2010a).

Scutellosporaceae comprised a single genus, Scutellospora. Two species of the

revised genus were found to have three spore walls and mono-lobed, orbital,

hyaline to subhyaline germination orbs on the inner, ‘germinal’ wall, with a single

germ tube initiation from where one to (rarely) two germ tubes emerge during

germination to penetrate the outer walls (Silva et al. 2008; Oehl et al. 2008). The

fact that such germ orbs and germination features were known previously only

162 ... Oehl & al.

for a few ancestral species in the Acaulosporaceae — e.g., Kuklospora colombiana

(Spain & N.C. Schenck) Oehl & Sieverd. 2006, K. kentinensis (C.G. Wu & Y.S.

Liu) Oehl & Sieverd. 2006, Acaulospora mellea Spain & N.C. Schenck 1984

(Spain 1992, Schenck et al. 1984, Sieverding & Oehl 2006, Silva et al. 2008) —

suggesting a close relationship between Acaulosporaceae and Scutellosporaceae

sensu Oehl et al. 2008 (e.g. Silva et al. 2008).

During the current study we investigated whether these two species,

Scutellospora pernambucana and S. projecturata, might be ancestral within

Scutellosporaceae. To test this hypothesis, partial LSU rDNA sequences were

generated for S. pernambucana, and the phylogenetic position of S. projecturata

was analyzed from almost complete 18S rDNA sequences obtained from public

databases. Because our hypothesis was confirmed and morphological and

phylogenetic analyses are congruent, we segregate the two species from the

other Scutellospora species that also have hyaline to subhyaline, but bi-lobed

and generally violin-shaped, germination shields, and we transfer both species

into a new genus within the Scutellosporaceae.

Material & methods

Morphological analyses

Oehl et al. (2008) and Silva et al. (2008) published the morphological analyses

relevant for this study. Scutellospora pernambucana (type species for the new genus) was

found in several sites of the Caatinga and Mata Atlantica biomes (e.g. Goto et al. 2010b)

in Igarassu, Araripina, Sao Bento do Una, and Abreu e Lima (all Pernambuco State),

Sao Luis (Maranhao State), and Mataraca (Paraiba State) in NE Brazil. Spores were

extracted from several recent collections between 2008 and 2010 at the type locality

in the ‘Zambana tropical forest fragment (Usina Sao José, Igarassu, Pernambuco State,

Silva et al. 2008) as described in Sieverding (1991). Although single species cultures

were established as described in Tchabi et al. (2010), Palenzuela et al. (2010), and Goto

et al. (2011), all trials failed so far. Thus, spores isolated directly from type collection

field samples were used for molecular analyses. Neither living spores nor type material

of S. projecturata were accessible to us.

Molecular analyses

DNA EXTRACTION: DNA was extracted from spores of S. pernambucana collected

from the type location as described in Goto et al. (2011). Spores were rinsed in distilled

water, sonicated 2-3 times for 1 min, crushed in 50 ul 10x Taq™ polymerase chain

reaction (PCR) buffer (750 mM Tris-HCl pH 8.8, 200 mM (NH,),SO,, 0.1% Tween 20;

Fermentas), centrifuged at 5,000 rpm for 5 min, and the supernatant incubated at 95°C

for 10 min. After extraction, the DNA was stored at —20°C.

AMPLIFICATION AND SEQUENCING: DNA extract was used as template for a semi-

nested PCR using primers ITS3 (White et al. 1990) - 28G2 (Silva et al. 2006) and LR1 (van

Tuinen et al. 1998) - 28G2 consecutively. PCR reactions were carried out in 50 ul batches

of 75 mM Tris-HCl pH 8.8, 200 mM (NH,),SO,, 0.01% Tween 20, 2mM MgCl, 200 uM

Orbispora gen. nov. (Scutellosporaceae) ... 163

each dNTPs, 1uM of each primer, and 2 units of Taq'™ DNA polymerase (Fermentas);

cycling parameters were 5 min at 95°C (1 cycle), 45s at 94°C, 1 min at 55°C, 1 min at

72°C (40 cycles), ending with a final 7-min elongation at 72°C. The amplified products

were purified with a PureLink PCR Purification Kit (Invitrogen), sequenced directly or

cloned with a CloneJET™ PCR Cloning Kit (Fermentas) following the manufacturer's

instruction, and sequenced. Sequencing was provided by the Human Genome Research

Center (Sao Paulo, Brazil).

SEQUENCE ALIGNMENTS: A BLASTn search of the National Center for Biotechnology

Information databases verified that the sequence obtained from S. pernambucana was

affiliated to the Scutellosporaceae (Glomeromycota). AM fungal sequences (partial LSU

rRNA) obtained in our laboratory were aligned with other glomeromycotean sequences

from GenBank using ClustalX (Larkin et al. 2007) and in BioEdit (Hall 1999) to obtain a

final alignment. The sequences were deposited at GenBank under the accession numbers

HQ871519, JF965445, and JF965446. Partial SSU rRNA sequences (obtained from

National Center for Biotechnology Information-NCBI) were also aligned as described

above.

PHYLOGENETIC ANALYSES: The phylogeny was reconstructed by independent LSU

and SSU rRNA analyses. The nucleotide substitution model was estimated using Topali

2.5 (Milne et al. 2004). Maximum likelihood (ML) analysis was performed in PhyML

(Guindon & Gascuel 2003), launched from Topali 2.5, using the GTR+G and GTR+G+I

models for LSU and SSU rRNA, respectively. Neighbor-joining analysis (established

with the same model used to construct the ML tree) was performed using PAUP*4b10

(Swofford 2003). Pacispora scintillans (S.L. Rose & Trappe) Sieverd. & Oehl ex C. Walker

et al. 2007 was used as outgroup.

Results

Molecular analyses

The LSU and SSU rRNA phylogenetic analyses (Fics. 1-2) support two

clear clades in Scutellosporaceae. The molecular trees show the clade containing

S. pernambucana and S. projecturata basal to the Scutellospora clade, supporting

the hypothesis that species with orb-like, coiled germination shields represent

an ancestral genus within the sporogenous cell-forming Glomeromycota.

New genus in Scutellosporaceae

Orbispora Oehl, G.A. Silva & D.K. Silva, gen. nov.

MycoBank MB 519533

Sporae terminaliter efformatae anguste adiacetae ad cellulas sporogeneas; cum tunicis

tribus; scutellum germinale coniunctum ad tunicam interiorem, orbiforme, hyalinum

ad alboflavum, mono-lobatum cum una depressione germationis; formans structuras

mycorrhizarum arbuscularum.

TYPE SPECIES: Orbispora pernambucana (Oehl et al.) Oehl et al.

ErymMo_oey: orbis (Latin: circle, orb), and spora (Latin: spore) referring to the mono-

lobed, coiled, orb-like germination shield of the spores.

164 ... Oehl & al.

Pacispora scintillans AJ619951

P. scintillans AJ619940

Orbispora projecturata AJ242729

oe Scutellospora aurigloba AJ276093

2 S. aurigloba AJ276092

Scutellospora sp. AJ306437

S. calospora AJ306445

S. calospora AJ306443

Gigaspora decipiens U96146

Tita G. margarita AM181143

2 1 | G gigantea EF014362

G. gigantea Z14010

G. albida 214009

G. albida AJ852599

G. rosea X58726

57 Dentiscutata colliculosa GQ376067

94 D. reticulata AJ871272

D. reticulata AJ871273

D. cerradensis AB041344

D. cerradensis AB041345

aa Fuscutata heterogama AJ852609

F. heterogama AY635832

Racocetra weresubiae AJ306444

83 R. fulgida AJ306435

R. castanea AF038590

R. gregaria AJ871274

R. gregaria AJ871275

R. tropicana GU385897

Cetraspora gilmorei AJ276094

81 C. pellucida Z14012

C. nodosa AJ306436

0.01

Fic. 1. Phylogenetic reconstruction of the Gigasporales obtained from partial SSU rDNA sequences

(~1800 bp). Bootstrap values (in %) are from neighbor-joining (NJ) and maximum likelihood

(ML) analyses (1000 bootstraps). Only topologies with bootstrap values of at least 50% are shown.

Orbispora gen. nov. (Scutellosporaceae) ... 165

Pacispora scintillans FM876832

P. scintillans FM876831

“ne Orbispora pernambucana JF965445

1400| O. pernambucana JF965446

58) O. pernambucana HQ871519

Scutellospora calospora FJ461864

83!_ S. calospora FJ461865

S. dipurpurescens FJ461868

S. calospora EU346867

S. calospora EU252109

Dentiscutata nigra AY900495

69 99] © D. nigra AY900497

93/2 Quatunica erythropus AM040354

son HT | Q. erythropus AM040357

400} 92 Fuscutata heterogama AY900503

400 76— F. heterogama FJ461877

78 F. heterogama FJ461871

ee F. heterogama DQ273792

Gigaspora albida FJ461861

G. rosea AM040350

100 G rosea Y12075

oop G. gigantea AY900506

G margarita AF396782

G. decipiens FJ461862

G. gigantea AY900504

67 G. margarita AF396783

Racocetra fulgida FJ461870

so0|—— ®. gregaria AJ510232

100/L_ R. coralloidea FJ461866

| R. persica FJ461880

R. tropicana GU385898

R. castanea Y12076

R. verrucosa AY900507

g8 R. verrucosa Ay900508

9 Cetraspora nodosa FM876833

100 400) C: NOdosa FM876836

100) C. helvetica HM565944

100#| C. helvetica HM565946

100 | C. helvetica HM565945

C. pellucida AY639261

ai C. pellucida AY639323

C. gilmorei FN547608

66| |" C. gilmorei FN547606

58 |, C. gilmorei FN547603

C. gilmorei FN547618

0.1

Fic. 2. Phylogenetic reconstruction of the Gigasporales obtained from partial LSU rDNA sequences

(~700 bp). Bootstrap values (in %) are from neighbor-joining (NJ) and maximum likelihood

(ML) analyses (1000 bootstraps). Sequences are labeled with their database accession numbers.

Only topologies with bootstrap values of at least 50% are shown.

166 ... Oehl & al.

KEY CHARACTERS: Sporocarps unknown. Spores formed on sporogenous cells

that form terminally on hyphae arising from mycelia in soil. Outer spore wall

(ow) generally (2-)3-layered and continuous with the wall of the sporogenous

cell. Two hyaline walls (middle wall ‘mw’ and inner wall ‘tw’) form de novo

during spore formation and have 1-2 and 2-3 layers, respectively. A germination

orb is formed on the outer Iw surface or between the outer and the subsequent

layer of tw. Germination orb is transparent, or hyaline to subhyaline, seldom

light yellow, mono-lobed; coiled and then, either circular or apparently broad

ellipsoid to rarely irregular; with one rounded germ tube initiation in the

outer periphery of the lobe. One (rarely two) germ tube arises from this gti to

penetrate the outer spore wall layers. Forming typical arbuscular mycorrhizae.

Orbispora pernambucana (Oehl, D.K. Silva, N. Freitas & L.C. Maia) Oehl,

G.A. Silva & D.K. Silva, comb. nov.

MycoBank MB 519535

= Scutellospora pernambucana Oehl, D.K. Silva, N. Freitas &

L.C. Maia, Mycotaxon 106: 363. 2009 [‘2008’].

Orbispora projecturata (Kramad. & C. Walker) Oehl, G.A. Silva & D.K. Silva, comb.

nov.

MycoBank MB 519550

= Scutellospora projecturata Kramad. & C. Walker, Annals Bot. 86: 22. 2000.

Dichotomous key to species in Scutellosporaceae

1. Spores with mono-lobed, coiled germination orbs ............... 00. e eee eee eee ye

1. Spores with non-coiled germination shields.............. 0c. cece eee eee eee 3

2. Spore wall non-ornamented, outer wall strongly expanding in lactic acid based

mountants; spores dark yellow to brown yellow (to yellow brown), 105-150 um

VORGEE Ts PVA ee Seb dee RUN ek ee ei Orbispora pernambucana

2. Outer spore wall with 2.0-4.0 um long protuberances formed by the structural,

laminated layer, spores golden yellow to ochraeous to sienna, 100-180 um in

AI 32 Sree EB, AG Bes x ital Pp tate oe tay oS iw ch weg Mita OG Ss als O. projecturata

3. Spores with bi-lobed, violin-shaped to oval germination shields.................. 4

3. Germination shield 1-(2)-lobed, oval to ellipsoid to irregular, with infolds of the

same lobe; spores with warted projections having central secondary projections

in the tip; spores cream to yellow, 100-170 um in diam............. S. crenulata

AUSporesvallssnolOrmanniented sy, g.4<c Mest cette hs letety ds toly et ia RR Al ty gaging 5

4. Spore walls with two types of ornamentation: small conical warts and blunt,

bacilliform larger projections; spores pale orange brown to dark orange brown,

130=180 pm diant: sos shies sos + las Pleas pln Fae Scutellospora dipapillosa

5. Spores light colored: subhyaline, pink, creamy, straw to greenish or brownish

SO Wirt he eee ee ee IN, ees en cite eka einstein Mette aaa 6

Orbispora gen. nov. (Scutellosporaceae) ... 167

6. Spores subhyaline to creamy to pale straw to greenish yellow..................0. 8

6. Spores dark yellow to golden yellow to brown yellow; spores golden yellow

to dull yellow, (130-)200-420(-520) um diam ................... S. aurigloba

7. Spores yellow brown to orange-brown, 160-360 um diam, outer and innermost

Walls‘stainine strongly in, Melzer’s reagent ..024 803i S. arenicola

7. Spores dark brown to black, 300-460 um diam .................2205- S. tricalypta

8. Spores subhyaline to pale straw to pale greenish-yellow, (100—)150-300(-500) um

diam, generally ellipsoid to oblong, with 2-layered middle wall...... S. calospora

8. Spores yellow to greenish-yellow; 140-240 um diam, generally globose to

subglobose, with one layered middle wall.................... S. dipurpurescens

Discussion

Orbispora species can easily be separated from other species in the

Scutellosporaceae by the germination shield that is coiled, orb-like, and mono-

lobed with one gti, in contrast to Scutellospora germination shields, which are

regularly bi-lobed, and generally violin-shaped to (rarely) oval. Morphological

analyses (Kramadibrata et al. 2000, Silva et al. 2008, Oehl et al. 2008) are

congruent with our phylogenetic analyses, supporting our hypothesis that

Orbispora is ancestral within Scutellosporaceae, and thus also ancestral within

all families of the sporogenous cell-forming Glomeromycetes, which have been

transferred in this volume to the new order Gigasporales (Oeh| et al. 2011).

This study renders the family Scutellosporaceae (sensu Oehl et al. 2008)

bi-generic based on molecular and spore morphological observations. The

phylogenetic position of some Scutellospora species, however, remains obscure

due to lack of molecular data, the rudimentary descriptions of the germ shields,

and the unavailability of good type material (e.g., for S. tricalypta (R.A. Herrera

& Ferrer) C. Walker & EE. Sanders 1986, S. arenicola Koske & Halvorson 1990,

and — above all — S. crenulata R.A. Herrera et al. 2001). We retain these species

in Scutellospora, as they all appear to have hyaline and bi-lobed germ shields.

However, this might be a mistake, at least for S. crenulata whose germination

shield, although still insufficiently described, appears to be mono- rather than

bi-lobed, but not orb-like or coiled (Oehl et al. 2008).

Acknowledgments

We acknowledge the valuable comments and revisions of several experts on the

manuscript, including presubmission reviews by Drs. Bruno Goto (Universidade Federal

de Rio Grande do Norte, Natal, Brazil) and Ewald Sieverding (University of Hohenheim,

Germany), and appreciate the corrections by Shaun Pennycook, Nomenclatural Editor,

and suggestions by Lorelei L. Norvell, Editor-in-Chief. This study was supported by

the Swiss National Science Foundation (SNSF, Project 315230_130764/1), Conselho

Nacional de Desenvolvimento Cientifico e Tecnolégico (CNPq, who provided a PhD

scholarship to D.K.A. da Silva, a fellowship to L.C.Maia and a grant through the INCT-

168 ... Oehl & al.

Herbario Virtual da Flora e dos Fungos), and by the Fundacao de Amparo a Ciéncia

e Tecnologia do Estado de Pernambuco (FACEPE) and the Universidade Federal de

Pernambuco (UFPE) which provided grants to F Oehl as ‘visiting professor.

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Goto BT, Silva GA da, Maia LC, Oehl F 2010a. Dentiscutata colliculosa, a new species in the

Glomeromycetes from Northeastern Brazil with colliculate spore ornamentation. Nova Hedwigia

90: 383-393. doi:10.1127/0029-5035/2010/0090-0383

Goto BT, Silva GA da, Yano-Melo AM, Maia LC. 2010b. Checklist of the arbuscular mycorrhizal fungi

(Glomeromycota) in the Brazilian semiarid. Mycotaxon 113: 251-254. doi: 10.5248/111.251

Goto BT, Silva GA da, Maia LC, Souza RG, Coyne D, Tchabi A, Lawouin L, Hountondji F, Oehl

EF, 2011. Racocetra tropicana, a new species in the Glomeromycetes from tropical areas. Nova

Hedwigia 92: 69-82. doi 10.1127/0029-5035/2011/0092-0069

Guindon S, Gascuel O. 2003. A simple, fast, and accurate algorithm to estimate large phylogenies

by maximum likelihood. Systematic Biol. 52: 696-704. doi: 10.1080/10635 150390235520

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NTT. Nucl. Acids Symp. Ser. 41: 95-98.

Kramadibrata K, Walker C, Schwarzott D, SchiiBler A. 2000. A new species of Scutellospora with a

coiled germination shield. Annals Bot. 86: 21-27. doi: 10.1006/anbo.2000.1146.

Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F,

Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG. 2007. Clustal W and

Clustal X version 2.0. Bioinformatics 23: 2947-2948. doi: 10.1093/bioinformatics/btm404

Milne I, Wright F, Rowe G, Marshal DF, Husmeier D, McGuire G. 2004. TOPALi: Software for

automatic identification of recombinant sequences within DNA Multiple Alignments.

Bioinformatics 20: 1806-1807. doi: 10.1093/bioinformatics/bth155.

Oehl F, Souza FA de, Sieverding E. 2008. Revision of Scutellospora and description of five new genera

and three new families in the arbuscular mycorrhiza-forming Glomeromycetes. Mycotaxon 106:

311-360.

Oehl £, Silva GA, Goto BT, Maia LC, Sieverding E. 2011. Glomeromycota: two new classes and a new

order. Mycotaxon 116: 365-379. doi: 10.5248/116.365

Palenzuela J, Barea JM, Ferrol N, Azcén-Aquilar C, Oehl E 2010. Entrophospora nevadensis, a

new arbuscular mycorrhizal fungus, from Sierra Nevada National Park (southeastern Spain).

Mycologia 102: 624-632. doi:10.3852/09-145.

Schenck NC, Spain J, Sieverding E, Howeler RH. 1984. Several new and unreported vesicular-

arbuscular mycorrhizal fungi (Endogonaceae) from Colombia. Mycologia 76(4): 685-699.

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

fungi (Glomales), is non-monophyletic. Mol. Phylogenet. Evol. 21: 190-197. doi:10.1006/

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Sieverding E. 1991. Vesicular-arbuscular mycorrhiza management in tropical agrosystems.

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Sieverding E, Oehl F. 2006. Revision of Entrophospora and description of Kuklospora and Intraspora,

two new genera in the arbuscular mycorrhizal Glomeromycetes. J. Appl. Bot and Food Qual. 80:

69-81.

Silva GA da, Lumini E, Maia LC, Bonfante P, Bianciotto V. 2006. Phylogenetic analysis of

Glomeromycota by partial LSU rDNA sequences. Mycorrhiza 16: 183-189. doi: 10.1007/s00572-

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Orbispora gen. nov. (Scutellosporaceae) ... 169

Silva DK da, Freitas NO, Maia LC, Oehl F. 2008. Scutellospora pernambucana, a new fungal species

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(Glomales). Mycotaxon 43: 331-339.

Swofford DL. 2003. PAUP”. Phylogenetic Analysis Using Parsimony (* and other methods), Version

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Tchabi A, Coyne D, Hountondji F, Lawouin L, Wiemken A, Oehl EF. 2010. Efficacy of indigenous

arbuscular mycorrhizal fungi for promoting white yam (Dioscorea rotundata) growth in West

Africa. Appl. Soil Ecol. 45: 92-100. doi:10.1016/j.apsoil.2010.03.001.

van Tuinen D, Zhao B, Gianinazzi-Pearson V. 1998. PCR in studies of AM fungi: from primers

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RNA genes for phylogenetics. 315-322, in: Innis MA, Gelfand D, Sninsky J, White T (eds.).

PCR protocols: a guide to methods and applications. Academic Press, San Diego, California.

MY COTAXON

Volume 116, pp. 171-174 April-June 2011

DOI: 10.5248/116.171

Uncispora sinensis, a new species from China

1GUANG ZHU YANG, JUN LUS, *ZEFEN YU, ‘KEQIN ZHANG & ? MIN QIAO

‘Laboratory for Conservation and Utilization of Bio-resources,

Key Laboratory for Microbial Resources of the Ministry of Education,

Yunnan University, Kunming, Yunnan, 650091, PR. R. China

School of Life Sciences, Yunnan University, Kunming, Yunnan, 650091, P. R. China

*CORRESPONDENCE TO: Qiaomin@ynu.edu.cn

‘These authors contributed to this work equally.

ABSTRACT — A second species of Uncispora, U. sinensis, is described and illustrated. The new

fungus was isolated from a submerged leaf collected in Xishuangbanna, Yunnan Province,

China. It is distinguished from the type species, U. harroldiae, by the shape and septation of

its conidia.

Key worps — dematiaceous hyphomycete, systematics, aquatic fungi

Introduction

Uncispora R.C. Sinclair & Morgan-Jones (Sinclair & Morgan-Jones 1979)

was described for dematiaceous hyphomycetes having brown, macronematous,

synnematous or fasciculate conidiophores and subhyaline to pale brown,

obclavate conidia that are curved at the apex and truncate at the base. It was

originally monotypic for U. harroldiae R.C. Sinclair & Morgan-Jones and is

similar to Sporidesmium Link. We add here a second, new species Uncispora

sinensis, which was isolated from submerged leaf.

Materials & methods

A culture was isolated from leaves of a dicotyledonous plant submerged in a river in

the Xishuangbanna Tropical Botanical Garden, Chinese Academy of Science (21°55'N,

101°16'E, elev. 567 m), Yunnan Province in southwest China, September 2010 (collected

by G. Z. Yang). A 3-4 x 6-7 cm rotten leaf was placed on the surface of CMA (20 g

cornmeal, 18 g agar, 40 mg streptomycin, 30 mg ampicillin, 1000 mL distilled water) for

ten days; single conidia were isolated using a sterilized toothpick while viewing with a

CX31 microscope and cultivated on CMA in Petri plates. Morphological observations

were made from CMA after incubation at 25°C for one week. Pure cultures and a

permanent slide are on deposit in the herbarium of the Laboratory for Conservation and

172 ... Yang & al.

Utilization of Bio-resources, Yunnan University, Kunming, Yunnan, P.R. China (YMF;

formerly Key Laboratory of Industrial Microbiology and Fermentation Technology of

Yunnan).

Taxonomy

Uncispora sinensis G.Z. Yang & Z.F. Yu, sp. nov. PLATE 1

MycoBank MB 519469

Coloniae in agaro pallide brunneae, pilose, effusae, post 10 dies 25°C ad 35 mm diam.

Mycelium plerumque in substrato immersum, ex hyphis septatis, ramosis, hyalinis vel

pallide brunneae compositum, 1.5-3.0 um latis. Conidiophora macronemata, synnemata

vel in fasiciculata aggregata, erecta, recta vel flexuosa, simplicia vel ramosa, pallide

brunnea vel brunnea, sursum pallidiora, laevia, modice crasse tunicata, septata, 60-112 x

2.7-4.1 um, cellae conidiogenae in conidiophoris incorporatae, monoblasticae, terminales,

determinatae, cylindricae. Conidia fasciculata, simplicia vel 1-7 septata, subhyalina vel

pallide brunnea, laevia sed verruculosa ubi matura, clavata, rostrata et hamata, ad basem

truncata, 67-89 um longa, 2-3.5 um crassa, basi 2.5-3 um apice, 0.8-1.2um lata.

Type: PR China, Yunnan province, Mengla County, Xishuangbanna Tropical Botanical

Garden, on submerged leaves of an unidentified dicotyledonous plant, Sep 2010,

G.Z.Yang. (Holotype: YMF 1.03683; ex-type culture YMF 1.03683).

ETyMOLoGy: ‘sinensis’ refers to China, the country in which this new species was

found.

Colonies pale brown to brown, attaining 35 mm diam after 10 days on CMA

at 25°C. Vegetative hyphae hyaline to pale brown, septate, smooth, 1.5-3.0

uum wide, aerial mycelium sparse, hyaline, septate, branched. Conidiophores

macronematous, synnematous, or sometimes in a fascicle of a few, very rarely

single, arising terminally or laterally on hyphae, erect, frequently branched,

pale brown to mid brown, slightly paler towards the apex, smooth, septate,

moderately thick-walled, 60-112 x 2.7-4.1 um. Conidiogenous cells integrated,

terminal, cylindrical, monoblastic, determinate, or proliferating percurrently to

produce several terminal and subterminal conidia in fascicles. Conidia clavate,

67-89 um long, 2-3.5 um at the broadest part, 0.8-1.2 um wide at the apex,

gently curved or hooked at the apex, truncate at base, 0-7 septate, subhyaline

to pale brown, smooth or with the wall of the lower cell minutely verruculose

at maturity.

DISTRIBUTION: Known only from the type collection.

Uncispora sinensis was referred easily to Uncispora based on its similarity to

U. harroldiae in having brown, macronematous, synnematous conidiophores

and clavate, apically curved or hooked conidia. However, conidia of

U. harroldiae are solitary, 3-4 septate, while those of U. sinensis are fasciculate

and 1-7 septate; moreover, conidia of U. sinensis are not so conspicuously

hooked as they are in U. harroldiae.

The role of U. sinensis in degradation of litter is the subject of continuing

research.

Uncispora sinensis sp. nov. (China)... 173

PLATE 1. Uncispora sinensis (holotype YMF1.03683).

A-E. Conidia. F. Conidiophores bearing conidia in clusters. G-H. Conidiophores.

Scale bar: A-E, G-H = 10 um, F = 25 um

174 ... Yang & al.

Acknowledgements

This work was jointly financed by National Natural Science Foundation Program

of PR China (30860004, 31060008), Grants from the Young Academic and Technical

Leader Raising Foundation of Yunnan Province (2010CI020, 2010CI106). We are very

grateful to Drs. G.J. Samuels and H.-O. Baral for critically reviewing the manuscript and

providing helpful suggestions to improve this paper.

Literature cited

Sinclair RC, Morgan-Jones G. 1979. Notes on hyphomycetes. XXVI. Uncispora harroldii [sic] gen.

et sp. nov. Mycotaxon 8: 140-143.

MY COTAXON

Volume 116, pp. 175-182 April-June 2011

DOT: 10.5248/116.175

Puccinia species new to Azad Jammu & Kashmir, Pakistan

N. S. AFSHAN’*, A. N. KHALID”, A. R. N1AzI® & S. H. IQBAL?

‘Centre for Undergraduate Studies & *Department of Botany, University of the Punjab,

Quaid-e-Azam Campus, Lahore, 54590, Pakistan

CORRESPONDENCE TO *: ** pakrust@gmail.com,* drankhalid@gmail.com,

’ mushroomniazi@gmail.com

ABSTRACT — Puccinia persistens subsp. agropyrina on Elymus semicostatus and P. variabilis

on Taraxacum officinale are newly recorded for Pakistan while P punctiformis on Cirsium

arvense and P. absinthii on Artemisia brevifolia are additions to the rust flora of Azad Jammu

& Kashmir.

Key worps — Muchal, Neelum valley, Sharda

Introduction

Azad Jammu & Kashmir (AJ & K) lies in northeast Pakistan. Due to the

wide topographic variations, plant species are highly diverse in this area.

Its flora ranges from the thorn bush type of the arid plains to the temperate

and alpine flora of higher altitudes. Prominent among the trees are Taxus

wallichiana, Cornus macrophylla, Diospyros lotus, Viburnum cylindricum, Acer

oblongum and Rhus succedanea. Common shrubs include Juniperus squamata,

Sageretia theezans, Dodonaea viscosa, Solanum verbascifolium, Lonicera

quinquelocularis, and Lyonia ovalifolia. Perennial herbs include Geranium

nepalense, Boenninghausenia albiflora, Oxalis acetosella, and Androsace

umbellate. About 10.6% of the total flora of Pakistan is represented in AJ & K

and adjacent northern areas of Pakistan (Ali & Qaiser 1986).

Although this floristically rich area has a number of host plants, it is still

poorly explored with respect to the occurrence of rust fungi. Until now, 25 rust

species have been described/reported from AJ & K (Afshan et al. 2010, 2011).

These figures indicate that the number of rust species collected from these areas

is small in relation to the vegetation.

In order to explore the diversity of Uredinales of this area, rusted plants

were collected from different localities in AJ & K. Among these, Puccinia

176 ... Afshan & al.

persistens subsp. agropyrina and P. variabilis are new records for Pakistan while

P. punctiformis on Cirsium arvense and P. absinthii on Artemisia brevifolia are

additions to the rust fungi of AJ & K.

Materials & methods

Freehand sections of infected tissue and spores were mounted in lactophenol and

gently heated to boiling. The preparations were observed under a NIKON YS 100

microscope and photographed with a digipro-Labomed. Drawings of spores and

paraphyses were made using a Camera Lucida (Ernst Leitz Wetzlar, Germany). Spore

dimensions were taken by an ocular micrometer. At least 25 spores were measured

for each spore stage. In addition to comparisons using light microscopy, images were

obtained of the rust spores using a scanning electron microscope (SEM). The rusted

specimens have been deposited in the herbarium of the Botany Department, University

of the Punjab, Lahore (LAH).

Taxonomy

Puccinia persistens subsp. agropyrina (Erikss.) Z. Urb. & J. Markova, in Markova,

Boln Soc. argent. Bot. 18(1-2): 180 (1977) (Figs. A-D)

MATERIAL EXAMINED: On Elymus semicostatus (Nees ex Steud.) Melderis (= Agropyron

semicostatum Nees ex Steud., Poaceae) with II and III stages, Pakistan, Azad Jammu &

Kashmir, Neelum valley, Sharda, at 1981 ma.s.l., 3 November 2006. NSA # 31106. (LAH

Herbarium No. NSA 1060).

SPERMOGONIA and AECIA unknown. UREDINIA on abaxial surface, brown,

0.04-0.06 x 0.07-0.1 mm. UREDINIOsPORES globose to subglobose or ovoid to

obovoid; 13-19 x 18-26 um; wall up to 2 um thick, echinulate, hyaline-light

yellow; germ pores up to 7, scattered, obscure; pedicel hyaline, not persistent,

short. TELIA on adaxial surface, black, covered by the epidermis, with brownish

stromatic paraphyses tending to divide the sorus into locules, striiform,

0.05-0.08 x 0.1-0.3 mm. TELIOSPORES 1-2-celled, 1-celled spores less common;

11-20 x 30-58 um; mostly nearly cylindrical or elongate to obovoid, constricted

at the septum; wall 0.5-1.5 um thick at sides, 2-6.5 um thick apically, apex

conical to smooth, sometimes with few digitations; brown to chestnut-brown,

paler basally. Pedicel brown, short, less than 15 um long.

CoMMENTs: Puccinia persistens was once considered closely related to

P. recondita and is still included by some in this broad species complex. Concepts

of species subspecific taxa differ considerably with respect to Puccinia recondita,

with some authors stressing a narrow species concept (and thus recognizing

many separate species, subspecies, or varieties) while others used a broad

species concept (Abbasi et al. 2005).

While researching the species complex Puccinia recondita s. lat. in detail,

Abbasi et al. (2005) morphologically and molecularly compared different

specimens in this complex with other Puccinia species on cereals and grasses.

Puccinia spp. from Azad Jammu and Kashmir (Pakistan) ... 177

WD | det | pressure HV | —— 20 um

9.9 mm ETD | 3.54e-6 Torr 00 kV | oa 3176-2007 EPM 120907 NSA#12

WD | det pressure HV = rie -5 um

9.8mm | ETD} 3.62e-6 Torr} 20.00 kV! 3176-2007 EPM 120907 NSA#

Fics. A-B. Puccinia persistens subsp. agropyrina: Scanning electron micrographs.

(A). Uredinium containing urediniospores. (B). Urediniospore showing echinulate

wall ornamentation.

178 ... Afshan & al.

Fics. C-D. Puccinia persistens subsp. agropyrina: Lucida drawings.

(C). Urediniospores (D). Teliospores. Scale bars: C = 8 um; D = 14 um.

The ITS sequence data supported a narrow species concept, so that Based upon

molecular and morphological comparisons, Abbasi et al. (2005) proposed that

the name P. persistens subsp. triticina be restricted to wheat leaf rust and P

persistens subsp. agropyrina be used to refer to leaf rust on Elymus spp.

Rust fungi previously reported on Elymus/Agropyron from Pakistan

include P. recondita and P. agropyri on Agropyron orientale, P. graminis on A.

semicostatum, and P. graminis subsp. graminicola on Agropyron sp. (Ahmad et

al,-1997,).

Puccinia persistens subsp. agropyrina is a new record for Pakistan.

Puccinia spp. from Azad Jammu and Kashmir (Pakistan) ... 179

Fics. E-F, Puccinia variabilis: Lucida drawings.

(E). Urediniospores. (F). Teliospores. Scale bars = 10 um.

Puccinia variabilis Grev., Scott. crypt. fl. (Edinburgh) 2: pl. 75 (1824) _— (Fies. E-F)

MATERIAL EXAMINED: On Taraxacum officinale (L.) Weber (Asteraceae), with II + III

stages, Pakistan, Azad Jammu & Kashmir, Neelum valley, Sharda, at 1981 m a.s.L, 3

November 2006. NSA # 902. (LAH Herbarium No. NSA 1099).

SPERMOGONIA and AECIA not found. UREDINIA amphigenous, but mostly

hypophyllous, brown to yellowish brown, scattered, rounded, 0.09-0.1 x 0.3-0.4

mm. UREDINIOSPORES subglobose to obovoid, pale brown to yellowish brown,

17-26 x 23-28.32 um (22 x 26 um on the average); germ pores 2-3, equatorial;

echinulate, wall 1-2 um thick; pedicel minute, hyaline, not persistent. TELIA

180 ... Afshan & al.

amphigenous, but chiefly hypophyllous, dark brown to black, scattered or

clustered, naked, irregular patches, 0.09-0.1 x 0.3-0.5 mm. TELIOSPORES

ellipsoid to broadly ellipsoid or obovoid, rounded at both ends, not or slightly

constricted at the septum, 19-26 x (23-)25-36 um (mean 24 x 32 um); wall

1.5-3 um thick, chestnut brown, finely verrucose, mostly smooth; germ pores

2, sub apical in distal cells and between the pedicel and septum in proximal

cells, with hyaline papilla; pedicel hyaline, about as long as the spore, 7-9 x

17-35 um.

ComMENTs: Puccinia variabilis is a new record for Pakistan. From Pakistan,

P. hieracii (= P. taraxaci) and P. sylvatica have previously been reported on

Taraxacum officinale from Swat, Kalam, Kaghan, and Batakundi by Ahmad

(1956a,b), Malik et al. (1968) and Malik & Virk (1968); and P. silvaticella on

Taraxacum sp. from Kaghan valley by Ono (1992).

Puccinia punctiformis (F. Strauss) R6hl., Deutschl. Fl., Edn 2 (Frankfurt) 3: 132

(1813) (Fic. G)

MATERIAL EXAMINED: On Cirsium arvense (L.) Scop. (= Cnicus arvensis (L.) Roth)

(Asteraceae), with II + III stages, Pakistan, Azad Jammu & Kashmir, Neelum valley,

Muchal, at 3000 m a.s.l., 3 November 2006. NSA # 789. (LAH Herbarium No. NSA

1082).

SPERMOGONIA and AECIA unknown. UReEpInIA hypophyllous, intermixed with

telia, dark brown, scattered, pulverulent, 0.09 x 0.2-0.3 mm. UREDINIOSPORES

globose to subglobose, 23-28 x 26-30 um, pale brown to chestnut brown,

uniformly echinulate; germ pores 3-4, mostly 3, equatorial; wall 2-2.5 um thick;

pedicel hyaline, minute. TeL1A hypophyllous, dark brown to black, scattered

Fic. G. Puccinia punctiformis: Lucida drawings of urediniospore and teliospores.

Scale bar = 10 um.

Puccinia spp. from Azad Jammu and Kashmir (Pakistan) ... 181

or aggregated in the form of colonies, 0.09-0.1 x 0.2-0.3 mm. TELIOSPORES

ellipsoid to broadly ellipsoid or obovoid, finely verrucose, rounded at both ends,

not or slightly constricted at the septum, pale yellow to chestnut brown, 20-28

x 26-38 um, apex 2-5 um thick, wall 1.6-3 um thick; germ pore in upper cell

apical or sub-apical, in lower cell equatorial; pedicel hyaline, mostly oblique,

5-9 x 4-16 um.

ComMENTs: Previously, P punctiformis has been reported on Cnicus arvensis

from Shahdara, Changa Manga, Dass, and Lahore by Ahmad (1956b), but it is

an addition to the Uredinales for AJ & K.

Fics. H-I. Puccinia absinthii: Lucida drawings.

(H). Urediniospores. (I). Teliospores. Scale bars = 8 um.

Puccinia absinthii DC., Fl. Franc. 6: 56 (1815) (Fics. H-I)

MATERIAL EXAMINED: On Artemisia brevifolia Wall. (Asteraceae), with II & III stages,

Pakistan, Azad Jammu & Kashmir, Neelum valley, Sharda, at 1981 m a.s.l., 3 November

2006. NSA # 901. (LAH Herbarium No. NSA 1097).

SPERMOGONIA and AECIA unknown. UREDINIA amphigenous, light brown,

scattered, rounded, 0.09-0.2 x 0.2-1.0 mm. UREDINIOSPORES ovoid or obovoid

to ellipsoid, light yellow to pale brown, 16-24 x 23-32 um (21 x 28 um on

the average); germ pores 2, equatorial, without a papilla; echinulate, wall 1.5-2

um thick at sides, 3-4 um thick apically; pedicel hyaline, short, not persistent.

TELIA amphigenous, on leaves, small pustules, roundish, dark brown to black,

scattered, 0.09-0.1 x 0.3-1.0 mm. TELIosPorEs ellipsoid to broadly ellipsoid

182 ... Afshan & al.

or obovoid, not or slightly constricted at the septum, attenuated towards base,

22-30(-33) x (38-)40-57 um (mean 27 x 46 um), chestnut brown, wall 3-3.5

uum thick at sides, 4-7.5 um thick apically, apex conical or rounded, verruculose

at the apex, smooth at lower side; germ pores 2, apical or sub apical in distal

cells and close to septum in proximal cells, with hyaline papilla; pedicel hyaline,

persistent, 4-12 x 84-142 um.

ComMENTSs: Previously, P absinthii has been reported on A. persica and

A. parviflora from Quetta, Chitral, and NWFP by Ahmad (1956a, b), Malik et

al. (1968) and Malik & Virk (1968), on A. dubia from Swat by Ono & Kakishima

(1992), and on A. dracunculus from Kaghan valley by Ono (1992). It is a new

record for AJ & K.

Acknowledgements

We sincerely thank Dr. Marcin Piatek, W. Szafer Institute of Botany, Polish Academy

of Sciences, Poland, and Dr. Mehrdad Abbasi, Associate Professor, curator of fungus

collection, Department of Botany, Iranian Research Institute of Plant Protection, Iran,

for their valuable suggestions to improve the manuscript and acting as presubmission

reviewers. We are highly obliged to Higher Education Commission (HEC) of Pakistan

for providing financial support.

Literature cited

Abbasi M, Ershad D, Hedharoude GHA. 2005. Taxonomy of Puccini recondita s. lat. Causing brown

rust on grasses in Iran. Iran. J. Plant Pat. 41: 245-253.

Afshan NS, Iqbal SH, Khalid AN, Niazi AR. 2010. A new anamorphic rust fungus with a new record

of Uredinales from Azad Kashmir, Pakistan. Mycotaxon 112: 451-456. doi:10.5248/112.451

Afshan NS, Iqbal SH, Khalid AN, Niazi AR. 2011. Some additions to the Uredinales of Azad Jammu

and Kashmir (AJ & K), Pakistan. Pak. J. Bot. 43(2): 1373-1379.

Ahmad S. 1956a. Uredinales of West Pakistan. Biologia 2(1): 29-101.

Ahmad S. 1956b. Fungi of Pakistan. Biological Society of Pakistan, Lahore Monograph 1: 1-126.

Ahmad §, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Nabiza Printing Press, Karachi, Pakistan.

Ali SI, Qaiser M. 1986. A phytogeographic analysis of the phanerogams of Pakistan and Kashmir.

Proceedings of the Royal Society of Edinburgh 89 B: 89-101.

Cummins GB. 1971. The rust fungi of cereals, grasses and bamboos. Springer Verlag Berlin-

Heidelberg-New York.

Malik SA, Virk. 1968. Contribution to the knowledge of parasitic fungi of Quetta-Kalat Region.

Biologia 14: 27-35.

Malik SA, Javaid MT, Ahmad M. 1968. Uredinales of Quetta-Kalat region of Pakistan. Biologia 14:

37-46.

Ono Y. 1992. Uredinales collected in the Kaghan Valley, Pakistan. Cryptogamic flora of Pakistan

1: 217-240.

Ono Y, Kakishima M. 1992. Uredinales collected in the Swat Valley, Pakistan. Cryptogamic flora of

Pakistan 1: 197-216.

MY COTAXON

Volume 116, pp. 183-189 April-June 2011

DOI: 10.5248/116.183

Hydnoid basidiomycetes new to Brazil

ALICE DA CruUZ LIMA GERLACH & CLARICE LOGUERCIO-LEITE

Departamento de Botanica, Centro de Ciéncias Biologicas, Universidade Federal de Santa Catarina,

Campus Universitario, 88040-900, Florianopolis, SC, Brazil

CORRESPONDENCE TO: alice_gerlach@yahoo.com.br & clleite@ccb.ufsc.br

ABSTRACT — A survey of wood-decaying fungi from an Araucaria forest in the state of

Santa Catarina in southern Brazil yielded numerous species of Agaricomycetes. ‘Three

hydnaceous species collected (Mycobonia brunneoleuca, Mycoacia aurea and Spongipellis

africana) represent first records from Brazil. Illustrations and keys to the Brazilian species of

Mycobonia, Mycoacia, and Spongipellis are provided.

Key worps — Polyporales, corticioid, fungal distribution

Introduction

The Atlantic Forest is still common in the state of Santa Catarina, where

secondary forest dominates much of the landscape and 23% of the original

vegetation cover remains (SOS Mata Atlantica/INPE 2009). The most

outstanding feature of this subtropical region is the large extent of mixed

Araucaria forests that cover the inner plateaus of southern Brazil and the

Misiones Province in Argentina (Oliveira-Filho et al. 2009). These forests are

easily recognized by their canopies, which are dominated by the chandelier-

like crowns of Araucaria angustifolia (Bertol.) Kuntze (Sonego et al. 2007), and

form complex mosaics with grasslands at higher altitudes Jarenkow & Budke

2009).

Previous surveys and reviews of hydnoid species from Brazil were published

by Rick (1932a,b, 1959), Bononi (1979, 1981), Bononi et al. (1981, 2008),

Hjortstam & Bononi (1986a,b, 1987), Capelari & Maziero (1988), Sdtao et al.

(1991), Jesus (1993), Nietiedt & Guerrero (2000), Gibertoni et al. (2007), and

Baltazar & Gibertoni (2009).

The aim of this study was to increase knowledge of hydnoid basidiomycetes

(Agaricomycetes) from Araucaria forests in Brazil.

184 ... Gerlach & Loguercio-Leite

Materials & methods

The study was made in the Reserva Particular do Patrimonio Natural Rio das Furnas

(RPPN Rio das Furnas) located in the Alfredo Wagner municipality, in Santa Catarina,

Brazil (27°40'45"S, 49°10'38.4"W). This is a 10 ha reserve within the Araucaria forest

region.

Microscopic characters were examined and measured using light microscopy, on

slides mounted with 1% aqueous phloxine solution (plus 1% or 5% KOH) and Melzer’s

reagent (Ryvarden 1991). Drawings were made with the aid of a drawing tube. Voucher

specimens are archived at FLOR (Thiers 2007).

Taxonomy

Mycobonia brunneoleuca, Mycoacia aurea, and Spongipellis africana represent

new records from Brazil and are described below based on the newly collected

material. Keys including related Brazilian taxa are also provided.

Gloeophyllaceae Jiilich

Mycobonia brunneoleuca (Berk. & M.A. Curtis) Pat., Essai Tax. Hyménomyc.:

23: V9OO: FIG. 1

VOUCHER MATERIAL: BRAZIL. Santa Catarina: Alfredo Wagner, RPPN Rio das

Furnas, 07.VII.2008, Gerlach & Giovanka 122, FLOR 32323.

BRIEF DIAGNOSIS: Mycobonia brunneoleuca is characterized by the longer

hyphal pegs (< 180 um outside the hymenium), robust basidia (< 80 x

13-20 um), and ovoid-elliptic basidiospores, 15-24(-25) x (6-)7-11 um, which

contain many oil droplets.

ADDITIONAL SPECIMENS EXAMINED — BRAZIL. SANTA CATARINA: Itapiranga, Scholz

s/n, 15.IV.1985, FLOR 0109; Trés Barras, Flona, Drechsler-Santos s/n, 22.X1.2003,

FLOR 31465; Mondai, Linha Uruguai, 27.XII.2006, Campos-Santana, Santana & Souza-

Rodrigues 188, FLOR 32226; 23.V.2007 Campos-Santana & Santana 223, FLOR 32227.

DISTRIBUTION: Neotropical.

COMMENTS: MyYCOBONIA BRUNNEOLEUCA AND M. FLAVA (Sw.) Pat. are

macroscopically indistinguishable except for their basidiospores, which are

wider and ovoid-elliptic in M. brunneoleuca. Dennis (1970), who regarded

these taxa as the same species, considered them common in cloud forests of

Venezuela (e.g., Sierra de la Costa) between 1600 and 2000 meters. According

to Reid (1976) it is possible that both species occupy the same geographical

region but live in different niches, with M. brunneoleuca occurring at higher

elevations. Mycobonia flava was previously collected within the Mondai (220 m

elev.) and Itapiranga (< 544 m) municipalities and the Floresta Nacional de Trés

Barras reserve (766 m) in Santa Catarina. Our material was collected between

750 and 900 m in Araucaria forest, which agrees with observations made by

Reid.

Hydnoids new to Brazil... 185

Fic. 1. Mycobonia brunneoleuca: A, hyphal pegs; B, basidiospores; C, basidia.

Scale bars = 10 um.

Key to Mycobonia species in Brazil

1. Spores ovoid-ellipsoid, (6-)7-10 um wide ................2.004. M. brunneoleuca

LeSpores fusoid-ellipsdid/5=7 wim wide. <5 seatie ice eatatlt aati a ev ewes M. flava

Meruliaceae P. Karst.

Mycoacia aurea (Fr.) J. Erikss. & Ryvarden, Corticiac. N. Europe 4: 877. 1976. Fic. 2

VOUCHER MATERIAL: BRAZIL. SANTA CATARINA: Alfredo Wagner, RPPN Rio das

Furnas, Reck, 17.VII.2009, Gutjahr 09, FLOR 32415.

BRIEF DIAGNOSIS: Mycoacia aurea has resupinate basidiomata that are very

soft when fresh and with a horny consistency when dried. The species is

characterized by a palisade hymenium (< 35 um thick), subiculum with thin to

slightly thick-walled, clamped generative hyphae, clavate 4-spored basidia with

basal clamps, and suballantoid basidiospores (4-5 x 1.5-2 um).

ADDITIONAL SPECIMEN EXAMINED: SWEDEN. Darina: Husby, Rankholmen i

Flinesjon, naturreservat, 6.5 Km SO Husby K:a. Lat/long: 60°22'N, 16°06 _'E, Grid:

186 ... Gerlach & Loguercio-Leite

Fic. 2. Mycoacia aurea: A, hymenium; B, basidiospores; C, generative hyphae.

Scale bars = 10 um.

RN 6694300 1517200, Alt. 80 m. Prunus-Laga i graalskog, alvplan, 18.III.1990, Janolof

Hermansson $253, Herb. Univ. Upsaliensis F-140874, 425667.

DISTRIBUTION: Cosmopolitan.

Comments: Eriksson & Ryvarden (1976) distinguish M. aurea from other

Mycoacia species by its small allantoid basidiospores, absence of cystidia,

and absence of a red KOH reaction. Hjortstam & Ryvarden (1996) noted that

M. aurea lacks incrusted hyphal endings in its spines and has suballantoid

basidiospores, which coincides with our material. The specimen from Sweden

(F-140874) has some encrusted hyphae but is otherwise very similar to the

Brazilian specimen.

Key to Mycoacia species in Brazil

(based on Hjortstam & Ryvarden 1996)

1. Hymenophore tuberculate, subulate cystidia rare .................0.000. M. livida

1. Hymenophore hydnoid, cystidia either not subulate or absent ................... 2

Hydnoids new to Brazil ... 187

2iCvestidia present ia 2s eae see anaes hein ck hen 28 hin SE ein OES ln BG dln See ala ces 3

2 AOU SUC AD SEU AAR RAR PRE PAPA cals Rac che Pi Gee Pg ts Rd wecott Pisin eR ccgals Bact OE 4

3. Cystidia subulate to acicular, encrusted hyphae present............... M. fuscoatra

3.,Cystidia-subfusiform,-ericrusted hyphae absent osioi0 oa nonce seas nonce ws M. uda

4. Basidiospores allantoid, 4-4.5 x 1-1.25 um ...............00000. M. subconspersa

4. Basidiospores suballantoid, 3.5-5.5 x 1.5-2 UM...... 6... eee eee M. aurea

Polyporaceae Corda

Spongipellis africana Ipulet & Ryvarden, Syn. Fung. 20: 97. 2005. FIG. 3

VOUCHER MATERIAL: BRAZIL. SANTA CATARINA: Alfredo Wagner, RPPN Rio das

Furnas, 17.XII.2008 ,Gerlach, Giovanka 155, FLOR 32416.

BRIEF DIAGNOSIS: Spongipellis africana has widely adhered to effuse reflexed

basidiomata that are soft when dried, clavate 4-spored basidia with basal

clamps, and basidiospores that are (sub)globose [4-5(-6) x 4-5 um], thick-

walled, smooth, and with positive cyanophilic reaction.

ADDITIONAL SPECIMEN EXAMINED: URUGUAY. Dto Treinta e tres Quebrada de los

Cuervos, on dicotyledonous dead wood, 1.VII.1993, C. Prigioni MVM 374, BAFC

34059.

DISTRIBUTION: Pantropical.

3 © V6

Fic. 3. Spongipellis africana: A, hymenium; B, basidiospores; C, cystidia.

Scale bars = 10 um.

ComMENts: According to Ipulet & Ryvarden (2005), S. africana is

macroscopically identical to S. pachyodon (Pers.) Kotl. & Pouzar but easily

separated by its smaller basidiospores. Ipulet & Ryvarden (2005) did not observe

188 ... Gerlach & Loguercio-Leite

cystidia in this species; however, our material has inconspicuous metuloid

cystidia in the hymenium. The specimen from Uruguay (BAFC 34059) was

previously identified as S. pachyodon but it has smaller basidiospores. Therefore,

we believe that this specimen is also S. africana, which extends the distribution

of S. africana to Brazil and Uruguay.

Key to Spongipellis species in Brazil

(based on Piatek et al. 2004, Rajchenberg & Meijer 1990, Gilbertson & Ryvarden 1986)

is Hyimenoplroredistincthy hydrioia: 1155.0 esse ENS sg Uae toe Pee eet oe eee te Nene ta Ne 2

1. Hymenophore poroid, or sometimes sinuous to daedaleoid ...................4. 3

2. Basidiospores globose to broadly ellipsoid, 5.5-7 x 4.5-5.5 um ....... S. pachyodon

2) Basidiospores'slobOse, 474.5 Ut, ssvavhs scvacta Hava sshe Mana a teva actiona sha ona S. africana

3. Pores subgyrose, 1-3/mm, compressed when dry, dentate to fimbriate,

hymenophore pinkish red to pinkish salmon when fresh, basidiospores

ellipsoid to broadly ellipsoid, 4.6-6 x 3.3-4.3 um.............0004. S. caseosus

3. Pores angular, 4-5/mm, hymenophore white to cream, basidiospores broadly

ellipsoid to globose (ovoid), 4.5-6 x 4-5 um... eee eee S. fractipes

Acknowledgments

The authors are grateful to Gabriela Giovanka, Renato Rizzaro, Matheus Reck and

Marcio Gutjahr, especially for their help with the fieldwork. We are also grateful to

the reviewers, Karen Nakasone (USA) and Maria Alice Neves (Brazil), as well Shaun

Pennycook (New Zealand) and would like to thank them for their suggestions and for

improving this article.

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from the Brazilian Atlantic Forest. Mycotaxon 109: 439-442. doi:10.5248/109.439

Bononi VLR. 1979. Basidiomicetos do parque Estadual da Ilha do Cardoso: III. Espécies hidndides.

Rickia 8: 63-74.

Bononi VLR. 1981. Alguns basidiomicetos hidndides da regiao Amazénica. Rickia 9: 13-30.

Bononi VLR, Trufem SFB, Grandi RAP. 1981. Fungos macroscdépicos do parque Estadual das

Fontes do Ipiranga, Sao Paulo, Brasil, depositados no herbario do Instituto de Botanica. Rickia

9037593:

Bononi VLR, Oliveira AKM, Quevedo JR, Gugliotta AM. 2008. Fungos macroscopicos do Pantanal

do Rio Negro, Mato Grosso do Sul, Brasil. Hoehnea 34(4): 489-511.

Capelari M, Maziero R. 1988. Fungos macroscépicos no estado de Rondé6nia, regiao dos rios Jaru

e Ji Parana. Hoehnea 15: 28-36.

Dennis RWG. 1970. Fungus flora of Venezuela and adjacent countries. Kew Bulletin Additional

Series 3: 1-530.

Eriksson J, Ryvarden L. 1976. The Corticiaceae of North Europe, vol. 4. Hyphodermella—Mycoacia.

Fungiflora, Oslo, Norway.

Gibertoni TB, Santos PJP, Cavalcanti MAQ. 2007. Ecological aspects of Aphyllophorales in the

Atlantic Rain Forest in Northeast Brazil. Fungal Diversity 25: 49-67.

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Gilbertson RL, Ryvarden L. 1986. North America Polypores, vol.1. Abortiporus—Lindtneria.

Fungiflora, Oslo, Norway.

Hjortstam K, Bononi VLR. 1986a. Fungos corticéides do Brasil (Aphyllophorales). Rickia 13:

113-125.

Hjortstam K, Bononi VLR. 1986b. Studies in tropical Corticiaceae (Basidiomycetes) VI: A new

species of Steccherinum from Brazil. Mycotaxon 25: 467-468.

Hjortstam K, Bononi VLR. 1987. A contribution to the knowledge of Corticiaceae s.l.

(Aphyllophorales) in Brazil. Mycotaxon 28: 1-15.

Hjortstam K, Ryvarden L. 1996. New and interesting wood-inhabiting fungi (Basidiomycotina-

Aphyllophorales) from Ethiopia. Mycotaxon 60: 181-190.

Ipulet P, Ryvarden L. 2005. New and interesting polypores from Uganda. Synopsis Fungorum 20:

87509:

Jarenkow JA, Budke JC. 2009. Padrées floristicos e andlise estrutural de remanescentes florestais

com Araucaria angustifolia no Brasil. 113-126, in: Fonseca CR, Souza AF, Leal-Zanchet

AM, Dutra T, Backes A, Ganade G (Org.). Floresta com araucaria: ecologia, conservagao e

desenvolvimento sustentavel. Ribeirao Preto, Holos Editora.

Jesus MA. 1993. Basidiomicetos lignoceluloliticos de floresta nativa e de Pinus elliottii Engelm. do

Parque Estadual das Fontes do Ipiranga, Sao Paulo, SP. Hoehnea 20(1/2): 119-126.

Nietiedt SA, Guerrero RT. 2000. Familia Corticiaceae (s.1.) - Novas citagoes para o Brasil. Iheringia

sér. Bot. Porto Alegre 54: 45-56.

Oliveira-Filho AT, Jarenkow JA, Budke JC. 2009. Patterns of floristic differentiation among Atlantic

forests and the influence of climate II: The southern scenario. In: Impacts of Global Changes on

Tropical Ecosystems Cross-cutting the Abiotic, Biotic and Human Spheres. Joint Meeting of the

Association for Tropical Biology and Conservation (ATBC) and Society for Tropical Ecology,

Marburg.: Philipps Universitat Marburg, p. 433-433.

Piatek M, Seta D, Szczepkowski A. 2004. Notes on Polish polypores 5. Synopsis of the genus

Spongipellis. Acta Mycologica 39(1): 25-32.

Rajchenberg M, Meijer AAR. 1990. New and noteworthy polypores from Parana and Sao Paulo

States, Brazil. Mycotaxon 38: 173-185.

Reid DA. 1976. Notes on polypores. 2. Memoirs of the New York Botanical Garden 28(1):

179-198.

Rick J. 1932a. Hidnaceaes Riograndenses. Egatea 17: 1-9.

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Rick J. 1959. Basidiomycetes eubasidii in Rio Grande do Sul - Brasilia. 3. Hypochnaceae,

Clavariaceae, Craterellaceae, Hydnaceae. theringia, Bot. 5: 125-192.

Ryvarden L. 1991. Genera of polypores. Nomenclature and taxonomy. Synopsis Fungorum 5:

1-363.

Sonego RC, Backes A, Souza AF. 2007. Tree community structure and composition in a mixed

ombrophilous forest in S40 Francisco de Paula, Rio Grande do Sul State, Brazil. Acta Botanica

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MY COTAXON

Volume 116, pp. 191-202 April-June 2011

DOI: 10.5248/116.191

Phylogenetic relationships and reclassification of

Spirosphaera lignicola, an enigmatic aeroaquatic fungus

HERMANN VOGLMAYR

Department of Systematic and Evolutionary Botany, University of Vienna,

Rennweg 14, A-1030 Vienna, Austria

CORRESPONDENCE TO: hermann.voglmayr@univie.ac.at

ABSTRACT — The enigmatic Spirosphaera lignicola is revised based on recent collections from

Austria. The taxon does not fit the generic circumscription of the genus Spirosphaera in its

morphological features but clearly belongs to Dendroclathra. Nuclear ITS rDNA sequence

data from recent collections and the type specimens indicate conspecificity of S. lignicola with

Dendroclathra caeruleofusca. Spirosphaera lignicola is transferred to the genus Dendroclathra,

and a recent collection with an ITS sequence identical to the type specimen is designated as

epitype. Morphology is illustrated with SEM and LM pictures. Phylogenetic analyses of LSU,

ITS and tef1 sequence data reveal a phylogenetic affinity of Dendroclathra to the Microascales

(Hypocreomycetidae), being therefore phylogenetically distant from Spirosphaera floriformis,

the generic type, which belongs to the Leotiomycetes. The recent collections are the first

records of this species for Austria. The distribution and aeroaquatic ecology of D. lignicola

are briefly discussed.

KEY worpDs — anamorph, ascomycetes, freshwater fungi, molecular phylogeny, taxonomy

Introduction

Spirosphaera lignicola was described from deciduous wood submerged in a

stream in Italy (Abdullah et al. 1998). Although the description and illustrations

indicated that S. lignicola does not fit the morphological generic circumscription

of Spirosphaera Beverw. due to the lack of distinct coiling of the conidial filament

and of the unilateral branching pattern typical of Spirosphaera (Hennebert

1998), no suitable alternative classification could be proposed with the data at

hand (Voglmayr 2004). No type culture was deposited in a culture collection,

and the sporulation on the holotype specimen (dried malt extract agar culture)

did not reveal features enabling an appropriate generic classification. It was

therefore considered an enigmatic fungus requiring additional studies.

Dendroclathra Voglmayr & G. Delgado is a monotypic anamorph genus,

with D. caeruleofusca as the type species (Voglmayr & Delgado-Rodriguez

192 ... Voglmayr

2001). The diagnostic features of the genus were compact multicellular

globose conidia, formed by a conidial filament first branching extensively tree-

like and subsequently many times irregularly dichotomously in a tangential

manner, the tips frequently fusing with neighboring cells upon mutual

contact. Its multicellular conidia are therefore morphologically quite complex.

Dendroclathra caeruleofusca was collected and isolated from a dead bamboo

stem submerged in a river in tropical Cuba. This habitat, together with its

conidial morphology, indicated an aeroaquatic ecology (Voglmayr & Delgado-

Rodriguez 2001). Although it could be easily isolated in pure culture and grew

vigorously on PDA and MEA, no sporulation was observed on agar cultures but

only on natural substrate. No connection with Spirosphaera lignicola was made

at that time despite some superficial similarity, and according to the description

and the SEM pictures, the latter apparently lacked the very prominent fusion

of the conidial cells. The conidia present on the holotype of S. lignicola, which

consists of a dried culture, showed a different, more irregular branching pattern

not in line with the more complex conidia of D. caeruleofusca, which were only

known from natural substrate.

During recent research on biodiversity and phylogeny of aeroaquatic fungi,

a fungus was repeatedly collected and isolated in the surroundings of Vienna

from wood submerged in various small streams, ditches, and oxbow lakes,

which closely matched Dendroclathra caeruleofusca in conidial ontogeny and

morphology. However, unlike the type culture of the latter, in some isolates

atypical but abundant conidia were formed on agar cultures, which closely

resembled those present on the dried holotype culture of Spirosphaera lignicola.

To elucidate whether S. lignicola and D. caeruleofusca are congeneric or even

conspecific and to which species the Austrian collections belong, detailed DNA

and morphological studies were performed.

Materials & methods

Sampling and documentation

Submerged, rotting twigs were taken from various small rivulets, ditches and oxbow

lakes, packed into plastic bags and returned to the laboratory. The twigs were cut into

pieces, rinsed in tap water and spread on Petri dishes lined with moist filter paper, kept

damp and exposed to natural light at room temperature. The Petri dishes were regularly

examined under a dissecting microscope for the presence of propagules of aeroaquatic

fungi. After spores had developed, they were transferred aseptically to 2% malt extract

agar (MEA) plates, where they readily germinated. Spores from both natural substrate

and agar cultures were examined under the light microscope. Twig fragments containing

spores and sporulating agar cultures were air-dried and deposited in the herbarium of

the Faculty Center of Biodiversity of the University of Vienna (WU). Living cultures

were deposited in CBS (Utrecht, The Netherlands). The collection data of the specimens

investigated are listed in the Taxonomy section.

Dendroclathra lignicola comb. nov. ... 193

Light microscopy

Conidia from fresh and dried specimens were mounted in tap water or L4 (general

mounting fluid after Clémencgon 1972) for light microscopy. Slides were examined

and photographed using a Zeiss Axio Imager.Al (Zeiss, Jena, Germany) microscope

equipped with a Zeiss AxioCam ICc3 digital camera.

Scanning electron microscopy

For scanning electron microscopy (SEM), spores grown on natural substrate were

prepared according to the method described in Halbritter (1998). After preparation, the

specimens were mounted on Cambridge stubs, sputter coated with gold, and examined

in a Jeol T 300 scanning electron microscope at 10 kV.

DNA extraction, PCR and sequencing

For sample preparation from living cultures, small pieces were cut out at the margin

of actively growing agar cultures, put into 2 ml reaction-tubes, immediately frozen,

and freeze-dried. Alternatively, liquid cultures were grown in 100 ml 1% aqueous malt

extract solution in 250 ml flasks, immediately frozen after harvesting, and freeze-dried.

After addition of several glass beads (ca. 2 mm diameter), the samples were ground in

a Retsch 200 mixer mill for 10 minutes. Subsequently, DNA was extracted using the

modified CTAB-protocol described in Riethmiiller et al. (2002).

For DNA extraction from the type specimen of Spirosphaera lignicola (IMI 375019),

a small piece (ca. 2x2 mm) containing only mycelium immersed in agar was cut from

the margin of the dried culture using a sterile scalpel and forceps and placed in a 1.5

ml reaction-tube. Then, 300 ul CTAB buffer and quartz sand were added, the samples

incubated in a thermo block for about 10 minutes at 65 °C and subsequently thoroughly

ground using a conical micro pestle. Afterwards, the sample was again incubated for

additional 50 minutes at 65 °C and further processed like the samples from living

cultures.

The complete ITS rDNA region was amplified with primers ITS4 and ITS5 (White

et al. 1990); the D1, D2 region of the nuLSU rDNA region with primers LROR (Rehner

& Samuels 1994) and TW14 (White et al. 1990). Alternatively, a ca. 1550 bp fragment

containing partial SSU rDNA, ITS1, 5-88 rDNA, ITS2 and partial nuLSU was amplified

with primers V9G (de Hoog & Gerrits van den Ende 1998) and LR5 (Vilgalys & Hester

1990). A ca. 1200 bp fragment of the tef1 (translation elongation factor 1 alpha) gene was

amplified with primers EF1728F (Chaverri & Samuels 2003) and TEF1LLErev (Jaklitsch

et al. 2006). The latter fragment includes the fourth and the fifth intron and a significant

portion of the last large exon. PCR products were purified either by gel electrophoresis

in combination with the QlIAquick Kit (Qiagen) according to the manufacturer's

instructions or an enzymatic PCR cleanup as described in Voglmayr & Jaklitsch (2008).

DNA was cycle-sequenced using the ABI PRISM Big Dye Terminator Cycle Sequencing

Ready Reaction Kit v. 3.1 (Applied Biosystems, Warrington) with primers ITS4, ITS5,

LROR, TW14, V9G, LR3 (Vilgalys & Hester 1990), or LR5, depending on the PCR

fragment, and an automated DNA sequencer (ABI 377 or 3130xl Genetic Analyzer,

Applied Biosystems). The GenBank accession numbers of the sequences obtained in the

present study are listed in the Taxonomy section.

194 ... Voglmayr

Data analysis

For the nuLSU rDNA analysis, first a blast search in the GenBank sequence

database was performed. As the sequences most similar to Dendroclathra all were

from Hypocreomycetidae, representative sequences were selected according to Zhang

et al. (2006) and downloaded from GenBank. The GenBank accession numbers of the

selected sequences are given in the tree, following the taxon names (Fic. 1). The nuLSU

sequence alignment was produced with Muscle version 3.7 (Edgar 2004) and visually

checked with BioEdit (Hall 1999), version 7.0.9.0.

Nucleotide blast searches were done with the complete ITS rDNA region as well as

the exon of the tefl gene the of Dendroclathra lignicola to reveal the closest sequences. A

representative taxonomic selection of sequences (TABLE 1) were aligned with Muscle to

obtain a representative matrix for phylogenetic analysis.

TABLE 1. Taxa and GenBank accession numbers of ITS and tefl sequences

selected for phylogenetic analyses.

SEQUENCES

SPECIES els. aerke

Ambrosiella xylebori Arx & Hennebert DQ471102

Aniptodera chesapeakensis Shearer & M.A. Mill. DQ471064

Balansia henningsiana (Moller) Diehl AY489610

Ceratocystis polonica (Siemaszko) C. Moreau AY233899

Chaetosphaeria tulasneorum Réblova & W. Gams AF178547

Claviceps purpurea (Fr.) Tul. AF543778

Coniochaeta sp. AM262406

Cordyceps bassiana Z.Z. Li et al. AB237657

Doratomyces stemonitis (Pers.) F.J. Morton & G. Sm. DQ836916

Elaphocordyceps capitata (Holmsk.) G.H. Sung et al. AY489615

E. inegoensis (Kobayasi) G.H. Sung et al. AB027368

E. ophioglossoides (Ehrh.) G.H. Sung et al. AY489618

Fimetariella rabenhorstii (Niessl) N. Lundq. EU781677

Glomerella cingulata (Stoneman) Spauld. & H. Schrenk AF543773

G. lindemuthiana Shear EU400135

G. tucumanensis (Speg.) Arx & E. Mill. AY944753

Graphium basitruncatum (Matsush.) Seifert & G. Okada EF165016

G. penicillioides Corda DQ471110

Hypocrea americana (Canham) Overton DQ471043

Lasiosphaeria sorbina (Nyl.) P. Karst. AY587934

Microascus longirostris Zukal DQ836913

M. trigonosporus C.W. Emmons & B.O. Dodge DQ471077

Myrothecium cinctum (Corda) Sacc. AJ302004

M. roridum Tode AY489603

Nectria cinnabarina (Tode) Fr. AF543785

N. mauritiicola (Henn.) Seifert & Samuels AY 138847

Niesslia exilis (Alb. & Schwein.) G. Winter AY489614

Peethambara spirostriata (Rossman) Rossman AY489619

Petriella setifera (Alf. Schmidt) Curzi DQ836911

Porosphaerella cordanophora E. Mill. & Samuels AF178563

Pseudallescheria boydii (Shear) McGinnis et al. AY228112

Scedosporium prolificans (Hennebert & B.G. Desai) E. Guého & de Hoog AY228120

Verticillium dahliae Kleb. AY 489632

Xylaria acuta Peck (outgroup) DQ471048

X. hypoxylon (L.) Grev. (outgroup) DQ471042

X. longipes Nitschke (outgroup) AY909016

X. mali Fromme (outgroup) AF163040

Dendroclathra lignicola comb. nov. ... 195

Maximum parsimony (MP) analyses were performed with PAUP*4.0b10 (Swofford

2002), using 1000 replicates of heuristic search with random addition of sequences

and subsequent TBR branch swapping (MULTREES option in effect, steepest descent

option not in effect, COLLAPSE=maxbrlen). Maximum parsimony bootstrap analyses

were done with the same settings, with ten rounds of random sequence addition and

subsequent TBR branch swapping during each bootstrap replicate. Gaps were treated

as missing data.

Results

Sequence similarity of Dendroclathra caeruleofusca and Spirosphaera lignicola

The complete ITS rDNA sequences of the types of Dendroclathra

caeruleofusca and Spirosphaera lignicola as well as of all Austrian accessions

isolated during the present study were highly similar (sequence similarities of

98.8-100%; sequence differences of 0-6 nucleotides including gaps). The ITS

sequence of the type of S. lignicola was identical to one Austrian collection

(A270), whereas the type of D. caeruleofusca from Cuba was most similar to

another Austrian collection (A266), from which it differed by two nucleotide

substitutions. The tefl sequences were also highly similar (sequence similarities

of 98.8-100%); however, the type of D. caeruleofusca differed from the Austrian

collections by 22-23 nucleotides (almost all in introns 4 and 5), whereas the

Austrian collections showed only 0-6 nucleotide differences from each other.

For the type of S. lignicola, no tef1 sequence was available. All nuLSU sequences

of Dendroclathra revealed were identical.

Phylogenetic relationships

After exclusion of ambiguously aligned regions, 1573 characters of the

resulting partial nuLSU alignment were included in the subsequent phylogenetic

analyses. Of these, 1030 characters were constant; of the variable characters

144 were parsimony-uninformative and 399 parsimony-informative. The MP

analyses of the nuLSU rDNA alignment revealed one most parsimonious tree

of score 1743 (Fic. 1). The topology of the tree is similar to that of Zhang et

al. (2006); however, most of the tree backbone lacked significant bootstrap

support. Dendroclathra was embedded within a paraphyletic Microascales, also

without support. Spirosphaera floriformis, the generic type, was placed within

Leotiomycetes with maximum support (Fie. 1).

Inthenucleotideblastsearch, noITS sequences highlysimilarto Dendroclathra

could be retrieved; the most similar taxonomically identified sequences belonged

to members of Hypocreales, Microascales and Glomerellaceae. The sequence

alignment of these most similar ITS sequences contained numerous indels and

ambiguously aligned positions within both the ITS1 and ITS2 regions. After

exclusion of these regions, 650 characters remained for a phylogenetic analysis,

of which 300 characters were constant, 117 variable characters parsimony-

196 ... Voglmayr

Aniptodera chesapeakensis U46882

Nimbospora effusa U46892

Nohea umiumi U46893

Halosphaeria appendiculata U46885

84 Lignincola laevis U46890

72 Corollospora maritima U46884

Halosphaeriales

100 Varicosporina ramulosa U44092

76 Ceriosporopsis halima U47844

97 ¢ Microascus longirostris AF400865

99 Microascus trigonosporus DQ470958

Doratomyces stemonitis DQ836907

51 Petriella setifera DQ470969

95 r— Graphium penicillioides AFO027384

Graphium pseudormiticum AM943880

100 - Ambrosiella xylebori DQ470979

Ceratocystis adiposa AY281101

Ceratocystis fimbriata U17401

99 Custingophora olivacea AF178566

Gondwanamyces capensis FJ176888

Dendroclathra lignicola A268

Dendroclathra lignicola A270

Dendroclathra lignicola A266

ie Dendroclathra lignicola A269

Dendroclathra caeruleofusca A53 (type)

Nectria cinnabarina U00748

Hypocrea citrina AY544649

Niesslia exilis AY489720

Myrothecium roridum AY489708

95 Peethambara spirostriata AY489724

75 Elaphocordyceps capitata AY489721

Elaphocordyceps ophioglossoides AY 489723

Balansia henningsiana AY489715

100

Microascales

Hypocreomycetidae

Hypocreales

Claviceps purpurea AF543789

100 Melanospora tiffanii AYO15630

Melanospora zamiae AY046579

= Bertia moriformis AY695260 |

100 Coronophorales

Chaetosphaerella phaeostroma AY346274

99 Glomerella cingulata AF543786

Glomerella acutata FJ588234

Verticillium dahliae AY489737

72 Leotia lubrica AY544644

Microglossum rufum DQ470981

Spirosphaera floriformis HQ696658

Botryotinia fuckeliana AY544651

Melanosporales

Glomerellaceae

Leotiomycetes

— 10 changes

Fic. 1. Phylogram of the MP tree revealed by PAUP from an analysis of the nuLSU rDNA matrix of

selected Hypocreomycetidae and Leotiomycetes, showing the phylogenetic affinities of Dendroclathra

to the Microascales. GenBank sequence accession numbers follow the taxon names. MP bootstrap

support above 50% is given above or below the branches. Labels in bold denote sequences obtained

during the present study.

uninformative and 233 parsimony-informative. The MP analysis revealed nine

most parsimonious trees; in all of these trees, Dendroclathra was sister group

of the Microascales clade (Scedosporium prolificans, Pseudallescheria boydii,

Dendroclathra lignicola comb. nov. ... 197

Ceratocystis polonica, Graphium basitruncatum), however, without bootstrap

support (data not shown).

For the tefl gene, a nucleotide blast search revealed members of Hypocreales,

Microascales and Glomerellaceae as closest relatives as well. Of the altogether

1018 nucleotide characters included in the MP analysis, 654 characters were

constant, 86 variable characters parsimony-uninformative and 278 parsimony-

informative. MP analysis revealed a single most parsimonious tree, which

revealed Dendroclathra as sister group to Graphium penicillioides (Microascales)

with moderate bootstrap support (78%; data not shown).

Morphology

The cultures of the Austrian isolates produced conidia very similar to those

of the dried type culture of Spirosphaera lignicola. Conidia in culture differed

from those on natural substrate in being simpler, i.e. the conidial filaments

are rather loosely branched tree-like and less tightly interwoven (Fic. 2a-d),

lacking the tangential branching of tightly packed conidial filaments typical for

older conidia on natural substrate. Therefore, also the typical fusion of conidial

cells upon mutual contact (Fics. 2g, 3j) is not observed in conidia produced in

pure culture. Anisotomous-dichotomous branching of the conidial filament is

present in conidia from pure culture (Fic. 2a) as well as from natural substrate

(Fic. 2e), but mostly less apparent and regular in conidia from pure culture

(Fic. 2b-d). The conidia of the Austrian collections from natural substrate

matched those of the type collection of Dendroclathra caeruleofusca (compare

Fics. 2e-g, 3 with Voglmayr & Delgado-Rodriguez 2001: figs. 1-14). Upon

higher magnification, in SEM the conidial cells are verruculose (Fic. 3k).

Taxonomy

Dendroclathra lignicola (Abdullah, Gené & Guarro) Voglmayr, comb. nov.

MycoBank MB 519527 FIGS. 2, 3

= Spirosphaera lignicola Abdullah, Gené & Guarro, Mycotaxon 66: 268 (1998).

= Dendroclathra caeruleofusca Voglmayr & G. Delgado, Can. J. Bot. 79: 997 (2001).

Type — ITALY. Siena: Monticiano, on submerged twig in a stream, 5 Nov. 1996

S.K. Abdullah (IMI 375019, holotype of Spirosphaera lignicola; ex-type ITS sequence:

EU873531). AUSTRIA. WiEN: Ottakring, Moosgraben, on wood of deciduous tree

submerged in small rivulet, 10 Apr. 2006 A270 (WU 31301 epitype here designated,

ex-epitype culture CBS 122534; ex-epitype sequences: ITS-LSU: EU873530, tefl:

EU873534)

Discussion

This is the first time that sequence data are available for Spirosphaera

lignicola and Dendroclathra caeruleofusca. The molecular data indicate that

both species are conspecific, as their LSU sequences are identical, and there are

only few nucleotide substitutions in the ITS. However, in the tefl sequences,

198 ... Voglmayr

the Cuban isolate of D. caeruleofusca deviates significantly from the European

isolates, which corroborates some genetic differentiation between European

and American isolates indicative of geographic isolation. Conspecificity of the

Austrian isolates with the type of S. lignicola from Italy is also corroborated

by the similarity of conidia produced in pure culture. However, the conidia

produced in pure culture deviate from those produced on natural substrate in

being less complex and atypical, which demonstrates that morphological species

identification based on conidia can be difficult in case of aberrant conidiation

in pure culture. Spirosphaera lignicola has been described solely from conidia

produced in pure culture. Conspecificity with D. caeruleofusca was therefore

not apparent, as the species description and illustrations of S. lignicola were

incomplete, and conidiation was absent in the type culture of D. caeruleofusca.

Conidial morphology of S. lignicola does not fit the genus Spirosphaera. In

S. lignicola, the conidial filaments are never coiled which in combination with

unilateral branching, is the main diagnostic feature of Spirosphaera (Hennebert

1998). In addition, the cells of the conidial filament of S. lignicola are different in

being mostly isodiametric and commonly fusing upon mutual contact, whereas

they are elongated and never fusing in Spirosphaera (Voglmayr & Delgado-

Rodriguez 2001). In addition, S. lignicola is placed within Hypocreomycetidae

(Fic. 1) and therefore phylogenetically distinct from the generic type of

Spirosphaera, which belongs to the Leotiomycetes (Fic. 1; Voglmayr 2004). As

S. lignicola is apparently conspecific with D. caeruleofusca and has priority, it

has to be transferred to Dendroclathra, D. caeruleofusca becoming a synonym

of D. lignicola.

Concerning conidial morphology, Dendroclathra is a typical member of the

aeroaquatic fungi (Voglmayr & Delgado-Rodriguez 2001) in having buoyant

multicellular conidia enclosing air between their cells. The ecology of the

Austrian collections is similar to the Italian and Cuban collections, which also

grew on wood submerged in small streams. This habitat is rather atypical for

aeroaquatic fungi, which are mostly found on litter submerged in stagnant

water bodies characterized by oxygen depletion (Michaelides & Kendrick 1981,

Webster & Descals 1982). Conversely, the habitats of the Austrian collections

appeared to be well aerated, lacking the sulfide smell typical of most aeroaquatic

habitats. This may be the reason why it has only rarely been collected and

not previously recorded for Austria despite the extensive investigations on

aeroaquatic fungi by the author. It may be a rather common species in some

areas, as it has been found at several localities in the vicinity of Vienna and

may have a wide distribution in suitable habitats, at least in the northern

hemisphere.

ADDITIONAL SPECIMENS EXAMINED — AUSTRIA. NIEDEROSTERREICH: Moédling,

Giefhiibl, on wood of deciduous tree submerged in small rivulet, 2 Apr. 2006 H.

Voglmayr A268 (WU 31302; culture deposited as CBS 122533; sequences: ITS-LSU:

Dendroclathra lignicola comb. nov. ... 199

Fic. 2. LM of Dendroclathra lignicola. a-d. Conidia produced on 2% malt extract agar, showing the

anisotomously dichotomous branching of the conidial filament, but no tangential branching and

without fusion of filament cells upon mutual contact. e-f. Immature conidia from natural substrate

in side view (e) and from above (f); in (e) showing the anisotomously dichotomous branching of

the conidial filament. g. Detail from squash mount of mature conidium from natural substrate,

showing densely packed conidial filament fusing upon mutual contact (white arrows). Sources:

a-d. IMI 375019 (type), e-g. WU 31304. Scale bars: a—c, e-g = 10 um, d = 20 um.

Dendroclathra lignicola comb. nov. ... 201

EU873529, tefl: EU873536); Baden, Helenental, Cholerakapelle, on wood of deciduous

tree submerged in small rivulet, 23 Apr. 2006 H. Voglmayr A269 (WU 31303, culture

deposited as CBS 122531; sequences: ITS-LSU: EU873528, tefl: EU873535). WIEN:

Donaustadt, Lobau, Panozzalacke, on wood of deciduous tree submerged in an oxbow

lake, 8 Apr. 2006 H. Voglmayr A266 (WU 31304, culture deposited as CBS 122532;

sequences: ITS-LSU: EU873527, tefl: EU873533); Donaustadt, Lobau, Mihlleitener

Furt, on wood of deciduous tree submerged in an oxbow lake, 21 Apr. 2006 H. Voglmayr

A267 (WU 31305, culture deposited as CBS 122530; sequence: tefl: EU873537). CUBA.

PINAR DEL Rio: Sierra del Rosario Biosphere Reserve, San Juan River, on submerged

bamboo twig, 20 Sep. 2000 G. Delgado-Rodriguez (WU 21361, isotype of Dendroclathra

caeruleofusca; ex-holotype sequences: ITS-LSU: EU873526, tefl: EU873532).

Acknowledgments

Eric McKenzie and Gregorio Delgado-Rodriguez are gratefully acknowledged

for their pre-submission reviews. The curator of IMI is thanked for loan of the type

specimen of Spirosphaera lignicola. Financial support was provided by the Hochschuljub

ilaumsstiftung der Stadt Wien (grant no. H-1162/2003) and by the KIOS of the Austrian

Academy of Sciences (grant P2008-02).

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mycologia.98.6.1076

MY COTAXON

Volume 116, pp. 203-216 April-June 2011

DOI: 10.5248/116.203

Spiroplana centripeta gen. & sp. nov., a leaf parasite

of Philadelphus and Deutzia with a remarkable

aeroaquatic conidium morphology

HERMANN VOGLMAYR’, MI-JEONG PARK? & HYEON-DONG SHIN”

"Department of Systematic and Evolutionary Botany, University of Vienna,

Rennweg 14, A-1030 Vienna, Austria

Division of Environmental Science and Ecological Engineering, College of Life Sciences

and Biotechnology, Korea University, Seoul 136-701, Korea

*CORRESPONDENCE TO: hermann.voglmayr@univie.ac.at & hdshin@korea.ac.kr

ABSTRACT — Spiroplana centripeta is described as a new genus and species from Korea.

Its spirally coiled conidia closely resemble those of the aeroaquatic genus Spirosphaera but

its ecology differs totally, as it is parasitic to living Philadelphus and Deutzia leaves causing

symptoms superficially similar to powdery mildew disease. No sexual state has been found,

but molecular phylogenies inferred from ITS1-5.8S-ITS2 rDNA and partial nuLSU sequences

support it within the Pleosporales (Dothideomycetes) and thus phylogenetically distinct

from the generic type, Spirosphaera floriformis in the Leotiomycetes. Molecular phylogenies

further show that Spirosphaera is polyphyletic and that generic diagnostic characters have

evolved multiple times as an adaptation to conidium dispersal in the aeroaquatic niche.

Morphologically, Spiroplana centripeta differs from Spirosphaera in its branching pattern,

characterised by a main coil of cells in the conidial filament that give rise to 1-2 daughter

filaments only on the inner side of the main coil. The daughter filaments then grow, coiling

inwards with occasional additional branching to produce a tightly interwoven propagule

enclosing air in a manner similar to aeroaquatic fungi. As primary branching takes place in

one plane, the conidia are laterally flattened. In light of these molecular, morphological and

ecological differences, a new genus is described. We believe the astounding similarity of the

Spiroplana and Spirosphaera conidia is related to dispersal on the surface of a water film.

Key worps — leaf pathogen, Philadelphaceae, taxonomy

Introduction

During an inventory on plant parasitic fungi of South Korea, a

remarkable fungal leaf parasite was found on living Philadelphus and Deutzia

(Philadelphaceae) leaves. Although superficially similar to a powdery mildew

by forming a whitish, cobweb-like mycelium on the lower leaf epidermis,

microscopical investigations revealed complex multicellular conidia most

204 ... Voglmayr, Park & Shin

similar to those of the aeroaquatic anamorph genus Spirosphaera Beverw.

However, the latter genus comprises solely saprobic aquatic species inhabiting

organic litter submerged in stagnant to slow-flowing water bodies (Voglmayr

2004) and is therefore ecologically distinct; in addition, the present fungus did

not fit any described Spirosphaera species.

Subsequently, detailed molecular phylogenetic and morphological

investigations were initiated to reveal the systematic and phylogenetic affiliations

of the Spirosphaera-like leaf pathogen. As a result, a new genus and species

are described, acknowledging its morphological, phylogenetic and ecological

distinctness from Spirosphaera.

Materials & methods

Sampling

During 2004-2010 in Korea, 23 samples of a Spirosphaera-like fungus were collected

from Philadelphus schrenkii and three from Deutzia parviflora. The fungus occurred

on host plants growing in shady humid sites located mostly near streams, and it was

collected from July to October, especially in the monsoon season. Herbarium samples

of dried infected leaves are deposited at KUS and WU. Type or authentic cultures of

Spirosphaera beverwijkiana Hennebert (CBS 469.66), S. floriformis Beverw. (CBS

402.52), and S. minuta Hennebert (CBS 476.66) were selected from the CBS (Utrecht,

The Netherlands) for sequencing.

Light microscopy

Descriptions and measurements of cellular structures were based mainly on dried

specimens removed directly from the infected leaves and mounted in tap water or

L4 (general mounting fluid after Cleémencon 1972) for light microscopy. Slides were

examined and photographed using a Zeiss Axio Imager.Al (Zeiss, Jena, Germany)

microscope equipped with a Zeiss AxioCam ICc3 digital camera. Measurements are

reported as maxima and minima in parentheses and the mean plus and minus the

standard deviation of a number of measurements given in parentheses.

Scanning electron microscopy

Fresh infected leaf samples were prepared for scanning electron microscopy (SEM)

according to the method described in Halbritter (1998). The prepared specimens were

mounted on Cambridge stubs, sputter-coated with gold, and examined in a Jeol JSM-

6390LV scanning electron microscope (Jeol Ltd., Japan) at 10 kV and a Zeiss Supra

55VP field-emission scanning electron microscope (Carl Zeiss, Germany) at 2 kV.

Cultures

Conidia were scraped off the leaf lesion into a drop of sterile distilled water to

prepare a conidial suspension, which was streaked onto potato dextrose agar (PDA)

supplemented with streptomycin sulphate (200 ppm). After germination, each

conidium was transferred on PDA and grown at 25°C under a 12-hr photoperiod. Five

monoconidial isolates were successfully obtained in the present study. All the isolates,

listed in TaBLE 1, were deposited at the KACC (Korean Agricultural Culture Collection,

Suwon, Korea).

Spiroplana centripeta gen. & sp. nov. ... 205

TABLE 1. Information on the Spiroplana centripeta isolates cultured and sequenced.

ITS, LSU GENBANK

Host ISOLATE NO. VOUCHER SPECIMEN NO.

ACC. NO,

Philadelphus schrenkii KACC42611 KUS-F22135, WU 31234 HQ696660, HQ696652

P. schrenkii KACC43022 KUS-F22730 HQ696661, HQ696653

P. schrenkii KACC43136 KUS-F22752, WU 31235 HQ696662, HQ696654

Deutzia parviflora KACC43189 KUS-F23006, WU 31237 HQ696663, HQ696655

D. parviflora KACC45741 KUS-F25316, WU 31238 HQ831441, HQ696656

DNA extraction, PCR and sequencing

Mycelia harvested from the colonies grown on PDA were used for genomic DNA

extraction which was carried out according to Lee & Taylor (1990). The ITS rDNA

region of Spiroplana was amplified using primers ITS1 and ITS4 (White et al. 1990),

and nuLSU rDNA region using primers LROR (Moncalvo et al. 1995) and LR7 (Vilgalys

& Hester 1990). The PCR products were purified using a LaboPass PCR purification

kit (COSMO Genetech, Seoul, Korea). For Spirosphaera, the partial nuSSU - complete

ITS - partial nuLSU rDNA region was amplified with primers V9G (De Hoog & Gerrits

van den Ende 1998) and LR5 (Vilgalys & Hester 1990); PCR products were purified

using an enzymatic PCR cleanup as described in Voglmayr & Jaklitsch (2008). DNA was

cycle-sequenced using the ABI PRISM Big Dye Terminator Cycle Sequencing Ready

Reaction Kit v. 3.1 (Applied Biosystems, Warrington) and the PCR primers; in addition,

for the SSU-ITS-LSU fragment the primers ITS4 (White et al. 1990) and LR3 (Vilgalys &

Hester 1990) were used. Sequencing was performed on a 3730xl DNA analyzer (Applied

Biosystems, Foster City, CA, USA). The sequence data were edited using the DNASTAR

computer package version 5.05 (Lasergene, Madison, WI). The newly obtained ITS and

nuLSU rDNA sequences of the five isolates of Spiroplana and of the three Spirosphaera

species were deposited in GenBank.

Data analysis

As a nucleotide BLast search did not reveal highly similar sequences for the ITS

rDNA region of Spiroplana, the ITS was not used for phylogenetic analyses. For the

phylogenetic analyses of the nuLSU rDNA, representative sequences from Leotiomycetes

and Dothideomycetes were selected from GenBank according to the results of nucleotide

BLAST searches of Spiroplana and Spirosphaera sequences. After a first rough phylogenetic

analysis, additional LSU sequences were selected according to the phylogenies of Schoch

et al. (2009) and Zhang et al. (2009) to obtain a representative sampling. The outgroup

included Leotia lubrica (Scop.) Pers., Microglossum rufum (Schwein.) Underw., and

Spirosphaera floriformis (Leotiomycetes). GenBank accession numbers of the selected

sequences are given in the tree, following the taxon names (Fic. 1). In addition, new and

GenBank LSU sequences of verified Spirosphaera species were included.

Sequence alignments were generated with Muscle version 3.6 (Edgar 2004) and

visually checked and refined with BioEdit (Hall 1999), version 7.0.9.0; excessive leading

and trailing gaps were removed. The final LSU matrix contained 1561 characters.

Maximum parsimony (MP) analyses were performed with PAUP* version 4.0

b10 (Swofford 2002), using 1000 replicates of heuristic search with random addition

of sequences and subsequent TBR branch swapping (MULTREES option in effect,

206 ... Voglmayr, Park & Shin

Alternaria alternata DQ678082

Pleospora herbarum DQ678043

T7293

Coniothyrium obiones DQ678054

Cucurbitaria elongata DQ678061

99/99/100

Ophiosphaerella herpotricha DQ767656

Phaeosphaeria eustoma DQ678063

Didymella cucurbitacearum AY293792

Byssothecium circinans AYO16357

100/100/100

91/98/100

Splanchnonema platani DQ678065

97/94/100

Katumotoa bambusicola AB524595

Wie Lentithecium fluviatile GU301825

100/100/100 a i, o

Bimuria novae-zelandiae AYO16356

Phaeodothis winteri DQ678073

Corynespora smithii GU323201

aaex100 fF Lophiostoma arundinis DQ782384

100/100/100

Lophiostoma crenatum DQ678069

Trematosphaeria heterospora AYO16369

74874100

Preussia terricola AY 544686

Westerdykella cylindrica AY004343

Spiroplana centripeta KACC42611

Pleosporales

90/98/97

Spiroplana centripeta KACC43136

Spiroplana centripeta KACC43022

Spiroplana centripeta KACC45741

84/84/-

100/100/100

Spiropiana centripeta KACC43189

ezi7anoo [ Herpotrichia juniperi DQ678080

Trematosphaeria pertusa DQ678072

77+] 10071004100

498

Pleomassaria siparia DQ678078

168/99

Ulospora bilgramii DQ678076

bata Verruculina enalia DQ678079

Spirosphaera cupreorufescens AY616236

Delitschia winteri DQ678077

Capnodium coffeae DQ247800

89/100/100 Davidiella tassiana DQ678074

00° Kae Dothideomycetidae

Mycosphaerella punctiformis DQ470968

Dothidea sambuci AY544681

s00/96/100 Botryosphaeria dothidea DQ678051

Botryosphaeria ribis DQ678053

Tubeufia cerea DQ470982

Tubeufia helicomyces DQ767654

Botryosphaeriaceae

170199

™*— Dothideomycetes

| Tubeufiaceae Microthyriaceae

100/100/100 Microthyrium microscopicum GU301846

100/100/100 Spirosphaera beverwijkiana HQ696657

168/99 Spirosphaera minuta HQ696659

Venturia inaequalis EU035460

Venturia tremulae EU035438

99/100/100 Leotia lubrica AY544644

2SEE GO Microglossum rufum DQ470981 Leotiomycetes

Spirosphaera floriformis HQ696658

100/100/100

| Venturiaceae

— 0.05 substitutions/site

Spiroplana centripeta gen. & sp. nov. ... 207

COLLAPSE=MAXBRLEN, steepest descent option not in effect). All molecular

characters were unordered and given equal weight; analyses were performed with gaps

treated as missing data. Bootstrap analysis with 1000 replicates was performed in the

same way, but using 10 rounds of random sequence addition and subsequent branch

swapping during each bootstrap replicate.

For maximum likelihood (ML) analyses, 500 rounds of random addition of

sequences as well as 500 bootstrap replicates were computed with RAxML version

7.0.4 (Stamatakis 2006) using the GTRMIXI and GTRCAT algorithms, respectively.

GTRCAT efficiently approximates the well-known general time reversible model of site

substitution combined with a gamma distribution (GTR+G) to accommodate among-

site substitution rate heterogeneity. GTRMIXI uses GTRCAT during heuristic search,

but the full GTR+I+G model for the final likelihood computation. Best rearrangement

settings were estimated by RAXML during tree search.

For Bayesian analyses, the GTR+I+G model was implemented. Bayesian analyses

were performed with the computer program MrBayes (version 3.1.2; Huelsenbeck &

Ronquist 2001). Three parallel runs of four incrementally heated, simultaneous Markov

chains were performed over 5 million generations from which every 200" tree was

sampled in each run. The first 1000 trees sampled were discarded, and a 90% majority rule

consensus of the remaining trees was computed to obtain estimates for the probabilities

that groups are monophyletic given the sequence data (posterior probabilities). To test

convergence of runs, the results were analysed using AWTY (Nylander et al. 2008); no

indication of lack of convergence was detected.

Results

The final alignments and the trees obtained were deposited in TreeBASE

(http://www.treebase.org) and are available at http://purl.org/phylo/treebase/

phylows/study/TB2:S11131.

Of the 1561 characters in the nuLSU alignment, 338 were parsimony-

informative. Parsimony analysis revealed 133 MP trees consisting of 1580 steps

(not shown). Topology of the 133 MP trees differed in the deeper nodes of

Dothideomycetes, which collapsed to a polytomy up to the Pleosporales clade in

the strict consensus tree, and within the Pleosporales, where several of the deeper

nodes of the backbone lacking significant support collapsed to a polytomy as

well. Fic. 1 shows the best ML tree (InL = -9699.653), which is fully compatible

with the MP strict consensus tree. Tree topologies of the Bayesian analyses were

fully compatible with the ML tree. The three Bayesian runs revealed almost

identical posterior probabilities (PP).

Fic. 1. Phylogram of the best ML tree revealed by RAXML from an analysis of the nuLSU rDNA

matrix of selected Dothideomycetes and Leotiomycetes, showing the phylogenetic position of

Spiroplana centripeta within the Pleosporales. GenBank sequence accession numbers or KACC

strain numbers (for Spiroplana centripeta) follow the taxon names. MP and ML bootstrap support

above 65% and Bayesian posterior probabilities above 90% are given at first, second and third

position, respectively, above or below the branches. Labels in bold denote sequences obtained

during the present study.

208 ... Voglmayr, Park & Shin

In all analyses, Spiroplana centripeta was placed in the Pleosporales with

high support (Fic. 1); however, its closest relatives within the order could

not be revealed. All species of the morphologically similar anamorph genus

Spirosphaera included in the analyses are unrelated to Spiroplana; in addition,

they are mostly unrelated to each other as well (Fic. 1). Whereas Spirosphaera

floriformis, the generic type, is placed in the Leotiomycetes, the other three

species fall into the Dothideomycetes. Spirosphaera beverwijkiana is closely

related to Microthyrium microscopicum Desm., whereas S. minuta is sister

species to the S. beverwijkiana/Microthyrium clade, but separated by wide

genetic distance (Fic. 1). Spirosphaera cupreorufescens Voglmayr is embedded

within the Pleosporales, representing the species of Spirosphaera most closely

related to Spiroplana.

Taxonomy

Spiroplana Voglmayr, M.J. Park & H.D. Shin gen. nov.

MycoBank MB 519376

Mycelium parasiticum in foliis vivis, partim epiphyllum, partim endoparasiticum. Hyphae

septatae, ramificantes. Conidiophora erecta, mononematosa, septata. Cellulae conidiogenae

integratae, holoblasticae, terminales. Conidia irregulariter globosa vel elongata, lateraliter

compressa, formantur filamentis spiraliter intertextis ramificantibusque; filamentum

primarium centripete gerens filamenta filialia circinata.

TyPE: Spiroplana centripeta Voglmayr, M.J. Park & H.D. Shin

Erymo ocy: Referring to the laterally flattened conidia made up of conidial filaments

branching and coiling in one plane and the morphological similarity to Spirosphaera.

Mycelium parasitic on leaves, partly superficial, partly endoparasitic.

Hyphae septate, branching. Conidiophores erect, mononematous, septate.

Conidiogenous cells terminally integrated, holoblastic. Conidia irregularly

globose to elongated, laterally flattened, formed by branched, tightly spirally

interwoven, septate conidial filaments; the primary conidial filament giving

rise to centripetally growing, coiled daughter filaments at the inner side of the

coil.

Fic. 2. Disease symptoms and LM of Spiroplana centripeta. a. Infected leaves of Philadelphus

schrenkii from above and below, showing the yellowish green to brownish lesions above and

the whitish surface mycelium with abundant sporulation below; black dots are holes caused

by insects. b. Detail of infected leaf underneath, showing whitish spores on conidiophores.

c. Septate hyaline hyphae in surface mycelium with brownish appressoria (white arrows).

d-g. Erect conidiophores arising from superficial mycelium; white arrows denote detachment scars

of conidia; d—f unbranched, g branched conidiophores. h-r conidia showing the inward branching

of the main filament in one plane and the tight coiling of the densely interwoven daughter filaments;

h-l. young, n-r. mature conidia; 1, n, r. with verrucose conidial filament; n. with uppermost cell

of conidiophore still attached. Sources: a. KUS-F22920, b-r. KUS-F23616. Scale bars: a = 1 cm,

b = 200 um, c, h-r = 10 um, d-g = 20 um.

Spiroplana centripeta gen. & sp. nov. ... 209

210 ... Voglmayr, Park & Shin

Spiroplana centripeta Voglmayr, M.J. Park & H.D. Shin sp. nov. Figs. 2-3

MycoBank MB 519377

Mycelium album, partim epiphyllum epidermidem inferiorem foliorum viventium

obtegens, partim endoparasiticum. Hyphae hyalinae vel subhyalinae, septatae, 2.5-10 um

latae. Conidiophora macronematosa, mononematosa, hyalina vel subhyalina, septata,

non vel rarim ramosa, ca 55-240 um longa, 3-8.5 um lata. Cellulae conidiogenae

terminales integratae, holoblasticae. Conidia hyalina, irregulariter subglobosa, diametro

ca 30-105 um, lateraliter compressa, formantur filamentis spiraliter intertextis centripete

ramificantibusque. Filamenta conidialia hyalina, circinata, distincte septata, ad septa

constricta, ca 5-10 um lata, septis ca 3-9 um latis; cellulis ca 7.5-30 um longis, curvatis,

saepe unum vel dua filamenta lateralia circinata centripetalia gerentia.

Type: Korea, Chuncheon, Bongmyeong-ri, 37°46'49"N, 127°48'55"E, 270 m a.s.l., on

Philadelphus schrenkii, 11 Sep. 2006, M.J. Park & H.D. Shin (KUS-F22135 Holotype,

WU 31234 Isotype); ex-type culture KACC42611; ex-type sequences HQ696660 (ITS),

HQ696652 (LSU).

ErymMo_oey: Referring to the primary coil of the conidial filament branching strictly

inwards, with daughter filaments growing towards the centre.

Lesions commonly present on leaves, irregularly polyangular, up to 3 cm in

diameter, with indistinct margins, confluent, causing yellowish to brownish

green discolouration of the host tissues. Mycelium partly superficial, partly

endoparasitic; surface mycelium growing cobweb-like on the abaxial (lower)

epidermis of the leaves, connected with the endoparasitic mycelium via the

stomata. Hyphae hyaline to subhyaline, septate; superficial hyphae frequently

branching, not constricted at the septa, smooth, hyphal cells 2.5-6.5 um wide,

occasionally with thick-walled, brownish, ellipsoid appressoria 8-11 um wide;

endoparasitic hyphae intercellular, branched, knobby, slightly constricted

at the septa, smooth, hyphal cells 2.5-10 um wide. Haustoria not observed.

Chlamydospores not observed. Colonies on PDA growing slowly, attaining

10-15 mm diam. after 30 d at 25°C, creamy white, later becoming pale creamy

brown with aging, raised, covered by dense, velvety aerial mycelium, with

irregular margin, reverse pale yellow, without sporulation. Conidiophores

erect, formed on surface hyphae at an angle of c. 90°, macronematous,

mononematous, hyaline to subhyaline, septate, not constricted at the septa,

mostly unbranched, rarely laterally branched, (56-) 79-160(-237) um long,

(2.8—)3.2-5.6(-8.4) um wide (n = 28), consisting of 3-5 cells. Conidiogenous

cells terminally integrated, holoblastic. Conidial secession schizolytic, scar

flat to slightly convex, without thickened cell wall, not pigmented. Conidia

white in mass, irregularly subglobose, laterally flattened, (36-)47-74(-106)

um long, (29-)34-54(-73) um wide (n = 42), formed by branched, tightly

spirally interwoven filaments consistently growing inwards. Conidial filaments

hyaline, coiled, septate, slightly to strongly constricted at the septa, cells

(7.4—)14-25.9(-30.2) um long, (4.7—)6.9-9.1(-10.2) um wide, (3.2-)4.3-7(-9.2)

Spiroplana centripeta gen. & sp. nov. ... 211

Fic. 3. SEM of conidia of Spiroplana centripeta (KUS-F25295). Arrows denote conidiophores. a,

b. Detached young conidia and surface mycelium. c-d. Immature conidia. e-i. Mature conidia.

Note the spiral conidial filaments branching inwards in one plane and the striate-verrucose surface

ornamentation of the conidial filament. Scale bars: 10 um.

212 ... Voglmayr, Park & Shin

tum at the septa (n = 50); primary filament hook-like, resuming growth after 2/3

to 3/4 of a full coil, giving rise to one or two daughter filaments unilaterally at

the inner side of the coil; daughter filaments growing inwards, strongly coiled

and tightly interwoven with their neighbour coils, retaining air between their

cells.

DISTRIBUTION: known thus far only from Korea.

HABITAT AND HOST RANGE: parasitic on living leaves of Philadelphus

schrenkii Rupr. and Deutzia parviflora Bunge (Philadelphaceae).

ADDITIONAL SPECIMENS EXAMINED: On Philadelphus schrenkii: KOREA. Chuncheon,

Bongmyeong-ri, 37°46'49"N, 127°48'55"E, 270 m a.s.L, 29 Jul. 2004, H.D. Shin (KUS-

F20531); 12 Aug. 2006, H.D. Shin (KUS-F21978); 22 Jul. 2007, M.J. Park & H.D. Shin

(KUS-F22730); 20 Aug. 2007, M.J. Park & H.D. Shin (KUS-F22778); 4 Sep. 2010, M.J.

Park & H.D. Shin (KUS-F25241); 15 Sep. 2010, M.J. Park & H.D. Shin (KUS-F25295);

Hoengseong, Hoengseong recreational forest, 37°32'09"N, 127°07'07"E, 230 m a.s.L., 4

Aug. 2004, H.D. Shin (KUS-F20566); 3 Aug. 2007, M.J. Park & H.D. Shin (KUS-F22752,

WU 31235); 21 Sep. 2007, M.J. Park & H.D. Shin (KUS-F22920); 27 Jul. 2009, M.J. Park

& H.D. Shin (KUS-F24377); Mt. Maebongsan, 37°29'18"N, 127°51'05"E, 220 m a.s.L,

15 Aug. 2008, M.J. Park & H.D. Shin (KUS-F23572); Hongcheon, Bukbang-myeon,

37°48'50"N, 127°50'56"E, 310 ma.s.l., 11 Aug. 2004, H.D. Shin (KUS-F20599); 24 Aug.

2004, H.D. Shin (KUS-F20645); Yeonhwasa temple, 37°48'02"N, 127°51'03"E, 290 m

a.s.l., 18 Sep. 2004, H.D. Shin (KUS-F20724); 21 Aug. 2008, M.J. Park & H.D. Shin (KUS-

F23599); Gangwon natural environment research park, 37°44'41"N, 127°51'59"E, 230 m

a.s.l, 4 Aug. 2006, H.D. Shin (KUS-F21958); Sutasa temple, 37°41'59"N, 127°57'43"E,

195 m a.s.l, 16 Jul. 2009, M.J. Park & H.D. Shin (KUS-F24322); Goesan, Ihwaryeong,

36°45'07"N, 128°01'56"E, 540 m asl, 20 Sep. 2009, M.J. Park & H.D. Shin (KUS-

F24642); Yanggu, Dong-myeon, 38°10'05"N, 127°03'23"E, 360 ma.s.l., 24 Sep. 2009, M.J.

Park & H.D. Shin (KUS-F24668); Pyeongchang, Jangjeon-ri, 37°30'02"N, 128°33'40"E,

460 m a.s.l., 26 Sep. 2010, M.J. Park & H.D. Shin (KUS-F25315); Jeongseon, Aesan-

ri, 37°22'19"N, 128°40'02"E, 340 m a.s.l., 26 Sep. 2010, M.J. Park & H.D. Shin (KUS-

F25328); Yangpyeong, Jungmisan recreational forest, 37°35'45"N, 127°28'08"E, 724

m a.s.l., 28 Aug. 2008, M.J. Park & H.D. Shin (KUS-F23616, WU 31236). On Deutzia

parviflora. KOREA. Hoengseong, Seowon-myeon, 37°31'33"N, 127°52'29"E, 285 m

a.s.l., 15 Oct. 2007, M.J. Park & H.D. Shin (KUS-F23006, WU 31237); 15 Aug. 2008, M.J.

Park & H.D. Shin (KUS-F23571); Pyeongchang, Jangjeon-ri, 37°30'02"N, 128°33'40"E,

460 ma.s.l., 26 Sep. 2010, M.J. Park & H.D. Shin (KUS-F25316, WU 31238).

Discussion

The conidia of Spiroplana most closely resemble those of the anamorph genus

Spirosphaera in having coiled, branched, tightly interwoven conidial filaments,

with usually only a single daughter filament per filament cell (unilateral

branching; Hennebert 1998). However, the conidial morphology of Spiroplana

differs in detail by the consistently centripetal growth of the daughter filaments

and the laterally flattened conidia, which is not observed in Spirosphaera.

Also the genus Clathrosporium Nawawi & Kuthub. shows some similarities to

Spiroplana in having coiled, branched, tightly interwoven conidial filaments;

however, the former differs by opposite (bilateral) branching of its conidial

filament, and by conidia disarticulating at age (Hennebert 1998).

Spiroplana centripeta gen. & sp. nov. ... 213

In addition, ecology is also markedly different: Whereas Spiroplana is

a pathogen of living leaves of Philadelphaceae, members of Spirosphaera

and Clathrosporium are aquatic saprotrophs. Ecologically, Spirosphaera and

Clathrosporium species belong to the aeroaquatic fungi (Voglmayr 2004), which

are characterised by growth on submerged litter in stagnant or slow-flowing

water bodies and by the production of buoyant propagules (Webster & Descals

1981, Michaelides & Kendrick 1982). Most members of this group produce

complex multicellular conidia only above the water level, which enclose air

between their cells, are hydrophobic and therefore buoyant. Dispersal of buoyant

conidia is on the water surface, where they attach to floating litter. Production

of such multicellular dispersal units has evolved multiple times independently

in various lineages of ascomycetes. This is evident also in Spirosphaera, the

members of which are not closely related but highly polyphyletic (Fic. 1), which

requires a revised taxonomy of the group. However, this will only be possible

after detailed morphological and molecular phylogenetic studies, which are not

yet available.

Theastounding similarities of the conidia of Spiroplana to those of aeroaquatic

species and the independent evolution of similar conidia indicate the presence

of similar functional constraints, despite their highly different habitats and

ecology. These are likely to be found in the dispersal of buoyant conidia on a

water film, and in an enhanced attachment of the large conidia on the substrate

enabling rapid colonisation by numerous germination hyphae. Conidia of

Spiroplana are evidently not adapted to wind-dispersal as they are comparatively

large, compact and do not have appendages. Spiroplana has been found in areas

characterised by humid climate and frequent rainfalls in summer in the vicinity

of streams, where dispersal on a water film, dew or via splash drops appears

likely, although this has not yet been observed in nature. A similar example of

a leaf parasite with multicellular complex conidia reminiscent of aeroaquatic

fungi concerns the anamorph genus Cristulariella depraedans (Cooke) Hohn.,

a widespread leaf spot pathogen of Acer spp. (Redhead 1975, Voglmayr &

Delgado-Rodriguez 2003, Narumi-Saito et al. 2006). Comparable cases of

spore adaptation to water dispersal have also been reported for some common

terrestrial foliicolous fungi producing tetraradiate conidia (e.g. Tripospermum

Speg., see Tubaki et al. 1985), which are morphologically similar to those of

Ingoldian aquatic fungi inhabiting leaf litter in rapidly flowing streams (G6ncz6l

& Révay 2006). In aquatic fungi, tetraradiate spores have been shown to be an

adaptation to improved and accelerated attachment to the substrate, a critical

issue for successful substrate colonisation in rapidly flowing water bodies

(Read et al. 1992). The same dispersal constraints may also apply to foliicolous

fungi in humid climates, resulting in the evolution of similar morphological

adaptations in spore shape. A recently detected example ofa leaf pathogen with

214 ... Voglmayr, Park & Shin

tetraradiate conidia similar to Ingoldian fungi is Miricatena prunicola Punith.

& Spooner, the agent of a leaf-spot disease of Prunus serotina (Punithalingam

& Spooner 2011). However, also in this case the dispersal mechanisms remain

to be investigated in detail.

Spiroplana centripeta is only known from a comparatively small area within

Korea, with Philadelphus schrenkii and Deutzia parviflora as only confirmed

hosts. Considering the distribution area of its hosts, which extends to central

China and south-eastern Russia, S. centripeta may have a wider distribution in

East Asia but may have escaped notice due to the rather inconspicuous disease

symptoms. It appears possible that additional hosts from Philadelphaceae are

susceptible to infection. As several species of Philadelphus and Deutzia are

commonly planted as ornamental shrubs in temperate climates world-wide,

the pathogen is of potential horticultural concern. However, considering the

limited disease symptoms observed in the present study, it is unlikely to become

a major threat.

Acknowledgements

We gratefully acknowledge Walter Gams and Roland Kirschner for pre-submission

reviews of the manuscript and Brian Spooner for providing information on Miricatena.

HV is grateful to SYNTHESYS grant NL-TAF-1915 for funding a research visit to CBS.

Financial support was provided by the KIOS of the Austrian Academy of Sciences

(grant P2008-02). This work was also supported by a grant from Regional Subgenebank

Support Program of Rural Development Administration, Republic of Korea.

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MY COTAXON

Volume 116, pp. 217-225 April-June 2011

DOI: 10.5248/116.217

On the variability of spore ornamentation in Laccaria tortilis

(Basidiomycota, Agaricales)

ALFREDO VIZZINI’*, MARCO CONTU?, KUULO KALAMEES;},

ENRICO ERCOLE’, ENZO MUSUMECI’, GABRIEL MORENO’,

Jose Luts MANJON? & PABLO ALVARADO?

‘Dipartimento di Biologia Vegetale - Universita degli Studi di Torino,

Viale Mattioli 25, I-10125, Torino, Italy

?Via Marmilla, 12 (I Gioielli 2), I-07026 Olbia (OT), Italy

*Institute of Agricultural and Environmental Sciences, Estonian University of Life Sciences,

181 Riia St., 51014 Tartu, Estonia

*Wiesendamm 10a, 4057 Basilea (CH), Switzerland

°Dpto. de Biologia Vegetal (Botanica), Univ. de Alcald, 28871 Alcala de Henares, Madrid, Spain

CORRESPONDENCE TO ”: alfredo. vizzini@unito.it

ABSTRACT — “Laccaria tortilis” sensu Clémencon 1984 is described as the new form, Laccaria

tortilis f. clemenconii, based on material from Kamchatka (Asiatic Russia), Switzerland,

Austria, and Italy. ITS rDNA analysis indicates that the new form is molecularly identical to

Laccaria tortilis, although smaller spores with shorter spines distinguish it morphologically.

Scanning electronic micrographs of the spores and a dichotomous key to European Laccaria

species with mono-bisporic basidia, are provided.

Key worps — Agaricomycetes, Hydnangiaceae, taxonomy, biodiversity

Introduction

Laccaria tortilis (Bolton) Cooke is the smallest representative of the genus

Laccaria Berk. & Broome. The species is widespread, known from Europe

(Mediterranean zone included) and both North and South America (Mueller

1992). Its tiny basidiomata usually grow in groups on poor, bare, and very

moist soil and are often found along the course of rivers and streams. It is easily

recognized due to its strongly plicate-striate and deformed pileus, very broad

lamellae, large globose spores [(9.2—)10-14.5(-16) x (8.3-)10-14.5(-16) um]

covered with very crowded, conic to pyramidal echinulae that are 1.4-—3.2(-4)

uum long and < 2.3 um wide at the base (see Mueller 1992: Figs 29a, 55c-d;

Vesterholt 2008: 659, Fig. C), and mono-bisporic basidia (Vellinga 1995;

Mueller 1987, 1992; Cacialli et al. 1996; Vesterholt 2008).

218 ... Vizzini & al.

Mueller designated a collection by P.D. Orton at Tanfield Lodge, Scotland, on

6.9.1969, as epitype for L. tortilis (Mueller 1997). Mueller (1987) had previously

described the collection as a neotype based on its globose spores averaging 13.4

x 13.4 um [“11.5-14.7(-17) x 11.5-14.7(-17) um”] and echinulae “2.3-4 um”

long and “1.3-1.8 um” wide at the base (see Mueller 1987: Fig. 2a).

In his key to the European Laccaria, Clemencon (1984) seems to have reported

a different taxon under the name “Laccaria tortilis’, which is distinguished

from the above collection by slightly smaller spores bearing much shorter

echinulae, only “1-1.5” um long. Clémengon (1984) referred the species with

elongate echinulae to the name “Laccaria echinospora (Speg.) Singer’, a species

concept followed by (among others) Kalamees & Vaasma (1993) and Pazmany

(1992, 1994). However, Agaricus echinosporus Speg. should be regarded as a

heterotypic synonym of Agaricus tortilis Bolton because Spegazzini’s original

material is conspecific with the type collection of Laccaria tortilis (Mueller

1992).

During fieldwork in Kamchatka (Asiatic Russia), K. Kalamees and M.

Vaasma had the opportunity to collect very small specimens of a Laccaria

whose micromorphological features fit Clémen¢gon’s and Pazmany’s concepts

of Laccaria tortilis rather well. The basidiomata were found in Kronok Nature

Reserve, Uzon Caldera on 21.8.1978, growing on a clayey moist grassland

area with pebbles, covered with mosses and sparse herbs. Kalamees &

Vaasma (1993) described this collection, characterized by globose spores with

1.3(-1.6) um long echinulae under the name “Laccaria tortilis (Bolt.) Boud.

sensu Clémengon, Z. Mykol. 50(1): 7, 1984’, thus noting that Clémencon’s

concept did not represent the typical L. tortilis.

While revising the Laccaria species housed in TAAM, we ascertained that

the taxon collected in Uzon Caldera by Kalamees and Vaasma (TAAM 120146)

represents the same taxon as Laccaria tortilis sensu Clémencon (1984), which

apparently differed from the Laccaria tortilis concept typified by Mueller

(19977).

Additional basidiomata representing the small-spored taxon were collected

by Enzo Musumeci, Irmgard Greilhuber, and Marco Contu from Switzerland,

Austria, and Sardinia (Italy), respectively.

In the present work we propose, for the pseudonomen “Laccaria tortilis”

sensu Clémencon, a new form, Laccaria tortilis f. clemenconii, named after

Heinz Clémencon and typified by a Swiss collection.

Materials & methods

Morphology

Macro- and micromorphological descriptions are based on both fresh and dried

material; dried material was reinflated with 3% KOH and mounted in Phloxin B in order

to detect the spore ornamentation. Additional data from the description published by

Spore ornamentation in Laccaria tortilis ... 219

Clémencgon (1984) are reported in brackets. Spore size (excluding ornamentation) is

expressed both as a range and mean value based on 30 randomly chosen spores. Author

citations follow the Index Fungorum website (http://www.indexfungorum.org/Names/

AuthorsofFungalNames.asp).

Herbarium abbreviations follow Thiers (2011). The type material is currently kept

in TO.

SEM photographs

Electronic micrographs were made under a Zeiss DSM 950 SEM following Moreno

et al. (1995).

DNA extraction, PCR amplification, and DNA sequencing

Genomic DNA was isolated from 1 mg of 4 herbarium specimens (Tas. 1) using

the DNeasy Plant Mini Kit (Qiagen, Milan Italy) according to the manufacturer's

instructions. Universal primers ITS1f£/ITS4 were used for the ITS region amplification

(White et al. 1990; Gardes & Bruns 1993). Amplification reactions were performed in

PE9700 thermal cycler (Perkin-Elmer, Applied Biosystems) in a 25 ul reaction mixture

using the following final concentrations or total amounts: 5 ng DNA, 1x PCR buffer (20

mM Tris/HCl pH 8.4, 50 mM KC]), 1 uM of each primer, 2.5 mM MgCl, 0.25 mM ofeach

dNTP, 0.5 unit of Taq polymerase (Promega). The PCR program was as follows: 3 min

at 95°C for 1 cycle; 30 s at 94°C, 45 s at 50°C, 2 min at 72°C for 35 cycles, 10 min at 72°C

for 1 cycle. PCR products were resolved on a 1.0% agarose gel and visualized by staining

with ethidium bromide. PCR products were purified and sequenced by DiNAMYCODE

srl (Turin, Italy). Sequence assembly and editing were performed using Geneious v5.1.6

(Drummond et al. 2009). The sequences are deposited in GenBank under the accession

numbers given in TABLE 1.

Sequence alignment and phylogenetic analysis

Sequences included in the phylogenetic analyses were either generated in this study

(Tas. 1) or retrieved from GenBank. Multiple sequence alignments for ITS fragments

TABLE 1. Laccaria tortilis samples used in this molecular study.

SPECIES : COUNTRY (LOCALITY) : HERBARIUM (ITS) Acc. Nos. : HABITAT

: : date — collector i

1—"L. tortilis Switzerland (Kanton Basel, TO-AVHL14 (JF284353) Under Picea abies

f.clemenconii : St. Chrischona) : 25.04.2008 — E. Musumeci

2—"L. tortilis : Switzerland (Kanton Basel, | TO-AVHL15 holotype (JF284354) : Under Picea abies

f. clemenconii St. Chrischona) 24.05.2008 — E. Musumeci i

3—*L. tortilis : Switzerland (Kanton i TO-AVHLI7 (JE284355) i Near Fagus

f.clemenconii : Solothurn, Rodersdorf) : 06.08.2008 — E. Musumeci : sylvatica, Quercus

4—*L, tortilis Deeg ae! : AH38997 (JE284356) ' Stream bank

f. clemenconii agree? : : 22.06.2008 — M. Contu i

Mee dl ee aU Cal a) ee BP ce Rede Wek EAS coe tind, ghee Waka Mins Manas taniod

5— L. tortilis USA: Colorado (Boulder DBGH20904 (DQ149872) Under Salix, Alnus

: Co. near Hwy. 72) i 23.06.2000 —

[Osmundson et al. 2005]

* = newly sequenced samples.

220 ... Vizzini & al.

were generated using ClustalX 2.0 (Larkin et al. 2007) with default conditions for gap

opening and gap extension penalty. The alignment was slightly edited using MEGA

4.0 (Tamura et al. 2007). Phylogenetic analyses were performed using both Bayesian

Inference (BI) and Maximum Likelihood (ML) approaches. The BI was performed

with MrBayes 3.1.2 (Huelsenbeck & Ronquist 2001) with four incrementally heated

simultaneous Monte Carlo Markov Chains (MCMC) ran over 10 millions generations,

under GTR+F evolutionary model. Trees were sampled every 1000 generations resulting

in an overall sampling of 10,001 trees; the first 2500 trees were discarded as “burn-in”

(25%). For the remaining trees, a majority rule consensus tree showing all compatible

partitions was computed to obtain estimates for Bayesian Posterior Probabilities (BPP).

Branch lengths were estimated as mean values over the sampled trees. ML was performed

with RAxML (Stamatakis et al. 2005) under GTRGAMMA model and using thorough

bootstrap with 20 runs and 1000 replicates. In both analyses a Laccaria laccata sequence

(Genbank accession FJ845416) was used as outgroup.

The ML consensus tree was used merely for comparison with the Bayesian tree

and to support the analysis. However the ML bootstrap and BPP (Bayesian posterior

probability) values over 50% are reported in the resulting tree.

Results

Molecular results

Maximum likelihood and Bayesian inferences were performed on a total of

19 samples, including 15 sequences available from GenBank and four newly

sequenced specimens. Final alignment length was 634 bp. Both maximum

likelihood and Bayesian analyses resulted in the same topology (Fic. 1). In both

analyses the four “L. tortilis” sensu Cleémencon sequences clearly cluster with

the L. tortilis sequence (the tortilis clade) with 100 percent of both BPP and

ML bootstrap values. Taking into account the robustness of the clade, the five

sequences can be considered conspecific.

Taxonomy

Laccaria tortilis f. clemenconii Contu, Vizzini, Kalamees & G. Moreno, f. nov.

MycoBank MB519740

“Laccaria tortilis” sensu Clémencon (1984: 7), Kalamees & Vaasma

(1993: 126), Pazmany (1994: 8) non Laccaria tortilis (Bolton)

Cooke, Grevillea 12: 70 (1884) [Mueller (1987: 306)]

A typo differt habitu regulariore, sporis minoribus spinisque sporalibus (1-) 1.5-2(-2.5)

um longis.

Type — Switzerland, Kanton Basel, St. Chrischona, 480 m a.s.l., 24.V.2008, leg.

E. Musumeci (holotype TO AVHL15).

EryMOLoGy — named in honour of Heinz Clémencon, leading specialist of Laccaria

and other Agaricales.

PiLEus 8-25 mm wide, not fleshy, convex, becoming plane to uplifted, often

depressed at the centre, plicate-striate, margin often undulate-distorted,

Spore ornamentation in Laccaria tortilis ... 221

Laccaria proxima Gus31707

#8 Laccaria proxima 1Q068958

- Laccaria proxima Gque7a77

56/41

Laccaria tortilis pqi4ss72

|— Laccaria tortilis f. clemenconii 1

100/100

Laccaria tortilis f. clemenconii 2 tortilis clade

— Laccaria tortilis f. clemenconii 3

—— Laecaria tortilis f. clemenconii 4

100/92 100/09 -— Lacearia amethystina 09499640

Lon—-—~-—-——= Lacearia amethystina ar4466s

ar x Laccaria montana DQ149865

—_—1_—_—__———\ Laccaria montana 8v486434

Laccaria lateritia rn68s%6

75/51 Laccaria murina ap2\1271

| 100/99 ~~ Laccaria pumila va149864

—- Laecaria pumila 0Q149873

= Laccaria pseudomontana 19149870

t———-—— Laccaria bicolor 4240531

Leanne nnnnnnnnennmmnnnn LaCCALIA laccata FI845416

FicurRE 1. Bayesian tree (GIR+G, 10mil-gen, burn-in 25%); BPP values and MLB values

(_/_). Numbers (1-4) refer to the collections reported in TaBLe I. The bar indicates number of

substitutions per site.

fulvous to salmon-pink or pale pink, fading buff, finely fibrillose. LAMELLAE

distant, thick, very broad, sinuate to adnate, occasionally subdecurrent to

decurrent, pink, white-pruinose in old basidiomata. STrpE 8-25 x 1-2(-3)

mm, rather short, gracile, mostly equal, polished or slightly fibrillose-sericeous,

concolorous with the pileus, basal mycelium white. CONTEXT very scarce, pink,

darker towards the stipe base, unchanging. SMELL indistinct to agreeable, at

times fruity. TasTE indistinct. SPORE-PRINT white.

Spores 10-12(-15) x 10-12(-14.5) um, mostly 11 x 11 um, globose,

hyaline, thick-walled, strongly echinulate, echinulae very crowded, pyramidal,

(1-)1.5-2(-2.5) um long, mostly 1.5 um long, 1-1.5 um wide at the base, apex

acute (Fic. 2a-f). Basip1A 45-60 x 9-12 um, clavate, stout, mostly monosporic

or bisporic, rarely intermixed with scarce tri- or tetrasporic ones, with basal

clamp-connection; SUBHYMENIUM ramose; HYMENOPHORAL TRAMA regular,

made up of cylindrical, subhyaline 4.5-15 um wide hyphae. MARGINAL CELLS

scarce, filamentous or subclavate, 4-6 um wide, hyaline, thin-walled. PILEUs

SURFACE consisting of a poorly differentiated cutis made up of rather compact,

cylindrical hyphae, 4.5-12 tm wide, rarely interwoven and with scattered

fascicles of perpendicular hyphae, with a pale brown intraparietal and plasmatic

pigment. CLAMP-CONNECTIONS abundant.

222 ... Vizzini & al.

HABITAT AND DISTRIBUTION. Gregarious on clayey moist areas covered

by mosses, often in small fascicles of 3-5 basidiomata. Autumn. Known from

Kamchatka (Asiatic Russia), Switzerland, Austria, and Sardinia (Italy).

ADDITIONAL MATERIAL STUDIED: RUSSIA, “Fungi Kamtschatici’, Tricholomataceae,

Laccaria tortilis (Bolt.) S.EGray ss Moser 1978, Regio kamtschatica, Kronoki

looduskaitseala, Uzoni kaldeera, j6elamm, 21.8.1978, leg. K. Kalamees, det. M. Vaasma,

TAAM 120146 (rev. a K. Kalamees 5.11.1992 atque rinominata Laccaria tortilis

ss Clémengon)”; SWITZERLAND: KANTON BasEL, St. Chrischona, 460 m a.s.l.,

25.04.2008, leg. E. Musumeci, (TO AVHL14); KANTON SOLOTHURN, Rodersdorf,

06.08.2008, leg. E. Musumeci (TO AVHL17); AUSTRIA: Braunau am Inn district, St.

Peter am Hart community, Hartwald, 370 maz.s.l., 20.09.2009, leg. R. Krisai (TO AVHL22

& WU); ITALY: Sarpin1a, Olbia-Tempio, Mount Limbara, Vallicciola, 22.06.2008, leg.

M. Contu (AH 38997).

Discussion

The spore ornamentation made up of very crowded pyramidal echinulae,

1.0-1.5 um wide at the base, the strictly globose spores, and the mono-bisporic

basidia undoubtedly place Laccaria tortilis sensu Clémencgon, Pazmany,

and Kalamees & Vaasma within the L. tortilis-complex. As pointed out in

the introduction, the new form and forma ftortilis are readily differentiated

based on spore size and ornamentation. Nonetheless, molecular studies

(Fic. 1) show that they represent a single species with rather variable spore

ornamentation. According to Mueller (1992), Laccaria tortilis var. gracilis Peck

is synonym of L. ohiensis (Mont.) Singer. There is another small-sized bisporic

Laccaria producing spores with equally crowded echinulae — L. pumila Fayod

(= L. altaica Singer; but see Sivertsen 1993 and Vesterholt 2008) — but it is

separated from L. tortilis by its ellipsoid spores with much shorter echinulae

(usually 0.5-0.8(-1) um long; Trimbach 1978, Bon 1983, Ballero & Contu

1989, Mueller 1992, Contu 2003) and ITS sequences (Fic. 1). Another small-

sized species, L. nana Massee, differs in dark cinnamon colouration, a non-

striate pileus, and globose spores with echinulae measuring (2.3-)2.8-4 um

long and 1.8-2.3 um wide at the base (Mueller 1992). In Laccaria, at least

one infraspecific taxon has been described with spores bearing shorter spines

than those of the type: Laccaria masoniae var. brevispinosa McNabb, from

New Zealand, has echinulae 1.2-1.5 um long, while the echinulae are 2-3.5

um long in L. masoniae G. Stev. var. masoniae (McNabb 1972). These last two

taxa, not known from Europe, clearly differ from L. tortilis in their violaceous

tinged basidiomata. Infraspecific variants characterised by spore echinulae

longer than those of the type have been described elsewhere, such as Laccaria

macrocystidiata var. longispinosa Contu (Contu 2003). Therefore, we do not

regard spore ornamentation as a good taxonomic marker, at least in some cases,

to delimit species concepts in Laccaria.

Spore ornamentation in Laccaria tortilis ... 223

FiGuRE 2. Laccaria tortilis f. clemenconii. Spores (SEM photographs). a-b. Collection TO AVHL15

(Switzerland, holotype). c-d. Collection TO AVHLI17 (Switzerland). e-f. Collection AH 38997

(Italy). Bars = 2 um.

224 ... Vizzini & al.

Key to the European bisporic species of the genus Laccaria

1 Mycelium at stipe base violaceous; in moist areas among bog mosses

(rare, known only from Spain) ........ L. violaceibasis Contu & Fernandez Sas.

Mycelium-atistipe base white + 2. 25-80 22 inc Wench lnc” bawje' Gale cS 2

ps Basidiomata reddish, with evident lilac hues, odour pronounced, fruity

CTEVOTUATIC im. 17 Fe, EF Pte Gp eitins beatin, abeaieheagteesehos oc lekan a L. vinaceorosea Contu

2" “Basidiomata lacking lilac hites;Hdour NOLIAMEy. «ies. ciitese dwn aateaotn a neot 3

3 DPOLES el MHL ONMA VOR ARE c:-< acest, esses ae cinnrneiete Se sN Gadd ashe Bye wd 4

3 Sporesdarsety 2-11 nto: AVOTACe Fence ee dae dats Schl eden ata hese 5

4 Spore echinulae 1.3-1.8-2 um long, quite spaced, pyramidal; pileus

scaly-areolate, striate only towards the margin, stipe fibrillose-striate;

TUOMAS WOODUS Ts sac ah se nrotan meretaen ee mene ye L. bisporigera Contu & Ballero

4 Spore echinulae 0.8-1 um long, conical, crowded; pileus almost smooth,

strongly striate, stipe slightly fibrillose; mainly in Mediterranean areas,

with Acacia, Eucalyptus and Cupressus ........+00.-000- L. lateritia Malencon

5 Spores subglobose, broadly ellipsoid to oblong ............... 00... e eee eee 6

5 Spores pertectly ClOD Uses fa. sety Pa. acters a Malahat mel alciee Bl pcre ba Athale soAE ae dale 7

6 Spores 9.5-15 x 8-12 um, echinulae 0.5-0.8 um long ............... L. pumila

6 Spores 10-12 x 9-10 um, echinulae 0.5-1 um long

PGA ee, Sine, Vivabcte Meottely: Mottin Wine tiee incest “L. vinaceorosea” [forma ined. |

7 Spores 13 um on average, with echinulae < 3.2(-—4) um long and

2S Wide at thease, 4) os vgtrteceed esa dns wa hiapes haha L. tortilis f. tortilis

7 Spores 11 um on average, with echinulae < 2(-2.5) um long and

<1 5 tanewideat-the-base.:? pt.ns Aid othe) wes! et L. tortilis f. clemenconii

Acknowledgements

We are grateful to Mr. A. Priego and Mr. J.A. Pérez of the Electron Microscopy

Service of the University of Alcala de Henares for their invaluable help with the SEM.

We also thank Luis Monje and Angel Pueblas of the Department of Drawing and

Scientific Photography at the Alcala University for his help in the digital preparation

of the photographs, Vladimir Antonin (Brno, Czech Republic) and Irmgard Greilhuber

(Wien, Austria) for their pre-submission reviews, and Dr. Shaun Pennycook for the

nomenclatural review.

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btm404

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MY COTAXON

Volume 116, pp. 227-229 April-June 2011

DOI: 10.5248/116.227

Validation of Kwoniella heveanensis, teleomorph of the

basidiomycetous yeast Cryptococcus heveanensis

SHENG SUN’, BANU METIN”“, KEISHA FINDLEY” ®,

ALVARO FONSECA’ & JOSEPH HEITMAN”

"Department of Molecular Genetics and Microbiology, Duke University Medical Center,

Durham, North Carolina 27710 USA

? Centro de Recursos Microbiologicos (CREM), Departamento de Ciéncias da Vida,

Faculdade de Ciéncias e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal

4 Current address: ATQ Biyoteknoloji, 7. Cad, No: 73/A,

Birlik Mahallesi 06610, Cankaya, Ankara, Turkey

® Current address: National Human Genome Research Institute,

National Institutes of Health, Bethesda, Maryland, USA

*CORRESPONDENCE TO: heitm001@duke.edu

ABSTRACT— Kwoniella heveanensis, recently published in an electronic journal and

without a designated holotype, is validated as the name of the newly discovered teleomorph

of Cryptococcus heveanensis.

Keyworps— Tremellales, mating, ex-type culture

Introduction

Cryptococcus heveanensis (Groen.) Baptist & Kurtzman is a basidiomycetous

fungus closely related to the human pathogenic fungus Cryptococcus neoformans

(Baptist & Kurtzman 1976, Findley et al. 2009, Metin et al. 2010). The sexual

life cycle of this species was recently discovered and the teleomorph named

as Kwoniella heveanensis (Metin et al. 2010). However, this name was invalid,

because no holotype was designated (McNeill et al. 2006: Art. 37), and the

publication was in an electronic-only journal (McNeill et al. 2006: Art. 29).

We here fulfill the requirements for valid publication of Kwoniella

heveanensis.

Kwoniella heveanensis Metin, K. Findley & Heitman, sp. nov. FIG. 1

MycoBank MB519319

“Species heterothallica. Hyphae dikaryoticae, fibulae fusae. Basidia aggregata, primum

submersa, deinde superficialia, globosa, 7.5-10.5 um diam, septis cruciatis divisa,

228 ... Sun & al.

”. : ae 2a .4-

Fic. 1 Different stages of the sexual cycle of Kwoniella heveanensis. A, differential interference

contrast (DIC) image showing hyphae with fused clamp connections (CC, indicated by black

arrows); B, fluorescence image of the same field as in A, stained with Sytox green, showing the

dikaryotic hyphae; C, globose basidia with cruciate septa (indicated by black arrows); D-F, DIC

images of basidial clusters: D, the early stage (~2 weeks) at which basidial clusters are embedded in

the agar; E, a young basidial cluster (~3 weeks) embedded in the agar; F, mature basidial clusters

(~4 weeks) on the surface of the agar.

sterigmatibus carentia. Basidiosporae subglobosae, 2.3-3.3 um diam, cito conidia

proferentia.” (Metin et al. 2010: 15).

Ho.LoryPe: Sexual product of mating between Cryptococcus heveanensis strains CBS

569 and BCC 8398, preserved as a permanent slide (BPI 881007).

“Heterothallic fungus. Hyphae dikaryotic, clamp connections fused. Basidia

clustered, submerged initially, finally on aerial hyphae, globose, 7.5-10.5 um,

cruciately septate, sterigmata not observed. Basidiospores subglobose, 2.3-3.3

um, germinating by conidia.” (Metin et al. 2010: 15).

COMMENTS— Strains CBS 569 and BCC 8398 (also deposited in CBS as CBS

12232) should be designated as the ex-type strain and the isotype strain,

respectively, for the teleomorph.

Acknowledgements

The authors thank Teun Boekhout, Jack Fell, Walter Gams, June Kwon-Chung and

Marelize van Wyk for pre-submission review of the manuscript, invaluable advice,

and encouragement. We also thank Dr. Jack Fell and Dr. Cletus Kurtzman for their

critical review of this paper. These studies were supported by National Institutes of

Health (NIH)/National Institute of Allergy and Infectious Diseases (NIAID) R37 award

AJ39115 and RO1 award AI50113 to Joseph Heitman.

Kwoniella heveanensis validated ... 229

Literature cited

Baptist JN, Kurtzman CP. 1976. Comparative enzyme patterns in Cryptococcus laurentii and its

taxonomic varieties. Mycologia 68: 1195-1203. doi:10.1016/S0953-7562(09)80616-3

Findley K, Rodriguez-Carres M, Metin B, Kroiss J, Fonseca A, Vilgalys R, Heitman J. 2009.

Phylogeny and phenotypic characterization of pathogenic Cryptococcus species and closely

related saprobic taxa in the Tremellales. Eukaryotic Cell 8: 353-361. doi:10.1128/EC.00373-08

McNeill J, Barrie FE Burdet HM, Demoulin V, Hawksworth DL, Marhold K, Nicolson DH, Prado J,

Silva PC, Skog JE, Wiersema J, Turland NJ. 2006. International Code of Botanical Nomenclature

(Vienna Code). Adopted by the Seventeenth International Botanical Congress, Vienna, Austria,

July 2005. Regnum Vegetabile 146. 568 p.

Metin B, Findley K, Heitman J. 2010. The mating type locus (MAT) and sexual reproduction of

Cryptococcus heveanensis: insights into the evolution of sex and sex-determining chromosomal

regions in fungi. PLoS Genet 6(5): e1000961. doi:10.1371/journal.pgen.1000961

MY COTAXON

Volume 116, pp. 231-245 April-June 2011

DOI: 10.5248/116.231

Observations on /nocephalus virescens comb. nov. and

Alboleptonia stylophora from northeastern Queensland

Davip L. LARGENT’ & SANDRA E. ABELL- Davis?

‘Biological Sciences, Humboldt State University, 1 Harpst St, Arcata CA 95521 U.S.A.

School of Marine and Tropical Biology, Australian Tropical Herbarium,

James Cook University, PO Box 6811, Cairns QLD 4870 Australia

* CORRESPONDENCE TO: mrp@humboldtl.com

ABSTRACT —Inocephalus virescens (= Leptonia virescens, = Entoloma virescens) is proposed

as a new combination and reported for the first time for northeastern Queensland;

Alboleptonia stylophora represents a first record for Australia. Macromorphological and

micromorphological features are provided for both species.

Key worps — Entolomataceae, Agaricales, Basidiomycota, new record

Introduction

Records of the Entolomataceae for Australia can be found in several

publications (Berkeley 1859, McAlpine 1895, Cooke 1892a, b, Massee 1898,

1899, Cleland 1927, 1931, 1933, Pegler 1965, Horak 1980, May & Wood 1997,

Grgurinovic 1997, Baroni & Gates 2006, Gates & Noordeloos 2007, Noordeloos

& Gates 2009, Gates et al. 2009). None of the cited collections, however, were

found in northeastern Queensland nor are any listed in the only regional

checklist published for Queensland (Bailey 1913).

Forster, Booth, Jensen, and Young, who made 18 collections representing

Entoloma sensu lato during several forays to the Wet Tropics rainforest region

in January-April 2001-2002 and November 2001, were the first to deposit

entolomatoid collections from northeastern Queensland into an herbarium

(Fechner pers. comm.). The first author (DLL), who examined the collections

in 2010, found that among the seven correctly referred to the Entolomataceae,

one represented a first record of Alboleptonia stylophora from Australia.

The information herein covers fungi collected during forays in northeastern

Queensland from the summer wet season through the early dry (autumn)

season in 2009 and 2010. A new combination, Inocephalus virescens, is proposed

for Entoloma virescens, previously known only from southeastern Queensland

232 ... Largent & Abell-Davis

(Fraser Island) to Victoria (Young 2005). Complete descriptions are provided

for I. virescens and Alboleptonia stylophora. ‘This constitutes a first record of

I. virescens for northeastern Queensland and of A. stylophora for Australia.

Materials & methods

Collections were made during February—April in 2009 and 2010 from various

northeastern Queensland localities in rainforests within the Wet Tropics bioregion.

Basidiomata collected in the field were carefully stored in plastic containers and

returned to the laboratory. GPS coordinates for each collection were taken in the field

using a Garmin GPSmap 60CSx. Macroscopical features were described from fresh,

recently collected materials. Colors were described subjectively and coded according to

Kornerup & Wanscher (1978) with color plates noted in parentheses. Color reference

abbreviations indicate page number, column(s), and row(s) (e.g., 8D-F5-6 = page 8,

columns D-F, rows 5-6).

Micromorphological features were examined from dried specimens with a

trinocular research-grade Nikon Labophot compound microscope fitted with bright

field optics following general protocols set forth in Largent (1994: 1-3) and techniques

used for measuring spores of the Entolomataceae set forth by Baroni & Lodge (1998:

681). Measurements of cuboid or rhomboid spores were in profile view with length

and width measured midway along the length and the width of each spore. Digitized

microphotographs were taken using a Nikon Coolpix 990 focused through the trinocular

head of the compound microscope.

All microscopic measurements were obtained using a GTCO Corporation Graphic

Digitizer, Model DP5A-111A connected to an IBM compatible Chem Book Laptop

computer. The software utilized was Measure Me 101 v 1.0 modified for laptop use from

a BASIC program (Metrics5) developed and provided by David Malloch, University

of Toronto. Factors determined using this program include: arithmetic means (x,) of

basidiospore lengths and widths + standard deviation measured for n objects; quotient

of basidiospore length by spore width (E) indicated as a range variation in n objects

measured; the mean of E-values (Q) + standard deviations. The sample size (n) = total

number of microscopic structures measured (x) divided by the number of basidiomata

studied (y), as shown in the formula n = x/y.

Taxonomy

Inocephalus virescens (Sacc.) Largent & Abell-Davis, comb. nov. PLATES 1-2

MycoBank MB 519313

= Agaricus virescens Berk. & M.A. Curtis, Proc. Amer. Acad. Arts

4: 116 (1860), nom. illegit. (non Schaeff. 1774).

= Leptonia virescens Sacc., Syll. fung. 5: 714 (1887).

= Entoloma virescens (Sacc.) E. Horak ex Courtec., Mycotaxon 27: 131 (1986).

BASIDIOMATA blue when damaged, becoming yellowish- or olive-green

(30A-D5-6) in places and when mature or drying becoming entirely grayish-

green (24-25E5); when very young pileus and stipe with whitish squamules.

PitEus 8-33 mm broad, 9-20 mm high, at first conic to conic-convex and

at times acutely umbonate, expanding to parabolic to conic campanulate or

Inocephalus virescens comb. noy. (Australia) ... 233

PraTE 1. Inocephalus virescens. A: Basidiomata, 3 hours old (DLL 9972); B: Basidiomata, 2.5 days

old (DLL 9972); C: Basidiomata changing to gray-green (DLL 9972); D: Off-white squamules and

fibrils (DLL 9972).

campanulate and at times broadly umbonate, blue (23E5) becoming quickly

yellowish-, olive- or grayish-green on exposure or handling, when very young

with whitish appressed squamules that become appressed to the surface then

appearing matted appressed fibrillose, dull, opaque; margin decurved at all

times, entire then eroded and eventually lobed-eroded and splitting radially.

TasTE bitter at first, quickly becoming acrid. Opor indistinct. CONTEXT blue

becoming greenish when exposed or damaged. LAMELLAE at first dark to dull

blue (23DE5-6) becoming greenish-blue (25D4) when bruised or damaged,

adnexed at all times, close to subdistant becoming subdistant to distant,

moderately broad then ventricose and sigmoid (8-20 mm long x 1.5-5.0 mm

high); margin lighter blue and eventually + concolorous. STIPE 32-62 mm

long, 1.5-2.0 mm broad at apex, 1.5-5.0 mm broad at base, equal to clavate,

often flattened and then 2.5-4.0 x 3.0-5.0 mm broad at the apex and 3-5 x

5-6 mm broad at the base, becoming longitudinally grooved when mature,

234 ... Largent & Abell-Davis

strongly pruinose at the apex, matted appressed-fibrillose elsewhere, when very

young and fresh with whitish squamules that quickly become appressed to the

surface, fibrils then bluish-gray (23B3) then nearly concolorous with the pileus

and becoming yellowish-, olive-, or grayish-green on aging or when damaged;

whitish towards the base.

Basip1ospores rhomboidal in profile or side views, mostly 4-angled

and rectangular, sometimes 5-angled (particularly in dorsi-ventral view),

isodiametric to heterodiametric, isodiametric and 5-angled in polar view,

6.9-12.2 x 6.9-9.9 um (x, = 9.31 + 1.05 x 8.84 + 0.75 um; E = 0.90-1.40;

Q = 1.06 + 0.11; n = 58/2). Basrpra cylindro-clavate, broad, tapering to a

narrow, curved foot, 42.7-53.1 x 10.8-13.5 um (E = 3.54-4.79; Q = 4.2;

n = 7/1). CHEILOCysTIpIA abundant forming a sterile layer in places, clavate

with brownish contents in 3% KOH and abundant lipoid bodies, 46.1-116.0

x 8.7-13.0 um (x, = 74.49 + 22.58 x 11.16 + 1.67; E = 3.98-12.48; Q = 6.87

+ 2.62; n = 10/1). PsEUDOCysTIDIA present on the lamellar edge and faces,

often cylindric, at times cylindro-clavate, mostly contorted and strangulated,

staining golden brown in 3% KOH, 26.7-72.7 x 2.0-6.0 um (x, = 47.5 x 4.0

um; E = 6.33-28.21; Q = 13.25; n = 9/1). HYPHAE OF LAMELLAR TRAMA with

abundant oleiferous hyphae that stain reddish-brown in 3% KOH, many with

light brown colored contents, some strongly pigmented, with two distinct

strands of the dense and crowded oleiferous hyphae in the subhymenium,

195-460 9.1-21.5 um. PILEIPELLISan entangled layer ofhyphae. PILEOCYSTIDIA

long and cylindro-clavate, 77.9-243.4 x 10.1-13.6 um. HYPHAE OF THE PILEAL

TRAMA with abundant oleiferous hyphae. STIPITIPELLIS similar to the pileipellis

except in places with clusters of hymenial elements. CauLocystip1< similar

in shape to the pileocystidia, 50.0-158.1 x 9.2-15.1 um. OLEIFEROUS HYPHAE

abundant in the pileal, lamellar, and stipe tramas. Lrporp BopieEs abundant

in the tramal hyphae and obscuring microscopic features. PIGMENTATION

cytoplasmic and yellowish-green in the cheilocystidia, pileipellis, stipitipellis,

and some hyphae of the lamellar trama, and exuding brownish color in spot

plate depression filled with 3% KOH; dark golden brown in the pseudocystidia.

CLAMP CONNECTIONS abundant at base of basidia, basidioles, cystidia and on

the hyphae of the pileipellis and stipitipellis.

HABIT, HABITAT, AND DISTRIBUTION Solitary amongst leaf litter in a simple

microphyll vine-fern forest beneath Balanops australiana F. Muell. and Agathis

atropurpurea B. Hyland (DL Largent 9772); or scattered on Calamus L. trunk

with Oraniopsis J. Dransf. & al. abundant in understory (Costion 2234); or

scattered to gregarious throughout CSIRO permanent plot EP30 (a very tall to

extremely tall closed forest that is a transitional simple notophyll to microphyll

vine-fern forest (SN/MiVFF), with a subcanopy dominated by Ceratopetalum

succirubrum C.T. White, Brombya platynema F. Muell. and Cyathea rebeccae

Inocephalus virescens comb. nov. (Australia) ... 235

PLATE 2. Inocephalus virescens. A: Basidiospores (DLL 9772); B: Basidiospores 4-angled, 5-angled

dorsi-ventral view (white arrow) (DLL 9772); C: Basidiospore in profile view (DLL 9771);

D: Basidiospore in dorsiventral view (DLL 9771); E: Pigmented cheilocystidia (DLL 9772);

F: Basidia and pseudocystidia originating from oleiferous hyphae (DLL 9772); G: Tramal hyphae

with lipoid bodies (DLL 9772); H: Stipitipellis showing caulocystidia as terminal cells (DLL 9772).

236 ... Largent & Abell-Davis

(F. Muell.) Domin conspicuous in the understory (DL Largent 9791). Species

in the Inocephalus virescens complex occur in the neotropics, the paleotropics,

in New Zealand, and along the eastern Australian coast from northeastern

Queensland to Victoria.

COLLECTIONS EXAMINED: AUSTRALIA. New SouTH WALEs, Hunter Region, Myall

Lakes National Park, Seal Rocks Area near Seal Rocks road, 2 May 2010, DL Largent

9972 (DAR) (32°25'05.3"S; 152°28'27.5"E; 50.9 m); QUEENSLAND, Cook Region,

Brooklyn Wildlife Sanctuary, Mt Lewis, 5 March 2010, DL Largent 9771 (BRI)

(16°36'05.16"S; 145°16'21"E; 1019.9 m); Mt Windsor National Park, 5 March 2010, DL

Largent 9772 (BRI, CNS) (16°16'00"S; 145°04'00"E, 1073 m); Daintree National Park,

Cape Tribulation Section, Mt Sorrow track, 12 December 2009, Costion 2234 (BRI)

(16°04'36.6"S; 145°25'55.2"E; 781.8 m).

DISTINCTIVE CHARACTERS Inocephalus virescens from northeastern Queensland

is distinctive in its 1) blue, moderate-sized basidiomes that become yellowish-

to grayish-green with age and on handling; 2) its rhomboidal, mostly 4-angled,

sometimes 5-angled basidiospores that average <10 um in length; 3) its clavate

cheilocystidia with yellowish-green cytoplasmic contents that are distinct from

the pseudocystidia; 4) its pileipellis and stipitipellis composed of entangled

hyphae with cylindro-clavate pileocystidia and caulocystidia; 5) its whitish to

off-white squamules on the pileus in young material; and 6) its bitter, then

latently acrid, taste.

Comments ‘The cuboid spores, long basidia, strangulated and contorted

pseudocystidia that originate from abundant oleiferous hyphae in the

subhymenium, conic-campanulate pileus with appressed-fibrils, and abundant

clamp connections (at least in the pileipellis) diagnose the collections from

northeastern Queensland as an Inocephalus species. Based on basidiomata

stature and colors, the 4-angled spores, the conic to campanulate blue pileus that

becomes grayish-green, and the pseudocystidia, our northeastern Queensland

and New South Wales collections would be identified as Rhodophyllus

holocyaneus Romagn. using the keys in Romagnesi (1941) and Romagnesi &

Gilles (1979) or as “Entoloma virescens” using the keys in Horak (1976, 1980).

Our collections, for the most part, also match descriptions of “Entoloma

virescens’ sensu Noordeloos & Hausknecht (2007) and Entoloma aeruginosum

Hiroe (= Rhodophyllus aeruginosus Hiroe) Hongo sensu Courtecuisse (1986).

Our collections also nearly match a description of I. virescens from the Lesser

Antilles (Baroni unpublished).

Rhodophyllus holocyaneus

Rhodophyllus holocyaneus from Madagascar and Gabon is characterized

by cylindro-clavate pileocystidia, clavate cheilocystidia, abundant oleiferous

hyphae that form pseudocystidia, abundant granules, and clamp connections

(Romagnesi & Gilles 1979). All those features are found in our collections.

Rhodophyllus holocyaneus can be distinguished from the Queensland

Inocephalus virescens comb. nov. (Australia) ... 237

collections by its mild taste, strictly cuboid (4-angled) basidiospores (9-11 um

in Romagnesi 1941, 10-11.5 um in Romagnesi & Gilles 1979) averaging >10

um, and only blue-green colors.

Romagnesi (1941) described two species that differed in color from

R. holocyaneus — R. psittacinus Romagn. with parrot-green colors that change

to yellow and R. cubisporus Pat. with uniformly indigo-blue colors without

green tinges. Romagnesi & Gilles (1979: 407), who found collections with

intermediate colors in Gabon, suggested that Rhodophyllus holocyaneus,

R. psittacinus, and R. cubisporus might be polychromatic forms of the same

species, recapitulating Romagnesi’s original hypothesis in 1941.

“Entoloma virescens” sensu Horak and sensu Noordeloos & Hausknecht

Agaricus virescens Berk. & M.A. Curtis from the Bonin Islands is the type-

bearing name. That species is characterized by blue colors that become green, a

centrally depressed pileus, and broad distant adnexed lamellae. After examining

the holotype of Agaricus virescens, both Romagnesi (1941) and Horak (1976)

determined the pileus to be conical rather than depressed and the basidiome

nolaneoid rather than leptonioid in stature. The holotype is in very poor shape

and only the basidiospores could be studied. Courtecuisse (1986) found the

cuboid spores to measure 8.8-11 um.

With respect to “Entoloma virescens” sensu Horak (1976, 1980), the

basidiospores average <10 um [(7—)8-10 um] and the pseudocystidia resemble

those from our Australian material, but Horak’s taxon differs in mild taste and

lack of clavate cheilocystidia. Horak also reported both the presence and the

absence of clamp connections for “Entoloma virescens” (Horak 1976: 202).

“Entoloma virescens’ sensu Noordeloos & Hausknecht (2007) from Seychelles

has cylindro-clavate cheilocystidia, pseudocystidia and similar macroscopic

features as our collections. However, the Noordeloos & Hausknecht taxon has

only 4-angled basidiospores (10-11.5 um, average 11.3 um) and a mild taste

contrasting with our material with 4-5-angled basidiospores and a bitter taste

that quickly becomes acrid.

Entoloma aeruginosum

Entoloma aeruginosum is macroscopically and microscopically similar to

“Entoloma virescens” sensu Noordeloos & Hausknecht (2007), but its holotype

no longer exists. Courtecuisse (1986), who examined two collections from

Japan with basidiospores averaging >10 um — one labeled Rhodophyllus

aeruginosus with basidiospores measuring 9.5-11.5(-12.0) um and the other

labeled Entoloma aeruginosum with basidiospores measuring 10-12(-12.5) um

— concluded that the basidiospores were more complex in their morphology

than those in the Agaricus virescens holotype. Entoloma aeruginosum differs

from Inocephalus virescens by 4-angled basidiospores that measure 9.5-12.5

um (average >10 um) and a mild taste.

238 ... Largent & Abell-Davis

Inocephalus virescens sensu Baroni

The bitter and latently acrid taste noted for the Lesser Antilles material

of I. virescens by Baroni (unpub. data) is a unique feature given that all other

descriptions of representatives of the I. virescens concept describe a mild

taste. Baroni plans to propose a new variety from the Caribbean based on the

acrid taste and production of a distinctive colored latex (Baroni pers. comm.).

Although our northeastern Queensland material had an acrid taste, colored

latex could not be verified, as all material was too mature when collected.

The 4- and sometimes 5-angled basidiospores and clavate cheilocystidia that

are distinct from the pseudocystidia differentiate our material of Inocephalus

virescens from the Caribbean material, in which all basidiospores appear

cuboid in all views and the broadly clavate cheilocystidia appear to represent

pseudocystidia.

In her poster on Inocephalus species from Brazil presented to the 2010

Australian Botanical Society meetings, Fernanda Karstedt emphasized the

4-5-angled basidiospores and uniform discoloration as characters to separate

I. virescens from other blue entolomatoid fungi that stain grayish-green. Both

features are either lacking or are not emphasized in any other publication

dealing with the I. virescens species complex.

Taxa related to /nocephalus virescens

Inocephalus virescens is closely related to Entoloma hochstetteri (Reichardt)

Stevenson from New Zealand and E. altissimum (Massee) E. Horak sensu

Horak (1976, 1980) from Singapore and Borneo. In both 1973 and 2008,

Horak recognized E. hochstetteri as a distinct species. Entoloma hochstetteri

sensu Horak (1973, 2008) can be easily separated from I. virescens by its larger

basidiospores (11-15 um in length, average >11.5 um), constricted fusoid

cheilocystidia, lack of pleurocystidia and pseudocystidia, non-distinctive taste,

dull blue basidiomata that change only to greenish-blue and then ochraceous

at the pileus apex, and stipe context that becomes bluish-green especially at the

base (Horak 2008; see also color illustrations by Horak at http//virtualmycota.

landcareresearch.co.nz/ webforms/vM_Species.aspx?pk=3353). | Entoloma

altissimum sensu Horak can be distinguished from I. virescens by its much

longer stipe (<150 mm long), lack of taste, and — most significantly — lack of

pseudocystidia and oleiferous hyphae (Horak 1976, 1980).

Conclusion

Clarification of taxa with blue-colored basidiomata with cuboid basidiospores

that become green on handling or maturity is hindered either by the absence of

holotype collections or by holotype material in poor condition. Elucidation of

taxa is also complicated when evanescent features present in young basidiomata

but not in mature specimens are inadvertently overlooked, such as the colorless

squamules on the pileus margin or pileus surface. Equally unhelpful are

Inocephalus virescens comb. nov. (Australia) ... 239

observations about the absence and presence of clamp connections within a

single description without noting which collections examined possessed or

lacked clamp connections.

We agree with Noordeloos & Hausknecht (2007) that thorough

documentation of material of Inocephalus virescens collected throughout its

entire geographical range is mandatory in order to determine morphological

and anatomical variation of this complicated species. Therefore, we consider,

as did Noordeloos & Hausknecht (2007) for their Mauritius material, that the

collections from northeastern Queensland and New South Wales fit the concept

of I. virescens sensu lato. We also conclude I. virescens sensu lato represents a

species complex for which molecular data would help determine species limits

and phytogeographical distributions in this group.

Alboleptonia stylophora (Berk. & Broome) Pegler, Kew Bull. 32: 199 (1977).

PLATES 3-4

= Agaricus stylophorus Berk. & Broome, J. Linn. Soc., Bot. 11: 537 (1871).

= Entoloma stylophorum (Berk. & Broome) Sacc., Syll. Fung. 5: 687 (1887).

= Rhodophyllus stylophorus (Berk. & Broome) Romagn.,

Prodr. Fl. Mycol. Madag. 2: 132 (1941).

= Leptonia stylophora (Berk. & Broome) Dennis, Kew Bull., Addit. Ser. 3: 78 (1970).

PitEus 6-32 mm broad, 2-15 mm high excluding umbo, umbo 1.5-3.0

mm high, typically cuspidate at times acutely umbonate, at first conic, then

convex and finally broadly convex to plane and undulate wavy, entirely

minutely appressed-fibrillose, white then pale yellowish-white or orange-white

(4-5A2-3), dull, faintly translucent-striate to the umbo, with age becomes

plicate-striate, hygrophanous when very mature and then grayish-orange

(5B3); margin minutely fibrillose fringed, incurved becoming decurved

then plane finally uplifted, eventually eroded and splitting radially; trama

white, unchanged, < 1.0 mm thick. TasTeE indistinct at first, then latently and

suggestively farinaceous. OpoR indistinct to somewhat pungent. LAMELLAE

adnexed to uncinate, rarely with a slight decurrent tooth, close to subdistant,

rarely narrow more commonly moderately broad and sigmoid (7-13 mm

long x 2.75-5.0 mm high), off-white (4-5A2-3); margin smooth then serrate

to eroded, concolorous. LAMELLULAE 3 (2 short, 1 medium to medium long)

between 2 lamellae. Stipe 27-52 mm long, 0.75-3.0 mm wide at apex, 0.75-4.5

mm broad at base, equal but more commonly tapered, white and + translucent,

becoming yellowish-white (4-5A2-3), at times pruinose at the apex, glabrous

but hygrophanous streaked with age, hollow, fragile and breaks easily; basal

tomentum scarce to absent.

BASIDIOSPORES nodulose-angular with 5-7 angles, isodiametric to

heterodiametric in side and profile views, isodiametric and 5-angled in polar

view, 8.9-13.9 x 6.8-11.4 um (x, = 11.8 + 0.95 x 9.0 + 0.84 um; E = 1.10-1.62;

240 ... Largent & Abell-Davis

Prate 3. Alboleptonia stylophora.

A: Basidiomata stature (DLL 9826); B: Pileal surface (DLL 9764).

Q = 1.32 + 0.11 (heterodiametric); n = 135/6). Basip1a cylindro-clavate, hardly

tapered, 1-, 2-, or 4-sterigmate, 24.2-36.8 x 7.8-11.3 um (x, = 29.3 + 3.07 x

9.7 + 0.88 um; Q = 2.14-4.20; E = 3.03 + 0.39; n = 27/5). CHEILOCYSTIDIA

typically absent in many sections; at times abundant, in clusters, and forming a

sterile edge in place in many gill sections, clavate to cylindro-clavate and thin-

walled in two collections (DL Largent 9826, DL Largent 9831), 19.9-53.3 x

Inocephalus virescens comb. nov. (Australia) ... 241

Prate 4. Alboleptonia stylophora.

A: Basidiospores (DLL 9751); B: 2-sterigmate basidium (DLL 9822);

C: Pileipellis (PIF27869); D: Cheilocystidia (DLL 9831).

4.5-9.9 um. PLEUROCYSTIDIA absent. HYPHAE OF LAMELLAR TRAMA felatively

short and easily separating from one another when squashed, 35.2-166.2 x

3.9-22.0 um. PILEIPELLIS an entangled layer in places, 24-120 um deep, often

collapsed and thus appearing to bea cutis. PILEOCYsTIDIA cylindric to cylindro-

clavate, 60-76 x 6-10 um. HYPHAE OF THE PILEUS TRAMA relatively short and

narrow, rarely broad, 56.3-155.9 x 5.4-10.8 um. STIPITIPELLIS a cutis, hymenial

elements absent. CAULOCYSTIDIA absent. OLEIFEROUS HYPHAE Scattered in the

pileal trama, rare in the stipe trama. Lipoid droplets absent. PIGMENTATION

none. CLAMP CONNECTIONS absent in all tissues.

HABIT, HABITAT AND DISTRIBUTION solitary, scattered, gregarious or

caespitose in soil amongst grasses or on rhizomes of Cyathea cooperi (F. Muell)

Domin; or in amongst leaf litter under Toona australis (Kuntze) Harms,

Ganophyllum falcatum Blume, Terminalia sericocarpa F. Muell., Argyrodendron

peralatum (Bailey) Edlin ex J.H. Boas, or Myristica insipida R. Br.; or in complex

mesophyll vine-forest throughout northeastern Queensland; also in Singapore,

242 ... Largent & Abell-Davis

Papua New Guinea, Solomon Islands (Horak 1980), Madagascar (Romagnesi

1941), Mauritius (Noordeloos & Hausknecht 2007), India (Manimohan et al.

1995), Sri Lanka (Petch 1917, Pegler 1977), the Hawaiian Islands (Horak &

Desjardin 1993), Puerto Rico (Baroni & Lodge 1998), Martinique (Pegler 1983),

Trinidad (Dennis 1953, Pegler 1983, Baroni & Lodge 1998) and Costa Rica

(Baroni & Lodge 1998). This is the first report of this species for Australia.

COLLECTIONS EXAMINED: AUSTRALIA. QUEENSLAND, Cook Region. In a garden of a

home in Redlynch, Cairns, 17 April 2009, Abell 377 (CNS) (16°57'27"S; 145°41'31"E;

87 m); Danbulla National Park, Kauri Creek Track, 28 February 2010, DL Largent

9751 (BRI, CNS) (17°07'50.4"S; 145°35'55.9"E; 730.6 m); 2 March 2010, DL Largent

9764 (BRI, CNS) (17°07'50.6"S; 145°35'55.8"E; 706.8 m); 24 March 2010, DL Largent

9822 (BRI, CNS) (17°07'50.4"S; 145°35'55.7"E; 715.7 m); Danbulla National Park,

Lake Euramoo Track, 21 March 2010, DL Largent 9805 (BRI, CNS), 24 March 2010,

DL Largent 9819 (BRI, CNS) (17°09'43.4"S; 145°37'40.7"E; 757.7 m); 24 March 2010,

DL Largent 9826 (BRI, CNS) (17°09'42.0"S; 145°37'45.0"E; 753.8 m); 25 March 2010,

DL Largent 9831 (BRI, CNS) (17°09'42.1"S; 145°37'40.3"E; 733.0 m); Saltwater Creek,

4km NW of Mossman National Park 29 November 2001, PIF27869 (BRI) (16°25'58"S;

145°20'04"E; 69.8 m); Smithfield Conservation Park, 5 June 2009, PN050609 (BRI, CNS)

(16°48'48.6"S; 145°40'51.9"E; 503.2 m).

Comments Alboleptonia stylophora is uniquely identifiable by the cuspidate

to acute umbo on a conic-campanulate to broadly convex, white, minutely

appressed-fibrillose pileus that quickly becomes pale yellow, the concolorous

stipe, the complex, nodulose angular large basidiospores, and the entangled

layer of hyphae that form the pileipellis. The absence of cheilocystidia and the

presence of clamp connections have been reported on hyphae in the context

in collections from Sri Lanka (Pegler 1977) and Lesser Antilles (Pegler 1983,

1986). Although Dennis (1970) and Romagnesi (1941) did not report on these

features for their A. stylophora collections from Trinidad and Madagascar,

respectively, the presence of cheilocystidia and lack of clamp connections were

reported by all other investigators. Cheilocystidia were absent in almost all of the

collections from northeastern Australia with the exception of DL Largent 9826

and DL Largent 9831 (collected from the same area at Lake Euramoo, Danbulla

National Park) and Abell 377 (collected on fern rhizomes in a garden). The

presence or absence of thin-walled cheilocystidia in different collections of the

same species is perplexing and has two explanations. Either these cheilocystidia

often collapse and are difficult to demonstrate (see Baroni & Lodge 1998: 692)

or they are present on only a part of the edges of the lamellae (more often the

lamellulae) and thus easily overlooked, as was the case in Abell 377.

Based on the appressed-fibrillose off-white pileus, the entangled pileipellis

layer, and the heterodiametric basidiospores, A. stylophora is considered a

good species of Alboleptonia. Furthermore, all features found in A. stylophora

are also those circumscribed for the genus by Largent & Benedict (1970) and

Largent (1994).

Inocephalus virescens comb. nov. (Australia) ... 243

However, nodulose angled basidiospores, lack of clamp connections, and

small basidia are not typically found in Alboleptonia and are not present in the

type species, A. sericella (Fr.) Largent & R. G. Benedict. Here also, molecular

sequence analyses should help determine the taxonomic significance of these

and other features.

Acknowledgments

We would like to thank Dr. John Kanowski for granting permission to collect in

the Brooklyn Wildlife Sanctuary adjacent to Mt. Lewis National Park. We gratefully

acknowledge the director of the Australian Tropical Herbarium, Prof. Darren Crayn,

for his support and encouragement for the first fungal foray (Mycoblitz) in the Wet

Tropics World Heritage forests of far North Queensland, Australia, 19-25 February

2009. It was at this meeting that contacts (* = attended the Mycoblitz) were made that

began a network of individuals without whom the study of the entolomatoid fungi

in northeastern Queensland would not have started. The existence of this network

is a testament to enthusiasm and support for research and mycology inherent in the

Australian people. We are extremely grateful to the following individuals who are

currently part of this network and responsible for site information for Inocephalus

virescens and Alboleptonia stylophora: Fay Adams; *Craig Costion; Nikki Bennetts; Alan

Curtis; *Dr. Sapphire McMullan-Fisher; *Skye Moore; *Pamela O’Sullivan; Dr. Bruce

Webber. We also gratefully acknowledge *Peter Newling, Landcare Volunteer for the

Trinity Inlet Catchment Management Association Inc, Queensland, for his enthusiasm

and identification of the coastal habitats of fungi in the Entolomataceae. Comments by

Dr. Andrew Methven, who reviewed the manuscript and by the nomenclature editor

Dr. Shaun Pennycook, were very helpful. We wish to thank Dr. Timothy Baroni for

sharing an unpublished manuscript and for also providing useful comments as a

reviewer. The fieldwork in Australia was supported by the Largent family trust and we

are particularly grateful for the support of Pamela Largent. Fieldwork and logistical

support was provided by the Australian Tropical Herbarium and the School of Marine

and Tropical Biology, James Cook University.

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244 ... Largent & Abell-Davis

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Société Mycologique de France 69:145-198.

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Series 3: 78.

Gates MG, Horton BM, Noordeloos M. 2009. A new Entoloma (Basidiomycetes, Agaricales) from

Tasmania. Mycotaxon 107: 175-179. doi:10.5248/107.175

Gates MG, Noordeloos ME. 2007. Preliminary studies in the genus Entoloma in Tasmania I.

Persoonia 19:157-226.

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Herbarium and the Flora and Fauna of South Australia Handbooks Committee: Adelaide.

Horak E. 1973. Fungi Agaricini Novaezelandiae. Entoloma (Fr.) and related genera. Beihefte zur

Nova Hedwigia 43: 1-86.

Horak E. 1976. On cuboid-spored species of Entoloma. Sydowia 28:171-236.

Horak E. 1980. Entoloma (Agaricales) in Indomalaya and Australasia. Beihefte zur Nova Hedwigia

65:1-352.

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

Zealand/Nga Harore o Aotearoa, Volume 5. Fungal Diversity Research Series 19.

Horak E, Desjardin D. 1993. Agaricales of the Hawaiian Islands, 2. Notes on some Entoloma species.

Mycologia 85(3): 480-489. doi:10.2307/3760708

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Largent DL. 1994. Entolomatoid fungi of the western United States and Alaska. Mad River Press

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Largent DL, Benedict RG. 1970. Studies in the rhodophylloid fungi II: Alboleptonia, a new genus.

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Inocephalus virescens comb. noy. (Australia) ... 245

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MY COTAXON

Volume 116, pp. 247-251 April-June 2011

DOT: 10.5248/116.247

The family Chaetomiaceae from China 4.

Two newly recorded species of Chaetomium

YUNZHONG Guo, MINGQI ZHU & GUANGYU SUN*

State Key Laboratory of Crop Stress Biology in Arid Areas and College of Plant Protection,

Northwest A&F University, Yangling, Shaanxi 712100, P. R. China

*CORRESPONDENCE TO: sgy@nwsuaf.edu.cn

ABSTRACT — Two species of the genus Chaetomium, C. virescens and C. senegalense, are

reported as newly recorded species in China. The characteristics of the two species are

described and illustrated from our materials. These specimens and living cultures examined

were deposited in Herbarium of Fungi in Northwest A&F University (HMUABO).

KEY worDs — saprophyte, ascomycetes, taxonomy

Introduction

Chaetomium (Chaetomiaceae, Sordariales) was introduced by Kunze based

on the type species Chaetomium globosum and characterized by superficial,

ostiolate ascomata, with hairs or setae and one-celled, brown or gray-olivaceous

ascospores. It is a species-rich genus, with about 95 currently accepted species

(von Arx et al. 1986, 1988; Kirk et al. 2008). In China, 33 species have been

recorded (Chen 1973, Tai 1979, Eriksson & Yue 1988, Sun et al. 2004, 2005;

Wang et al. 2005a,b). In a survey of Chaetomiaceae species from different

regions in China, numerous isolates were obtained from soil and plant debris,

and two new records, Chaetomium virescens and C. senegalense were found.

‘These species are described and illustrated.

Materials & methods

The isolates were from plant debris, which are dry branches of Pinus tabuliformis in

Nyingchi, Tibet Autonomous Region, dead twigs of Caragana microphylla in Chifeng,

Inner Mongolian Autonomous Region and dry tissues from a kind of graminaceous

weed in Zhangye, Gansu Province. These were cultivated on Martin Agar (MA: 5 g

peptone, 10 g dextrose, 1 g KH,PO,, 0.5 g MgSO, 7H,O, 15 g agar, 1000 ml distilled

water). Subcultures were grown on corn meal agar (CMA: 30 g cornmeal, 15 g agar,

248 ... Guo, Sun & Zhu

1000 ml distilled water). Colony characteristics, including colony diameter after 3 to

5 days of incubation on CMA at 28°C and 37°C in the darkness, were measured and

photographed. The plant samples and living cultures are deposited in Herbarium of

Fungi in the Northwest A&F University (HMUABO).

a &

Fic. 1 Chaetomium virescens: A. ascoma; B. asci; C. ascospores; D. textura.

Scale bars: B, C = 2 um; A, D = 10 um

Chaetomium virescens and C. senegalense new to China ... 249

Taxonomy

Chaetomium virescens (Arx) Udagawa, Trans. Mycol. Soc. Japan 21: 34, 1980.

Fic. 1

SPECIMENS EXAMINED—CHINA. TIBET AUTOMOUS REGION: Nyingchi (29°35’N

94°15'E), in dry branches of Pinus tabuliformis Carriere, H.M. Yue, HMUABO 61252.

INNER MONGOLIAN AUTOMOUS REGION: Chifeng (42°15’N 118°53’E), in dead twigs of

Caragana microphylla Lam., Y.L. Liu, HMUABO 61235.

Cotonlies with a daily growth rate of 4-8 mm on CMA at 28°C in the dark, with

abundant aerial mycelia and orange color or yellow green exudates; ASCOMATA

maturing within 16 days, superficial, black in reflected light, ellipsoid, 195-215

x 130-145 um, ostiolate, with a conical beak, wall dark brown, cells of textura

angularis; ASCOMATAL HAIRS sparse, straight, tapering seta-like, 220 um long,

2.5 um thick at the base; asci fasciculate, clavate, stalked, 8-spored, evanescent,

27.5-32.5 x 11.25-13.75 um; ascospores fusiform or ellipsoidal, often

inaequilateral, brown when mature, 6.25-7.5 x 9.4-12.5 um, with a distinct

apical germ pore, occasionally with a germ pore at both ends.

CoMMENTs: The strains had a broad range of growth temperature (15-45°C),

and ascoma formed abundantly at 28-37°C. Ascomata maturing within 6 days

at 37°C in the darkness. Colonies were pale at beginning, then a light yellow.

These features differed from C. virescens, since the exudates were also orange,

and the asci were shorter, and maximum length was only 32.5 um.

Chaetomium senegalense L.M. Ames, Monogr, Chaetomiaceae: 36, 1963. Fic. 2

SPECIMEN EXAMINED—CHINA. GANSU PROVINCE: Zhangye (38°54’N 100°27’E), in

dry tissues of a graminaceous weed, Y.Z. Guo, HMUABO 62070.

Co.tonizs with a daily growth rate of 7-8 mm on CMA at 28°C in the

darkness, with a white or pale aerial mycelium and pale yellow exudates;

ASCOMATA maturing within 18 days at 28°C, superficial, grayer in reflected

light, ellipsoidal-spherical, 210-250 x 225-290 um, ostiolate, peridium with a

brown wall and cells of textura angularis; AscOMATAL HAIRS mainly growing

in the upper part of ascomata, 650-850 um long, undulate, septate, branched

or unbranched, tapering, 2.5 um thick at the base; asci fasciculate, cylindrical,

with short stalks, 8-spored, evanescent, 37.5-50 x 6.25-8.75 lum; ASCOSPORES

ovate or nearly spherical, brown or dark brown when mature, 8.75-11.25 x

6.25-7.5 um, with nearly apical germ pores.

ComMENTs: Growth of these strains exhibited a broad temperature range

(15-45°C) and ascoma formed abundantly at 28-37°C. But compared with

those seen by Ames (1963), these asci were 20 um shorter at maximum length.

250 ... Guo, Sun & Zhu

Fic. 2 Chaetomium senegalense: A. ascoma; B. asci; C. ascospores; D. textura.

Scale bars: A = 10 um; B-D = 2 um

Acknowledgments

This work was supported by National Natural Science Foundation of China

(30771735), the 111 Project from Education Ministry of China (B07049), Program for

Changjiang Scholars and Innovative Research Team in University (IRT0748) and Top

Talent Project of Northwest A&F University. The authors wish to thank Dr. Tom Hsiang

(University of Guelph, Guelph, Ontario, Canada) and Professor Zhongyi Zhang (College

of Plant Protection, Yunnan Agricultural University, Kunming, Yunnan, China) for

reviewing the manuscript.

Chaetomium virescens and C. senegalense new to China... 251

Literature cited

Ames LM. 1963. A monograph of the Chaetomiaceae. Research and Development Service of United

States Army.

Chen QT 1973. The new species of Chaetomium and Septoria. Journal of Microbiology 13:

124-128.

Eriksson OE, Yue JZ. 1988. The pyrenomycetes of China, an annotated checklist. The University

of Umea, Sweden.

Kirk PM, Cannon PF, David JC, Stalpers JA. 2008. Dictionary of the fungi. 10 ed. Wallingford,

CAB International.

Sun GY, Tan YJ, Zhang R. 2004. The family Chaetomiaceae from China I. Species of the genus

Chaetomium. Mycosystema 23: 333-337.

Sun GY, Tan YJ, Zhang R. 2005. The family Chaetomiaceae from China III. Species of Chaetomium

and Thielavia. Mycosystema 24: 318-321.

Tai FL 1979. Sylloge fungorum sinicorum. Beijing: Science Press. 1527 p.

von Arx JA, Guarro J, Figueras MJ. 1986. The ascomycete genus Chaetomium. Beheifte zur Nova

Hedwigia 84: 1-162.

von Arx JA, Figueras MJ, Guarro J. 1988. Sordariaceous ascomycetes without ascopore ejaculation.

Nova Hedwigia 94: 1-104.

Wang XW, Zheng RY. 2005a. Chaetomium acropullum sp. nov. (Chaetomiaceae, Ascomycota),

a new psychrotolerant mesophilic species from China. Nova Hedwigia 80: 413-418.

doi: 10.1127/0029-5035/2005/0080-0413

Wang XW, Zheng RY. 2005b. Chaetomium ampulliellum sp. nov. (Chaetomiaceae, Ascomycota)

and similar species from China. Nova Hedwigia 81: 247-255. doi: 10.1127/0029-

5035/2005/0081-0247

MY COTAXON

Volume 116, pp. 253-264 April-June 2011

DOT: 10.5248/116.253

Saxicolous species of Claudopus (Agaricales, Entolomataceae)

from Australia

Davin L. LARGENT*’?, SANDRA E. ABELL-DAVIS’, GRIFFIN A. CUMMINGS?,

KATHRYN L. RYAN? & SARAH E. BERGEMANN?

‘Biological Sciences, Humboldt State University, 1 Harpst St, Arcata CA 95521 U.S.A.

*School of Marine and Tropical Biology, Australian Tropical Herbarium,

James Cook University, PO Box 6811, Cairns QLD 4870 Australia

3Biology Department, Middle Tennessee State University,

PO Box 60, Murfreesboro, TN 37132, U.S.A.

*CORRESPONDENCE TO: mrp@humboldtl.com

ABSTRACT — Claudopus rupestris, C. viscosus, and C. minutoincanus are described as new

to science. All three species produce abundant, white basidiomata on the underside of

granitic rocks and have a pileus that is sticky to the touch. Claudopus rupestris has unusual

basidiospores, some of which are 4-angled. Claudopus viscosus and C. minutoincanus possess

subcapitulate to capitulate pileocystidia and are morphologically similar to Entoloma jahnii

of Europe.

Key worps — Basidiomycota, mtSSU, LSU, RPB2, novel species

Introduction

The genus Claudopus Gillet is considered a subgenus within the genus

Entoloma (Fr.) P. Kumm. by some researchers (Noordeloos 1981, 1992, 2004,

Manimohan etal. 2006). Others consider it an independent genus (Dennis 1970,

Horak 1980, 2008, Largent 1994, Pegler 1983), a concept favored in this report.

Because of the ease with which these species can be identified macroscopically,

those who do not recognize Claudopus as either an autonomous genus or as

a subgenus within Entoloma still place species into a key group based on the

distinct stature of the basidiomata (Hesler 1967, Knudsen & Vesterholt 2008).

Claudopus can be distinguished from pleurotoid species of Clitopilus (Fr.

ex Rabenh.) P. Kumm. or Rhodocybe Maire by their basidiospores, which are

angular in all views. Basidiospores are angular only in polar view in Clitopilus

and angular-pustulate in all views in Rhodocybe.

Claudopus species, which are easily recognized by their pleurotoid stature,

possess a centrally attached stipe that typically becomes eccentric, lateral, or

254 ... Largent & al.

disappears completely as the pileus expands and a pileus that is either matted-

fibrillose or has a fibrillose covering.

Although Claudopus species are cosmopolitan, basidiomata are often

overlooked by researchers because of their small size or unusual habitats.

As a consequence, researchers typically describe only one or two species per

publication (e.g. Horak 2008, Manimohan et al. 2006, Pegler 1977, 1983) and

often include none in floristic studies (Romagnesi & Gilles 1979). Nonetheless,

Noordeloos (2004) reported ten species from Europe, and Horak (1980)

included five from Indomalaya and Australasia.

Only two species, Claudopus byssisedus (Pers.) Gillett and C. depluens

(Batsch) Gillet, have been reported from Australia (May & Wood 1997).

Although authentic Australian material does exist for C. byssisedus, all

Australian C. depluens material has been re-identified as a species of Crepidotus

(Fr.) Staude (Horak 1980, May & Wood 1997).

We report here three Claudopus species that possess features different

from any other described species of Claudopus, namely abundant (10-75)

basidiomata on the underside of granitic rocks and a pileus that is sticky to the

touch. Claudopus rupestris, C. viscosus, and C. minutoincanus are described as

new to science and are reported for the first time from Australia.

Materials & methods

Macromorphological and micromorphological features

Collections were made during February—April in 2009 and 2010 within the Wet

Tropics Bioregion throughout northeastern Queensland and in April 2010 from various

localities in central New South Wales. GPS coordinates for each collection were taken

in the field using a Garmin GPSmap 60CSx. Basidiomata were carefully collected in

the field, stored in plastic containers, and returned to the laboratory. Macroscopic

descriptions were made from recently collected fresh material. Colors were described

both subjectively and coded according to Kornerup & Wanscher (1978), with color plates

noted in parentheses. Abbreviations of color plates used in specific descriptions indicate

the page number, column(s), and row(s) [e.g., 8D-F5-6 indicates page 8, columns D-F,

rows 5-6.]

Micromorphological features of dried specimens were examined with a trinocular

research-grade Nikon Labophot compound microscope fitted with bright field light

contrast optics following general protocols set forth in Largent (1994: 1-3) and

techniques used for measuring spores of the Entolomataceae set forth by Baroni &

Lodge (1998: 681). Digitized microphotographs were made using a Nikon Coolpix 990

focused through the trinocular head of the compound microscope. All microscopic

measurements were obtained using a GITCO Corporation Graphic Digitizer, Model

DP5A-111A connected to an IBM compatible Chem Book Laptop computer. The

Measure Me 101 v 1.0 software used was modified for laptop use from a BASIC program

(Metrics5) developed and provided by David Malloch, University of Toronto. Factors

determined using this program include: arithmetic means (x, ) of basidiospore lengths

Saxicolous Claudopus spp. nov. (Australia) ... 255

and widths + standard deviation measured for n objects; quotient of basidiospore length

by spore width (E) indicated as a range variation in n objects measured; the mean of

E-values (Q) + standard deviations. The sample size (n) = total number of microscopic

structures measured (x) divided by the number of basidiomata studied (y), as shown in

the formula n = x/y.

Molecular data

Dried basidiomata tissues were pulverized with glass beads in a FastPrep FP120

homogenizer (QBiogene, Carlsbad CA USA) at 4 mps for 20s. Genomic DNA was

extracted using a 2 x cetyl trimethylammonium bromide (CTAB) modified method,

and then purified on GeneClean Turbo columns (MP Biomedicals, Solon OH USA)

following the Baumgartner et al. (2010). Three loci were PCR (Polymerase Chain

Reaction) amplified including a portion of the mitochondrial small subunit ribosomal

DNA (mtSSU) using primers MS1 and MS2 (White et al. 1990) following the PCR and

cycling protocols of Gomes et al. (2000). The variable domains D1 and D2 of the 28S

ribosomal DNA (LSU) were PCR amplified with the primers ctb6 and tw13 (White et

al. 1990) following the PCR and cycling protocols outlined in Bergemann & Garbelotto

(2006). The RNA polymerase II subunit II (RPB2) was amplified with primers rpb2i6f

or rpb2i7r following protocols described in Co-David et al. (2009).

PCR products were cleaned using 1 uL of ExoSAP-IT (GE Healthcare, Pittsburgh,

PA) and incubation of 37°C for 15 min followed by 80°C for 45 min. Sequencing

reactions were carried out in 10 uL reactions containing 1 uL of ABI BigDye ver. 3.1

(Applied Biosystems, Carlsbad CA USA), 0.4 uM primer, 0.875 Sequencing Buffer

(Applied Biosystems, Carlsbad CA USA) and 1 pL PCR water. Cycling conditions

were as follows: 39 cycles of 96°C for 10 s, 50°C for 5 s, 60°C for 4 min. Reactions were

precipitated with 1 uL of 3 M NaOAc, 1 wL 250 mM EDTA (pH 8) and 25 uL of 100%

EtOH by centrifugation at 2500 G for 30 min. Reactions were rinsed with 70% EtOH

and centrifuged at 2500 G for 15 min. Precipitated products were stored at -20°C. Bi-

directional sequences were generated on an Applied Biosystems 3130xl Genetic Analyzer

at Middle Tennessee State University. The contigs of bi-directional sequences were

edited in Sequencher 4.8 (GeneCodes Corp., Ann Arbor, MI USA). Sequences for each

new species from three loci were deposited in GenBank (HQ731511-731518), with the

exception of the RPB2 sequence from DL Largent 9624 (Claudopus rupestris), omitted

after repeated unsuccessful attempts to obtain sequences from weak PCR amplifications.

The GenBank designators for each sequence of all three loci are provided at the end of

each holotype description together with type habitat data and GPS co-ordinates. Actual

sequences can be obtained from GenBank using the designator number. All of the

sequences will be used in a later phylogenetic study.

Taxonomy

Claudopus rupestris Largent & Abell-Davis, sp. nov. PLATE 1

MycoBank MB 519408

Ab aliis congeneris combinatione characterum sequentium differt: habitatio saxatilis; pileus

lucens viscidusque; basidiosporae plerumque quadrangulae, interdum quinqangualae,

subisodiametricae, 6.5-9.2 x 5.9-8.0 um, longitudine mediale < 8.5 um; basidia

sterigmatibus 5-7; cystidia hymenialia cylindrica vel anguste obclavata; fibulae absens.

256 ... Largent & al.

Type — Australia, Queensland, Cook Region, Wooroonooran National Park, Josephine

Falls Track, within 50 m of 17°26'00.9"S, 145°51'33.8"E, 26 February 2009, DL Largent

9624 (mtSSU HQ731512, LSU HQ731515) (holotype BRI; isotype CNS).

EryMoLocy — derived from Latin word rupestris, ‘rock-dwelling.

BASIDIOMATA very abundant (50-75) and most often sessile to laterally stipitate.

PitEus 1-4 mm broad, 1-3 mm deep, flabelliform to dimidiate, white with very

pale yellowish tinges, minutely matted-fibrillose, glistening and sticky, even,

opaque, not hygrophanous; margin decurved, even to irregularly lobed and

not striate. TASTE indistinct. ODOR mild. LAMELLAE 5-8 per basidiome, 1-3

mm long, less than 1 mm high, white, adnexed, minutely ventricose, distant.

LAMELLULAE 1 between lamellae; margin smooth and concolorous. STIPE

at first centrally stipitate, then laterally stipitate, and then most often absent,

1 mm or less long, less than 0.75 mm broad, equal white and at first minutely

pruinose then glabrous; stipe base or basidiomata attachment with minute

hair-like rhizoids.

BASIDIOSPORES 4-5-angled in profile view, 5-angled in side view,

angles distinct to indistinct but not rounded in any view, isodiametric to

subisodiametric, rarely heterodiametric, on the average subisodiametric,

6.5-9.2 x 5.9-8.0 um (x. = 8.1 + 0.62 x 6.8 + 0.44 um; E = 1.0-1.36;

Q = 1.18 + 0.10; n = 27/1). BAsip1a commonly 2 or 4-sterigmate, uncommonly

5-7-sterigmate, 20.8-31.8 x 8.0-11.5 um (x, = 27.19 + 2.92 x 9.7 + 1.02 um;

E =2.45-3.10; Q = 2.81 + 0.24; n= 15/1). HYMENIAL cysTIpIA not readily visible

on edge of entire gills, rare to scattered in squash mounts, cylindric, cylindro-

clavate, or narrowly obclavate, 31.1-48.2 x 3.8-7.8 um; n = 8/1. PILEIPELLIS an

entangled layer of inflated hyphae. Prteocystip14 cylindro-clavate to broadly

cylindro-clavate, 25.2-81.5 x 6.2-13.9 um, (x, = 42.0 + 15.3 x 9.2 + 2.0 um;

E = 2.7-5.4; Q = 4.55 + 1.32; n = 13/1). PILEAL TRAMA 26.5-41.6 um deep.

TRAMAL HYPHAE in pileus, lamellae and stipe not measured because of minute

size of basidiomata. STIPITIPELLIS not examined. OLEIFEROUS HYPHAE absent.

LIPoID BODIES absent. CLAMP CONNECTIONS absent in all tissues.

HABIT, HABITAT AND DISTRIBUTION gregarious on undersurface of small

(25 x 25 cm) granitic rocks growing on 1 mm thin to nearly non-existent soil

layer with minute mosses; basidiomata apparently attached to the rhizoids of

the moss gametophyte; complex mesophyll forest. Known only from the type

locality.

DIAGNOSTIC CHARACTERS: basidiomata 1-4 mm diameter, growing on soil

with minute mosses on the undersurface of small granitic rocks; basidiospores

in profile view commonly 4-angled, sometimes 5-angled, on the average

subisodiametric (6.5-9.2 x 5.9-8.0 um; x_ = < 8.5 um; E= 1.18). Basrp1a small,

occasionally 5-7-sterigmata, hymenial cystidia rare to scattered, cylindric to

cylindro-clavate to narrowly obclavate, clamps absent.

Saxicolous Claudopus spp. nov. (Australia) ... 257

* Pci

, NGS a ae -. : AS Oy

A ieee aes Pe

. q 4 \

= \ 7

a is y

~ *) “se

> at P ;

PiatE | - Claudopus rupestris (DL Largent 9624: HoLotyPe). A: Basidiomata (9x); B: Pileocystidia

(400x); C: Pileipellis (100x); D: Basidiospores (1000x); E: Cheilocystidia and Basidia (400x).

ComMENTS— In Australia, Claudopus rupestris is macroscopically similar to

C. viscosus and C. minutoincanus, both also found in saxicolous habitats. The

4-5-angled basidiospores and absence of subcapitulate to capitulate pileocystidia

distinguish the C. rupestris from C. viscosus and C. minutoincanus, both of which

lack 4-angled basidiospores and have subcapitulate to capitulate pileocystidia.

Taxa related to Claudopus rupestris

Entoloma albotomentosum Noordel. & Hauskn. (from central Europe and

Norway) and Claudopus pandanicola E. Horak (from Papua New Guinea)

morphologically resemble C. rupestris in their pleurotoid, white basidiomata

258 ... Largent & al.

with a fibrillose pileus, mild taste or odor, occasionally 4-angled or similar sized

basidiospores, and absence of clamp connections. Entoloma albotomentosum

can be differentiated by its longer (6 mm) stipe, non-sticky, larger pileus (2-10

mm broad), adnate or decurrent, very distant lamellae, translucent-striate pileus

when expanded, longer (9-12.5 um) 4-6-angled basidiospores, and habitat

on leaves of grasses and sedges in marshes (Noordeloos 1992). Claudopus

pandanicola can be distinguished by the larger (5-15 mm broad) non-sticky

pileus, habitat on rotting Pandanus leaves, 5-6-angled basidiospores (none

of which are rectangular), and pigment-encrusted pileipellis hyphae (Horak

1980).

Claudopus viscosus Largent & Abell-Davis, sp. nov. PLATE 2

MycoBank MB 519409

Ab Entolomate (subg. Claudopo) jahnii habitatione saxatili, pileo viscido margine striato

minute sulcato, sapore odoreque subtiliter farinaceo, basidiosporis 7.7-12.0 x 5.3-7.9 um

(mediane 9.8 x 6.7 um), fibulis absentibus differt.

Type — Australia, Queensland, Cook Region, Danbulla National Park, Kauri Creek

Track, 17°07'56.3"S, 145°35'54.3"E, 17 March 2010, DL Largent 9788 (mtSSU HQ731513,

LSU HQ731516, RPB2 HQ731518) (holotype BRI; isotype CNS).

EryMoLocy — derived from the Latin word viscosus, referring to the ‘sticky’ surface of

the basidiomata.

BASIDIOMATA shiny and sticky to fingers, forceps, knives, leaves and debris;

attached by minute rhizomorphs to rhizoids of moss gametophytes. PILEUS

1.5-11.0 mm broad, 2-10 mm deep, 1-1.5 mm high, broadly convex from

side view, not hygrophanous, at first white and opaque, entirely matted-

fibrillose and minutely petaloid to flabelliform in top view with an incurved to

decurved margin, upon expansion and maturity becoming more flabelliform

and + dimidiate with a decurved, plane, or uplifted margin that is eroded and

crenulate and in some of the broader specimens the fibrillose layer disappears

with age revealing a + glabrous undersurface that is minutely sulcate,

suggestively striate and off white to pinkish-white; striate in dried specimens.

TASTE and Opok latently farinaceous. LAMELLAE up to 6 mm long and 2.5 mm

high, adnexed at the point of attachment, white at first, moderately broad and

close, quickly becoming broad to ventricose and subdistant to distant; margin

even and concolorous. LAMELLULAE 1-3 (1-2 short, 1 medium long) between

lamellae. Stipe 1.0 x 0.5 mm and covered with minute, white hairs in very

young specimens, with maturity and expansion of the pileus, the hyphae of the

stipe quickly merge with the hyphae of the pileus or pileus surface and thus is

absent in older specimens.

BASIDIOSPORES 5-6-angled, subisodiametric to more __ typically

heterodiametric in profile and side views, 6-angled and isodiametric in polar

view, 7.7-12.0 x 5.3-7.9 um (x, = 9.8 + 0.76 x 6.7 + 0.57 um; E = 1.22-1.88;

Saxicolous Claudopus spp. nov. (Australia) ... 259

x. ——_>e . . ~

- ~~ - p

é be, )

= oe p ~ - . =

- . = ~ .

a ee ee . .

PLATE 2 - Claudopus viscosus (DL Largent 9788: HoLotyPe). A: Basidiomata (4x); B: Pileipellis at

point of attachment (100x); C: Capitulate pileocystidia (400x); D: Basidia and basidioles (400x);

E: Basidiospores, all profile view (1000x).

Q = 1.47 + 0.13 (heterodiametric); n = 16/4). Basip1A small, clavate to

cylindro-clavate, 23.6 x 33.6 x 8.0-11.5 um (x, = 29.5 + 2.64 x 10.0 + 0.84 um;

E = 2.48-3.79; Q = 2.97 + 0.30; n = 16/2); 1-, 2-, 3- or 4-sterigmate. HYMENIAL

260 ... Largent & al.

CYSTIDIA absent. HYPHAE OF LAMELLAR TRAMA thin-walled, subparallel,

relatively short, 90.0-122.6 x 15.7-18.7 um. PILEIPELLIs an entangled layer of

hyphae, 80-224 um deep. PILEOCysTip1A with a granular surface, frequently

subcapitulate to capitulate, 35.66-79.64 x 2.31-8.15 um, capitulum 4.5-10.9

um broad. OLEIFEROUS HYPHAE absent. LIPOID BODIES absent. PIGMENTATION

non-existent. CLAMP CONNECTIONS absent in all tissues.

HABIT, HABITAT AND DISTRIBUTION scattered to gregarious in a thin layer

of soil in amongst and attached to the rhizoids of few, tiny moss gametophytes

on the underside of granitic rocks in simple to complex notophyll vine forest,

northeastern Queensland.

ADDITIONAL COLLECTIONS EXAMINED — AUSTRALIA, QUEENSLAND, Cook Region,

Dinden National Park, near end of Davies Creek Road, 17°02'13.1"S, 145°36'47.8"E,

23 February 2010, DL Largent 9733; Danbulla National Park, Kauri Creek Track,

17°07'56.3"S, 145°35'54.3"E 9840 (three topotypes) - 17 March 2010, DL Largent 9789;

24 Mar 2010, DL Largent 9828; 27 Mar 2010, DL Largent. (All collections BRI; split

collections CNS.)

DIAGNOSTIC CHARACTERS: Basidiomata shiny and sticky to fingers, forceps,

knives, leaves and debris; attached by minute rhizomorphs to rhizoids of moss

gametophytes on the underside of granitic rocks. Pileus at first white, opaque,

and entirely matted fibrillose then, in some, + glabrous towards the margin and

minutely sulcate and suggestively striate. Taste and odor latently farinaceous.

Basidiospores 5-6-angled, measuring 7.7-12.0 x 5.3-7.9 um (average = 9.8 x

6.7 um). Cheilocystidia and clamp connections are absent.

CoMMENTS— In Australia, C. viscosus and C. minutoincanus produce pleurotoid,

sticky basidiomata with a white fibrillose pileus and subcapitulate to capitulate

pileocystidia, and both are found in a saxicolous habitat. The mild taste and

odor, sulcate striate mature pileus lacking yellowish tinges, smaller basidia, and

longer narrower heterodiametric (E = 1.47) basidiospores distinguish C. viscosus

from C. minutoincanus with its latently farinaceous taste and odor, yellow-

tinged opaque even mature pileus, longer larger basidia (33-38 x 10-13 um),

and shorter and broader subisodiametric basidiospores (E = 1.22) averaging

9.1 x 7.4 um. Claudopus rupestris (Wooroonooran National Park, Queensland)

is also saxicolous but lacks subcapitulate to capitulate pileocystidia and has

4—5-angled basidiospores.

Taxa related to Claudopus viscosus

Entoloma jahnii Wolfel & Winterh. (from Germany and Belgium), which

produces pleurotoid basidiomata with white fibrillose pilei and capitulate

pileocystidia, is differentiated from C. viscosus and C. minutoincanus by its mild

taste and odor, larger basidiospores (9.7-12.9(-15) x 7.6-11(-15) um), presence

of clamp connections, and habit on rotting wood and bark (Noordeloos 2004).

Saxicolous Claudopus spp. nov. (Australia) ... 261

Claudopus minutoincanus Largent & Abell-Davis, sp. nov. PLATE 3

MycoBank MB 519410

Ab Entolomate (subg. Claudopo) jahnii habitatione saxatili, pileo viscido luteoalbo suffuso,

basidiosporis 7.4-11.4 x 6.3-9.6 um (mediane 9.01 x 7.4 um), fibulis absentibus differt.

Type — Australia, New South Wales, Central Hunter District, Barrington Tops National

Park, Williams Day Use Area, end of Blue Gum Loop, 32°09'11.0"S, 151°31'38.9"E, 14

April 2010, DL Largent 9871 (mtSSU HQ731511, LSU HQ731514, RPB2 HQ731517)

(holotype DAR).

EryMoLocy — derived from a combination of the Latin minutus + incanus, referring to

the ‘minutely’ ‘hoary’ pileal surface.

BASIDIOMATA attached by minute stipe as well as the basal portion of pileus.

PILEus in side view convex, 1-3 mm high, in top view minutely petaloid then

dimidiate, 6 mm broad, 2-5 mm deep, whitish with a hint of yellowish-white

(3A2), matted fibrillose 2/3 way to margin, minutely tomentulose-canescent

towards the margin and with an incurved then decurved and eventually

uplifted, minutely canescent margin, glistening and sticky, dull, even, opaque,

not hygrophanous. Taste and Opor indistinct. LAMELLAE adnexed, 3 mm

long, 0.5 mm high, narrow, close, yellowish-white (3A2); margin smooth and

concolorous. STIPE lateral and minute < 1 mm long, < 0.5 mm broad, white,

covered with fine hairs that disappear when dried, equal; basal tomentum

absent.

BASIDIOSPORES 6-angled, isodiametric to heterodiametric in profile and side

views, 6-angled and isodiametric in polar view, 7.4-11.4 x 6.3-9.6 um (x, =

9.01 + 0.94 x 7.4 + 0.77 um; E = 1.08-1.44; Q = 1.22 (subisodiametric) + 0.09; n

= 30/1). Basrp1a clavate, medium in length, 4-sterigmate, 32.7-38.3 x 9.8-13.2

um (x, = 34.9 x 11.5 um; E = 2.75-3.411]; Q = 3.05; n = 8/1). HYMENIAL CYSTIDIA

absent. PILEIPELLIs a superficial entangled layer of hyphae, scant at the point of

attachment to the substrate and then transitioning into an erect palisadoderm

of slender pileocystidia. PiLEocystTip1a cylindric and subcapitulate to

capitulate, suggestively covered with a thin layer of material, 14.4-68.3 um

long, 2-3 um broad, capitulum 3.6-6.3 um near point of attachment, at the

margin particularly abundant, 39.5-53.1 um long, 2-3 um broad, capitulum

4.1-6.4 um. STIPITIPELLIS not examined. OLEIFEROUS HYPHAE absent. Lrporp

BODIES absent. PIGMENTATION non-existent. CLAMP CONNECTIONS absent in

all tissues.

HABIT, HABITAT AND DISTRIBUTION scattered on thin layer of soil and

mosses beneath a rock, subtropical gallery rainforest. Known only from type

locality.

DIAGNOSTIC CHARACTERS Basidiomata glistening, sticky, and laterally

stipitate; pileus white with a hint of yellowish-white, matted fibrillose, minutely

canescent near or at the margin, taste and odor mild; basidiospores 6-angled,

262 ... Largent & al.

PLaTE 3 - Claudopus minutoincanus (DL Largent 9871: HoLorype). A: Basidiomata (7x);

B: Pileipellis near point of attachment (120x); C: Pileipellis at pileus margin (250x); D: Basidiospores

(1000x); E: Subcapitulate to capitulate pileocystidium left (400x).

Saxicolous Claudopus spp. nov. (Australia) ... 263

on the average subisodiametric (E = 1.22), and with an average length < 9.01

um, subcapitulate to capitulate pileocystidia; lack of hymenial cystidia and

clamp connections.

ComMMENTS— Please refer to the comments under Claudopus viscosus.

Discussion

All the basidiomata collected for the species described in this report stuck

to the surface of the collecting implements as well as fingers, the surface of

the collecting boxes, leaves, and pieces of paper. The surface hyphae of the

basidiomata glistened as if covered by a thin gelatinous material. A very thin

layer of colorless, non-staining, amorphous material was observed on the

capitulate to subcapitulate pileocystidia of Claudopus minutoincanus and

C. viscosus. No discernable material could be observed on any other hyphae

of the pileipellis of these two species and on any hyphae of any part of the

basidiomata of C. rupestris. There currently is no viable explanation for the

stickiness of the basidiomata.

Acknowledgements

We would like to thank Dr. Peter Wilson of the Botanic Gardens Trust in Sydney

Australia for the Latin translations that are included in our descriptions. The DNA

sequences generated are based upon work supported by the National Science Foundation

under Grant No. DRI 0922922 awarded to SE Bergemann. ‘The fieldwork in Australia was

supported by the Largent family trust and we are particularly grateful for the support

of Pamela Largent. Fieldwork was also supported and logistical support was provided

by the Australian Tropical Herbarium and the School of Marine and Tropical Biology,

James Cook University. Comments by the two reviewers, Dr. Timothy Baroni and

Dr. Andrew Methven, and by the nomenclature editor, Dr. Shaun Pennycook, were very

helpful.

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MY COTAXON

Volume 116, pp. 265-281 April-June 2011

DOI: 10.5248/116.265

Notes on Trametes (Basidiomycota) in China

Hat-Jrao LI & SHUANG-Hu!1 HE*

Institute of Microbiology, P.O. Box 61, Beijing Forestry University, Beijing 100083, China

*CORRESPONDENCE TO: heshh1981@yahoo.cn

ABSTRACT —Four Trametes species, T. ellipsospora, T.: maxima, T. mimetes, and T. tephroleuca,

are reported as new for the Chinese fungal flora. Illustrated descriptions of these species are

given according to the Chinese materials. A key is provided to the twenty-six species recorded

in China. The relationships of Chinese Trametes spp. and related genera are discussed.

Key worps — Polyporaceae, taxonomy, wood-decaying fungi

Introduction

Trametes Fr. was established based on T! suaveolens (L.) Fr. as type species

by Fries (1836). The genus is characterized by pileate basidiocarps, a trimitic

hyphal system with clamp connections on generative hyphae, hyaline and

thin-walled basidiospores that are negative in Melzer’s reagent, and species

that cause a white rot (Gilbertson & Ryvarden 1987, Nufiez & Ryvarden 2001,

Ryvarden & Gilbertson 1994). Trametes was considered as one of the most

complicated genera in the Polyporaceae. Many species in the genus share similar

basidiospores. Many taxonomic studies have been carried out during the last

thirty years (Corner 1989, Dai et al. 2007a, Gilbertson & Ryvarden 1987, Hattori

2005, Ko & Jung 1999a,b, Leessoe & Ryvarden 2010, Li & Cui 2010, Nufiez &

Ryvarden 2001, Quanten 1996, Roy & De 1996, Ryvarden 2004, 2009, Ryvarden

& Iturriaga 2003, Ryvarden & Johansen 1980, TomSovsky 2008, TomSovsky et

al. 2006, Zhang et al. 2006, Zhao 1998, Zhao et al. 1983), and around 50 species

in the genus are accepted worldwide (Kirk et al. 2008). Twenty-two species

were recorded in China previously (Cui et al. 2008, Dai 2009, Dai & Penttila

2006, Dai & Yuan 2010, Dai et al. 2003, 2004, 2007a,b,c,d, 2009a, Li & Cui 2010,

Li et al. 2007, 2008, Teng 1996, Wang et al. 2009, Yuan & Dai 2008, Zhao 1998,

Zhao & Zhang 1991). Among them, six species are considered medicinal fungi

(Dai & Yang 2008, Dai et al. 2009b).

During our study of Trametes in China, four species were newly identified

and are briefly described here based on the Chinese materials. A key to the

266 ... Li & He

Chinese species of Trametes is provided, and relationships among the related

species are discussed.

Materials & methods

The studied specimens were deposited in herbaria as cited below. All the materials

were examined under microscope Nikon ECLIPSE 80i. Sections were studied under

magnification up to x 1000. In presenting the variation in the size of the spores, 5% of the

measurements were excluded from each end of the range, and are given in parentheses.

The width of a basidium was measured at the thickest part, and the length was measured

from the apex (sterigmata excluded) to the basal septum. In the text the following

abbreviations were used: 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 among the specimens studied (quotient of the mean spore length and the mean

spore width of each specimen), n = number of spores measured from given number of

specimens. IKI = Melzer’s reagent, IKI- = both inamyloid and nondextrinoid; KOH

= 5% potassium hydroxide; CB = Cotton Blue, CB- = acyanophilous, CB+ = cyano-

philous. Drawings were made with the aid of a drawing tube. Special color terms are

from Petersen (1996).

Taxonomy

Trametes ellipsospora Ryvarden, Mycotaxon 28(2): 539, 1987. FIG. 1

FruiTBopy — Basidiocarps annual, pileate to effused-reflexed, usuallyimbricate,

without odour or taste when fresh, soft coriaceous and light in weight when

dry. Pilei semicircular to almost circular with a umbilicate base, projecting up

to 2.5 cm long, 3 cm wide, 2 mm thick at base; pileal surface cream, clay-buff

to pale gray after drying, velutinate, slightly concentrically zonate and sulcate,

irregular warts or protuberances occasionally present near the base; margin

thin and sharp. Pore surface cream to straw-colored, glancing; pores round to

angular, 4-6 per mm; dissepiments thin, entire. Context cream, coriaceous, up

to 1.3 mm thick. Tube layer cream to pale straw-colored, coriaceous, up to 0.7

mm long.

HyPHAL STRUCTURE — Hyphal system trimitic; generative hyphae bearing

clamp connections; skeletal and binding hyphae dominant, thick-walled to

subsolid, IKI-, CB-; tissue unchanged in KOH.

CONTEXT — Generative hyphae scanty, hyaline, thin-walled, branched,

2-3 um in diam; skeletal hyphae dominant, hyaline, thick-walled to subsolid,

branched, sometimes collapsed, interwoven, 2.8-5 um in diam; binding hyphae

hyaline, thick-walled to almost solid, branched, interwoven, 1.5-3.2 um in

diam.

TuBEs — Generative hyphae infrequent, hyaline, thin-walled, rarely

branched, 1.5-3 um in diam; skeletal hyphae dominant, hyaline, thick-walled

to subsolid, branched, interwoven, 2-3.5 um; binding hyphae hyaline, flexuous,

in China... 267

Trametes

AN IEe S

ESS

ISS

——

>> oy, \ <> w=

of Trametes ellipsospora (drawn from Cui 6259).

idia and basidioles. c: Cystidioles.

. b: Bas

| cystidia. e: Hyphae from tube. f: Hyphae from context

sidiospores

268 ... Li & He

thick-walled to almost solid, branched, 1.6-3.2 um. Facial cystidia occasionally

present, globose, pyriform, clavate to fusoid, hyaline, thin-walled, 11-25 x

7-10.5 um; fusoid cystidioles occasionally present in the hymenium, hyaline,

thin-walled, 13-19 x 3-4.5 um; basidia clavate, with four sterigmata and a basal

clamp connection, 9-17 x 3.7-5 um; basidioles in shape similar to basidia, but

slightly smaller.

Spores — Basidiospores ellipsoid, hyaline, thin-walled, smooth, IKI-,

CB-, (3-)3.2-4.6(-5.1) x (2.1-)2.6-3.2(-3.5) um, L = 3.86 um, W = 2.98 um,

Q = 1.29-1.31 (n = 90/3).

SPECIMENS EXAMINED — CHINA. YUNNAN PROVINCE, Mengla County, Xishuangbanna

Botanical Garden, on fallen angiosperm trunk, 31.X.2009 Cui 8343 (BJFC). HAINAN

PROVINCE, Chengmai County, on fallen angiosperm trunk, 6.V.2009 Cui 6259 (BJFC);

Wanning County, Tianmao, on fallen angiosperm trunk, 14.V.2009 Cui 6665 (BJFC).

REMARKS — Trametes ellipsospora is characterized by its small ellipsoid

basidiospores. Our collections fit all the characters except its upper surface,

which was originally described as glabrous (Ryvarden 1987). After studying

one authentic specimen collected by Ryvarden, however, we found that it

also is velutinate. One specimen collected from Yunnan Province, Southwest

China, seems unique, because although cystidia are usually absent in Trametes,

different kinds of facial cystidia were observed on or near the pore surface.

Trametes ellipsospora resembles T. marianna (Pers.) Ryvarden, but the latter

species has cylindrical basidiospores (6-7 x 2-2.5 um; Ryvarden & Johansen

1980). Trametes pavonia (Hook.) Ryvarden, which resembles T: ellipsospora in

velutinate pileus and small pores (5-6 per mm), can be separated by larger

basidiospores (5-6 x 3-4 um; Gilbertson & Ryvarden 1987).

Trametes maxima (Mont.) A. David & Rajchenb., Mycotaxon 22(2): 315, 1985. Fie. 2

FruiTBopy — Basidiocarps annual, pileate, solitary or imbricate, without odour

or taste when fresh, corky and light in weight when dry. Pileus semicircular to

dimidiate, projecting up to 2.5 cm long, 3.4 cm wide, 2 mm thick at base; pileal

surface buff-yellow to cinnamon-buff after drying, glabrous, concentrically

zonate and sulcate; margin thin, entire or slightly wavy. Pore surface cinnamon-

buff to cinnamon; pores angular, 3-4 per mm; dissepiments thin, slightly

lacerate. Context cream, corky, up to 1.2 mm thick, a black line present towards

upper surface. Tube layer concolourous with pore surface, corky, up to 0.8 mm

long.

HyPHAL STRUCTURE — Hyphal system trimitic; generative hyphae bearing

clamp connections; skeletal and binding hyphae dominant, thick-walled to

subsolid, IKI-, CB-; tissue unchanged in KOH.

CONTEXT — Generative hyphae scanty, hyaline, thin-walled, rarely branched,

1.9-3 um in diam; skeletal hyphae dominant, hyaline, thick-walled to subsolid,

(

0 0 lf

ie : \ y

"s d

som 101m

Fic. 2. Microscopic structures of Trametes maxima (drawn from Dai 6865).

a: Basidiospores. b: Basidia and basidioles. c: Hyphae from tube. d: Hyphae from context.

branched, interwoven, 3-5.5 um in diam; binding hyphae hyaline, thick-walled

to subsolid, frequently branched, interwoven, 1.2-3.2 um in diam.

270 ... Li & He

TuBEs — Generative hyphae infrequent, hyaline, thin-walled, rarely

branched, 1.6-2.4 um in diam; skeletal hyphae dominant, hyaline, thick-walled

to subsolid, branched, interwoven, 2.5-3.8 um; binding hyphae hyaline, thick-

walled to subsolid, frequently branched, interwoven, 0.8-2.5 um. Cystidia and

cystidioles absent; hyphal pegs occasionally present; basidia clavate, with four

sterigmata and a basal clamp connection, 10-15 x 3-5 um; basidioles in shape

similar to basidia, but slightly smaller.

Spores — Basidiospores oblong ellipsoid, hyaline, thin-walled, smooth,

IKI-, CB-, (4.1-)4.2-5.1(-5.8) x 2-2.4(-2.5) um, L = 4.78 um, W = 2.18 um,

Q =2.19 (n = 30/1).

SPECIMEN EXAMINED — CHINA. YUNNAN PROVINCE, Menglun County, Xishuangbanna

Botanical Garden, on fallen angiosperm trunk, 6.VIII.2005 Dai 6865 (BJFC).

REMARKS — Trametes maxima is characterized by its buff-yellow to cinnamon-

buff, glabrous, concentrically zoned, sulcate pileus, its lacerate pore surface,

and the black line in the context towards the upper surface.

Trametes cystidiolophora B.K. Cui & H.J. Li resembles T. maxima in an

uneven pore surface and pore size (2-3 per mm), but T. cystidiolophora differs

by its pale grayish brown to pale cinnamon-buff pileal surface with distinctly

concentric zones and radial veins and larger basidiospores (6.6-9.2 x 2.4-3 um;

Li & Cui 2010).

A black zone is also present in T: hirsuta (Wulfen) Pilat and T. versicolor (L.)

Lloyd, but they have distinctly cylindrical basidiospores.

Trametes mimetes (Wakef.) Ryvarden, Norweg. J. Bot. 19: 236, 1972. FIG. 3

FruiTBopy — Basidiocarps annual, pileate to effused-reflexed, usually

imbricate, without odour or taste when fresh, coriaceous to corky and light in

weight when dry. Pileus semicircular to dimidiate, projecting up to 1.1 cm long,

2.7 cm wide, 3.5 mm thick at the base; pileal surface cream to buff after drying,

glabrous, narrowly concentrically zonate and sulcate and radially wrinkled;

margin thin and sharp, entire or slightly lobed and incised. Pore surface buff

to buff-yellow, glancing; pores round, 3-4 per mm; dissepiments thin, entire.

Context cream, corky, up to 0.8 mm thick. Tube layer buff to buff-yellow, corky,

up to 2.7 mm long.

HyPHAL STRUCTURE — Hyphal system trimitic; generative hyphae bearing

clamp connections; skeletal and binding hyphae dominant, thick-walled to

subsolid, IKI-, CB-; tissue unchanged in KOH.

CONTEXT — Generative hyphae scanty, hyaline, thin-walled, rarely branched,

2.5-3.5 um in diam; skeletal hyphae dominant, hyaline to pale yellowish, thick-

walled with a wide lumen, branched, interwoven, 2.5-5 um in diam; binding

hyphae hyaline to pale yellowish, thick-walled to almost solid, frequently

branched, interwoven, 1.2-3.5 um in diam.

n China... 271

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272 ... Li & He

TuBESs — Generative hyphae infrequent, hyaline, thin-walled, rarely

branched, 1.8-2.5 um in diam; skeletal hyphae dominant, hyaline to pale

yellowish, thick-walled with a wide to narrow lumen, frequently branched,

interwoven, 2-3.3 um; binding hyphae hyaline to pale yellowish, flexuous,

thick-walled to almost solid, frequently branched, interwoven, 1-3 um. Cystidia

and cystidioles absent; hyphal pegs occasionally present; basidia clavate, with

four sterigmata and a basal clamp connection, 16-20 x 6-7 um; basidioles in

shape similar to basidia, but slightly smaller.

SporES — Basidiospores cylindrical, hyaline, thin-walled, smooth, IKI-,

CB-, (10—)10.2-12.8(-14) x 3-4 um, L = 11.56 um, W = 3.53 um, Q = 3.27

(n = 30/1).

SPECIMEN EXAMINED — CHINA. JIANGXI PROVINCE, Jinggangshan County,

Jinggangshan Nature Reserve, on fallen angiosperm trunk, 23.1X.2008 Dai 10608

(BJFC).

REMARKS — Trametes mimetes is characterized by its glabrous, concentrically

zoned, sulcate, radially wrinkled pileus and large cylindric basidiospores.

Ryvarden & Johansen (1980) describe the pileal surface as ochraceous or fulvous

to dark brown; the Chinese specimen is paler, its pores are slightly smaller (2-3

per mm), and its basidiospores are larger than the 8-11 x 3.2-4 um given by

Ryvarden & Johansen (1980).

Trametes tephroleuca Berk., Hooker's J. Bot. Kew Gard. Misc. 6: 165, 1854. Fic. 4

FruiTBopy — Basidiocarps annual, pileate, solitary or imbricate, corky when

dry; pilei semicircular, dimidiate to flabelliform; projecting up to 5 cm, 7.8 cm

wide, and 1.2 cm thick at the base; margin obtuse. Pileal surface cream to pale

brown when dry, concentrically sulcate or not, strigose to hirsute; pore surface

cream to pale buff, becomes gray with age; pores round to angular, 1-2 per mm;

dissepiments moderately thick, entire; context cream, corky when dry, up to

7 mm thick, a black line occasionally present towards the upper surface; tubes

cream and become gray with age, corky when dry, up to 5 mm long.

HYPHAL STRUCTURE — Hyphal system trimitic; generative hyphae bearing

clamp connections; skeletal and binding hyphae IKI-, CB-; tissue unchanged

in KOH.

CONTEXT — Generative hyphae hyaline, thin-walled, 1.5-4.8 um in diam;

skeletal hyphae hyaline, thick-walled to subsolid, most with a narrow lumen,

usually collapsed when with a wide lumen, branched, more or less regularly

arranged, 2.3-6.5 um in diam; binding hyphae hyaline, thick-walled to subsolid,

frequently branched, strongly interwoven, 1.2-3 um in diam.

TuBEes — Generative hyphae hyaline, thin-walled, 1.3-3.5 um in diam;

skeletal hyphae hyaline, thick-walled to subsolid, branched, interwoven,

sometimes projecting into the hymenium with or without a thin-walled and

Trametes in China... 273

YW) FI

ae DOL ROE

10 yum

a TN

10 ym

Fic. 4. Microscopic structures of Trametes tephroleuca (drawn from Cui 7977).

a: Basidiospores. b: Basidia and basidioles. c: Cystidioles.

d: Hyphae from trama. e: Hyphae from context.

274 ... Li & He

inflated tip, 2-5 um in diam; binding hyphae hyaline, thick-walled to subsolid,

frequently branched, strongly interwoven, 1-2.3 um in diam; cystidia absent,

fusoid cystidioles occasionally present, 13-17 x 4-6 um; basidia clavate to

barrel-shaped, with four sterigmata and a basal clamp connection, 12-16 x

4-5.5 um; basidioles in shape similar to basidia, but slightly smaller.

Spores — Basidiospores cylindrical to oblong ellipsoid, hyaline, thin-walled,

smooth, IKI-, CB-, (4.5-) 4.8-7(-9) x (2-) 2.5-3.3 um, L = 5.87 um, W = 2.88

um, Q = 1.90-2.19 (n = 60/2).

SPECIMENS EXAMINED — CHINA. YUNNAN PROVINCE, Tengchong County, Gaoligong

Mountains, on fallen angiosperm trunk, 23.X.2009 Cui 7977, 7981 & 7987 (BJFC).

REMARKS — Trametes tephroleuca is characterized by its strigose to hirsute

pileus and pores that turn grayish with age. Berkeley (1854) described it from

Nepal and Roy & De (1996) reported it from India. The species resembles

T: hirsuta, characterized by similar basidiocarps and pores that age grayish to

blackish, but differs in smaller pores (3-4 per mm) and narrower basidiospores

(6-9 x 2-2.5 um; Nufiez & Ryvarden 2001).

Key to species of Trametes in China

(spore dimensions are provided after species names)

1. Basidiocarps with sterile cup-shaped structures....... T. conchifera (Schwein.) Pilat

6.1-8 x 2-2.9 um, L = 7.2 um, W = 2.2 um, Q = 3.3 (n = 30/1)

1. Basidiocarps without sterile cup-shaped structures............. 0. cece eee eee 2

2. Pileus cream, with strong anise odour when fresh................... T. suaveolens

6.5-9 x 3-3.7 um, L = 7.3 um, W = 3.2 um, Q = 2.1-2.4 (n = 60/2)

2 Pileus cream to brown to gray, without anise odour when fresh. .................. 3

3, Pores 1=3-permiipor even larger. eso. seu x sete saw ie ele see oe ea le Oe ae od 4

.. POLES io <1 OND OT BNL. «Fats faa son Maes en Bays Paes Payee Bese Be te ne Lys

4, Hymenophore become grayish with age ..................000e eee T. tephroleuca

4.8-7 x 2.5-3.3 um, L = 5.9 um, W = 2.9 um, Q = 1.9-2.2 (n = 60/2)

4 TIyMENOph Ore ne ver DECOM OAVISH <i, ote ely Beayculs Hebets lates tee Beene wMaadehe opaudle tears 5

5. Pileus persistently velutinate, strigose to tomentose ............ 0.00. cece eee eee 6

5. Pileussmiore.or lessalabrous: 265 2. ctw 22m Senco Gch lence law chy Cina bly aincbry ee 9

Gi BasichGspores 55; pit LON Gis. < oes ac ee pa ies ace gahyaee Badin eer ad sherew dian Dace « 7

Ga Basidiospores? > S25 Lil Onkyo aco sln te gala ee glenda palace aed adios obainesa dct acebeotte 8

7. Pileus velutinate to glabrous, pores radially elongated ......... T. gibbosa (Pers.) Fr.

3.1-4.9 x 1.9-2.5 um, L = 4.1 um, W = 2.1 um, Q =2 (n = 60/2)

7. Pileus strigose to tomentose, pores angular.............. T. pocas (Berk.) Ryvarden

4.2-5.5 x 2.4-3 um, L = 4.8 wm, W = 2.7 um, Q = 1.8 (n = 30/1)

8. Pileus velutinate, context duplex............. T. subsuaveolens B.K. Cui & Y.C. Dai

(5.2-)5.6-7.2(-8.2) x (2.3-)2.3-3(-3.1) pm, L = 6.38 pm, W = 2.56 pm, Q = 2.46-2.52 (n = 60/2)

8. Pileus strigose to tometose, context homogeneous .......... T. villosa (Sw.) Kreisel

5.5-8.5 x 2.5-3.5 um (Gilbertson & Ryvarden 1987)

Trametes in China... 275

9. Hymenophore irregular, often different from the base to the margin

Leite +. sina ena, SM Cee Fe, Heeatienh, oie taremre ane: cine fs T. elegans (Spreng.) Fr.

5.8-7 x 2.3-3 um, L = 6.3 um, W = 2.8 um, Q = 2.3 (n = 60/2)

9. Hymenophore:regular, angular to:circtlar’. «5 0.te seas + vines 6 heed 6 Oneida ele ons 10

10.

Vil

hz,

1 e%,

hs:

13.

14.

lich

LS;

16.

iy.

Ly.

18.

19:

20.

Zl,

Pore surface distinctly dentate to form a hydnoid surface ....... T. cystidiolophora

6.6-9.2 x 2.4-3 um, L = 8.1 um, W = 2.8 um, Q = 2.8-3.0 (n = 60/2)

POLE SUTAGE OVER co Face Mage ese Rage bess Raat eet gt Ree Be Ms ates oma 11

Distributed in warm temperate China, pores 1.5-2 per mm T: lactinea (Berk.) Sacc.

5.2-6.7 x 2.1-2.8 um, L = 6 um, W = 2.3 um, Q = 2.6 (n = 30/1)

Distributed in tropical to subtropical China, pores 2-3 per mm

Vttengonh oasis, 5 ote da. ken dealan.s seas ened ands eds T. orientalis (Yasuda) Imazeki

6-8 x 2.6-3.1 um, L = 7.1 wm, W = 2.9 um, Q = 2.5 (n = 30/1)

Pileus.persistently velutinates-tometose tO iPS te slates glass gee anaes aace tata sedate 13

BileusmOre sor Less; STADTOUGS. vel, lina he ofan aly whancbag tole t wautbes heute abate okt tbe oll 18

Hymenophore become grayish or even darker with age................ T. hirsuta

5.5-8 x 2.5-3.3 um, L = 6.5 um, W = 2.9 um, Q = 2.1-2.6 (n = 270/9)

Eymenophorenever become grayishior blake # ie eof in wale ee w gle wie Wiehe, Win e's Rg ecel 14

Basidiospores ellipse 0225 ces Baca. ce Big Rage gh a Rg lo RR a lo Baie le vn ot 15

Basi @sporesiey rtd rial te 7 scart: temvarats tina ak Sank cee teaacshe toarkcote tearkcot eakerteat saa e Roti « 16

Pileus multizonate, more or less ochraceous. ..........0--. eee eee eens T. pavonia

4.4-5.2 x 2.9-3.3 um, L= 4.9 um, W = 3 um, Q = 1.6 (n = 30/1)

Pileus slightly concentrically zonate and sulcate, cream to clay-buff T. ellipsospora

3.2-4.6 x 2.6-3.2 um, L = 3.9 um, W = 3 um, Q = 1.3 (n= 90/3)

Pileus with a black layer between context and upper tomentum....... T. versicolor

4-5 x 1.7-2 um, L= 4.4 um, W = 1.9 um, Q = 2.3 (n = 30/1)

Pileus without a black layer between context and upper tomentum............. 17

Pileal surface cream to warm buff................. T. pubescens (Schumach.) Pilat

6-7 x 1.8-2.1 um, L = 6.5 um, W = 1.9 um, Q = 3.4 (n = 30/1)

Pileal surface ochraceous to reddish brown ..... T. ochracea (Pers.) Gilb. & Ryvarden

5-7 x 2-2.8 um, L = 6 um, W = 2.4 um, Q = 2.2-2.8 (n = 180/6)

BASTCIOS POLES O 4 ATe WIE eset 1cbe 08.8 laden laa Sata ataeh aba rina tates Wass lone ake heat nue 19

Basidiospores:<y Se prik Wide: eos eos eae eee te meow en OF eset Solna ene t Salk 20

Basidiospores“ellipsoid. 32 s6ceg shove eae Vie nsey eee Ee oe Ee T. Ijubarskyi Pilat

4.7-5.8 x 3-4 wm, L = 5.2 um, W = 3.5 um, Q = 1.5 (n = 30/1)

Dasidiospores ey lind fival. Ay xem Fgh d Batak hast Ra! Mattel ak Sats bet | T. mimetes

10.2-12.8 x 3-4 um, L = 11.6 um, W = 3.5 um, Q = 3.3 (n = 30/1)

Basidiospores < 4 um long, basidiocarps pinkish brown to cocoa-colored,

usually covered with cream to ochraceous outgrowth spreading from the

| ey orca RNG A OG ABE Dv, PBI ol Seo T. modesta (Kunze ex Fr.) Ryvarden

3.1-4 x 1,8-2.2 um, L = 3.7 um, W = 2.1 um, Q = 1.6-2 (n = 210/7)

Basidiospores > 4 um long, basidiocarps different.................... 0.0 ee eee val

Basidiospores ntogthy 1.52 pit Wide sw tee ee Glace Seah ethno ea nce h aly ts dle 22.

ASIC OSDO Tes: NIOStY 2 Sa MP a Ped ale oe egal bg ao Wig dnt tects Rg el 23

276 ... Li & He

22;

23.

23%

24.

25.

26;

Pileus applanate, white to buff, dissepiments rather thin, context unchanged in

1566 in Nene este nhc Font «Or nt. rt ata Cod ert at COE T. velutina (Pers.) G. Cunn.

5-6 x 1.8-2.1 um, L = 5.7 um, W = 1.9 um, Q = 2.7-3.3 (n = 180/6)

. Basidiocarps usually sub-stipitate, pileus in different gray shades, context turns to

SO WETTED Scan dts bite eth iktme dey aaa DE ht ne teeth g T. menziesii (Berk.) Ryvarden

5-6.3 x 1.8-2.1 um, L = 5.6 um, W = 2 um, Q = 2.8 (n = 60/2)

PORES? S sO PICTON. Bhs aalucndhy Salucnths, ilucke 6 Btlycke y Alucke § alucds wAhtiucks sAdeuahs duo sa 24

POPES D> POM AME Is Ne oes thee Sp hg Ro halt Sethe redial tetigale ol kale oa alt oe Salt eg 2

Pileus flabelliform, thin and flexible, cream to pale tan, usually radial wrinkled

SA TROTNREE VAs ce Ra atone st aehee Re oy tn tle edi duly? T. membranacea (Sw.) Kreisel

4.5-6 x 2-2.5 um (Gilbertson & Ryvarden 1987)

Pileus applanate, multizonate in tan to brown and sulcate, often turn to reddish

brown from the base......... T. ectypa (Berk. & M.A. Curtis) Gilb. & Ryvarden

4-5.8 x2-2.3 um, L = 4.4 um, W = 2.1 um, Q = 2.1 (n = 26/1)

Pileus cinnamon, basidiospores mostly < 5 um long.................. T. maxima

4.2-5.1 x 2-2.4 um, L= 4.8 um, W = 2.2 um, Q = 2.2 (n = 30/1)

Pileus glabrous, cream to pinkish-buff to gray, basidiospores mostly >5 um long 26

Pileal surface more or less grayish, distinctly tuberculate

FOOD Bs Na AON Lear hay ately ahr ry ae T. manilaensis (Lloyd) Teng

5-7.8 x 2.2-3 um, L = 6.3 um, W = 2.5 um, Q = 2.5 (n = 30/1)

Pileal surface cream-ochraceous to brown, more or less smooth ...... T. ochracea

5-7 x 2-2.8 um, L = 6 um, W = 2.4 um, Q = 2.2-2.8 (n = 180/6)

OTHER SPECIMENS EXAMINED — Trametes conchifera CHINA. HEILONGJIANG

PROVINCE, Yichun, Fenglin Nature Reserve, on fallen trunk of Ulmus, 8.1X.2002 Dai

3670 (IFP); Ning’an County, Jingpohu Park, on fallen trunk of Ulmus, 8.1X.2007 Dai

8359 (IFP). INNER MonGcoLia AUTONOMOUS REGION, Tongliao, Daqinggou Nature

Reserve, on fallen angiosperm trunk, 24.IX.2002 Dai 3966 (IFP). JILIN PROVINCE, Antu

County, Changbaishan Nature Reserve, on fallen angiosperm trunk, 12.XII.2007 Dai

9069 (IFP).

T. cystidiolophora. CHINA. YUNNAN PROVINCE, Baoshan, Gaoligongshan Nature

Reserve, on dead angiosperm tree, 25.X.2009 Cui 8084 & 8087 (BJFC).

T. ectypa. CHINA. ZHEJIANG PROVINCE, Linan County, Tianmushan Nature

Reserve, on fallen angiosperm trunk, 9.X.2005 Cui 2580 (IFP).

T. elegans. CHINA. HAINAN PROVINCE, Danzhou, on fallen angiosperm trunk,

7.V.2009 Cui 6293 (BJFC). GUANGDONG PROVINCE, Shixing County, Chebaling Nature

Reserve, on fallen angiosperm trunk, 13.1X.2009 Cui 7398 (BJFC).

T. gibbosa. CHINA. ZHEJIANG PROVINCE, Linan County, Tianmushan Nature

Reserve, on fallen trunk of Liquidambar formosana, 8.X.2005 Cui 2524 (IFP). JILIN

PROVINCE, Antu County, Changbaishan Nature Reserve, on fallen trunk of Tilia,

12.XII.2007 Dai 9026 (IFP). GUANGDONG PROVINCE, Shixing County, Chebaling

Nature Reserve, on fallen angiosperm trunk, 12.1X.2009 Cui 7390 (BJFC).

T. hirsuta. CHINA. SHANDONG PROVINCE, Taian, Tai Mountains, on fallen trunk of

Populus, 26.1X.2005 Cui 2496 (IFP); Linyi, Linyi Botanical Garden, on fallen angiosperm

trunk, 17.VII.2009 Cui 6774 (BJFC). HAINAN PROVINCE, Chengmai County, on fallen

angiosperm trunk, 6.V.2009 Cui 6238 & 6241 (BJFC). FuyIAN PROVINCE, Wuyishan

County, Wuyi Palace, on angiosperm stump, 29.VIII.2006 Cui 4190 (IFP). BEIJING,

Trametes in China... 277

Beijing Botanical Garden, on living tree of Prunus, 9.V1I.2008 Cui 5547 & 5549 (BJFC).

ANHUI PROVINCE, Huangshan County, Huangshan Mountains, on fallen trunk of

Prunus, 10.X.2004 Dai 6034 (IFP). GUANGDONG PROVINCE, Ruyang County, Nanling

Nature Reserve, on fallen trunk of Castanopsis, 14.V.2009 Dai 10883 & 10889(BJFC).

T. lactinea. CHINA. JILIN PROVINCE, Hunchun, Hadamen, on fallen trunk of

Quercus, 7.VIII.2009 Cui 7084 (BJFC). JIANGxI PROVINCE, Jiujiang, Nanhu Park, on

fallen angiosperm trunk, 10.X.2008 Cui 6097 (BJFC).

T. [jubarskyi. CHINA. HENAN PROVINCE, Neixiang County, Baotianman Nature

Reserve, on fallen angiosperm branch, 28.VIII.2005 Li 286 (IFP).

T. manilaensis. CHINA. HAINAN PROVINCE, Danzhou County, on fallen trunk of

Acacia, 7.V.2009 Dai 10747 (BJFC).

T. menziesii. CHINA. GUANGDONG PROVINCE, Ruyang County, Nanling Nature

Reserve, on fallen angiosperm trunk, 16.1X.2009 Cui 7563, 7583, 7603 & 7613 (BJFC);

Shixing County, Chebaling Nature Reserve, on fallen trunk of Castanopsis carlesii,

12.1X.2009 Cui 7360 (BJFC); on fallen angiosperm trunk, 13.1X.2009 Cui 7419 (BJFC);

14.1X.2009 Cui 7465 (BJFC).

T. modesta. CHINA. FUJIAN PROVINCE, Jianou County, Wanmulin Nature Reserve,

on fallen angiosperm trunk, 30.VII.2006 Cui 4210 (IFP). HAINAN PROVINCE, Ledong

County, Jianfengling Nature Reserve, on fallen angiosperm trunk, 18.X1.2007 Dai 9290

(BJFC); on fallen trunk of Cyclobalanopsis, 11.V.2009 Dai 10844 (BJFC); on fallen branch

of Syzygium, 8.V.2009 Cui 6400 (BJFC); Changjiang County, Bawangling Nature Reserve,

on fallen angiosperm trunk, 8.V.2009 Cui 6406 (BJFC); on living tree of Cyclobalanopsis,

8.V.2009 Cui 6407 (BJFC); on fallen trunk of Sapium discolor, 9.V.2009 Cui 6452 (BJFC).

YUNNAN PROVINCE, Mengla County, Xishuangbanna Tropical Botanical Garden, on

fallen angiosperm trunk, 31.X.2009 Cui 8364 (BJFC).

T. ochracea. CHINA. HEBEI PROVINCE, Xinglong County, Wulingshan Nature

Reserve, on fallen angiosperm trunk, 29.VII.2009 Cui 6848 (BJFC); on fallen trunk of

Populus, 30.V1I.2009 Cui 6888 (BJFC). JIANGSU PROVINCE, Nanjing, Zijin Mountains,

on fallen trunk of Xylosma racemosa, 3.V1.2005 Dai 6571 (IFP). JILIN PROVINCE,

Antu County, on fallen trunk of Betula, 11.III.1993 Dai 898 (IFP); on Stump of Betula,

9.1X.1995 Dai 2005 (IFP). SHAANXI PROVINCE, Zhouzhi County, Taibai Mountains, on

angiosperm stump, 24.X.2006 Yuan 2695 (IFP). SHANXI PROVINCE, Qinshui County,

Lishan Nature Reserve, on fallen trunk of Quercus, 18.X.2004 Yuan 964 (IFP); Jiaocheng

County, Pangquangou Nature Reserve, on fallen trunk of Betula, 22.1X.2006 Yuan

2477 (IFP). ZHEJIANG PROVINCE, Lin'an County, Tianmushan Nature Reserve, on

angiosperm stump, 12.X.2005 Cui 2757 (IFP).

T. orientalis. CHINA. HAINAN PROvINCE, Danzhou County, on fallen trunk of

Acacia, 7.V.2009 Cui 6300 (BJFC). GUANGDONG PROVINCE, Ruyang County, Nanling

Nature Reserve, on fallen angiosperm trunk, 17.1X.2009 Cui 7642 (BJFC).

T. pavonia. CHINA. Tarwan PROVINCE, Taichung, on fallen angiosperm trunk,

VUI.1997 Dai 11596 (IFP).

T. pocas. CHINA. HAINAN PROVINCE, Changjiang County, Bawangling Nature

Reserve, on fallen angiosperm branch, 7.XII.2009 Dai 11577 (BJFC). YUNNAN

PROVINCE, Tengchong County, Gaoligong Mountains, on fallen angiosperm trunk,

25.X.2009 Cui 8074 (BJFC).

T. pubescens. CHINA. HEBEI PROVINCE, Xinglong County, Wulingshan Nature

Reserve, on fallen angiosperm trunk, 30.VII.2009 Cui 6902 (BJFC). JILIN PROVINCE,

Antu County, Changbaishan Nature Reserve, on fallen trunk of Populus, 19.1X.2002 Dai

3824 (IFP). GUANGDONG PROVINCE, Shixing County, Chebaling Nature Reserve, on

fallen angiosperm trunk, 14.[X.2009 Cui 7464 (BJFC).

278 ... Li & He

T. suaveolens. CHINA. SHANXI PROVINCE, Ningwu County, Fengxiang Mountains,

on fallen trunk of Salix, 8.IV.2009 Dai 10729 (BJFC). HEILONGJIANG PROVINCE,

Ning’an County, Jingpohu Park, on fallen trunk of Betula, 10.[X.2007 Dai 8888 (IFP).

T. subsuaveolens. CHINA. INNER MONGOLIA AUTONOMOUS REGION, Chifeng,

Baiyinaobao Nature Reserve, on fallen trunk of Picea, 19.[X.2003 Cui 269 & 258 (IFP).

T. velutina. CHINA. SHAANXI PROVINCE, Zhouzhi County, Taibai Mountains, on

fallen angiosperm trunk, 24.X.2006 Yuan 2628 (IFP); Foping County, Foping Nature

Reserve, on stump of Quercus, 27.X.2006 Yuan 2774 (IFP). JILIN PROVINCE, Antu

County, Changbaishan Nature Reserve, on fallen trunk of Betula, 1.[X.1993 Dai 971

(IFP); 11.1X.1995 Dai 2135 (IFP); on fallen decorticated trunk of Acer, 11.IX.1995 Dai

2008 (IFP). LIAONING PROVINCE, Kuandian County, Tianhuashan Nature Reserve, on

fallen branch of Betula, 29.VII.2008 Cui 5625 (BJFC).

T. versicolor. CHINA. JILIN PRovINCE, Antu County, Changbaishan Nature

Reserve, on fallen angiosperm trunk, 13.[X.2007 Dai 9156 (IFP). HEBEI PROVINCE,

Xinglong County, Wulingshan Nature Reserve, on stump of Betula, 29.VIII.2009 Cui

7182 (BJFC).

Discussion

Trametes species are not well known in China, and more samples and

molecular data are needed to define them. Trametes cervina (Schwein.) Bres.

(Bresadola 1903) was transferred to Funalia Pat. based on its cyanophilous

skeletal hyphae (Dai 1996), but nuclear LSU and mtSSU rDNA sequence analyses

support its transfer to the new genus Trametopsis TomSovsky, as Trametopsis

cervina (Schwein.) TomSovsky (TomSovsky 2008). Based on its unusual sterile

disc- or cup-shaped structures T. conchifera was recognized as Poronidulus

conchifer (Schwein.) Murrill (Murrill 1904). However, its hyphal structure and

spores are typical of Trametes, where we retain it. One specimen (HMAS 23451)

representing T: drummondii (Klotzsch) Ryvarden, previously was recorded in

China (Zhao 1998) was found to be sterile, making that species uncertain in

the country. Trametes elegans was considered to represent Lenzites (Nunez &

Ryvarden 2001) because of its irregular, partly lamellate pores, but molecular

data indicate it is closely related to Trametes (TomSovsky et al. 2006), where we

also place it. Nufiez & Ryvarden (2001) transferred T. scopulosa (Berk.) Bres. to

Whitfordia Murrill based on its laterally stipitate or sub-stipitate basidiocarps

with a blackish cuticle near the base and coloured vegetative hyphae; molecular

analyses are needed to support this placement. Trametes thujae J.D. Zhao was

described on Cupressaceae from southwestern China (Zhao & Zhang 1991);

Dai & Yuan (2010) place it in Funalia based on its coarsely tomentose to hirsute

pileus and cyanophilous skeletal hyphae.

Funalia is usually treated as a synonym of Trametes. Both genera share

trimitic hyphal system and more or less similar basidiospores, but the skeletal

hyphae are cyanophilous in Funalia and acyanophilous in Trametes (Dai 1996,

Niemela et al. 1992). Earliella Murrill is undoubtedly related to Trametes by

sharing the same type of hyphal system and basidiospores (Nunez & Ryvarden

Trametes in China... 279

2001). Earliella has resupinate to effused-reflexed basidiocarps with a reddish

upper surface and sinuous pores. However, effused-reflexed basidiocarps and

sinuous pores are also present in some Trametes species. Therefore, these

characters may not be sufficient to separate them, and further studies are

needed to revise the relationships of the two genera.

Lenzites Fr., which is closely related to Trametes, has lamellate pores and a

characteristic catahymenium with pointed skeletal hyphae. Coriolopsis Murrill,

Fomitella Murrill, and Hexagonia Fr. deviate from Trametes by their brown

colours (Cui et al. 2010, Hattori 2005, Nufiez & Ryvarden 2001). Further studies

are needed to delimitate the above-mentioned genera from Trametes.

Microporus P. Beauv. seems closely related to Trametes because both have

similar spores and hyphal structure, but Microporus is characterized by stipitate

basidiocarps, small pores (5-10 per mm), and (for some species) coralloid

dendrohyphidia in the dissepiments (Nufiez & Ryvarden 2001).

Pycnoporus P. Karst. is similar to Trametes in all characters except its bright

reddish-orange colour (Nufez & Ryvarden 2001, Ryvarden & Johansen 1980).

According to our experience, its bright colour does not last and fades with age.

Although analyses of LSU and ITS (nrDNA) sequences by TomSovsky et al.

(2006) place Pycnoporus inside the paraphyletic Trametes clade, for the time

being we retain Pycnoporus as an independent genus.

Acknowledgements

We express our gratitude to Dr. Michal TomSovsky and Dr. Hai-Sheng Yuan who

reviewed the manuscript. The research was financed by the Specific Programs in Graduate

Science and Technology Innovation of Beijing Forestry University (No. BLYJ2011-02),

the Fundamental Research Funds for the Central Universities (No.YX2010-22) and the

National Natural Science Foundation of China (No. 31000006 and No. 30900006).

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MY COTAXON

Volume 116, pp. 283-293 April-June 2011

DOI: 10.5248/116.283

Notes on Amylocorticiellum (Amylocorticiales, Basidiomycota),

with some new combinations

SERGIO P. GorJON™?, ALINA G. GRESLEBIN’” & MARIO RAJCHENBERG’”

‘Centro de Investigacion y Extension Forestal Andino Patagoénico, Area de Proteccion.

CC 14, 9200 Esquel, Chubut, Argentina

?Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET) Argentina

*CORRESPONDENCE TO: spgorjon@usal.es

AxpstTRAcT — A review of the known species in Amylocorticiellum is provided.

Amylocorticiellum molle is reported as new to Argentina as well to the Southern Hemisphere.

Three combinations in Amylocorticiellum, A. iaganicum, A. luteolum, and A. oblongisporum,

are proposed, and a key to all accepted species is provided.

KEY worps — corticioid fungi, Hypochniciellum, Patagonia, spore amyloidy, taxonomy

Introduction

Amylocorticiellum Spirin & Zmitr. was described to accommodate species

with corticioid basidiomes, smooth thick-walled basidiospores, and amyloid

spore walls (greyish to bluish in Melzer’s reagent), with Corticium subillaqueatum

as the generic type species (Zmitrovich & Spirin 2002). In addition, two

other species formerly placed in Hypochniciellum Hjortstam & Ryvarden,

Corticium cremeoisabellinum and Thelephora mollis, have been transferred to

Amylocorticiellum, and A. sinuosum was described as a new species (Zmitrovich

& Spirin 2002). Hypochniciellum originally included only Leptosporomyces

ovoideus Jiilich, characterized by thick-walled and cyanophilous basidiospores

that are negative in Melzer’s reagent (Hjortstam & Ryvarden 1980). Hjortstam

(1981) later emended the generic circumscription to include species with thick-

walled basidiospores with greyish walls that are not distinctly blue in Melzer’s

reagent. There are few reliable characters separating Amylocorticiellum and

Hypochniciellum except for the basidiospore amyloidy reaction. The fact that

some corticioid genera contain species with both amyloid and non-amyloid

basidiospores (e.g., Scytinostroma Donk, Vararia P. Karst.) while elsewhere

spore amyloidy is reliably diagnostic at the generic level has been supported

by molecular analysis (e.g., both Aphanobasidium Jilich and Amyloxenasma

284 ... Gorjén, Greslebin & Rajchenberg

(Oberw.) Hjortstam & Ryvarden produce smooth basidiospores that are amyloid

in the latter). It is noted that Hypochniciellum ovoideum (Jiilich) Hjortstam &

Ryvarden grows on both hardwood and conifer wood while almost all species

in Amylocorticiellum show a strong association with conifers.

Molecular studies support Amylocorticiellum subillaqueatum (type species

of Amylocorticiellum) in the Amylocorticiales K.H. Larss.et al., closely related

to the monotypic genus Podoserpula D.A. Reid (Binder et al. 2010). A BLAST

search of the LSU rRNA partial sequence of Amylocorticiellum molle available

in Genbank suggests that it is closely related to Amyloathelia crassiuscula

Hjortstam & Ryvarden (96% coincidence) and Corticium subillaqueatum (95%).

Hypochniciellum ovoideum, is still not molecularly analysed (Larsson 2007).

The recently proposed order Amylocorticiales has been shown to be sister to

the Agaricales s.l., suggesting that the radiation of pileate stipitate mushrooms

resulted from the elaboration of resupinate ancestors (Binder et al. 2010).

Most taxa referred to the Amylocorticiales (cf. Binder et al. 2010) have amyloid

spore walls. As several genera in this order appear to be non-monophyletic, a

sound generic classification within the Amylocorticiales awaits more intensive

sampling, especially from species that have not yet been sequenced.

To date, four species have been placed in Amylocorticiellum. Three more

species with smooth, thick-walled, amyloid basidiospores are still referred

to Hypochniciellum: H. luteolum from Colombia (Hjortstam & Ryvarden

2000), H. iaganicum from the Patagonian Andes forest (Greslebin 2002),

and H. oblongisporum from South Argentina and New Zealand (Greslebin

& Rajchenberg 1999). It has not been possible to perform molecular studies

of the South American species because we had no success in getting pure

cultures and/or DNA extraction. Nonetheless, we transfer these three species to

Amylocorticiellum, based on their morphology, leaving Hypochniciellum in its

restricted sense of the generic type. We also review the known Amylocorticiellum

species and provide a key for their identification.

Material & methods

For light microscopic studies, samples were mounted in 3% potassium hydroxide

(KOH), Melzer’s reagent (IKI), and 0.1% cotton blue in 60% lactic acid to determine

basidiospore cyanophily. Line drawings were made with a camera lucida attachment.

Specimens are deposited in the Centro de Investigacién y Extension Forestal Andino

Patagoénico (CIEFAP, Esquel, Argentina) herbarium. For species distribution in the

Southern Hemisphere we follow Hjortstam & Ryvarden (2007).

Taxonomy

Amylocorticiellum Spirin & Zmitr., Mikol. Fitopatol. 36(1): 22, 2002.

TYPE SPECIES: Corticium subillaqueatum Litsch.

Amylocorticiellum combs. nov. ... 285

Basidiome resupinate, effused, pellicular to membranaceous, margin fibrillose.

Hymenophore smooth to merulioid or hydnoid, light coloured. Hyphal system

monomitic, hyphae with evident clamps, thin- to thick-walled in the subiculum.

Cystidia usually absent, or present often poorly differentiated. Basidia clavate,

sinuous, with a basal clamp. Basidiospores cylindric to ellipsoid, smooth, thick-

walled, with a greyish to bluish reaction in Melzer’s reagent, cyanophilous. On

coniferous wood, causing a brown rot.

Amylocorticiellum cremeoisabellinum (Litsch.) Spirin & Zmitr., Mikol. Fitopatol.

36(1): 22, 2002.

= Corticium cremeoisabellinum Litsch., Ann. Mycol. 39(2/3): 117, 1941.

= Hypochniciellum cremeoisabellinum (Litsch.) Hjortstam, Mycotaxon 13(1): 125, 1981.

= Leucogyrophana cremeoisabellina (Litsch.) Parmasto, Eesti

NSV Tead. Akad. Toim., Biol. seer 16(4): 385, 1967.

ComMENTs — Amylocorticiellum cremeoisabellinum is characterized by a white

to cream-brown pellicular basidiome, ellipsoid basidiospores that are inamyloid

or slightly greyish in Melzer’s, and the lack of cystidial elements. For a detailed

description and illustration see Eriksson & Ryvarden (1976) and Kotiranta &

Saarenoksa (1990).

Known from continental Europe and Russia. Two specimens from Tierra

del Fuego (Argentina) previously reported (Hjortstam & Ryvarden 1985) have

since been referred to H. iaganicum (Greslebin 2002).

neal DADAD QDaGDA

apopo90 AVOVO GNDID

qdgq00. GODQG, 0QUDD.

OQO0QIBADH

“QO00C 0000

PLaTE 1. Basidiospores. a) Amylocorticiellum luteolum LR 15952;

b) A. molle SPG 2991; c) A. oblongisporum SPG 2933; d) A. iaganicum AG 1507.

286 ... Gorjén, Greslebin & Rajchenberg

S Cae

Cj @G

PLaTE 2. Amylocorticiellum iaganicum (coll. AG 1507)

Amylocorticiellum iaganicum (Speg.) Gorjon, Gresleb. & Rajchenb., comb. nov.

MycoBank MB519219 PLATES 1,2

= Corticium iaganicum Speg., Bol. Acad. Nac. Cs. Cordoba 11(2): 172, 1888.

= Hypochniciellum iaganicum (Speg.) Rajchenb. & J.E.

Wright, Mycologia 79(2): 247, 1987.

Amylocorticiellum combs. nov. ... 287

DESCRIPTION — Basidiome resupinate, byssoid, detachable from the substrate,

130-400 um thick; hymenial surface smooth, white, cream, ocassionaly with

ochraceous tints; margin thinning out or abrupt. Hyphal system monomitic,

hyphae with clamps, 3-5 um in diam., thin- to thick-walled in the subiculum,

asterocrystals sometimes present. Basidia clavate, with constrictions or sinuous,

25-35 x 5-7 um, guttulate, with 4 sterigmata, and a basal clamp. Basidiospores

ellipsoid, 6-9 x 4-6 um, thick-walled, smooth, amyloid, the outer part of the

wall reacting in Melzer’s reagent while the inner one remains unchanged,

cyanophilous.

REPRESENTATIVESPECIMENSEXAMINED— ARGENTINA. CuuBut: Tehuelches, Senguerr

river, La Plata lake, on dead branches or logs of Nothofagus pumilio (Nothofagaceae),

21-22.X.1993, leg. M. Rajchenberg & A. Greslebin, MR 10785, MR 10789. Los Alerces

National Park, Futalaufquen lake, Irigoyen waterfall stream, on Nothofagus dombeyi

(Nothofagaceae), 19.V1.1997, leg. M. Rajchenberg, MR 11269. Languifieo, Enganio lake,

on N. pumilio, 2.V.1997, leg. A. Greslebin, AG 917, 947. NEUQUEN: Maipu, Lote 42 y 43

ca. Laguna Rosada, on N. pumilio, 2-3.X1.1997, leg. A. Greslebin, AG 1187, 1268, 1311.

Rio NEGRO: Nahuel Huapi National Parkl, Challhuaco Valley, on N. pumilio, 29.X.1997,

leg. A. Greslebin, AG 1094. TIERRA DEL FUEGO: Rio Grande, Estancia Esperanza,

close to Esperanza lake, on Nothofagus antarctica (Nothofagaceae), 24.11.1998, leg. A.

Greslebin, AG 1507, 1508. Ushuaia, Estancia Moat, Chico river, on Nothofagus betuloides

(Nothofagaceae), 26.111.1998, leg. A. Greslebin, AG, 1524, 1527. CHILE. TIERRA DEL

FugGo: on Maytenus magellanica (Celastraceae), LPS 3725, holotypus.

ComMENtTs - Amylocorticiellum iaganicum is morphologically close to

A. cremeoisabellinum but differs in its larger and distinctly amyloid basidio-

spores.

Known from southern Argentina and Chile.

Amylocorticiellum luteolum (Hjortstam & Ryvarden) Gorjon, Gresleb.

& Rajchenb., comb. nov. PLATES 1,3

MycoBank MB519220

= Hypochniciellum luteolum Hjortstam & Ryvarden, Mycotaxon 74(1): 247, 2000.

DESCRIPTION - Basidiome resupinate, pellicular, soft, easily detachable,

hymenophore smooth, pale yellowish, margin indeterminate. Hyphal system

monomitic, hyphae with clamps, subhymenial hyphae thin-walled, 2-3 um in

diam., subicular hyphae thin-walled, 2-4 um wide. Cystidia absent. Basidia

subcylindric, (10-)12-20 x 3.5-4 um, with four sterigmata, and a basal clamp.

Basidiospores short ellipsoid to subglobose, 2.5-3(-3.25) x 2-2.25 um, smooth,

with distinct to thickened walls, amyloid, acyanophilous or with a weak reaction

in cotton blue.

SPECIMEN EXAMINED — COLOMBIA. MAGDALENA, Sierra Nevada de Santa Marta,

Reserva Forestal San Lorenzo, on wood, 17-19.VI.1978, leg. L. Ryvarden, LR 15952

(isotypus GB-008 9325).

ComMENTs - Amylocorticiellum luteolum is characterized by a pale yellowish

pellicular basidiome, the absence of cystidial elements, and small basidia and

‘

yee)

; |

Amylocorticiellum combs. nov. ... 289

PiatE 4. Amylocorticiellum molle (coll. SPG 2999)

Amylocorticiellum molle (Fr.) Spirin & Zmitr., Mikol. Fitopatol. 36(1): 23, 2002.

= Thelephora mollis Fr., Syst. mycol. (Lundae) 1: 443, 1821.

= Hypochniciellum molle (Fr.) Hjortstam, Mycotaxon 13(1): 125, 1981.

= Leucogyrophana mollis (Fr.) Parmasto, Eesti NSV Tead.

Akad. Toim., Biol. seer 16(4): 385, 1967.

SPECIMENS EXAMINED — ARGENTINA. CuuBvtT: Futaleuft, Valle 16 de Octubre, close

to Los Rifleros river, on dead wood of Austrocedrus chilensis (Cupressaceae), 15.X.2010,

leg. S.P. Gorjon, SPG 2991.

290 ... Gorjén, Greslebin & Rajchenberg

Comments - Amylocorticiellum molle is characterized by the narrowly ellipsoid

basidiospores that are greyish in Melzer’s reagent and the distinct cylindrical

cystidia (Eriksson & Ryvarden 1976, Bernicchia & Gorjon 2010).

Known from Europe, Canary Islands and North America. New to the

Southern Hemisphere.

Amylocorticiellum oblongisporum (G. Cunn.) Gorjon, Gresleb. & Rajchenb.,

comb. nov. PLATES 1,4

MycoBank MB519221

= Odontia oblongispora G. Cunn., Trans. Roy. Soc. New Zealand 86(1-2): 95, 1959.

= Hypochniciellum oblongisporum (G. Cunn.) Gresl. &

Rajchenb., Mycotaxon 73: 14, 1999.

DESCRIPTION - Basidiome resupinate, hypochnoid, detachable from the

substrate, 100-250 um thick. Hymenial surface at first smooth, then irpicoid to

clearly hydnoid with smooth areas between the aculei, whitish to pale yellow or

yellowish with chestnut zones; aculei conical or slightly flattened, 0.5-1.5(-2)

mm long. Subiculum cottony, white, 150-200 um thick. Margin white or paler

than the hymenial surface, loosely myceliar or pellicular, detachable from the

substrate. Hyphal system monomitic, hyphae with clamps, 2-4 um in diam.,

thin-walled in the subhymenium, thick-walled in the subiculum and in the

core of the aculei, parallel and compact in the aculei, loosely arranged in the

subiculum. Septocystidia little differentiated, scarce, arising from the subiculum

and projecting up to 30 um. Basidia clavate, 15-25 x 4-5 um, with 4 sterigmata

and a basal clamp. Basidiospores ellipsoid, 4-5(-5.5) x 2.5-3 um, thick-walled,

smooth, amyloid, cyanophilous.

SPECIMENS EXAMINED — ARGENTINA. Cuusut: Los Alerces National Park, southern

arm of Menéndezlake, on Fitzroya cupressoides (Cupressaceae), 8.V.1998, leg. A.Greslebin,

AG 1625. NEUQUEN: Los Lagos, Laguna Rosales, Lote 43, on Nothofagus pumilio stand,

2.X1.1997, leg. A. Greslebin, AG 1214. Nahuel Huapi National Park, Puerto Blest, Frias

river, on Pilgerodendron uviferum (Cupressaceae), 30.V.2010, leg. S.P. Gorjon, SPG

2933. NEW ZEALAND. AUCKLAND, Piha Valley, on Agathis australis (Araucariaceae),

VIII.1953, leg. J.M. Dingley, PDD 17980. Upper Piha Valley, Waitakeres, VIII.1953, leg.

J.M. Dingley, PDD 17981, holotypus. Waitakere, Waitakere Ranges, Spragg track, on

Agathis australis, 2.VIII.1998, leg. M. Rajchenberg s.n. Rangitoto Is., on Metrosideros

excelsa (Myrtaceae), VII.1950, leg. J.M. Dingley, PDD 17984. Oraco, Alton Valley,

Tuatapere, on Podocarpus hallii (Podocarpaceae), 11.1954, leg. J.M. Dingley s.n.

Comments - The combination of a hypochnoid basidiome, a monomitic

hyphal system with clamped generative hyphae, and small, smooth, thick-

walled, amyloid spores supports combination in Amylocorticiellum. The

hydnoid to irpicoid hymenophore with rather long aculei is a unique feature

in the genus. However, some Amylocorticiellum species (e.g. A. molle) show a

more or less merulioid hymenial configuration and, as in most other cases in

Amylocorticiellum combs. nov. ... 291

100 pm ‘\ |

— ww

Nit

Hee

vps

AAX

LAE

Pais!

PiaTE 5. Amylocorticiellum oblongisporum (coll. SPG 2933)

corticioid homobasidiomycetes, hymenophore configuration is variable at the

generic level (e.g. Phlebia Fr., Hyphodontia J. Erikss., Phanerochaete P. Karst.).

Known from New Zealand and Argentina (Patagonia).

Amylocorticiellum sinuosum Spirin & Zmitr., Mikol. Fitopatol. 36(1): 24, 2002.

ComMENtTs - Amylocorticiellum sinuosum is characterized by sinuous to

constricted leptocystidia, long, sinuous to clavate basidia, and weakly amyloid

ellipsoid basidiospores (Zmitrovich & Spirin 2002).

292 ... Gorjon, Greslebin & Rajchenberg

Known growing on Pinus sylvestris from the type locality in western Russia

(Nizhegorod).

Amylocorticiellum subillaqueatum (Litsch.) Spirin & Zmitr., Mikol. Fitopatol.

36(1): 23, 2002.

= Corticium subillaqueatum Litsch., Annls mycol. 39(2/3): 128, 1941.

= Hypochniciellum subillaqueatum (Litsch.) Hjortstam, Mycotaxon 13(1): 126, 1981.

= Leucogyrophana subillaqueata (Litsch.) Julich, Persoonia 8(1): 56, 1974.

Comments - Amylocorticiellum subillaqueatum is characterized by whitish to

cream coloured or yellowish basidiomes, chlamydospores, and small ellipsoid

basidiospores with a greyish Melzer’s reaction. For a detailed description and

illustration see Eriksson & Ryvarden (1976) and Bernicchia & Gorjon (2010).

Known from continental Europe, Great Britain, the Canary Islands, Russia,

and North America.

Key to Amylocorticiellum

la. Cystidia present (but generally few and little differentiated) .................... 2

LB Ey stidiarcOmipletely: ADSOINC ‘tay 0%F saactartencte toavnits scant seavneshe omnes or hee okra and 4

2a. Hymenophore hydnoid to irpicoid, with septocystidia.......... A. oblongisporum

2b. Hymenophore smooth to slightly merulioid, with non-septate leptocystidia...... 3

3a. Leptocystidia distinct, ca. 80-100 x 6-10 tum, spores 6-7 x 2.5-3.5 um ... A. molle

3b. Leptocystidia slender 20-35 x 3-4 um, spores 5.5-8 x 3-3.7 um...... A. sinuosum

Hay S Oto aga soe Tl hOIN Ge ART gM aba ladon rad el ad A EA Pa a aaa 5

4bespores sinaller,-upto: 44.5 fimo ssn ra tlene uactya tsmah a neenet tah acetone ota cae 6

5a. Spores:6-9 4-6. tin, Blaishiain: WD ot, syeey)s peruete x aeette x sect see ea A. iaganicum

5b. Spores 5-7 x 3-4 um, invariable or greyish in IKI........... A. cremeoisabellinum

6a. Spores 3.5-4.5(-5) x (2-)2.5-3 um, greyish in IKI, chlamydospores present in

SOME SPS CUINETIS oes Fin Mau, ota ne Seth aries an atl tertiles ett Maa st A. subillaqueatum

6b. Spores 2.5-3.25 x 2-2.5 um, bluish in IKI, chlamydospores absent .... A. luteolum

Acknowledgments

Masoomeh Ghobad-Nejhad and Viacheslav Spirin acted as pre-submission reviewers,

making most valuable suggestions to improve the text. Karl-Henrik Larsson and Ellen

Larsson (Herbarium GB) are thanked for the loan of Hypochniciellum luteolum. The

Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET, Argentina)

supported this research by PIP 80101000. Sergio Pérez Gorjoén is a postdoctoral research

fellow of the Agencia Esparfiola de Cooperacion Internacional (MAEC-AECID). Alina

Greslebin and Mario Rajchenberg are researchers of CONICET.

Amylocorticiellum combs. nov. ... 293

Literature cited

Bernicchia A, Gorjon SP. 2010. Corticiaceae s.]. Fungi Europaei 12. Ed. Candusso. Italia.

Binder M, Larsson KH, Matheny PB, Hibbett DS. 2010. Amylocorticiales ord. nov. and Jaapiales

ord. nov.: early diverging clades of Agaricomycetidae dominated by corticioid forms. Mycologia

102(4): 865-880. doi:10.3852/09-288

Eriksson J, Ryvarden L. 1976. The Corticiaceae of North Europe. Vol. 4. Hyphodermella-Mycoacia.

Fungiflora, Oslo, Norway.

Greslebin AG. 2002. Fungi, Basidiomycota, Aphyllophorales: Coniophoraceae, Corticiaceae,

Gomphaceae, Hymenochaetaceae, Lachnocladiaceae, Stereaceae, Thelephoraceae. Tulasnellales:

Tulasnellaceae. Buenos Aires, CONICET.

Greslebin AG, Rajchenberg M. 1999. Corticioid Aphyllophorales (Basidiomycota) from the

Patagonian Andes Forests of Argentina 5. Some new taxa. Mycotaxon 73: 9-17.

Hjortstam K. 1981. Notes on Corticiaceae (Basidiomycetes) IX. Three new combinations in

Hypochniciellum. Mycotaxon 13(1): 124-126.

Hjortstam K, Ryvarden L. 1980. Studies in tropical Corticiaceae (Basidiomycetes) Il. Mycotaxon

12(1): 168-184.

Hjortstam K, Ryvarden L. 1985. New and noteworthy Basidiomycetes (Aphyllophorales) from Tierra

del Fuego, Argentina. Mycotaxon 22(1): 159-167.

Hjortstam K, Ryvarden L. 2000. Corticioid species (Basidiomycotina, Aphyllophorales) from

Colombia II. Mycotaxon 74(1): 241-252.

Hjortstam K, Ryvarden L. 2007. Checklist of corticioid fungi (Basidiomycotina) from the tropics,

subtropics and the southern hemisphere. Synopsis Fungorum 22: 27-146.

Kotiranta H, Saarenoksa R. 1990. Reports on Finnish corticolous Aphyllophorales (Basidiomycetes).

Karstenia 30: 43-69.

Larsson KH. 2007. Re-thinking the classification of corticioid fungi. Mycological Research 111(9):

1040-1063. doi:10.1016/j.mycres.2007.08.001

Zmitrovich IV, Spirin VA. 2002. A contribution to the taxonomy of corticioid fungi. II. The genera

Serpula, Serpulomyces gen. nov., Amylocorticiellum gen. nov. Mikologiya i Fitopatologiya 36(1):

11-26.

MY COTAXON

Volume 116, pp. 295-302 April-June 2011

DOT: 10.5248/116.295

Melanoderma microcarpum gen. et sp. nov. (Basidiomycota)

from China

Bao-Kal Cur !, CHANG-LIN ZHAO 2 & YU-CHENG DAI *

Institute of Microbiology, Beijing Forestry University, RO. Box 61, Beijing 100083, China

CORRESPONDENCE TO *: ' baokaicui@yahoo.com.cn, * giangzi717@163.com,

& 3* yuchengd@yahoo.com

ABSTRACT — A new genus Melanoderma (Polyporales, Basidiomycota), collected from Hainan

and Hunan Provinces in southern China, is proposed and typified by M. microcarpum. The

new species is characterized by a perennial habit, pileate to effused-reflexed basidiocarps with

a black crust at the pileal surface, small round pores, a dimitic hyphal system with clamped

generative hyphae, dextrinoid and cyanophilous skeletal hyphae, cylindrical, thin-walled,

non-amyloid, non-dextrinoid and acyanophilous basidiospores, the presence of apically

encrusted cystidia, and lack of both hyphal pegs and dendrohyphidia. The new genus is

compared with related genera of Australoporus, Datronia, Dichomitus, Megasporoporia, and

Perenniporia. rDNA sequence analysis supports Melanoderma as a monophyletic clade that

is closely related to Vanderbylia.

KEY worps — polypore, lignicolous fungi, poroid fungi, phylogeny, taxonomy

Introduction

During investigations on lignicolous and poroid fungi in China, three

specimens were collected from Hainan and Hunan Provinces in southern

China. These collections are microscopically similar to Perenniporia Murrill in

having a dimitic hyphal system with clamped generative hyphae and dextrinoid

and cyanophilous skeletal hyphae, but differ from Perenniporia in having thin-

walled, non-dextrinoid and acyanophilous basidiospores. These collections

may also be confused with species of Australoporus P.K. Buchanan & Ryvarden,

Datronia Donk, Dichomitus D.A. Reid and Megasporoporia Ryvarden & J.E.

Wright, but have distinct differences. Since the species cannot be accommodated

in any known genera, we propose a new genus for these collections. To support

our proposal as well as to evaluate the position of the new genus and related

taxa, phylogenetic analysis was performed using 28S rDNA sequence data.

296 ... Cui, Zhao & Dai

Materials & methods

MoRPHOLOGICAL STUDIES. — The studied specimens are deposited at the herbarium

of Beijing Forestry University (BJFC) and the herbarium of Institute of Applied

Ecology, Chinese Academy of Sciences (IFP). The microscopic procedure follows Cui

et al. (2007). In presenting the variation in the size of the spores, 5% of measurements

were excluded from each end of the range, and given in parentheses. In the text the

following abbreviations are used: IKI = Melzer’s reagent, IKI- = negative in Melzer’s

TABLE 1. Species and sequences database accession numbers used in this study.

SPECIES NAME SAMPLE NO. GENBANK NO.

Abundisporus sclerosetosus Decock & Laurence MUCL 41438 FJ393868

A. violaceus (Wakef.) Ryvarden MUCL 38617 FJ393867

Antrodia malicola (Berk. & M.A. Curtis) Donk TFRI 349 EU232296

FP 104329 EU232297

Antrodiella americana Ryvarden & Gilb. L 3468 EU232269

HHB 4100 EU232270

Byssomerulius corium (Pers.) Parmasto KHL 8593 AY586640

Ceriporia reticulata (Hoftm.) Domanski KHL 11981 EU118614

Donkioporia expansa (Desm.) Kotl. & Pouzar MUCL 35116 FJ393872

Fomitopsis pinicola (Sw.) P. Karst. TFRI 513 EU232290

Ganoderma applanatum (Pers.) Pat. BCRC 36091 EU232274

Gloeocystidiellum aspellum Hjortstam LIN 625 AF506432

G. porosum (Berk. & M.A. Curtis) Donk FCUG 2661 AF310099

FCUG 2768 AF310100

Melanoderma microcarpum DAI 9811 HQ678175

DAI 8116 HQ678176

Perenniporia corticola (Corner) Decock DAI 7330 HQ654108

P. cystidiata Y.C. Dai et al. CUI 8459 HQ654113

P. detrita (Berk.) Ryvarden MUCL 42649 FJ393866

P. martia (Berk.) Ryvarden CUI 7992 HQ654114

MUCL 41677 FJ393859

MUCL 41678 FJ393860

P. medulla-panis (Jacq.) Donk MUCL 43520 FJ393875

MUCL 49581 FJ393876

P. truncatospora (Lloyd) Ryvarden CUI 6987 HQ654112

Perenniporiella chaquenia Robledo & Decock MUCL 47648 FJ393856

MUCL 49758 FJ393857

Phellinus alni (Bondartsev) Parmasto TW 162 AF311025

Pyrofomes demidoffii (Lév.) Kotl. & Pouzar MUCL 41034 FJ393873

Trametes elegans (Spreng.) Fr. BCC 23750 FJ372713

BCC 23751 FJ372714

Vanderbylia vicina (Lloyd) D.A. Reid MUCL 44779 FJ393862

Wrightoporia lenta (Overh. & J. Lowe) Pouzar KN 150311 AF506489

Melanoderma microcarpum gen. et sp. nov. (China) ... 297

reagent, KOH = 5% potassium hydroxide, CB = Cotton Blue, CB+ = cyanophilous,

CB- = acyanophilous, L = mean spore length (arithmetic average of all spores),

W = mean spore width (arithmetic average of all spores), Q = variation in the L/W ratios

between the specimens studied, n = number of spores measured from given number of

specimens. Sections were studied at magnification up to x1000 using a Nikon Eclipse E

80i microscope and phase contrast illumination. Drawings were made with the aid of a

drawing tube. Special colour terms follow Petersen (1996) and Anonymous (1969).

MOLECULAR PROCEDURES AND PHYLOGENETIC ANALYSES. — DNA was extracted

from dried herbarium materials following the methods of Gardes & Bruns (1993) and

Cui et al. (2008) with some modifications. In the study, nuclear ribosomal RNA genes

were used to determine the phylogenetic position of the new species. Primers for PCR

amplification were LROR and LR7 described by White et al. (1990). LSU sequences

of the new taxa were deposited in GenBank, and used as references to blast against

GenBank, similar sequences were downloaded, taxa used in the phylogenetic analyses

are listed in TaBLE 1. Maximum parsimony and Bayesian analysis were applied to the

nLSU dataset. All characters were treated as unordered and equal weight. Maximum

parsimony analysis (PAUP* version 4.0) was used (Swofford 2002), based on strict

heuristic searches with a tree-bisection reconnected (TBR) branch swapping algorithm,

stepwise addition and collapse of branch if maximum length is zero. Bootstrap values

were calculated from 1000 replicates. Bayesian analysis with MrBayes3.1.2 (Ronquist &

Huelsenbeck 2003) implementing the Markov Chain Monto Carlo (MCMC) technique

and parameters predetermined with MrMODELTEST2.3 was performed. When four

simultaneous Markov chains were run with 100,000 generations, the average standard

deviation of split frequencies was 0.00849.

Taxonomy

Melanoderma B.K. Cui & Y.C. Dai, gen. nov.

MycoBank MB 519872

Carpophorum perenne, pileatum vel effuso-reflexum, pileatum nigrum, crustum.

Contextum cremea bubalina; facies pororum nivea vel cremea, pori rotundi. Systema

hypharum dimiticum, hyphae generatoriae fibulatae, hyphae skeletales dextrinoideae;

cystidia pallida, ventricosa, incrustata. Sporae hyalinae, cylindricae, IKI-, CB-.

Type Species: Melanoderma microcarpum B.K. Cui & Y.C. Dai

ErymMo_oey: from Greek melano- (= black), derma (= skin); referring to the black crust

at pileal surface.

Basidiocarps perennial, pileate to effused-reflexed, pileus circular to

irregularly formed. Pileal surface black when fresh, colour unchanged when

dry, concentrically zonate, glabrous; margin obtuse. Pore surface white when

fresh, cream buff when dry; pores circular, dissepiments thick, entire. Context

cream-buff, woody hard. Tubes cream-buff, woody hard, stratified. Hyphal

system dimitic; generative hyphae bearing clamp connections; skeletal hyphae

dextrinoid, CB+; tissue unchanged in KOH. Generative hyphae clamped, scanty,

hyaline, thin-walled, unbranched; skeletal hyphae dominant, thick-walled to

subsolid, usually branched, strongly interwoven. Cystidia clavate to ventricose,

298 ... Cui, Zhao & Dai

hyaline, thin-walled, usually apically encrusted; cystidioles clavate, hyaline,

thin-walled. Basidia clavate, with four sterigmata and a basal clamp connection;

basidioles similar in shape to basidia, but slightly smaller. Rhomboid crystals

frequently present in trama and hymenium. Basidiospores cylindrical, hyaline,

thin-walled, smooth, IKI-, CB-.

The genus Melanoderma is characterized by a perennial growth habit, pileate

to effused-reflexed basidiocarps with a black crust at pileal surface, a white

to cream-buff pore surface, and small, round pores. A dimitic hyphal system

with clamped generative hyphae, dextrinoid and cyanophilous skeletal hyphae,

cylindrical, thin-walled, non-amyloid, non-dextrinoid and acyanophilous

basidiospores, apically encrusted cystidia, and lack of hyphal pegs and

dendrohyphidia also distinguish the genus.

Melanoderma microcarpum B.K. Cui & Y.C. Dai, sp. nov. Fic. 1

MycoBank MB 519873

Carpophorum perenne, pileatum vel effuso-reflexum. Contextum cremea bubalina; facies

pororum nivea vel cremea, pori rotundi, 7-9 per mm. Systema hypharum dimiticum,

hyphae generatoriae fibulatae, hyphae skeletales dextrinoideae, hyphae skeletales

contexti 1.5-5.7 um; cystidia frequentia, pallida, ventricosa, incrustata. Sporae hyalinae,

cylindricae, IKI-, CB-, 5.1-6.4 x 1.9-2.7 um.

Type: China. Hunan Province, Yizhang County, Mangshan Forest park, on fallen

angiosperm trunk, 25.V1.2007 Dai 8116 (holotype in BJFC; isotype in IFP).

ErymMo.oey: from Greek micro- (= small), -carpus (= fruited); referring to the small

basidiocarps.

FruITBoDY — Basidiocarps perennial, pileate to effuse-reflexed, narrowly

attached, woody hard upon drying, without odour or taste, pileus circular to

irregularly formed, projecting up to 1.6 cm, 1.5 cm broad and 5 mm thick at

base. Pileal surface black when fresh and dry, concentrically zonate, glabrous;

margin obtuse, cream-buff when juvenile, becoming black when mature. Pore

surface white when fresh, cream to cream-buff when dry; pores distinctly

circular, 7-9 per mm, dissepiments thick, entire. Context cream-buff, woody

hard, up to 1 mm thick, upper surface with a black crust. Tubes cream-buff,

woody hard, stratified, about 1mm long in each layer.

HyPHAL STRUCTURE — Hyphal system dimitic; generative hyphae bearing

clamp connections; skeletal hyphae dextrinoid, CB+; tissue unchanged in

KOH.

CONTEXT — Generative hyphae scanty, hyaline, thin-walled, rarely

branched, 1.5-3.6 um in diam; skeletal hyphae dominant, hyaline, thick-walled

to subsolid, often branched, interwoven, 1.5-5.7 um in diam.

TuBES — Generative hyphae scanty, hyaline, thin-walled, unbranched,

1.2-3 um in diam; skeletal hyphae dominant, thick-walled to subsolid, usually

branched, strongly interwoven, 1.2-5 um in diam. Cystidia clavate to ventricose,

Melanoderma microcarpum gen. et sp. nov. (China) ... 299

Fic. 1. Microscopic structures of Melanoderma microcarpum (drawn from the holotype).

a: Basidiospores. b: Basidia and basidioles. c: Cystidia and cystidioles. d: Hyphae from tubes.

e: Hyphae from context.

hyaline, thin-walled, usually apically encrusted, 20-32 x 4.8-7.6 um; cystidioles

clavate, hyaline, thin-walled, 17.6-24 x 4-6.2 um. Basidia clavate, with four

sterigmata and a basal clamp connection, 9.8-16 x 5-6.5 um; basidioles similar

300 ... Cui, Zhao & Dai

in shape to basidia, but slightly smaller. Rhomboid crystals frequently present

in trama and hymenium.

Spores — Basidiospores cylindrical, hyaline, thin-walled, smooth, IKI-,

CB-, (5-)5.1-6.4(-7.2) x (1.8-)1.9-2.7(-3) um, L = 5.92 um, W = 2.28 um,

Q = 2.25-2.79 (n = 90/3).

TYPE OF ROT — White rot.

ADDITIONAL SPECIMENS EXAMINED: CHINA. Har Province, Lingshui County,

Diaoluoshan Forest Park, on fallen angiosperm trunk, 29.V.2008 Dai 9811 (BJFC & IFP).

Ledong County, Jianfengling Nature Reserve, on fallen angiosperm branch, 11.V.2009

Cui 6582 (BJFC & IFP).

Phylogeny

A primarily phylogenetic analysis on the position of the new species was

done, and the parsimony consensus tree (Fic. 3) was made up based on the 28S

rDNA sequences, in the tree, sequences of Melanoderma microcarpum were

grouped together as a monophyletic cluster with strong support by Bayesian

analysis and Maximum Parsimony analysis.

Discussion

Melanoderma microcarpum is characterized by a perennial growth habit,

pileate to effuse-reflexed basidiocarps with a distinct black crust at pileal

surface, small and round pores, dimitic hyphal system with clamped generative

hyphae, dextrinoid and cyanophilous skeletal hyphae, cylindrical, thin-walled,

non-amyloid, non-dextrinoid and acyanophilous basidiospores, presence of

apically encrusted cystidia, and lack of both hyphal pegs and dendrohyphidia.

Melanoderma microcarpum has dextrinoid and cyanophilous skeletal hyphae,

which may be confused with species of Perenniporia. However, Perenniporia

species have thick-walled, cyanophilous basidiospores (Dai et al. 2002), while

M. microcarpum has thin-walled, acyanophilous basidiospores.

Melanoderma microcarpum is similar to Megasporoporia in sharing a dimitic

hyphal system with clamped generative hyphae, dextrinoid and cyanophilous

skeletal hyphae, cylindrical and thin-walled basidiospores, and rhomboid

crystals in trama and hymenium. Megasporoporia species, however, usually

have resupinate basidiocarps with big pores, large basidiospores, numerous

hyphal pegs and dendrohyphidia, lack cystidia (Ryvarden et al. 1982, Dai &

Wu 2004, Dai & Cui 2008, Du & Cui 2009). Melanoderma microcarpum has

small pores (7-9 per mm), relatively small basidiospores, apically encrusted

cystidia, and lacks hyphal pegs and dendrohyphidia. Furthermore, it has pileate

basidiocarps with a black crust at the upper surface.

Australoporus has a dimitic hyphal system, dextrinoid skeletal hyphae,

apically encrusted cystidia and cylindrical and thin-walled basidiospores,

similar to features found in Melanoderma, but Australoporus has an ungulate

basidiocarp without crust at pileal surface, pale pink to orange pore surface,

Melanoderma microcarpum gen. et sp. nov. (China) ... 301

65/0. 72 [(—__ Perenniporia martia HQ654114

erenniporia martia FJ393860

Perenniporia martia FYJ393859

Ganoderma appianatum EU232274

Perenniporia cystidiata HQ654113

7oPerenniporia truncatospora HQ654112

Perenniporia corticola HQ654108

Fy eomes demidoffti FJ393873

00; Perenniporia medulla-panis F¥393875

Perenniporia medulla-panis FJ393876

Perenniporia detrita FJ393866

Abundisporus sclerosetosus FJ393868

Abundisporus violaceus FJ393867

Donkioporia expansa ¥J393872

/. 00, Perenniporielia chaquenia F3393857

Perenniporiella chaquenia FJ393856

100/1.00 - Melanoderma microcarpum HQ678175

Melanoderma microcarpum HQ678176

Vanderbylia vicina FJ393862

Trametes elegans FJ372714

100/1. 00' Trametes elegans ¥FJ372713

00/1. O@Antrodia malicola EU232297

Antrodia malicola EU232296

Fomitopsis pinicola EU232290

1000/1, 00 Antrodielia americana EU232270

Antrodiella americana EU232269

Byssomerulius corium AY586640

Ceriporia reticulata EU118614

100/1. 00; Giceocystidiellum porosum AF310099

Gloeocystidiellum porosum A¥F310100

Wrightoporia lenta AF506489

Gioeocystidiellum aspeilum AF506432

Phellinus aint AF311025

60/0. 71

100/1. 00

Fic. 2. Strict consensus tree illustrating the phylogeny of Melanoderma microcarpum generated by

Bayesian and Maximum Parsimony analyses based on 28S rDNA sequences. Posterior probabilities

and bootstrap values (250%) are shown the above or below branches.

and finely encrusted skeletal hyphae in dissepiments (Buchanan & Ryvarden

1988, Ryvarden 1991).

Datronia and Dichomitus have a dimitic hyphal system with clamped

generative hyphae, cyanophilous skeletal hyphae, and cylindrical and thin-

walled basidiospores, but those genera differ from Melanoderma in having non-

dextrinoid skeletal hyphae and lacking apically encrusted cystidia (Masuka &

Ryvarden 1999; Ryvarden 1991; Nufiez & Ryvarden 2001).

The phylogenetic analysis confirmed Melanodermaasa distinct monophyletic

cluster closely related to the genus Vanderbylia D.A. Reid, which, however, has

distinctly thick-walled, dextrinoid, and cyanophilous basidiospores (Decock &

Masuka 2003). Based on both morphological and molecular data, a new genus

is proposed for the new species.

Acknowledgements

We express our gratitude to Prof. Kevin D. Hyde (Thailand) for revising English text,

and to Drs. Mario Rajchenberg (Argentina) and Peter Buchanan (New Zealand) who

reviewed the manuscript. The research was financed by the National Natural Science

Foundation of China (Project No. 30900006).

302 ... Cui, Zhao & Dai

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Cui BK, Dai YC, Decock C. 2007. A new species of Perenniporia (Basidiomycota, Aphyllophorales)

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Cui BK, Wang Z, Dai YC. 2008. Albatrellus piceiphilus sp. nov. on the basis of morphological and

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taxa IV. Perenniporia mundula and its presumed taxonomic synonym, Vanderbylia ungulata.

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294X.1993.tb00005.x

Masuka AJ, Ryvarden L. 1999. Dichomitus in Africa. Mycological Research 103: 1126-1130.

Nujiez M, Ryvarden L. 2001. East Asian polypores 2. Polyporaceae s. lato. Synopsis Fungorum 14:

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Ryvarden L. 1991. Genera of polypores. Nomenclature and taxonomy. Synopsis Fungorum 5:

1-363.

Ryvarden L, Wright JE, Rajchenberg M. 1982. Megasporoporia a new genus of resupinate polypores.

Mycotaxon 16: 172-182.

Swofford DL. 2002. PAUP*, Phylogenetic analysis using parsimony (*and other methods). Version

4.0b10 Sinauer Associates, Sunderland, Massachusetts.

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MY COTAXON

Volume 116, pp. 303-312 April-June 2011

DOI: 10.5248/116.303

Zygomycetes from “Reserva Bioldgica de Mogi Guacu’,

Sao Paulo State, Brazil

JOSE IVANILDO DE SOUZA’, CARMEN LIDIA AMORIM PIRES-ZOTTARELLI,

JAILSON FRANCISCO DOS SANTOS & JANAINA PINHEIRO COSTA

Instituto de Botanica, Nucleo de Pesquisa em Micologia

Avenida Miguel Stefano 3687, 04301-902, Sado Paulo, SP, Brazil

*CORRESPONDENCE TO: jisouza@yahoo.com.br

AsBsTRACT — The taxonomic composition of zygomycetes from a reserve of Brazilian

Cerrado was analyzed. Soil and leaf litter samples were collected at five sampling dates.

Cultures of Absidia, Backusella, Circinella, Rhizopus and Conidiobolus were obtained from

canopy- and soil-plates. Due to the scarcity of detailed taxonomic information for Brazilian

zygomycetes, additional information such as descriptions, standard colony colour codes,

illustrations, geographical coordinates and vouchers are provided for Absidia spinosa var.

spinosa, Backusella lamprospora, Circinella simplex, Rhizopus stolonifer var. stolonifer and

Conidiobolus coronatus.

Key worps — Entomophthorales, Mucorales, taxonomy

Introduction

Zygomycetes are an ecologically heterogeneous assemblage of fungi that are

generally saprobes but may also act as pathogens or parasites of plants, other

fungi, invertebrates, and vertebrates, including humans (Alexopoulos et al.

1996). They have commonly been isolated from soil, herbivore dung, almost

all plant materials, mushrooms and insects (O’Donnell 1979), and also from

diverse types of habitats and substrates such as bee brood chamber (Hesseltine

et al. 1990), cave crickets (Zalar et al. 1997) and from Antarctic mosses (Tosi

et al. 2004). Although they are not the most numerous and combative fungi in

soil (Dix & Webster 1995), they are morphologically and ecologically one of

the most diversified groups of fungi with highly successful survival strategies,

although still little studied (Benny et al. 2001).

The “Reserva Bioldgica de Mogi Guacu” (RBMG), constitutes one of

the few remnants of Cerrado vegetation (Brazilian savanna) in Sao Paulo

304 ... de Souza & al

State and is being conserved by the Instituto de Botanica since 1970. Due

to the high diversity and endemism of species, the Cerrado is listed among

the world’s 25 hotspots (Durigan et al. 2004). Originally it covered ca. 23%

of Brazil (including ca. 14% of Sao Paulo State), but the intense demand of

agricultural areas for sugar cane, Pinus, Eucalyptus, Citrus, and pastures led to

destruction and fragmentation of this vegetation and decreased it to less than

1% of its natural distribution in Sao Paulo State (Durigan et al. 2003). In RBMG

taxonomic studies on Glomeromycota (Bononi & Trufem 1983, Carrenho et

al. 1997, Carrenho & Trufem 2001), terrestrial and aquatic anamorphic fungi

(Grandi 1985, Schoenlein-Crusius 2002, Grandi & Silva 2006), Agaricomycetes

(Bononi 1984, Gugliotta 1997, Baseia & Milanez 2001, 2002a, b, Baseia 2005),

and lichens (Jungbluth 2006, Marcelli et al. 2007) have been already carried out,

but no detailed studies on zygomycetes.

In order to study the taxonomy and ecology of zygomycetes from leaf

litter and soil and of zoosporic organisms from soil and water (Nascimento

et al. 2011), a research project is being carried out in RBMG. The aim of this

paper is to provide descriptions, including standard colony colour codes, and

illustrations for Absidia spinosa var. spinosa, Backusella lamprospora, Circinella

simplex, Rhizopus stolonifer var. stolonifer, and Conidiobolus coronatus obtained

from leaf litter and soil samples of RBMG. ‘These taxa generally have only

been cited in lists of fungi from Brazil, with little or no detailed taxonomic

information provided.

Material & methods

STUDY AREA — The RBMG is composed by two remnants of Cerrado, named areas A

(343 ha) and B (127 ha), totalling 470 ha, which are located in a region of savannah-like

tropical moist climate (Aw) with dry winter (Sparovek et al. 2007). Area A has 585-635

m of altitude, its vegetation types were characterized as wet field, gallery forest, Cerrado

sensu stricto, rapanean Cerrado, transition zone, Cerrado field and Cerrado burnt field,

surrounded by deforested areas, Pinus spp. plantations and annual crops (Mantovani

& Martins 1993), but there is no recent information about alterations on its vegetation.

Area B is located ca. 5 km from area A has similar altitude and is mainly covered by

cerradao (forested savanna).

SAMPLING, ISOLATION AND SLIDES PREPARATION — On five sampling dates, leaf litter

and soil samples were collected from 12 sites (TABLE 1), established either in gallery

forest (GF) < 10 m from watercourses (streams) or in Cerrado (CE, savanna, and

forested savanna) = 10 m from watercourses. Samples of moist and decomposing leaf

litter and soil were collected with trowels and plastic bags. Soil samples were collected at

< 10 cm in depth immediately below the sites of leaf litter collection. Canopy- and soil-

plates were prepared with CMYA (cornmeal agar 10 g, malt extract 10 g, yeast extract

2 g, agar 10 g in 1 L distilled water). Chloramphenicol (CP) 500 mg and rose Bengal

(RB) 50 mg were added in order to avoid bacterial growth but also because RB restricts

Zygomycetes from Sao Paulo State (Brazil) ... 305

fungal colony sizes (Bills & Foster 2004). The pH of CMYA plus RB ranged from

5.6-5.9. Leaf litter from each site was fragmented in a Petri dish with two scalpels, mixed

with water agar medium (0.7%, w v') and spread over ca. 60% of the internal surfaces

of three CMYA plates supplemented with CP and RB (modified canopy-plate method,

Drechsler 1952). Small aliquots of soil (0.11 + 0.07 g, n = 22) were sprinkled over three

plates of the same medium (modified soil-plate method, Warcup 1950). Dishes were

incubated in the dark at 25°C. Colonies were isolated after 5-30 days from canopy-

plates and after 3-7 days from soil-plates, transferred to SMA plates (Hesseltine 1954),

incubated and re-isolated until obtaining pure cultures, which were identified at generic

level and preserved by the Castellani method (Smith & Onions 1994). Cultures from

water storage were re-incubated on SMA during 5-16 days while its mycelial fragments

were removed for slide preparations with distilled water plus glycerol or glycerin (ca.

10 drops in 30 mL). Twenty measurements of fungal structures were taken from each

specimen. Colour names more appropriate as possible were attributed to the colonies

whereas its respective standard colour codes, in percentage combinations of black (N,),

cyan (C,), magenta (M,) and yellow (Y,), were adopted according to Kiippers (2002).

Cultures were deposited at CCIBt culture collection, “Colecao de Culturas de Algas,

Cianobactérias e Fungos do Instituto de Botanica’, at Sao Paulo City, Sao Paulo State,

Brazil.

TABLE 1. Soil and leaf litter collection sites in RBMG, Mogi Guacu Municipality, Sao

Paulo State, Brazil.

SITES COORDINATES VEGETATION AREAS

S1 21449" SATO ZW :

ee, en

$3 22°15'35"S 47°11'35"W A

Eble OP raat nto CORN Arey A el |

S5 22°14'59"S 47°09'57"W PL CE

Pan odes Ei gage mgs R ES EN usa

S7 22°11'58"S 47°08'40"W Pleats taietecima

Gare sg Sergi apsosisaw ee

Lean cee Tammi Sh OL unp ro Pa

samy SLO PE cet) AAA RRA eee IO oh oll d ‘

ees SU nnn ee LL 27'S 47°08 SAW ad CE

$12 22°11'26"S 47°08'54"W

Vegetation types: GF = gallery forest; CE = Cerrado

Results & discussion

Cultures of Absidia, Backusella, Circinella, Conidiobolus, Cunninghamella,

Mucor, Piptocephalis, Rhizopus, and Zygorhynchus were obtained from canopy-

and soil-plates. Five taxa were selected for description and illustration here,

because previous reports of Brazilian material have been incomplete and

vouchers of them are scarce or nonexistent. Vouchers of different specimens

and provenances will be valuable for subsiding molecular taxonomic studies.

306 ... de Souza & al

Absidia spinosa Lendn., Bull. Herb. Boissier, Sér. 2, 7: 250. 1907.

var. spinosa. Fics 1-6

SPECIMEN EXAMINED: BRAZIL. SAo PauLo STATE, Mogi Guacu, RBMG, $5, from soil-

plate; J.I. de Souza, 12.1X.2007; CCIBt 2307.

COLONIES 6-15 days old on SMA at 25°C, obverse reddish chestnut (N,, Y,, M.,)

at 15"" day, reverse greyish beige milky, 87 x 71 mm in size, 12 mm high touching

Petri dish lid. SPORANGIOPHORES single or 2-6 in each verticil, greenish hyaline

to pale chestnut, 22-225 um long, 2.7—-7.7(-8.9) um in basal and (2.4—)3.1-6.5

lum in apical diameter. SPORANGIA pyriform, with deliquescent walls, greenish

chestnut, 9.0-41.6 x 10.6-44 um. COLUMELLAE hemispheric with or without

3.3-10.6 um erect projections, or almost spherical lacking projections, some with

granular content and/or collar, greenish hyaline to grayish, (4.0—)5.0-43 x 6.1-

43 um. SPORANGIOSPORES cylindrical, smooth, pale olive green, (2.6-)3.0-5.5

x 2.0-3.5 um. CHLAMYDOSPORES rare in feeding mycelium. YEAST CELLS

absent. ZYGOSPORANGIA globose to applanate, smooth, hyaline to greyish when

young, pale chestnut to dark brown when mature, 42-69(-81) x 37-69(-80)

um, scarse; suspensors opposite, unequal, majority of major suspensors with

pale brown granular content whereas the minor not having it, finger-like

appendages arising from both or only one suspensor.

Notes. Identified using Hesseltine & Ellis (1964). This is the first description

of A. spinosa var. spinosa from Brazil, but A. spinosa var. biappendiculata was

already reported by Trufem (1981a). Other named varieties are A. spinosa var.

madecassensis and A. spinosa var. azygospora (Mycobank 2010).

Backusella lamprospora (Lendn.) Benny & R.K. Benj., Aliso 8(3): 320. 1975.

Figs 7-11

SPECIMEN EXAMINED: BRAZIL. SAO PAULO STATE, Mogi Guacu, RBMG, S1-S3, S5-S7,

from leaf litter canopy-plates; J.I. de Souza, J.F. Santos & J.P. Costa, 12.1X.2007, 28.1.2008,

20.X.2008; CCIBt 2308 and 2334.

Co.ontgs 5-15 days old on SMA at 25°C, obverse beige (N,, Y,, M,,) at 15"

day, reverse beige with pale orange centre, 90 mm in size, 14 mm high touching

Petri dish lid. SPORANGIOPHORES straight or slightly curved, recurved near apex

when young, mono- or sympodially branched, some inflated, some constricted

at apex, septate near branches and/or sporangia and columellae, hyaline to

greyish, with chestnut granular content and olive green droplets, (38-)50-2473

um long, 4.9-20.3 um in basal and 3.9-15.4 um in apical diameter. SPORANGIA

globose, with deliquescent walls, olive green to greenish/dark chestnut,

25.2-82 x 25.2-85 um. COLUMELLAE globose, cylindrical, ellipsoidal, cuneiform,

conical and hemispherical, collar present or absent, hyaline to greyish, with or

without yellowish chestnut to chestnut granular content, 7.3-46 x 8.2-44 um.

Zygomycetes from Sao Paulo State (Brazil) ... 307

SPORANGIOLAR PEDICELS slightly curved to curved, some lightly constricted

at apex, single or having 1-3 lateral branches, coenocytic, hyaline, greenish

to greyish, with or without olive green granular content, 11-190 um long,

2.2-13.2(-17.3) um in basal and 1.6-8.6(-11.4) um in apical diameter.

UNISPORED SPORANGIOLA globose, lightly spiny, pale chestnut to yellowish

chestnut, 8.4-33 x 8-33 um, rare. MULTISPORED SPORANGIOLA globose,

with persistent and undulate walls, olive green to yellowish, 11-50 x 13.5-53

um, abundant, containing 2-38 sporangiospores. SPORANGIOSPORES ovoid,

rounded, some ellipsoidal, or irregular, greyish, pale olive green to greyish

chestnut, with homogeneous or granular content, 4.0-24.6(-29) x 3.5-21.2 um.

FEEDING MYCELIUM with inflated regions, pale chestnut to olive green content,

ending in thin rhizoid-like filaments; having oidia-like cells, rounded, ellipsoidal

or irregular, hyaline, olive green to dark chestnut, abundant in whitish-dense

spots in colony regions that can be seen at naked-eye. CHLAMYDOSPORES and

ZYGOSPORANGIA absent.

Notes. Identified using Benny & Benjamin (1975). Some multispored

sporangiola may be detached and remain intact in slide preparations. ‘This

description complements the information of B. lamprospora from Brazil; Trufem

(1978) reported Mucor lamprosporus (= B. lamprospora) but the specimen was

destitute of the typical unispored and multispored sporangiola.

Circinella simplex Tiegh., Annls Sci. Nat., Bot., Sér. 6, 1: 92. 1875. Figs 12-16

SPECIMEN EXAMINED: BRAZIL. SAO PAULO STATE, Mogi Guacu, RBMG, S3, S9, from

soil-plates; J.J. de Souza, J.F. Santos & J.P. Costa, 28-V1I-2008; CCIBt 2312.

CoLonigs 7-15 days old on SMA at 25°C, producing baker's yeast odour,

obverse yellowish (N,, C,, Y,,) to reddish beige (N,, Y., M,,) with white to ivory

margin at 15" day, reverse pale beige to greyish white milky, 83 x 81 mm in size,

up to 4 mm high. VEGETATIVE MYCELIUM hyaline, containing yellowish green

granules and septa along hyphae. SPORANGIOPHORES sympodially branched,

lightly to very circinate, septate near sporangia and columellae, hyaline to

greyish containing olive green to pale chestnut droplets, (3.6-)6.0-17.5 um in

basal and 3.0-14 um in apical diameter. SPoRANGIA globose, with smooth and

deliquescent walls, or having undulated surface when minor, yellowish chestnut

to dark chestnut, 14.5-82 x 14.5-84 um. COLUMELLAE globose, ovoid, conical

and obpyriform, generally with collars, hyaline to greyish, sometimes with olive

green content, 7.7-39(-51) x 8.9-34(-43) um. SPORANGIOSPORES irregular-

shaped, rounded or elongated (minority), greyish to dark grey, 2.6-15.4(-19.8)

x 2.4-6.7 um. CHLAMYDOSPORES, YEAST CELLS and ZYGOSPORANGIA absent.

Notes. Identified using Hesseltine & Fennell (1955). Trufem (1981b) reported

two specimens isolated from soil in Brazil.

308 ... de Souza & al

Rhizopus stolonifer (Ehrenb.) Vuill., Revue mycol. Toulouse, 24: 54. 1902.

var. stolonifer Figs 17-21

SPECIMEN EXAMINED: BRAZIL. SAo PAULo StaTE, Mogi Guacu, RBMG, S10, from

soil-plate; J.J. Souza, J.E Santos & J.P. Costa, 28.V11.2008; CCIBt 2329.

Cotonigs 7-13 days old on SMA at 25°C, with scarce mycelium, obverse

greenish chestnut (N,, Y., M,,) dotted with great black sporangia at 13" day,

reverse greyish white, 90 mm in diameter, 7 mm high. SPORANGIOPHORES

short to long, monopodially branched, coenocytic, hyaline to chestnut (amber),

with olive green granular content, 108-2228 um long, 3.7—28 um in basal and

4.9-24.6 um in apical diameter. SPORANGIA globose, with deliquescent walls,

chestnut to black, 45-333 x 45-345 um. COLUMELLAE globose, subglobose and

ellipsoidal (majors), hemispheric or applanate (minors), with or without collar,

hyaline, with olive green content, 11.2-160 x 18.4-166 um. SPORANGIOSPORES

ovoid to rounded, striate (hazelnut-like), pale chestnut, with homogeneous

content, (6.5-)8.2-16.7 x 6.1-16.3 um. CHLAMYDOSPORES, YEAST CELLS and

ZYGOSPORANGIA absent.

Notes. Identified using Schipper (1984). This description complements the

information of R. stolonifer var. stolonifer from Brazil; Trufem (1981a) reported

R. nigricans (= R. stolonifer var. stolonifer). Other named varieties are R. stolonifer

var. luxurians and R. stolonifer var. lyococcus (Mycobank 2010).

Conidiobolus coronatus (Costantin) A. Batko, Entomophaga, Mémoires hors série

2: 129. 1964 [51962’]. Fics 22-26

SPECIMENS EXAMINED: BRAZIL. SAO PAULO STaTE, Mogi Guacu, RBMG, S4, $11, from

leaf litter canopy-plates; J.J. de Souza, 20.X.2008; CCIBt 2335 and 2336.

CoLoniges 7-16 days old on SMA at 25°C, obverse white (Y,, M,, C,,) to

yellowish (Y,, M,, C,,) and reverse yellowish (Y,,_,, My) C,,) at 16" day, 90 mm

in diameter, or composed of up to 10 mm colonies occupying almost all the

plate surfaces, flat, aerial mycelium absent. FEEDING MYCELIUM filamentous,

branched, rarely septate, greenish to pale brown containing olive green droplets,

5-20.8 um in diameter, releasing or not HYPHAL BODIES (segments) of 37-180

um long. CONIDIOPHORES undifferentiated from feeding mycelium. PRIMARY

CONIDIA spherical to globose, pale olive green to greenish chestnut, 23.5-65

um in diameter. PAPILLAE applanate or cone-shaped with blunt or sharp ends,

respectively, 4.9-19 x 8.3-20.9 um. VILLOSE CONIDIA spherical, rarely papillate,

similar to primary conidia in size. REPLICATIVE CONIDIA rare. MICROCONIDIA

ovoid, lemon-shaped to rounded 14.5-28 x 14-24 um (CCIBt 2335) or absent

(CCIBt 2336). CHLAMYDOSPORES and ZYGOSPORANGIA absent.

Notes. Identified using King (1976a,b, 1977). Both specimens showed poor

growth on CMYA when compared to growing on SMA. This description

Zygomycetes from Sao Paulo State (Brazil) ... 309

J as >, @

i 1s— a 2 ~8 of 16 7

Fx. me . . >. 2

OOD ames

Fics 1-26. Absidia spinosa var. spinosa (1-6): 1, verticil with five sporangiophores bearing

sporangia; 2, sporangium; 3, columella with erect projection; 4, sporangiospores; 5-6, zygosporangia

with finger-like appendages from suspensors. Backusella lamprospora (7-11): 7-8, sympodially

branched sporangiophores; 9, 10. sporangiophores bearing columellae; 11, sporangiospores and

detached sporangium. Circinella simplex (12-16): 12-13, circinate sporangiophores bearing

sporangia; 14-15, circinate sporangiophores bearing columellae; 16, sporangiospores. Rhizopus

stolonifer var. stolonifer (17-21): 17, sporangiophore bearing sporangium; 18-20, columellae;

21, sporangiospores. Conidiobolus coronatus (22-26): 22, primary conidium bearing microconidia;

23-24, replicative conidia; 25, primary conidia, hyphal bodies and microconidia; 26, villose

conidium.

Bars: 1, 5-8, 13 = 50 um; 2, 3, 9-12, 15, 19-24, 26 = 20 um; 4, 16 = 10 um; 14, 18 = 40 um;

17, 25 = 100 um.

complements the information on C. coronatus from Brazil; Porto et al (1987)

isolated C. coronatus from soil with and without vegetal detritus.

310 ... de Souza & al

Acknowledgements

We thank Dr Matias J Cafaro, Department of Biology, University of Puerto Rico,

Dr Sandra FB Trufem, Universidade Sao Marcos, Brazil, and Dr. Shaun R Pennycook,

Landcare Research, New Zealand, for the critical reviewing of the manuscript, Dr

Agostina V Marano, Instituto de Botanica Carlos Spegazzini, Argentina, for valuable

comments and suggestions, Ms Janaina da Rocha for laboratorial support, Mr Edson

Pires for field assistance, Fapesp (project no. 2008/53146-4) for financial support given

to CLA Pires-Zottarelli and CNPq (PIBIC) for scholarship given to JF Santos.

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$0102-33062002000100007

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MY COTAXON

Volume 116, pp. 313-316 April-June 2011

DOT: 10.5248/116.313

A new species and a new record of Herpothallon

(lichenized Ascomycota) from India

T.A.M. JAGADEESH RAM & G.P. SINHA*

Botanical Survey of India, Central Regional Centre, Allahabad-211002, India

CORRESPONDENCE TO *: tamjagadeesh@yahoo.co.in & * drgpsinha@yahoo.co.in

ABSTRACT — Herpothallon sticticum, a new lichen species is described from the Eastern

Himalaya, India. Herpothallon echinatum is also reported for the first time from India.

Key worps — Arthoniales, Arthoniaceae, lichens

Introduction

A number of species in the lichen genus Herpothallon Tobler (Arthoniales)

have recently been reported from India (Jagadeesh Ram et al. 2009, Jagadeesh

Ram & Sinha 2009) since its reinstatement by Aptroot et al. (2009). Further

examination of additional specimens from Arunachal Pradesh and Sikkim

in the eastern Himalayan ranges revealed the presence of an additional new

species and a new record for India. Now, 11 Herpothallon species are known

from India. The new species is described below and a brief note on the new

record is provided.

Materials & methods

Specimens collected from Eastern Himalaya and deposited in BSA were

investigated. External morphological features were observed with an Olympus SZ61

dissecting microscope. Thin hand-cut sections of thalli were mounted in water,

10% KOH solution, Lugol's iodine solution and lactophenol cotton blue (LCB) and

examined with a Leica DM 2500 compound microscope. The thallus colour reactions

were carried out by 10% aqueous potassium hydroxide solution (K), aqueous calcium

hypochlorite solution (C), Steiner’s stable solution (P) and long wavelength UV.

Calcium oxalate crystals in the thallus were identified by 25% H,SO, mounts (Aptroot

et al. 2009). The lichen substances were identified by thin layer chromatography

(Orange et al. 2001).

314 ... Jagadeesh Ram & Sinha

New species

Herpothallon sticticum Jagadeesh & G.P. Sinha, sp. nov. Fic. 1

MycoBank MB 519528

Thallus corticola, epiphloeodes, byssoideus, glauco-griseus ad luteo-griseus; prothallus et

hypothallus albidus; pseudisidiis granularibus ad globosis, 0.05-0.1(-0.25) x 0.05-0.1

(-0.2) mm; apothecia et pycnidia ignota; acidum sticticum et consticticum continens.

Type - India, Arunachal Pradesh, West Kameng District, Jameri Road side, on Bombax

trunk, alt. 1200 m, 11 October 2001, G.P Sinha & T.A.M. Jagadeesh Ram 11331

(Holotype ASSAM-BSA).

EryMo_oey: the species epithet refers to the major secondary metabolite, stictic acid.

THALLUS corticolous, epiphloeodal, tightly attached, suborbicular to irregular,

3-7 cm across, glaucous grey to pale yellow-grey, sometimes with an

ochraceous tinge, thinly pruinose, minutely felty, up to 65-95(-140) um thick,

with calcium oxalate crystals; crystals many, scattered, 3-8(-16) um wide.

HYPOTHALLUS white, below the entire thallus, closely attached, hyphae 1-2 um

wide. PROTHALLUS whitish, sometimes indistinct, closely attached, byssoid, of

closely interwoven hyphae, up to 2 mm wide. PSEUDOISIDIA numerous, granular

to globose or irregular wart-like, sometimes slightly flattened and elongated,

dense, minutely felty with projecting hyphae, of the same colour as the thallus,

0.05-0.1(-0.25) x 0.05-0.1(-0.2) mm, merging or bursting into minute soredia-

like granules throughout. PHotosiont Trentepohlia, cells single or in short,

irregular threads, 8-15 x 5-9 um. AsciI and PYCNIDIA not seen.

CuHEMIstTrY - Thallus K+ yellow, C-, P+ orange, I+ and KI+ pale blue in

patches (section), UV-; stictic acid (major), constictic acid (minor), hypostictic

and norstictic acids (trace) detected by TLC.

ADDITIONAL SPECIMENS EXAMINED — INDIA, Sikkim, East Sikkim: near Setipool, 900

m, 20 Apr. 2010, G. Swarnalatha 5657 (BSA); near Samdong, on the way to Dikchu,

940 m, 21 Apr. 2010, G. Swarnalatha 5726, 5728 (BSA); 6 km to Pakyong, on the way

to Rhenok, 537 m, 22 Apr. 2010, G. Swarnalatha 5816 (BSA); Gangtok, BSI - SHRC

campus, 1724 m, 17 Apr. 2010, G. Swarnalatha 6201, 6204 (BSA). North Sikkim: on the

way to Phodong, near Panchong, 1434 m, 24 Apr. 2010, G. Swarnalatha 5832 (BSA); 1

km to seven sister water falls from Gangtok, 1578 m, 24 Apr. 2010, G. Swarnalatha 5859

(BSA); Tangla, 1689 m, 24 Apr. 2010, G. Swarnalatha 5889 (BSA).

REMARKS - Herpothallon sticticum is characterized by the thallus with granular

to globose, irregular wart-like pseudoisidia that merge or burst into minute

soredia-like granules and the stictic acid complex as the secondary metabolites.

Herpothallon furfuraceum G. Thor, H. granulare (Sipman) Aptroot & Liicking,

and H. granulosum Jagadeesh & G.P. Sinha are other species where the

pseudoisidia merge into soredia-like granules, but all differ in having alternative

chemistry (Aptroot et al. 2009, Jagadeesh Ram et al. 2009). Chemically

H. sticticum resembles H. isidiatum Jagadeesh & G.P. Sinha, a common species

known from India but distinguished by the long cylindrical pseudoisidia in

Herpothallon sticticum sp. nov. (India) ... 315

Fic. 1. Habit of Herpothallon sticticum (holotype): A. thallus with prothallus, B. enlarged

pseudoisidiate region, C. enlarged pseudoisidiate-sorediate region. Scale = 1 mm.

the latter (Jagadeesh Ram et al. 2009). At present H. sticticum is known from

Arunachal Pradesh and Sikkim in the Eastern Himalaya.

New record

Herpothallon echinatum Aptroot, Liicking & Will-Wolf

This species was previously known from Australia, Indonesia, Norfolk

Island, Papua New Guinea, Taiwan, and Thailand (Aptroot et al. 2009) and has

316 ... Jagadeesh Ram & Sinha

now been found in Sikkim. It is characterized by the loosely attached thallus

lacking calcium oxalate crystals, white hypothallus and prothallus, cylindrical

pseudoisidia up to 0.5 x 0.1 mm, and the presence of psoromic acid.

SPECIMEN EXAMINED - INDIA, Sikkim, Gangtok, Pengala, 27°22'24.7"N, 88°35'56.6"E,

alt. 1948 m, 24 May 2008, T'A.M. Jagadeesh Ram 4480 (BSA).

Acknowledgments

We thank Dr. M. Sanjappa, Director, Botanical Survey of India, Kolkata and

Dr. A.A. Ansari, Scientist ‘E’ and Head of Office, Botanical Survey of India, Central

Regional Centre, Allahabad for facilities. The authors also gratefully acknowledge

Emeritus Prof. J.A. Elix, Research School of Chemistry, Australian National University,

Canberra, Australia and Prof. Gdran Thor, Department of Ecology, Swedish University

of Agricultural Sciences, Uppsala, Sweden for presubmission review. We are grateful to

Dr. K.P. Singh, Co-ordinator, AICOPTAX - Lichens for encouragement and Ministry

of Environment & Forests, New Delhi for financial assistance under the AICOPTAX

scheme.

Literature cited

Aptroot A, Thor G, Licking R, Elix JA, Chaves JL. 2009. The lichen genus Herpothallon reinstated.

Bibliotheca Lichenologica 99: 19-66.

Jagadeesh Ram TAM, Sinha GP. 2009. New species and new records of Herpothallon (lichenized

Ascomycota) from India. Mycotaxon 110: 37-42. doi:10.5248/110.37

Jagadeesh Ram TAM, Sinha GP, Singh KP. 2009. New species and new records of Cryptothecia

and Herpothallon (Arthoniales) from India. Lichenologist 41: 605-613. doi:10.1017/

$0024282909008123

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society, UK.

MY COTAXON

Volume 116, pp. 317-328 April-June 2011

DOT: 10.5248/116.317

Cortinarius xanthodryophilus sp. nov. -

a common Phlegmacium under oaks in California

DIMITAR BOJANTCHEV™ & R. MICHAEL DAVIS?

'MushroomHobby.com, 345 Shipwatch Lane, Hercules, CA 94547 USA

*Department of Plant Pathology, University of California, Davis, CA 95616 USA

*CORRESPONDENCE TO: dimitar@pontix.com

ABSTRACT -- A new Cortinarius species (subgenus Phlegmacium) associated with oaks

in California is described. Cortinarius xanthodryophilus is a commonly encountered

representative of the fulvoid bulbopodiums and is characterized by predominantly light to dark

yellow colors, distinctly emarginated bulb, weak alkali reaction, and association with oaks.

Based on phylogenetic analysis of nrITS sequences, it is placed in the /pseudoglaucopodes/

humolens clade.

Key Worbps -- Cortinariaceae, fungal taxonomy

Introduction

The state of California, USA spans several distinct ecological regions with

diverse biota and climatic patterns. The genus Cortinarius is widespread

throughout the state with primary symbionts ranging from conifers along the

Northern coastal belt and the Sierra/Cascade mountain ranges to hardwoods in

the warmer coastal zone and the xeric Mediterranean-type climate areas.

Historically, the major contributions to the study of the genus in the state

were initiated by A. H. Smith, who described a number of taxa from the Pacific

Northwest and California (Smith 1939, 1944; Smith & Rea 1944). Ammirati,

who studied Dermocybe, also published new species from the area (Ammirati

& Gilliam 1975, Ammirati & Smith 1977, 1978, 1984). In the 1990’s, Moser and

Ammnirati, who published a series of studies on Pacific Northwest taxa, described

a number of Cortinarius species from California and surrounding areas (Moser

2002; Moser & Ammirati 1996, 1997, 1999, 2000). Thiers & Smith (1969) and

Fogel (1994) made important contributions to the study of hypogeous cortinarii

in the region, and a few Cortinarius taxa have been described from California

by other authors. Environmental sampling is becoming an important tool for

detecting Cortinarius species from root tips that have yet to be collected or

reported in the literature.

318 ... Bojantchev & Davis

During our study of Cortinarius in California for the past six years, we

have collected a number of undescribed taxa. The description of Cortinarius

xanthodryophilus is our first installment in a planned series of contributions to

the study of Cortinarius in California and the Pacific Northwest.

Materials & methods

MORPHOLOGICAL STuDIES: All collections have been observed and described by

the authors. Emphasis was placed on describing basidiomata in all developmental

stages in order to analyze such fleeting characters as the original lamellar/context colors

and bruising reactions. All fresh material was processed on the day of the collection.

In all cases we tested the fresh material for odor, taste, macrochemical reactions, and

UV reflection. High-resolution photographs with bracketed exposures (+1, 0, -1) EV

were made for each fresh collection. Taxonomically important macromorphological

features were carefully depicted. High-resolution photographs are available online (see

www.mushroomhobby.com under genus Cortinarius). KOH,,, and Melzer’s were the

primary agents for macrochemical analysis in Cortinarius taxonomy. KOH was applied

to all external surfaces and context of the pileus, stipe and bulb. Color codes follow the

Munsell soil color charts (2000). Spore prints were collected directly on microscopic

slides in order to evaluate the precise shade of the spore color and to obtain a rich set

of mature spores for study. UV fluorescence was tested with a 400 nM Ultra Violet

Blacklight Flashlight. All examined collections are preserved in the first author's private

herbarium, designated as DBB, and (where noted) in the University of California,

Berkeley Herbarium (UC).

MICROSCOPIC STUDIES were conducted using a light compound microscope.

Basidiospores (aberrant spores excluded) were studied at magnifications of 1000-1600x

under immersion oil, with two slides with spores from two different basidiomata

examined for each collection. A minimum of 32 spores were measured in each case. The

spores were mounted in H,O, KOH and Melzer’s reagents. The following abbreviations

are used: Q for quotient of length and width and Q_, for average quotient. Cell structures

were studied at 640-1600x magnification and observed in KOH, Melzer’s, and H,O.

Congo Red was used for the study of cuticular morphology, cell structures, cell walls,

and incrustations. All microstructure measurements reflect data from all examined

collections, including the holotype.

MOLECULAR EXTRACTION: The universal primers, ITS4 and ITS5, were used to

amplify the internal transcribed regions (ITS 1, ITS 2), the 5.8 gene nuclear ribosomal

subunit, and part of the large and small ribosomal subunits by polymerase chain

reaction (White et al. 1990). Amplification was carried out in 50 ul reactions containing

3 ul DNA, 50 mM KCl, 10 mM Tris-HCl (pH 9), 1% Triton X-100, 2.5 mM each of

dATP, dCTP, dGTP, and dTTP, 25 mM MgCl, 50 mM of each primer, 1 unit of Taq

polymerase, and 32.6 ul of milliQ water. The PCR reaction was conducted using a PTC-

100 thermocycler (MJ Research, Watertown, MA) with the following parameters: 40

cycles of 1 min at 94°C, 2 min at 55°C, and 2.5 min at 72°C, and a final extension time of

10 min at 72°C. Negative controls (no template DNA) were included in every assay. PCR-

amplified DNA was visualized on 1.5% agarose gels (Invitrogen corp., Carlsbad, CA) by

staining with ethidium bromide (0.25 ug/ml) and photographed under UV light. The

Cortinarius xanthodryophilus sp. nov. (U.S.A.) ... 319

remaining PCR products were purified using the Qiagen QIAquick PCR Purification Kit

(Qiagen Inc., Valencia, CA) according to the manufacturer's protocol. The purified DNA

fragments were sequenced in both directions at the UC Davis DBS Automated DNA

Sequencing Facility. The chromatograms were processed with Chromas Lite v2.01 and

visually inspected for correctness. The forward and backward sequences were visually

reconciled using MEGAS5 (Tamura et al. 2011, unpublished).

Phylogenetic analysis

We have downloaded and reviewed all Cortinarius ITS1/5.8s/ITS2

sequences available on GenBank (http://www.ncbi.nlm.nih.gov) and UNITE

(http://unite.ut.ee/). During the initial analysis, we selected several hundred

Phlegmacium sequences from the northern hemisphere based on the quality

and representation of well-supported taxa. Added were approximately two

hundred sequences from our Phlegmacium collections, mainly from California,

but also from western North America and Europe. The phylogenetic analysis of

that large dataset (not shown) clusters Cortinarius xanthodryophilus within the

/pseudoglaucopodes/humolens clade.

A phylogenetic analysis of 19 sequences within the /pseudoglaucopodes/

humolens clade (shown in Fic 1) contains 12 sequences from public databases

and seven from our collections — three of C. xanthodryophilus, three collections of

\C. pseudoglaucopus TUB011872 Garnica et al, Germany EUROPE

ry | C. pseudoglaucopus TUBO12731 Garnica et al, Switzerland EUROPE

c. pseudogiaucopus AB01-09-91 Frosiev at al, Denmark, EUROPE

—C. pseudoglaucopus, DBB19789, Pirin Mtn, Bulgaria, EUROPE

- , |C- “lilacbeauty" DBB27249 Casper, California

NE i C. “lilacbeauty" DBB09552 Mendocino, California

C. "“lilacbeauty" DBB09434 Jackson State Forest, California

C. flavaurora 1B19890187, Wyoming, USA

54% | C. flavaurora JFA9913, Wyoming, USA

C. elotoides JFA9983, Wyoming, USA

al yee ae praetermissus MES-4312, Garnica et al, Spain EUROPE

pet 100% C. praetermissus MES-4294, Garnica et al, Spain EUROPE

Pe Cc. xanthodryophilus DBB26451 *holotype

4 Tate + Cc. xanthodryophilus DBB27933

Cc. xanthodryophilus DBB29113

|\C. humolens CFP1281 *isotype, Froslev at al, France, EUROPE

10 |C. humolens DBO5-1 0-05, Frosiev af ai, France, EUROPE

Fata ie; humolens TUBO12722 Garnica et al, Germany EUROPE

C. humolens TUB012723 Garnica et al, Germany EUROPE

Cortinarius calochrous var. coniferarum TUBO12691, Germany, EUROPE

Fic 1. The single most parsimonious tree derived from partial nrDNA ITS sequence data

showing the position of Cortinarius xanthodryophilus in relation to the other members of the

Pseudoglaucopodes/Humolens clade. Branch lengths are shown above and the jackknife consensus

numbers below the branches. The Bayesian inference (BI) analysis produced the same tree topology

with very strong posterior probability support.

320 ... Bojantchev & Davis

TABLE 1. List of the Cortinarius collections referenced in the phylogram (Fie 1).

TAXA Host TREE & LOCATION VOUCHER No. GENBANK No.

C. calochrous vat. Picea abies, Hinterstein, Germany TUB 012691 EU056956

coniferarum

C.elotoides Pengelmannii, Wyoming, USA | JFA9983._ | EU056948

NEA De tan OER OEE Se a desk Gee en ete eee ae ee

Ua aN aon ae E ergelimarini, Wyoming USA ado Ds cucbt OOOO hia

C. xanthodryophilus Quercus agrifolia, California, USA DE Beets HQ441244

1 AEN oa et BE Rae et es CON, MRD MNS TL es SOLO) Lt AOR Sin, MNCS ere... ek

raeisteia cate Recency <neecitte et Oe OLMIA AST ee bas eA a Re PEG Mas ee

: Notholithocarpus densiflorus, : DBB27933 i JF273636

? California, USA : :

: P sitchensis, JSE, California, USA ? DBB09434 ? HQ997909

C. praetermissus : Q. ilex subsp. rotundifolia, Spain : MES-4294 : EU684534

sa RU a a a ae ep Os ee oe AN the

: halepensis, Spain

Ly pseudoglaucopus |: Germany a csssatneuamatnatatneint UB 011872 02 AY669573

i P. abies, La Chaux-de-Fonds, i TUB012731 —? _EU056952

LAM, Pee ee Susi atl sr ye ee en A Pe Be

ccc Denmark 00 ABOI-09-91 | DQ663394

: P abies, Pirin Mountain, Bulgaria : DBB 19789 i JE273633

an undescribed species from California (provisionally labeled C. “lilacbeauty’),

and one European collection of C. pseudoglaucopus (M.M. Moser) Quadr. from

Bulgaria. Cortinarius calochrous var. coniferarum (M.M. Moser) Nezdojm.

was selected as the outgroup because it is fairly representative of the overall

calochroid super-clade but falls outside of the /pseudoglaucopodes clade.

Multiple sequence alignments were generated with both ClustalX2 2.0.12

(Thompson 1997) and MAFFT v6.821b (Katoh 2002) with the G-INS-i global

alignment iterative refinement strategy. The results were compared and visually

inspected for areas of ambiguous alignment. The alignment and the molecular

phylogenetic tree are available in TreeBase (http://purl.org/phylo/treebase/

phylows/study/TB2:S11184)

Maximum parsimony (MP) analysis was performed with PAUP* 4.0b10

(Swofford 2003) utilizing a heuristic search with tree-bisection-reconnection

(TBR) branch swapping and 10000 random addition sequence replicates and

maxtrees set at 1000. The analysis resulted in a single most parsimonious tree

(Fic 1). To test branch length support, a Jackknife (JK) consensus tree from

1000 replicates was calculated with 50% majority rule.

Cortinarius xanthodryophilus sp. nov. (U.S.A.) ... 321

In addition, Bayesian inference (BI) was run with Mr.Bayes v.3.1.1

(Huelsenbeck & Ronquist 2003) with the General Time Reversible substitution

model plus gamma distribution (GTR + I) as the best fit recommended by

MrModeltest v.2.3 (Posada & Crandall 1998). The BI ran two independent

analyses with four chains for 1000000 generations with sampling frequency for

every 100th generation and a burnin ratio set at 2500 (25%). The 50% majority

rule consensus tree showed high posterior probabilities (PP) and produced the

same topology as the most parsimonious tree (Fic 1).

Fic 2. Cortinarius xanthodryophilus (collection DBB26491) - the most typical form.

Taxonomy

Cortinarius xanthodryophilus Bojantchev & R.M. Davis, sp. nov. FIGs 2-4

MycoBank MB 519109

Pileo 60-100 mm lato, hemispherico, dein plano-convexo, glutinoso, margine involuto,

flavo, flavobrunneo, interdum rufo-brunneo maculato, Lamellis emarginatis, pallide

luteis, Stipite 50-100 mm longo, bulbo marginato 30-50 mm lato, Velo universale albido.

Carne albida, cortina copiosa, rufo-brunnea, sapore miti. Sporis 10-12 x 5.5-7 um,

amygdaliformibus usque limoniformibus, grosse verrucosis, basidiis 30-40 x 7-10 um,

tetrasporigeris, fibulis praesentibus.

Type: “USA, California, Contra Costa County, Tilden Park, Berkeley, 2009/11/15 col.

Dimitar Bojantchev DBB26451 UCB Herbarium: Holotype UC 1860808 Genbank

nrITS HQ441244”

ErymMo_oey: from the Greek: xanthos = yellow, drys = oak, philios = loving

STATURE pileocarpous bulbopodium, very variable in aspect ratio. PILEUS

60-100 mm diam, hemispherical to convex to plano-convex to uplifted in age.

Margin persistently involute. Colors rather uniform, predominantly in yellow

322 ... Bojantchev & Davis

Fic 4. Cortinarius xanthodryophilus. a) DBB26451 (*holotype), b) DBB40412, c) DBB11176,

d) DBB40412 showing KOH,,, reaction on pileus and context, e) DBB27933, f) DBB29113.

All collections shown in Fic 4 have matching ITS1/5.8s/ITS2 sequences.

shades, starting pale straw- to sulphur-yellow (2.5Y 8/6) becoming yellow-

brown (1OYR 8/6-8/8), darker near the center (LOYR 6/6-8/8), frequently

with reddish-brown discolorations. Surface glutinous when wet, glabrous to

dull glossy when dry, at age developing cracks and areolations near the disk,

remaining smoother near the margin. LAMELLAE L=80-120, crowded, 8-15

mm broad, pale sulphur-yellow (2.5Y 8/6-8/8) to off-white when young,

turning various shades of yellow-brown to brown (7.5R 6/6-5/6) as the spores

mature. Edges even to occasionally slightly wavy, frequently eroded with age.

Attachment notched. Lamellulae abundant, with widely varying extensions,

15-75%, series of 3-5. StrPE 50-100 mm long, 15-30 mm wide, cylindrical

to subclavate above the bulb. Mostly white, but occasionally with light bluish

tinges in the upper part. BuLB 30-50 mm diam at the widest point, always

well-developed, abruptly emarginated, tapering below, the subterrestrial part

Cortinarius xanthodryophilus sp. nov. (U.S.A.) ... 323

with a white cottony mycelial felt. CoNTExT mostly white, slowly bruising

brownish, few of the basidiomata feature a strong bluish cast in the upper stipe

context, more pronounced near the surface. A watery grayish-blue cast above

the lamellae is present in almost all basidiomata, rather faint in dry conditions,

but persisting deep into maturity. UNIVERSAL VEIL white, frequently leaving

floccose patches on the pileus surface, occasionally forming volva-like extensions

on the bulb margin in recently expanded basidiomata. Cortina white to pale

yellow, turning rusty brown due to mature spore drop, copious, persistent,

leaving an annular zone of dense fibrils on the stipe and frequently forming

a hairy appendiculate zone on the pileal margin. The cortina deposits form a

distinct brownish belt on the bulb edge, which can be fused into a gelatinized

matrix on the periphery. MACROCHEMICAL REACTIONS KOH,, light reddish

brown on the pileus surface, stronger on the brownish spots near the disk. On

the context the reaction varies from little to none on very young material, to

yellowish-brown on mature basidiomata, stronger in the lower stipe and near

the surfaces. No reaction was observed on the basal mycelium. UV no reaction

was detected with both fresh and dry material. Opor mild, leafy, and earthy.

TASTE mild, earthy.

Fic 3. Cortinarius xanthodryophilus. a) Basidiospores b) Gelatinous cuticle.

BASIDIOSPORES (9.5-)10-12(-13) x (5-)5.5-7(-7.5) um (mean 11.2 x

6.3 um) Q = 1.66-1.85, Q. = 1.78 (N = 213, 7 collections, 14 basidiomata),

amygdaliform to citriform, distinctly and coarsely verrucose, deep rusty brown

in deposit, slightly dextrinoid. Basip1a 30-40 x 7-10 um, 4-spored, cylindro-

clavate, clamped. Hymenial layer not reacting to alkaline or iodine solutions.

CystTip1A none observed. PILEIPELLIS a cutis, simplex, no hypodermium

detected, composed of parallel to interwoven hyphae in a dense gelatinous

matrix 150-250 um thick. The outer 10-15 layers of hyphae 2-4 um diam,

entangled, some erect, irregularly shaped, strangulated to twisted, commonly

with non-parallel walls, mostly with refractive cytoplasmic pigment. Lower

layer of cuticle hyphae 3-7 um diam, mostly parallel, with thicker yellow walls

and hyaline content. The yellow pigmentation is emphasized when mounted

in KOH. No distinct reactions to Melzer’s reagent were observed. CLAMP

324 ... Bojantchev & Davis

CONNECTIONS common in all parts. TRama composed of cylindrical cells

10-15(-20) um diam, hyaline with pale yellow walls. Occasional oleiferous

hyphae present.

HABITAT AND DISTRIBUTION - Solitary to gregarious under oaks. This

species is common in California under live oak (Quercus agrifolia). In the Sierra

Nevada foothills we have collected it under interior live oak (Q. wislizenii) and

canyon live oak (Q. chrysolepis). In Mendocino Co. it was collected under

tanoak (Notholithocarpus densiflorus). Based on a molecular data match there is

a collection (Genbank DQ974721) under blue oak (Q. douglasii) in the Central

Valley. There is also a collection (Genbank GQ159771) from Vancouver Island,

British Columbia, under garry oak (Q. garryana).

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: Contra Costa Co., Tilden

Park, under Quercus agrifolia, 3 Dec 2008 (coll. D. Bojantchev DBB11492); 23 Nov

2009 (coll. D. Bojantchev DBB28181); 8 Dec 2008 (coll. D. Bojantchev DBB26491); San

Mateo County, Huddart Park, under Quercus agrifolia, 8 Dec 2008 (coll. D. Bojantchev

DBB11176, UCB Herbarium: UC 1860807); 8 Dec 2009 (coll. D. Bojantchev DBB29113);

Marin County, Point Reyes, under Quercus agrifolia, 28 Nov 2006 (coll. D. Bojantchev

DBB26128); San Mateo County, San Francisco Watershed, under Quercus agrifolia,

3 Dec 2010 (coll. D. Bojantchev DBB40412); Mendocino County, Casper Cemetery

under Notholithocarpus densiflorus 22 Nov 2009 (coll. D. Bojantchev DBB27933).

Discussion

Phylogenetically, C. xanthodryophilus belongs to the /pseudoglaucopodes

clade (Garnica et al. 2009) where it holds a well delineated position as shown

in Fic 1. Cortinarius xanthodryophilus can easily be distinguished from species

closely related to C. pseudoglaucopus (Jul. Schaff. ex M.M. Moser) Quadr.

(Fic 5), which are typically associated with conifers and possess lilac veils

(more or less obvious) and significantly larger spores than C. xanthodryophilus.

Cortinarius “lilacbeauty” (Fic 6), the only clade member known to occur in

California, is distinctly lilac-gray and thus easily separated in the field.

Amongst the hardwood-associated members of the clade, C. xanthodryophilus

bears the closest resemblance to C. humolens Brandrud. Although they share

many morphological features and the spores are very similar in size, shape and

ornamentation, only C. xanthodryophilus has frequent bluish tinges on the

context and upper stipe. For a good treatise and iconography of C. humolens,

refer to Brandrud et al. (1998), Bidaud et al. (2004), and Consiglio et al.

(2007).

Another closely related species, C. praetermissus Bergeron ex Reumaux,

was collected from Morella, Eastern Spain, under evergreen oaks. Cortinarius

praetermissus is not well known, but it was originally described as a beech-

associated, pale bluish species. Its spore shape and size, which fit quite

well with the species close to C. pseudoglaucopus, are larger than those of

C. xanthodryophilus.

Cortinarius xanthodryophilus sp. nov. (U.S.A.) ... 325

Fic 5. Cortinarius pseudoglaucopus collections DBB19789 and DBB19822 from the Pirin Mountain

Bulgaria, Europe, under Picea abies, showing the full range of coloration in younger and older

basidiomata.

Fic 6. Cortinarius “lilacbeauty” collections. DBB 09434 and DBB 09552 from Northern California,

under Picea sitchensis. Based on the partial nrITS sequences this species is very close to Cortinarius

pseudoglaucopus (Fic 5).

Other yellow bulbopodiums in the northern California mixed woods add to

the challenge of identifying C. xanthodryophilus in the field. The local species

most likely to be confused with C. xanthodryophilus is “Cortinarius fulmineus”

sensu Moser & Ammirati 1997 (Fic 7). This species, which shares the same

general colors with C. xanthodryophilus, tends to oxidize red-brown on the pileus,

particularly on the disk. It also has a strong alkaline reaction, which is purple

red on the pileal surface and pinkish on the context, as well as smaller spores

(8-10 um). Cortinarius elegantior var. americanus M.M. Moser & McKnight

(Fic 8), a conifer-associate that can be confused with C. xanthodryophilus and

frequently occurs with it in mixed woods, can be differentiated by the strong

red alkaline reaction on the pileus and bulb and the much larger spores.

There are several other common and undescribed yellow bulbopodiums in

Northern California that can be confused with C. xanthodryophilus. Genetically,

they fall in the clades around C. citrinus P.D. Orton, C. elegantissimus

Rob. Henry, C. flavovirens Rob. Henry, C. fulvocitrinus Brandrud, C. platypus

326 ... Bojantchev & Davis

Fic 7. “Cortinarius fulmineus” sensu Moser & Ammirati 1997. This species is the most likely to

be confused with C. xanthodryophilus in the field. The pronounced tendency of the pileus of even

young basidiomata to oxidize red-brown and the strong KOH,,, reaction (lower right photo) aids

field identification.

Fic 8. Cortinarius elegantior var. americanus, col. DBB27264.

KOH,,, reaction on the right.

(M.M. Moser) M.M. Moser, C. xanthophyllus (Cooke) Rob. Henry, and others.

A complete treatment of these species is beyond the scope of this article, but they

all differ from C. xanthodryophilus by the combination of macromorphology,

macrochemical reactions, and microscopic detail. In a future publication we

will provide a comprehensive key to the Phlegmacium species of Northern

California with an emphasis on field level identification.

A complete iconography of Cortinarius xanthodryophilus and a comparative

image study is available on the website http://www.mushroomhobby.com.

Cortinarius xanthodryophilus sp. nov. (U.S.A.) ... 327

Acknowledgements

We thank Prof. Dennis Desjardin and Prof. Joseph Ammirati for their reviews and

comments. We are very grateful to Dr. Else Vellinga for her wise counsel on a broad

array of subjects concerning the preparation of this manuscript. Dr. Boris Assyov,

who reviewed the Latin diagnosis and the rest of the paper in depth, offered several

key corrections and recommendations. Special acknowledgement is directed to the

community of informed amateur collectors in California, whose observations and data

have been influential in solidifying the species concepts, ecology, and distribution of

genus Cortinarius in California.

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MY COTAXON

Volume 116, pp. 329-333 April-June 2011

DOT: 10.5248/116.329

Zwackhiomyces turcicus sp. nov. (Ascomycota, Xanthopyreniaceae)

from Turkey

MUuSTAFA KOCAKAYA', MEHMET GOKHAN HALICcr’’ & AHMET AKSOY?

‘Bozok Universitesi, Fen Edebiyat Fakiiltesi, Biyoloji Boliimti, 66200 Yozgat, Turkey

23 Erciyes Universitesi, Fen Fakiiltesi, Biyoloji Boliimii, 38039 Kayseri, Turkey

CORRESPONDENCE TO*: ‘mustafa.kocakaya@bozok.edu.tr, *mghalici@erciyes.edu.tr,

& -aksoy@erciyes.edu.tr

AxBstRAcT — Zwackhiomyces turcicus is described as new from the thallus of Physcia

magnussonii from southern Turkey. The new species produces one of the largest ascomata in

the genus and is easily differentiated from Z. physciicola (described from Physcia caesia) by its

larger verruculose ascospores and thinner interascal filaments.

Key worps — biodiversity, lichenicolous fungi, lichens

Introduction”

Grube & Hafellner (1990) established the genus Zwackhiomyces for

Arthopyrenia coepulona and eight related lichenicolous taxa. Since then, several

new species have been described or transferred to this genus, and Calatayud et

al. (2007) provided a key to the known 22 Zwackhiomyces species. ‘Thereafter,

several authors have added new species (Hawksworth & Iturriaga 2006, Brackel

2008, Diederich & Zhurbenko 2009, Diederich & Schultz 2009, Roux 2009),

bringing the total described to 28.

Six Zwackhiomyces species (Halici 2008, Halici & Candan 2009, Halici et

al. 2009) are included among the 160 species of lichenicolous fungi reported

from Turkey (Halici et al. 2010). Here we describe a new one from the region,

Zwackhiomyces turcicus on Physcia magnussonii.

Material & methods

Type material is deposited in the herbarium of Erciyes University, Science & Art

Faculty, Biology Department, Kayseri. Specimens were examined with a Leica DM-1000

research microscope. Photomicrographs were prepared using a Leica DFC 420. The

perithecia were examined by handmade sections prepared by razor blade and examined

in I (Lugol's iodine with [KI] and without [I] pre-treatment in 10% KOH), 10%

KOH, and water. Ascus and ascospore measurements were made in water. Ascospore

330 ... Kocakaya, Halici & Aksoy

measurements are given as: the arithmetic mean, flanked by the mean + standard

deviation and parenthetical minimum and maximum values. The length/breadth ratio

of ascospores is indicated as 1/b and given in the same way. The abbreviation n indicates

the number of measurements.

Taxonomy

Zwackhiomyces turcicus Kocakaya, Halici & Aksoy, sp. nov. Fic. 1

MycoBank MB 519612

Fungus lichenicola in thallo Physciae magnussonii. Perithecia nigra, superficialia,

200-450 um diam.. Asci (65-)70-92(-95) x (18-)19-21(-22) um. Ascosporae 1-septatae,

hyalinae, verruculosae, (17-)18-22(-23) x (7-)8-9.5(-10) um, I/b = (2-)2.15-2.47

(-2.57). Filamenta interascalibus 1-1.5 ym lata.

TYPE COLLECTION: Turkey, Konya, Taskent, Gevne Vadisi, Pinus nigra forest, 36°45’

N, 32°25’ E, alt. 1700 m, on thallus of Physcia magnussonii Frey on siliceous rocks, 6

August 2010, M. Kocakaya, M.G. Halici 0.6317 & A. Aksoy (Erciyes University, Lichen

herbarium - holotype).

Erymo.oey: The epithet “turcicus” refers to Turkey, where the type specimen was

collected.

Lichenicolous on the lobes of Physcia magnussonii. AscoMaTA perithecioid,

black, subglobose to globose, c. 200-450 um diam., generally scattered on the

margins of the lobes of the host thallus, arising singly, superficial. Ascomatal wall

pseudoparenchymatous, of textura angularis, dark reddish brown, uniformly

coloured, not distinctly separated into two layers, in section 30-60 um wide,

with 6-10 layers of the cells, with the inner layers slightly paler; cells + rounded

to somewhat irregular in surface view, mostly 4-7 um diam. HYMENIAL GEL I-,

K/I-. Interascal filaments abundant, branched and anastomosing, c. 1-1.5 um

wide. Asci (4-)6-(-8)-spored, (65-)70-81-92(-95) x (18-)19-20.5-21(-22)

um (n = 20), the wall is thickened at the apex, with a distinct ocular chamber,

elongate-clavate, stipitate, ascospores distichously arranged in the upper

part, uniseriately arranged in the lower part, endoascus I+ orange-red and

K/I+ orange-red, dehiscence fissitunicate. AscosporeEs 1-septate, markedly

constricted at the septum, lower cell narrower than the upper cell, hyaline, with

a verruculose surface, usually one big oil droplet present in each cell, (17-)18-

20-22(-23) x (7-)8-8.5-9.5(-10) um, 1/b ratio: (2-)2.15-2.31-2.47(-2.57) (n

= 50). CONIDIOMATA not seen.

ECOLOGY AND DISTRIBUTION: The new species is known only from the

type locality in southern Turkey, growing on Physcia magnussonii on siliceous

rocks. It is commensalistic or weakly parasitic, as slight bleaching occurs on

the infected part of the host thallus. The type specimen was collected in a

Pinus nigra forest with a typical Mediterranean climate. As the host species

is abundant in the northern hemisphere, the species should be searched for

elsewhere, especially on epilithic Physcia species.

Zwackhiomyces turcicus sp. nov. (Turkey) ... 331

Fic. 1. Zwackhiomyces turcicus (holotype). A, Perithecia on the host thallus; B, Ascomata wall

in K; C, 6-spored ascus in I; D, 8-spored ascus in I; E, Interascal filaments in K; EK, Ascospores

showing verruculose surface in I; G, Ascospores showing verruculose surface in I with higher

magnification.

332 ... Kocakaya, Halici & Aksoy

OBSERVATIONS: Zwackhiomyces turcicus and Z. cladoniae (C.W. Dodge)

Diederich have the largest ascomata (c. 200-450 um diam.) in the genus (Grube

& Hafellner 1990, Hawksworth & Iturriaga 2006, Calatayud et al. 2007 ).

Another Zwackhiomyces species that also occurs on Physcia (P. caesia) is

Z. physciicola Alstrup, described from Denmark with only a few mature

ascospores (Alstrup 1993). Zwackhiomyces physciicola clearly differs in having

much smaller (100-150 um) ascomata, nonverruculose ascospores that are

clearly narrower (5.5-6.5 um) and constricted at the septa, and thicker (2

um diam.) interascal filaments (Alstrup 1993). According to Diederich (pers.

comm.), a careful examination of ascospores in Luxembourg material of

Z. physciicola revealed the presence of three pairs of setulae fixed on the larger

ascospore cell at c. 2 um from the septum (Sérusiaux et al. 1999). The same

kind of ascospore setula is known in most Lichenopeltella species (Aptroot et al.

1997) but has never been reported for Zwackhiomyces.

Zwackhiomyces berengerianus (Arnold) Grube & Triebel described on

Mycobilimbia berengeriana has similarly sized ascospores, but they are slightly

constricted at the septa and the perispore becomes brownish at maturity.

Furthermore, Z. berengerianus has much smaller (125-180(-200) um)

ascomata than Z. turcicus (Grube & Hafellner 1990). Zwackhiomyces kiszkianus

D.Hawksw. & Miadl. described on Peltigera canina also has verruculose

ascospores, but the spores are longer and wider (19.5-25.5x 8.5-13 um) than

Z. turcicus and the ascomata are immersed (Hawksworth & Miadlikowska

1997). Another related species, Z. coepulonus (Norman) Grube & R.Sant.

(known on Xanthoria elegans and Caloplaca spp.), has smaller ascomata which

are immersed to superficial (150-250 um) and slightly smaller ascospores

((15-)16-20(-21) x 5.5-8.5(-9)) (Grube & Hafellner 1990). As stated above,

Z. cladoniae, which is known on Cladonia spp., is another species in the genus

with very large ascomata, but it clearly differs from Z. turcicus in having much

narrower and shorter (12-19 x 3-5 um) ascospores (Hawksworth & Iturriaga

2006).

Acknowledgements

Paul Diederich and Ave Suija are thanked for reviewing this paper. Harrie Sipman

is thanked for checking the Latin diagnosis. This study was financially supported by

Erciyes University Scientific Research Project (FBD-10-3241 coded project).

Literature cited

Alstrup V. 1993. News on lichens and lichenicolous fungi from Nordic countries. Graphis Scripta

5: 96-104.

Aptroot A, Diederich P, Sérusiaux E, Sipman HJM. 1997. Lichens and lichenicolous fungi from

New Guinea. Bibliotheca Lichenologica 64: 1-220.

Brackel W. 2008. Zwackhiomyces echinulatus sp. nov. and other lichenicolous fungi from Sicily,

Italy. Herzogia 21: 181-198.

Zwackhiomyces turcicus sp. nov. (Turkey) ... 333

Calatayud V, Triebel D, Pérez-Ortega S. 2007. Zwackhiomyces cervinae, a new lichenicolous fungus

(Xanthopyreniaceae) on Acarospora, with a key to the known species of the genus. Lichenologist

39: 129-134. doi:10.1017/S002428290700583X

Diederich P, Schultz M. 2009. Zwackhiomyces namibiensis, a new lichenicolous ascomycete

(Xanthopyreniaceae) on Psorotichia from Namibia. Herzogia 22: 173-176.

Diederich P, Zhurbenko M. 2009. Sphaerellothecium phaeorrhizae and Zwackhiomyces sipmanii

sp. nov. on Phaeorrhiza sareptana from north-eastern Asia, with a key to the species of

Sphaerellothecium. Bibliotheca Lichenologica 99: 113-122.

Grube M, Hafellner J. 1990. Studien an flechtenbewohnenden Pilzen der Sammelgattung Didymella

(Ascomycetes, Dothideales). Nova Hedwigia 51: 283-360.

Halici MG. 2008. A key to the lichenicolous Ascomycota (including mitosporic fungi) of Turkey.

Mycotaxon 104: 253-286.

Halici1 MG, Candan M. 2009. New lichenicolous fungi from Turkey. Nova Hedwigia 88: 483-490.

doi:10.1127/0029-5035/2009/0088-0483

Halici1 MG, Candan M, Ozdemir Tiirk A. 2009. Notes on some lichenicolous fungi from Turkey II.

Turkish Journal of Botany 33: 389-392.

Halic1 MG, Akata I, Kocakaya M. 2010. New records of lichenized and lichenicolous fungi from

Turkey. Mycotaxon 114: 311-314.

Hawksworth DL, Iturriaga T. 2006. Lichenicolous fungi described from Antarctica and the sub-

Antarctic islands by Caroll W. Dodge (1895-1988). Antarctic Science 18: 291-301. doi:10.1017/

$0954102006000344

Hawksworth DL, Miadlikowska J. 1997. New species of lichenicolous fungi occurring on Peltigera in

Ecuador and Europe. Mycological Research 101: 1127-1134. doi:10.1017/S0953756297003778

Roux C. 2009. Naetrocymbe saxicola, Likeno kun Trentepohlia. Bulletin de la Société Linnéenne de

Provence 60: 127-142.

Sérusiaux E, Diederich P, Ertz D, van Den Boom P. 1999. New or interesting lichens and lichenicolous

fungi from Belgium, Luxembourg and northern France. IX. Lejeunia 173: 1-48.

MYCOTAXON

Volume 116, pp. 335-339 April-June 2011

DOI: 10.52.48/116.335

Arthonia anatolica sp. nov. (Arthoniaceae) on

Aspicilia contorta subsp. hoffmanniana,

a new lichenicolous species from Turkey

MEHMET GOKHAN HALIcCI”* & MEHMET CANDAN?

'Erciyes Universitesi, Fen Edebiyat Fakiiltesi, Biyoloji Boliimti, 38039 Kayseri, Turkey

Anadolu Universitesi, Fen Fakiiltesi, Biyoloji Béliimii, Eskigehir, Turkey

CORRESPONDENCE TO *: “mghalici@erciyes.edu.tr & *mecandan@anadolu.edu.tr

AsBstRAct —Arthonia anatolica sp. nov. is described on Aspicilia contorta subsp. hoffmanniana

on calcareous rocks from central and eastern Turkey. The new species clearly differs from

the other Arthonia species reported on Aspicilia sp. and most closely resembles Arthonia

amylospora, a species known on Porpidia rugosa. The differences are discussed.

KEY worps — Ascomycota, lichenicolous fungi, lichens

Introduction

The genus Arthonia Ach. includes c. 500 species (Hawksworth et al. 1995),

the majority of which are lichenized. About 83 species are lichenicolous fungi

(Grube & Matzer 1997, Candan & Halici 2009, Halici 2008a, Halici1 & Candan

2009, Zhurbenko & Grube 2010; Etayo 2010), and 38 lichenicolous species

were included in the key to Arthonia by Clauzade et al. (1989). Recently,

27 Arthonia species were included in the annotated key to the lichenicolous

fungi of Sweden (Ihlen & Wedin 2008), while 9 species were included in the

key to the lichenicolous fungi of Turkey (Halici 2008b). Grube & Matzer (1997)

provided useful taxonomical characters for Arthonia, focusing primarily on

lichenicolous taxa.

Currently 160 lichenicolous fungi species are known from Turkey (Halici

et al. 2010), with at least 250 species total expected from the country (Halici

et al. 2007). Of the 16 lichenicolous Arthonia species reported from Turkey

(Hafellner & John 2006, Candan & Halici 2009, Halici 2008a,b, Halici & Candan

2009), four — A. aysenae Halici & Candan on Acarospora cervina, A. epitoninia

Halici & Candan on Toninia sp., A. hawksworthii Halici on Dimelaena oreina,

A. rinodinicola Candan & Halici on Rinodina gennarii — have been originally

336 ... Halici & Candan

described from Turkey (Halici 2008a, Candan & Halici 2009, Halici & Candan

2009). We propose here a new species, A. anatolica on Aspicilia contorta subsp.

hoffmanniana, based on two collections from central and eastern Turkey.

Material & methods

The type material and the additional examined specimen of the new species are

deposited in the lichen herbarium of Erciyes University (Kayseri, Turkey) and ANES.

Specimens were examined with Leica DM 1000 research microscope. Microphotographs

were taken with Leica DFC 420 digital microscope camera with c-mount interface and

with a 5 megapixel CCD. Sections were prepared by hand and examined in I (Merck

Lugol's iodine) and water. Ascospore measurements were made in water. Ascospore and

asci measurements were given as: the arithmetic mean, flanked by the mean + standard

deviation and parenthetical minimum and maximum values. All measurements and

ratio includes the halo in ascospores. The length/breadth ratio of ascospore is indicated

as l/b and given in the same way.

Taxonomy

Arthonia anatolica Halici & Candan, sp. nov. FIGURE 1

MycoBank MB 519953

Fungus in thallo Aspicilia contortae incolens. Apothecia nigra, 200-300 um diam.

Epithecium 10-20 um altum, olivaceo-brunneum. Hymenium 100-110 um altum,

olivaceum. Hypothecium 10-15 um altum, hyalinum. Asci (49-)53-64-74(-90) x

(20-)21.5-25-28(-29) um, clavati, octospori. Ascosporae 1-septatae, hyalinae, halonatae,

(15-)18-19-21(-23) x (7-)8-9-10(-11) um.

Type: Turkey, Kayseri, Develi, Bakirdag, vicinity of Kale Village, steppe area with

calcareous mother-rock, 38°06’N, 35°42’E, alt. 1550 m, on thallus of Aspicilia contorta

subsp. hoffmanniana S.Ekman & Fréberg ex R.Sant. on calcareous rocks, 3 September

2010, leg. M. G. Halici (0.6799) (Erciyes University, Lichen herbarium - holotype).

Erymo oey: The specific name refers to Anatolia, the name of the geographical area of

Asian Turkey.

DESCRIPTION: Lichenicolous, on the areoles of Aspicilia contorta subsp.

hoffmanniana, causing slight bleaching in the infected parts, weakly parasitic.

VEGETATIVE HYPHAE below the hypothecium I+ blue. Ascomata apothecioid,

ageregated, numerous, 200-300 um diam., black, epruinose, superficial,

lacking an exciple, arthonioid. Epithecium olivaceous brown, K- 10-20 um

high; hymenium olivaceous green, I+ red, KI-, 100-110 um high; hypothecium

colourless, 10-15 um high. HAMATHECIUM of paraphysoids, abundant, septate,

branched and anastomosed, 2-2.5 um wide. Ascr broadly clavate, shortly

stalked, bitunicate in structure, 8-spored, inner part I+ red, KI-, without a

Fic. 1 (to right). Arthonia anatolica (holotype). A, Hymenium in KI; B, Hymenium in J];

C, Ascospores in water; D, Ascospores in I, showing the I + purple-red gelatinous sheath;

E, EF, Asci.

Arthonia anatolica sp. nov. (Turkey) ... 337

338 ... Halici & Candan

KI+ blue apical ring, (49-)53.2-63.8-74.4(-90) x (20-)21.6-24.8-28(-29) um

(n = 20). AscosporeEs irregularly arranged in the asci, ellipsoid, hyaline, 1-

septate, rounded to somewhat broadly pointed at the apices, slightly constricted

at the septa, the upper cell larger and longer than the lower one, halonate, halo

~1 um, smooth, I+ purple-red, (15-)18-19-21(-23) x (7-)8-9-10(-11) um

(n = 44), I/b = (1.45-)1.91-2.20-2.49(-3.0). CONIDIOMATA not observed.

ECOLOGY AND DISTRIBUTION: Arthonia anatolica seems to be parasymbiotic

or weakly parasitic on the thalli of Aspicilia contorta subsp. hoffmanniana, as

slight bleaching occurs in the infected areoles of the host. The new species is

known from two localities in central and eastern Turkey, where steppe vegetation

is pre-dominant. As the host species has a wide distribution in the northern

hemisphere on calcareous rocks, the species should be sought elsewhere.

ADDITIONAL SPECIMEN EXAMINED: TURKEY, ELaziG, southwest of Hidirbaba Village,

steppe area with calcareous mother-rock, 38°45'N, 39°00'E, alt. 1120 m, on thallus of

Aspicilia contorta subsp. hoffmanniana on calcareous rocks, 4 August 2004, leg. M.

Candan (ANES 11635).

OBSERVATIONS: Following Grube & Matzer (1997), Arthonia anatolica belongs

to the A. radiata group because of its olivaceous brown epithecium and thick

interascal filaments.

Several species of Arthonia are known or reported on Aspicilia spp. Arthonia

aspiciliae Alstrup & E.S. Hansen is known on Aspicilia elevata from Greenland

(Alstrup & Hansen 2001) and has much smaller (11-13 x 5-6 um) ascospores

than A. anatolica. Similarly, Arthonia hertelii (Calat. et al.) Hafellner & V. John,

which is known on vagrant Aspicilia spp. mainly in the steppe (Calatayud et

al. 2004), clearly differs from A. anatolica in having smaller ((12-)14-15-17 x

(4-)5-5-6 um)) and non-halonate (or inconspicuously halonate) ascospores,

a pale brown hymenium with an I+ purple reaction, and a deep brown

hypothecium. Arthonia urceolata (Elenkin) V.J. Rico et al., also a common

species on vagrant Aspicilia species in the Eurasian steppe (Calatayud et al.

2004), has clearly smaller ((12-)13-14-16(-18) x (4.5-)5-5-6 um) ascospores

and a hymenium with an I+ purple reaction. Arthonia oligospora Vézda was

reported on Aspicilia sp. from the eastern part of Turkey (Candan & Halici

2008) and from Iran (Valadbeigi & Sipman 2010); A. oligospora has 4-spored

asci and much smaller ((12.5—) 14-16 x 6.5-7 um) ascospores than A. anatolica.

Arthonia lobothalliae Etayo, a species described on Lobothallia alphoplaca, also

has much smaller (11-12.5 x 4.5-5 um) ascospores (Etayo 2010).

Arthonia amylospora Almq. on Porpidia rugosa appears most similar to

A. anatolica (Triebel 1989, Ihlen & Wedin 2008). Besides having a different host,

the hymenium is brownish and shorter (45-55 um), the hypothecium is dark

brown, and the asci are much smaller (36-38 x 19 um) than in A. anatolica.

Ascospore sizes of A. amylospora are given as 18-21 x 6-8 um in Triebel

Arthonia anatolica sp. nov. (Turkey) ... 339

(1989) and 16-24 x 6-10 um in Ihlen & Wedin (2008) whereas A. anatolica has

ascospores in range of (15-)18-19-21(-23) x (7-)8-9-10(-11) um.

Acknowledgements

Kerry Knudsen and Jana Kocourkova are thanked for reviewing this paper. This

study was financially supported by Erciyes University Scientific Research Project

(FBA-09-908 coded project).

Literature cited

Alstrup V, Hansen ES. 2001. New lichens and lichenicolous fungi from Greenland. Graphis Scripta

12: 41-50.

Calatayud V, Barreno E, Rico VJ. 2004. Two lichenicolous species of Arthonia on vagrant Aspicilia

species. Bibliotheca Lichenologica 88: 67-78.

Candan M, Halici MG. 2008. Seven new records of lichenicolous fungi from Turkey. Mycotaxon

104: 241-246.

Candan M, Halici MG. 2009. Two new lichenicolous Arthonia species from Turkey. Mycotaxon

107: 209-213. doi:10.5248/107.209

Clauzade G, Diederich P, Roux C. 1989. Nelikenigintaj fungoj likenlogaj. Bulletin de la Société

linnéenne de Provence, numéro spécial 1: 1-142.

Etayo J. 2010. Liquenes y hongos liquenicolas de Aragon. Guineana 16: 1-501.

Grube M, Matzer M. 1997. Taxonomic concepts of lichenicolous Arthonia species. Bibliotheca

Lichenologica 68: 1-17.

Hafellner J, John V. 2006. Uber funde lichenicoler nicht-lichenisierter Pilze in der Tiirkei, mit einer

Synopsis der bisher im Land nachgewiesenen Taxa. Herzogia 19: 155-176.

Halici MG. 2008a. Arthonia hawksworthii sp. nov. (Ascomycota, Arthoniaceae) on Dimelaena oreina

from Turkey. Mycotaxon 105: 89-93.

Halici MG. 2008b. A key to the lichenicolous Ascomycota (including mitosporic fungi) of Turkey.

Mycotaxon 104: 253-286.

Halici1 MG, Candan M. 2009. New lichenicolous fungi from Turkey. Nova Hedwigia 88: 483-490.

doi:10.1127/0029-5035/2009/0088-0483

Halic1 MG, Candan M, Ozdemir Tiirk A. 2007. New records of lichenicolous and lichenized fungi

from Turkey. Mycotaxon 100: 255-260.

Thlen PG, Wedin M. 2008. An annotated key to the lichenicolous Ascomycota (including mitosporic

morphs) of Sweden. Nova Hedwigia 86: 275-365. doi:10.1127/0029-5035/2008/0086-0275

Hawksworth DL, Kirk PM, Sutton BC, Pegler DN. 1995. Ainsworth & Bisby’s dictionary of the

fungi. 8 edn. Wallingford.

Triebel D. 1989. Lecideicole Ascomyceten. Eine Revision der obligat lichenicolen Ascomyceten auf

lecideoiden Flechten. Bibliotheca Lichenologica 35: 1-278.

Valadbeigi T, Sipman HJM. 2010. New records of lichens for the flora of Iran and their biographical

significance. Mycotaxon 113: 191-194.

Zhurbenko M, Grube M. 2010. Arthonia pannariae (Arthoniaceae, Arthoniales), a new lichenicolous

fungus from northern Holarctic. Graphis Scripta 22: 47-51.

MY COTAXON

Volume 116, pp. 341-347 April-June 2011

DOT: 10.5248/116.341

A new species and a new record of Marasmius from China

CHUN-YING DENG”, Tat-Hut L1”’, & BIN SONG?

‘School of Bioscience and Biotechnology, South China University of Technology,

Guangzhou, 510641, China

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

Guangdong Institute of Microbiology Guangzhou 510070, China

*CORRESPONDENCE TO: mycolab@263.net

ABSTRACT —Marasmius pseudoconfertus is described and illustrated as a new species of

section Sicci series Leonini. It has a reddish brown pileus, subdistant lamellae, a fuscous to

dark brown stipe, basidiospores 13-16 x 4-5 um, and lacks pleurocystidia and caulocystidia.

Marasmius suthepensis is reported for the first time from China.

KEY worps — morphology, Marasmiaceae, taxonomy

Introduction

Taxonomic studies of Marasmius have been extensively carried out in the

world (e.g. Singer 1964, 1976; Desjardin & Horak 1997, Desjardin & Ovrebo

2006, Antonin 2003, Antonin & Buyck 2006, Antonin & Noordeloos 2010).

Recently the morphological and molecular studies of Asian Marasmius species

were extensively conducted (Tan et al. 2009, Wannathes et al. 2009, Antonin

et al. 2010a, b), whereas the species of Marasmius from China are still poorly

known. Although Karsten (1892) reported the first records of Marasmius from

China, the reported species M. dryophilus (Bull.) P. Karst. and M. butyraceus

(Bull.) P. Karst. are currently accepted as Gymnopus dryophilus (Bull.) Murrill

and Rhodocollybia butyracea (Bull.) Lennox. Teng (1963) recorded 6 species,

Tai (1979) provided a comprehensive account of Marasmius with a list of 23

Chinese taxa, and only the species of Marasmius from Guangdong and Hainan

provinces were studied relatively intensively (Bi et al 1985, 1993; Bi & Li 1987,

Li et al. 1994), although a few other records have also been reported from

other areas in China. During a recent critical re-examination on the specimens

deposited in the Herbarium of Cryptogams, Kunming Institute of Botany of

the Chinese Academy of Sciences (KUN, with HKAS numbers) and Herbarium

of Guangdong Institute of Microbiology (GDGM), a new species and a new

record for China were discovered. They are described and illustrated herein.

342 ... Deng, Li, & Song

Materials & methods

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

drier, and then preserved in the herbarium. Color terms and notations follow those of

Kornerup & Wanscher (1978). Fungal tissues were mounted in 5% KOH for microscopic

examination. The terms used to describe lamellae spacing refer to the number of lamellae

reaching from the stipe to the pileus margin, and the spacing of lamellulae is indicated

by the number of series present. Spore statistics include: x, , the arithmetic mean of the

spore length by spore width (+ standard deviation) for n spores measured in a single

specimen; Q, the quotient of spore length by spore width in any one spore, indicated as a

range of variation in n spores measured; Qm, the mean of Q-values in a single specimen;

n, the number of spores measured per specimen; s, the number of specimens involved.

Specimens are deposited in the Herbarium of Cryptogams, Kunming Institute of Botany

of the Chinese Academy of Sciences (KUN) and Herbarium of Guangdong Institute of

Microbiology (GDGM). Authors of fungal names are cited according to the International

Plant Names Index Authors website (http://www.ipni.org/ipni/authorsearchpage.do).

Genomic DNA was isolated from dried specimens and the ITS1-5.8S-ITS2 segment

from the ribosomal DNA (rDNA) was amplified with primer sets ITS1 (5’-TCC GTA

GGT GAA CCT GCG G-3’) and ITS4 (5’-TCC TCC GCT TAT TGA TAT GC -3’) by

polymerase chain reaction (PCR) techniques (White et al. 1990). Amplified products

were examined with agarose gel electrophoresis using a 2kb DNA marker. The amplified

PCR products were directly sequenced and deposited in GenBank.

Taxonomy

Marasmius pseudoconfertus T.H. Li & Chun Y. Deng, sp. nov. Fries 1-2

MycoBank MB 519525

Pileus 15-35 mm latus, hemisphaericus vel plano-convexus, non-striatus, glaber,

aurantiacus. Lamellae adnexed, albae, distantes vel subdistantes. Stipes 10-70 x 2-5 mm,

apicem albidus, brunneus ad atrobrunneus, basi non-insititius. Basidiosporae 13-16 x

4-5 um, clavatae vel sublacrymoideae, hyalinae, inamyloideae. Cheilocystidia 12-22 x

4-8 um, clavata vel vesiculosa. Pleurocystidia nulla. Pileipellis hymeniformis, e cellulis typi

Sicci 7-22 x 5-8 um, clavatis vel irregularibus, setulosis. Caulocystidia nulla. Trama pilei

et lamellarum dextrinoidea.

Type: CHINA, SICHUAN, Jiulong County. 21 July 2005, 29°16'36"N, 101°28'17"E, alt.

3900 m, Z. W. Ge 593 (Holotype, HKAS 49088)..

ETyMoLocy: pseudoconfertus (Latin), refers to the basidiome’s macroscopic resemblance

to the species Marasmius confertus.

PiteEus 15-35 mm, hemispherical, convex to plano-convex, with distinct

obtuse umbo at centre, orange (6A7, 6B7) to brownish orange (6C7), paler

towards margin, smooth. LAMELLAE adnexed, subdistant (18-20), white, non-

marginate. STIPE 10-70 x 2-5 mm, cylindrical, glabrous, non-insititious, nearly

white at apex, becoming grayish orangish (6B4), fuscous to dark brown (7E6-7,

7F6-7) towards base.

BASIDIOSPORES 13-16(-18) x 4-5 um [x, = 13.643.4x 4.5 £0.15 um, Q=2.3-

3.5, Qm = 3.05 + 0.28, n = 20 spores, s = 1 specimen], clavate to fusoid, smooth,

Marasmius pseudoconfertus sp. nov. (China) ... 343

SW OM

Fics 1-2: Marasmius pseudoconfertus (HOLOTYPE, HKAS 49088). 1. Basidiomes (Photo: Z.W. Ge);

2. a. Basidiospores, b. Cheilocystidia, c. Pileipellis. Bars: 1 = 1 cm; 2 = 10 um.

hyaline, inamyloid, thin-walled. Basidia 21-28 x 6-10 um, clavate, 4-spored.

BASIDIOLES 25-30 x 5-7 um, fusoid to clavate. CHEILOCYSTIDIA numerous, in

form of Siccus-type broom cells; main body 12-22 x 4-8(-10) um, cylindrical

to clavate or irregular in outline, rarely lobed, hyaline, inamyloid, thick-

walled; apical setulae 5-12 x 1-2(-3) um, narrowly cylindrical, rarely forked,

light yellow, inamyloid, thick-walled. PLEUROcystTiIp1A absent. PILEIPELLIS

hymeniform, mottled, composed of Siccus-type broom cells; main body 7-22 x

5-8(-10) um, cylindrical to clavate or irregular in outline, rarely lobed, hyaline,

344 ... Deng, Li, & Song

inamyloid to weakly dextrinoid, thick-walled; apical setulae 5-12 x 1-2(-3) um,

cylindrical, obtuse, rarely forked, light yellow, inamyloid, thick-walled. PILEus

TRAMA Of interwoven, inflated hyphae; hyphae 5-13 um in diam., cylindrical,

hyaline, strongly dextrinoid, thin-walled, non-gelatinous. LAMELLAE TRAMA

regular, hyphae 5-8 um in diam., cylindrical, hyaline, strongly dextrinoid,

thin-walled, non-gelatinous. Stipe tissue monomitic; cortical hyphae 5-8 um

in diam., parallel, cylindrical, smooth, light yellow (at apex) to yellow or yellow

brown (at base), dextrinoid, thick-walled, non-gelatinous; medullary hyphae

3-5 um wide, parallel, cylindrical, smooth, hyaline, inamyloid, thin-walled.

CAULOCYSTIDIA absent. CLAMP CONNECTIONS present.

ECOLOGY AND DISTRIBUTION— Scattered to gregarious on debris in broad-

leaved forest in July; China (Sichuan).

ComMMENTS— The new species has a Siccus-type pileipellis, but does not possess

any hymenial setae, pileosetae, and pleurocystidia. It belongs to sect. Sicci series

Leonini based on a lack of pleurocystidia, a central well-developed stipe, and

a pileipellis composed of Siccus-type broom cells. This fungus is most similar

to M. confertus Berk. & Broome, which differs in the smaller basidiospores

(8-10(12) x 4—5(6) um) and presence of pleurocystidia (Pegler 1977, Wannathes

et al. 2009).

The new species also resembles M. corneri Wannathes et al., M. occultatus

Har. Takah., and M. rubricosus Mont., all belonging to sect. Sicci ser. Leonini

and sharing moderately large basidiomes and absence of pleurocystidia and

setae. Marasmius corneri, however, can be distinguished easily by its orange

striate to sulcate pileus and larger basidiospores (18-20 x 4-5 um) (Wannathes

et al. 2009). Marasmius occultatus differs in having light colored pileus, a

more slender stipe (0.8-1.3 mm) and a lignicolous habitat (Takahashi 2000).

Marasmius rubricosus differs in its densely striate pileus and slightly longer

basidiospores (15-18 x 4-5.5 um) (Singer 1976).

The rDNA-ITS (ITS1-5.8S-ITS2 segment) sequence of 692 bps of the new

species (HQ832733) differs from any known Marasmius sequences. Through

a Blast search against the GenBank DNA database, only 160 bps of 5.8S of the

sequence match those of M. bondoi (EU935476, EU935474, EU935472, and

EU935477). The remaining parts (ITS1 and ITS2, occupying about 76.8% of the

whole segment) of the sequence are so different that they are not comparable

with any known sequences. Therefore, M. pseudoconfertus is considered distinct

based on a combination of morphological and molecular characters.

Marasmius suthepensis Wannathes, Desjardin & Lumyong, Fungal Diversity 37:

288. 2009. Figs 3-4

PILEus 8-22 mm broad, obtusely conical to convex, with or without an umbo,

smooth to striate, glabrous or pruinose, dull, disc brownish orange (7C6-8)

Marasmius pseudoconfertus sp. nov. (China) ... 345

Fics 3-4: Marasmius suthepensis (GDGM 27277). 3. Basidiomes (Photo: C.Y. Deng);

4. a. Basidiospores, b. Basidia, c. Basidioles, d. Pleurocystidia, e. Cheilocystidia, f. Pileipellis.

Bars: 3 = 1 cm; 4 = 10 um.

346 ... Deng, Li, & Song

to light orange (6A4,5), fading to light yellow (4A4), margin orange white

(5A2) to light yellow (4A4-5). ConTExT thin, white. LAMELLAE adnexed to

free, subdistant to close (20-22) with 1-3 series lamellulae, 1-2 mm broad,

non-intervenose, non-marginate. Stipe 20-55 x 1mm, central, cylindrical,

glabrous, apex yellowish white, base reddish brown (8E5-8, 8D4-8), basal

mycelium strigose, yellow. Odor and taste not distinctive.

BASIDIOSPORES 10-13(-15) x 3.1-4 um [x = 11 + 1.2 x 3.6 + 0.2 um,

Q = 2.8-3.1, Q. = 3.0, n= 25 spores, s = 2 specimen], ellipsoid, smooth,

hyaline, inamyloid, thin-walled. Basidia 21-24 x 5.5-7 clavate, 2- or 4- spored.

BASIDIOLES cylindrical to clavate. CHEILOCysTIDIA abundant, of Siccus-type

broom cells; main body 10-20 x 5-11 um, cylindrical to clavate, hyaline,

inamyloid, thin-walled; apical setulae 2-8 x 1-1.5 um, cylindrical to conical,

obtuse to subacute, yellow to yellowish brown, thin walled. PLEUROCYSTIDIA

common, 27-40 x 5-7 um, cylindrical to fusoid, wavy to constricted and

sometimes lobed at the apex, hyaline, inamyloid, thin-walled. PILEIPELLIs

hymeniform, mottled, composed of Siccus-type broom cells; main body 7-18

x 4-10 um, cylindrical to clavate or pyriform, hyaline to pale yellow, inamyloid,

thin-walled; apical setulae 3-8 x 1(-1.5) um, crowded, cylindrical, subacute,

brown to light brown, thick-walled. PrILEus TRAMA hyphae 6-15 um diam.,

interwoven, cylindrical to inflated, smooth, hyaline, strongly dextrinoid, thin-

walled. LAMELLAE HYPHAE 3-10 um in diam., cylindrical to inflated, smooth,

hyaline, dextrinoid, thin-walled, non-gelatinous; trama interwoven to regular.

STIPITIPELLIS hyphae 3-7(-9) um in diam., subparallel, cylindrical, brown

to dark brown, smooth, inamyloid to weakly dextrinoid, thin-walled, non-

gelatinous. STIPE TRAMA hyphae 3-7 um in diam., parallel, cylindrical, hyaline,

smooth, dextrinoid, thin-walled, non-gelatinous. CAULOCysTIDIA absent.

Clamp connections present in all tissues.

HABIT, HABITAT, DISTRIBUTION — Scattered to gregarious on dicotyledonous

leaves or on wood, Northern Thailand and China.

COLLECTION EXAMINED — CHINA, GUANGDONG, Dinghushan National Nature

Reserve. 9 August, 2010, C.Y.Deng (GDGM 27277); YUNNAN, Xishuangbanna Tropical

Botany Garden, 26 July, 2006, J.F. Liang 385 (HKAS 50101).

CoMMENTS— Marasmius suthepensis is common in China. The Chinese

specimens are quite similar to those from Thailand except for a paler pileus

and slightly smaller basidiospores (Thai material with mean 12.6 x 4 um;

Wannathes et al. 2009).

Acknowledgments

The authors thank Vladimir Antonin (Moravian Museum, Brno, Czech Republic)

and Dennis E. Desjardin (San Francisco State University, San Francisco, USA) for

reviewing the manuscript. Thanks are also to Dr. Z. W. Ge and L. S. Wang, Kunming

Institute of Botany of CAS, for their assisting the first author to study the specimens.

Marasmius pseudoconfertus sp. nov. (China) ... 347

This study was supported by the National Natural Science Foundation of China (No.

30870019, 30970023).

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tropical Africa I. Sect. Epiphilli, Fusicystides, Globulares, Hygrometrici and Neosessiles.

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Antonin V, Buyck B. 2006. Marasmius (Basidiomycota, Marasmiaceae) in Madagascar and the

Mascarenes. Fungal Diversity 23: 17-50.

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Antonin V, Ryoo R, Shin HD. 2010a. Marasmioid and gymnopoid fungi of the Republic of Korea. 2.

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Antonin V, Ryoo R, Shin HD. 2010b. Marasmioid and gymnopoid fungi of the Republic of Korea.

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Guinea, New Caledonia and New Zealand taxa. Part 1: Papua New Guinea and New Caledonia

taxa. Part 2: New Zealand taxa. Bibliotheca Mycologica 168: 1-152.

Desjardin DE, Ovrebo CL. 2006. New species and new records of Marasmius from Panama. Fungal

Diversity 21: 19-39.

Karsten PA. 1892. Mycetes aliquot in Mongolia et China boreali a clarissimo C. N. Potonin lecti.

Hedwigia 31: 38-40.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3 ed. Eyre London: Methuen.

252. p:

Li TH, Bi ZS, Zheng GY, Zhang WM. 1994. Species of Marasmius from Guangdong and Hainan

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Botanic Gardens, Kew: 190-191.

Singer R. 1964. Marasmius. Flore Iconographique des Champignons du Congo, Fasc. 14: 253-278.

Singer R. 1976. Marasmieae (Basidiomycetes, Tricholomataceae). Flora Neotropica 17: 1-348.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing, China. 1-1527. (in Chinese)

Takahashi H. 2000. Three new species of Marasmius section Sicci from eastern Honshu, Japan.

Mycoscience 41: 313-321. doi:10.1007/BF02463944

Tan YS, Desjardin DE, Perry BA., Vikineswary S, Noorlidah A. 2009. Marasmius sensu stricto in

Peninsular Malaysia. Fungal Diversity 37: 9-100.

Teng SC. 1963. Fungi of China. Beijing: Science Press. 1-808. (in Chinese)

Wannathes N, Desjardin DE, Hyde KD, Perry BA, Lumyong S. 2009. A monograph of Marasmius

(Basidiomycota) from Northern Thailand based on morphological and molecular (ITS

sequences) data. Fungal Diversity 37: 209-306.

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

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PCR protocols: a guide to methods and applications. Academic Press Inc., San Diego.

MY COTAXON

Volume 116, pp. 349-354 April-June 2011

DOI: 10.5248/116.349

Two new species of Tuber from China

Li FAN’ , JIN-ZHONG CAo?, YAN-YUN Liu’, & Yu Li”

" 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 & * yuli966@126.com

ABSTRACT — Two new species of the genus Tuber are described from China. Tuber lijiangense

is differentiated from other brown species mainly by its globose ascospores with a lower

reticulated ornamentation, and T. sinoexcavatum is distinguished from T. excavatum by its

globose ascospores.

Key worps — Ascomycota, Tuberaceae, truffle

Introduction

The genus Tuber FH. Wigg. is an important group of fungi, both ecologically

and commercially. All species of the genus form ectomycorrhizal (EM)

symbiosis with many important timber and nut tree species (Gregory et al.

2009), and some species are frequently sought after for culinary and medical

purposes around the world. Tuber is a relatively large genus, containing around

100 species and being widely distributed throughout the northern hemisphere,

of which near half appear to be endemic to North America and about 33 species

are recognized from Europe (Gregory et al. 2009).

Since the first description of the genus in China by Liu (1985) with

T. taiyuanense, more than 20 Tuber species have been reported from China

(Cao 2010). Some of these species have already become a good business on

both domestic and international markets. The evident importance of the genus

in both biodiversity and economy has stimulated more investigations Tuber in

China. In this paper two new Tuber species are described and illustrated.

Materials & methods

One Tuber species that had been collected in Yunnan Province was discovered after

the dried specimens had been deposited in HKAS (Herbarium of Cryptogams, Kunming

Institute of Botany, Chinese Academy of Science). The specimens of the other samples

350 ... Fan & al.

studied were newly collected in Sichuan Province and are deposited in BJTC (Herbarium

Biology Department, Capital Normal University). The macroscopic characteristics were

described from both fresh and rehydrated specimens. The microscopic characteristics

were described from razor-blade sections mounted in 3%KOH, Melzer’s reagent, or

cotton blue. For scanning electron microscopy (SEM), ascospores were scraped from

the dried gleba of fruitbodies and mounted in distilled water on a cover glass. After

air drying the cover glasses were directly attached to a SEM stub with doubled-sided

tape, and then coated with gold-palladium. The treated materials were examined and

photographed with a HITACHI S-4800 SEM.

Taxonomy

Tuber lijiangense L. Fan & J.Z. Cao, sp. nov. Figs 1-5

MycoBank MB 519470

Ascomata ochracea vel bruneola, 0.5-3 cm diam, subglobosa vel globosa irregulara lobata,

primitus brevi-pubeula. Peridium 250-350 um crassum, stratis duobus: stratum exterius

pseudoparenchymaticum, stratum interius hyphi intertextis. Gleba solida, bruneola vel

purpureobruneola, veins albis. Asci spori 1-2(-3), 60-90 x 50-80 um. Ascosporae globosa,

ochraceae, 25-42.5 ym, reticulatae.

Type: Juan Chen 404 (HKAS 52005, holotype), 29 Oct. 2006, Yongsheng county, Lijiang

city, Yunnan Province, China, under Pinus yunnanensis Franch. (Pinaceae).

ErymMo_oey: lijiangense (Lat.), refers to the type locality.

AscomMatTa hypogeous, subglobose to irregularly lobed, with shallow to deep

furrows, 0.5-3 cm in diam., pale yellow or light brown when fresh, pubescent.

Odor faint when fresh. PERrpIUM 250-350 um thick with two layers; outer

layer 60-100 um thick, pseudoparenchymatous, composed of subglobose or

subangular, light brown cells 7.5-25 um in diam.; inner layer 190-250 um

thick, hyaline, of intricately interwoven hyphae with thin-walled cells, 2.5-5(-

7.5) um broad. Hairs abundant, composed of hair-like hyphae, arising from the

superficial cells, 60-100 um long and 2.5-5 um broad at base, tapered, usually

acute at apex, 1-2 septate. GLEBA whitish when young, brown or purple brown

at maturity, veins white, narrow and numerous. Asci globose to subglobose,

60-90 x 50-80 um, sessile or with a short stalk, 1-2(-3) spores. ASCOSPORES

globose, hyaline at first, brown or yellow brown at maturity, alveolate reticulum,

25-42.5 um excluding the ornamentation, the walls up to 2.5-5 um thick, the

meshes complete, five or six sided, regular to irregular, lower, usually less than

2 um deep, and fewer ascospores trending to be smooth walled, (3-)4-8(-12)

meshes across the spore width.

Eco_Locy, DIsTRIBUTION — Hypogeous, under Pinus yunnanensis. Known

from Yunnan Province, China.

ComMENTs — ‘The most distinctive morphological characteristics of

T. lijiangense that separate it from the other Tuber species are the ornamen-

tation of its ascospores, which are typically reticulate but with meshes that are

Tuber spp. nov. (China) ... 351

Fics 1-5. Tuber lijiangense ( HKAS 52005, holotype). 1. Ascocarps. 2. Ascospores. 3-4. Asci and

ascospores observed under light microscope. 5. Ascospores observed under the scanning electronic

microscope. Bars: 1 = 1 cm; 2, 4, 5 = 10 um; 3 = 20 um.

much lower (normally < 2 um in depth) and walls that are sometimes almost

smooth in some spores. To our knowledge, no other Tuber species has this type

of ascospore. Furthermore, the number of ascospores per ascus in this species

is also peculiar, usually 1-2 spores in most asci, instead of 1-4(-5) spores in

most other species.

352 ... Fan & al.

Globose ascospores are not common in Tuber, although one North

American species with globose spores, T: californicum Harkn., seems to

be common in Northern China (Gilkey 1954a,b; Wang 1988). It can easily

be distinguished from T. lijiangense by the distinctive and regular reticulate

ascospore ornamentation.

Tuber sinoexcavatum L. Fan & Yu Li, sp. nov. Fics 6-9

MycoBank MB 519471

Ascomata bruneola, excavatum, 1.5-3 cm diam, subglobosa vel globosa irregulara. Peridium

200-300 um crassum, stratis duobus: stratum exterius pseudo-parenchymaticum, stratum

interius hyphi intertextis. Gleba solida, purpureo-rufo-bruneola, veins albis. Asci spori

(1-)2-3(-4), 75-125 x 62.5-100 ym. Ascosporae subglobosa vel lato-ellipsoideae,

ochraceae, 35-50 x 30-45 um, reticulatae.

Type: De-fu Liu (BJTC FAN130, holotype), 20 Dec. 2007, China, Sichuan Province,

Panzhihua city, under the soil of Pinus sp.

ETYMOLOGY: sinoexcavatum (Lat.) = the endemic Chinese species that is similar to T.

excavatum.

AscoMatTa hypogeous, subglobose or depressed, with a typical basal cavity

(excavated ascomata), 1.5-3 cm in diam., pale yellow brown to brown when

fresh, smooth or minutely papillose, distinctly verrucose at the surface of cavity

area. PERIDIUM 200-300 um thick with two layers; outer layer 40-100 um thick,

pseudoparenchymatous, composed of subglobose or subangular, light brown

cells 5-15 um in diam.; inner layer 160-200 um thick, hyaline, of intricately

interwoven hyphae with thin-walled cells 2.5-5 um broad. GLEBA yellow brown

or red brown at maturity, veins white, narrow and numerous. Asci globose

or subglobose, 75-125 x 62.5-100 um, with a short stalk, (1-)2-3(-4) spores.

Ascospores globose, hyaline at first, brown or yellow brown at maturity,

alveolate reticulum, 35-50 x 30-45 um excluding the ornamentations, meshes

5 um deep and 7.5-15 um in diam., 3-5 meshes across the spore width.

Eco_Locy, DISTRIBUTION — Hypogeous, under the soil of Pinus sp. Known

from Sichuan Province, China.

ComMENTS — This species is very similar to T. excavatum Vittad., which is

very common in Europe (Lange 1956). It has globose to subglobose ascospores,

and its gleba is usually dark at maturity. Another European species, T: fulgens

Quel. (Lange 1956), also has the same type of ascospores, but it differs from

T. sinoexcavatum by ascomata that are orange-brown and hollowed with a

small hole leading to the outside. Tuber pseudoexcavatum, a popular species in

southwestern China (Wang et al. 1998) which also produces typically excavated

ascomata, is easily distinguished from T. sinoexcavatum by the brown or dark

brown, verrucose ascomatal surface and the ascospores with clearly distinct

spino-reticulate ornamentations.

Tuber spp. nov. (China) ... 353

Fics 6-9. Tuber sinoexcavatum (BJTC FAN 130, holotype). 6. Ascocarps. 7. Ascospores. 8. Ascospore

observed under the scanning electronic microscope. 9. Asci and ascospores observed under light

microscope. Bars: 6 = 1 cm; 7, 8 = 10 um; 9 = 20 um.

Acknowledgments

The study was supported by the National Natural Science Foundation of China (Nos.

30770005, 30870008). We are grateful to Dr. Guo-Zhong Lii and Professor Anthony

Walley for reviewing the pre-submitted manuscript. We also thank Dr. Cheng-Lin Hou

for critically correcting the manuscript.

Literature cited

Cao JZ. 2010. The genus Tuber in China [Ph.D. dissertation]. Changchun: Jilin Agricultural

University. 86 p.

Gilkey HM. 1954a. Tuberales. N. Amer. FI., Ser. II, 1: 1-36.

Gilkey HM. 1954b. Taxonomic notes on Tuberales. Mycologia 46: 783-793.

Gregory B, Trappe JM, Rytas V. 2009. North American truffles in the Tuberaceae: molecular and

morphological perspectives. Acta Bot. Yunnanica, Suppl. 16: 39-51.

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

354 ... Fan & al.

Liu B. 1985. New species and new records of hypogeous fungi from China (I). Acta Mycologica

Sinica 4(2): 84-89.

Wang Y. 1988. First report of study on Tuber species from China. Atti del II Congresso Internazionale

sul Tartufo. Spoleto, Nov 24-27: 45-50.

Wang Y, Moreno G, Riousset LJ, Manjon JL, Riousset G, Fourre G, Di Massimo G, Garcia- Montero

LG, Diez J. 1998. Tuber pseudoexcavatum sp. nov., a new species from China commercialized in

Spain, France and Italy with additional comments on Chinese truffles. Cryptogamie Mycologie

19; 113-120.

MY COTAXON

Volume 116, pp. 355-364 April-June 2011

DOI: 10.52.48/116.355

AFLP molecular characterizations of some Saccobolus species

(Ascomycota, Pezizales)

ARACELI M. RAMos , ISABEL E. CINTO, LUIS FRANCO TADIC &

MariA ESTHER RANALLI

Lab 9 (PRHIDEB-CONICET), Departamento Biodiversidad y Biologia Experimental,

Facultad de Cs. Exactas y Naturales, Universidad de Buenos Aires,

Int. Gitiraldes 2620 - C1428EHA - Buenos Aires - Argentina

*CORRESPONDENCE TO: araceli@bg.fcen.uba.ar

ABSTRACT — Amplified restriction fragment length polymorphisms (AFLP) were used to

assess the genetic diversity among species and isolates of the genus Saccobolus. Monosporic

strains of five Saccobolus species were used throughout. The dendrogram obtained from the

analysis of grouping (UPGMA) showed four groups of species. The ordination of isolates

through the principal coordinates method exhibited nearly the same relations between

isolates as the phenogram. Analysis of six samples identified as Saccobolus versicolor using

morphological characters indicated the same diagnostic fingerprints as S. verrucisporus

with the primer combinations tested. The isolates identified as S. versicolor may represent

an intraspecific variant of S. verrucisporus. The results support the use of AFLP markers

to delimit Saccobolus species. This methodology constitutes an additional tool to study the

taxonomy of the genus, which has previously used only morphological characters.

Key worps — molecular markers, coprophilous fungi, systematic

Introduction

The genus Saccobolus (Pezizales, Ascobolaceae) was established by Boudier

(1869). The presence of a common hyaline sheath around the ascospores was

considered one of the major criteria. Brummelen (1967) characterized the

genus within the subfamily Ascoboloideae as producing clustered ascospores,

comparatively short and broad, clavate, truncate asci, and rather short

paraphyses.

Saccobolus species are coprophilous fungi. They have a cosmopolitan

distribution and their spores usually are transported over long distances.

Most of these fungi are homothallic, probably originated as an adaptation

to the substrate, which determines their sexual isolation (Wicklow 1981).

356 ... Ramos & al.

Sexual reproduction enables ascospores to be dispersed by effective discharge

methods.

Different concepts have been used to define the fungal species. The

phenotypic concept is the classic approach based on morphological characters.

The polythetic concept circumscribes species based on a combination of

characters (Kohn 1992, Guarro et al. 1999). Saccobolus species have traditionally

been classified based on cytological and morphological characters (Brummelen

1967) and supported by the phenotypic concept. Even though such characters

as ascal structure and dimensions, type of dehiscence, spore ornamentation,

and the general ascocarp developmental pattern are clearly important to the

classification of the Saccobolus species, identification is often uncertain because

these characteristics are shared by two or more species or may vary according

to culture conditions. Here use of the polythetic concept could prove helpful in

characterizing the genus.

In an earlier study, Ramos et al. (2000) performed an isozyme analysis to

confirm morphology-based identification of Saccobolus species and to detect

intraspecific variability. In that study, isozymes showed little intraspecific

variability and confirmed the previous identification of the strains using

morphological characters, although only few species-specific bands were

found.

Molecular techniques have proved to be the appropriate tools to advance in

the study of genetic diversity of the intra- and inter-specific variation allowing

an insight on the phylogenetic relationships

For instance, the amplified fragment length polymorphism (AFLP), (Vos et

al. 1995) is a DNA-based fingerprinting technique frequently used in a diverse

array of organisms (Briard et al. 2000, Roa et al. 1997, Zeller et al. 2000) that

requires no prior sequence information on the organism under scrutiny.

Althoughthemorphological, physiological,andbiochemicalcharacterizations

of Saccobolus species have been the aim of many studies, molecular study is still

being developed.

In the present study we used the AFLP methodology to characterize five

Saccobolus species so as to investigate the efficiency of these molecular markers

in determining genetic diversity, to obtain diagnostic bands for species

recognition, and to compare them with the results of a previous isozyme

study.

Material & methods

Monosporic strains

Mature apothecia of Saccobolus citrinus Boud. & Torrend (Boudier & Torrend 1911),

S. saccoboloides (Seaver) Brumm. (Brummelen 1967), S. pseudodepauperatus Gamundi

& Ranalli (Ranalli & Gamundi 1976), S. verrucisporus Brumm. (Brummelen 1967), and

S. versicolor (P. Karst.) P. Karst. (Karsten 1885) were obtained by placing cow and horse

AFLPs in Saccobolus ... 357

dung from different geographical locations in Petri dishes with a layer of filter paper.

Thirty-two monosporic isolates of five species of the genus Saccobolus were obtained

from individual ascospore germination. TABLE 1 lists the monosporic isolates of each

species with their geographical location, substrate, and BAFC culture collection number.

All strains were deposited in the Herbarium and Culture Collection of the Departmento

de Biodiversidad, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos

Aires (BAFC). Cultures of all monosporic strains were regularly kept in PF medium

(yeast extract 3 g; agar 18 g; distilled water 1000 ml; a slice of filter paper) (Ranalli &

Forchiassin 1974) at 5°C.

TABLE 1. List of Saccobolus isolates employed in this survey with their geographical

location and BAFC number.

SpEATES Locality MONOSPORIC ISOLATES

(Argentina) (BAFC numbers)

S. verrucisporus Gualeguaychu, ER 3403-3405, 3449, 3450

S. verrucisporus Gobernador Castro, BA 3410-3412, 3451

S. versicolor Gualeguaychu, ER 3407-3409, 3452, 3453

S. saccoboloides Pergamino, BA 3454-3460

S. citrinus V. Tuerto, SF 3461-3463

S. pseudodepauperatus Arrecifes, BA 3464-3471

BA = Buenos Aires Province; ER = Entre Rios Province; SF = Santa Fe Province.

Identification of species

Morphological and cultural studies were carried out in order to identify the

species. The characters used to identify the Saccobolus species were the same used by

Brummelen (1967) that included diverse characteristics of apothecia, ascospores, asci,

and paraphyses. Also used were the distribution and appearance of apothecia in PF

medium (yeast extract 3 g; agar 18 g; distilled water 1000 ml; a slice of filter paper;

Ranalli & Forchiassin 1974) and ascospore germination in AA medium (Bacto agar 20g;

distilled water 1000 ml). Studies of spore germination and ascocarp development were

performed on AA plates and in PF medium plates respectively.

DNA extraction

Mycelium was obtained according to Ramos et al. (2000) procedure and ground to

powder in liquid nitrogen using a sterile pestle. Genomic DNA was extracted following

Gottlieb & Lichtwardt (2001). Quality control and quantification of genomic DNA was

carried out by agarose gel (0.8% w/v) electrophoresis and by comparison with a DNA

molecular-size standard (Lambda EcoRI/HinDIII, Promega Corp.). Ethidium bromide

gels were photographed under UV light.

AFLP

The AFLP methodology was carried out on 250 ng of genomic DNA using the

AFLP® Analysis System for Microorganisms Primer Kit (Invitrogen) as described in the

instructions manual with minor modifications (Gottlieb et al. 2005). Selective primers

were combined as in TABLE 2.

358 ... Ramos & al.

TABLE 2. AFLP Primers used in this study.

PRIMERS SEQUENCE 5-3”

E+AAC GAC TGC GTA CCA ATT CAA C

E+ACC GAC TGC GTA CCA ATT CAC C

M+A GAT GAG TCC TGA GTA AA

M+G GAT GAG TCC TGA GTA AG

M+C GAT GAG TCC TGA GTA AC

M+T GAT GAG TCC TGA GTA AT

E= EcoRI; M = Msel.

All PCR amplifications were performed in a TECHNE PROGENE thermal cycler.

Polyacrylamide gel electrophoresis conditions followed Gottlieb et al. (2005). A

30-330 bp AFLP DNA Ladder (Invitrogen) size marker was included twice in each

electrophoresis. Thus, the size of AFLP bands scored ranged from 90 to 330 bp. AFLP

bands were visualized using the SILVER SEQUENCE ™ DNA Sequencing System

(Promega). Air-dried gels were digitalized and visually analyzed using the Adobe

Photoshop TM (Adobe Systems, Mountain View, Ca, USA).

Statistical methods

Each AFLP band was considered as a dominant allele at a unique locus. TABLE 3

shows the total number of AFLP bands, species-specific bands, and polymorphic and

monomorphic bands detected for each selective primer combination from each fungal

species and percentage of polymorphic loci (P%). Monomorphic bands (bands present in

all individuals of a species) were discriminated within each species and across the entire

data set. The binary matrix was analyzed with the program NTSYS-PC version 2.02

(Rohlf 1993). The unweighted pair-group arithmetic mean method (UPGMA) cluster

analysis was performed based on the simple matching (SM), association coefficient

(Sneath & Sokal 1973). The phenogram showing similarity relations was generated by

the same program. The distortion produced during the grouping analysis was calculated

using the correlation cophenetic coefficients (r) (Sokal & Rohlf 1962) using the NTSYS-

PC version 2.02 (Rohlf 1993) program. A three-dimensional graphic was obtained with

the principal coordinated ordination method (Gower 1966).

The percentage of polymorphic loci (P = number of polymorphic loci/ number of

loci analyzed) x 100 for each primer combination was calculated.

Results

Characterization of AFLP markers

A total of 895 fragments in a range of 90 to 330 bp were generated for five

Saccobolus species using six selective primer combinations. As a result, the

number of reliable bands scored varied from 22 to more than 60, depending on

the primer pair combination used.

Monomorphic AFLP band patterns were identified within each species.

The diagnostic fingerprints (species-specific bands) of Saccobolus citrinus,

AFLPs in Saccobolus ... 359

S. saccoboloides, S. pseudodepauperatus and S. verrucisporus were obtained and

are described in TABLE 3.

TABLE 3. Total number of AFLP bands, species-specific bands (%), polymorphic and

monomorphic bands detected for each selective primer combination in each

fungal species assayed and percentage of polymorphic loci (P%).

BANDS

PRIMER won SPECIES- P%

COMBINATION SPECIES Nat POLYMORPHIC MONOMORPHIC een

M+G/E+ACG

S. citrinus 26 2; 24 26.9 7.7

S. saccoboloides 32 1 31 12.5 3.1

S. pseudodepauperatus 67 47 20 10.4 70.0

S. verrucisporus 32 4 28 i 12.5

S. versicolor 30 2 28 ¥ 6.7

M+C/E+AAG

S. citrinus 30 2 28 36.6 6.7

S. saccoboloides 33 1 33 39 3.0

S. pseudodepauperatus 49 39 10 10 79.6

S. verrucisporus 26 1 25 z 3.8

S. versicolor 18 1 17 * 5.6

M+T/E+ACG

S. citrinus 57 27 30 22.8 47.4

S. saccoboloides 58 13 45 27.6 22.4

S. pseudodepauperatus 86 62 24 4.5 72.1

S. verrucisporus 71 30 4] * 42.2

S. versicolor 51 5 46 * 9.8

M+A/E+ACG

S. citrinus 66 36 30 4.5 54.5

S. saccoboloides 72 11 61 12.5 15.3

S. pseudodepauperatus 114 74 40 4.4 64.9

S. verrucisporus 69 35 34 ‘' 50.7

S. versicolor 52 11 41 = 21.1

M+C/E+ACG

S. citrinus 68 34 34 17.6 50.0

S. saccoboloides 69 20 49 15.9 29.0

S. pseudodepauperatus 104 94 10 21.9 90.4

S. verrucisporus 52 20 32 % 38.5

S. versicolor 47 11 36 = 23.4

M+A/E+AAG

S. citrinus 72 30 42 6.9 41.7

S. saccoboloides 86 27 59 10.5 31.4

S. pseudodepauperatus 149 108 41 1.34 72.5

S. verrucisporus 72 28 44 * 38.9

S. versicolor 61 29 32 “ 47.5

* Data not shown (see TABLE 4).

Primers MA/EAAG and M/EACG were more effective in detecting

polymorphisms (TABLE 3). Genetic variability within species, measured

in terms of percentage of polymorphic loci was variable (P = 83.8% for

S. pseudodepauperatus, P = 41.0% for S. citrinus, P = 20.8% for S. saccoboloides,

P = 22.8% for S. versicolor, and P = 39.8% for S. verrucisporus); i.e.,

360 ... Ramos & al.

S. pseudodepauperatus was the most variable species, and S. saccoboloides and

S. versicolor the least variable.

Analysis of six samples from Gualeguaychu identified as S. versicolor using

morphological characters displayed a similar diagnostic fingerprint to that of

S. verrucisporus with all primer combinations used as is shown by the number

of shared bands and the species-specific bands obtained between these

two species (TABLE 4). Of the 330 bands obtained for S. verrucisporus and

S. versicolor, 253 (77%) were shared between them and 54 were species-specific

bands/diagnostic bands (present in both species). Of the 259 bands present in

S. versicolor, only 6 were absent from S. verrucisporus.

TABLE 4. Total number of AFLP bands, species-specific bands and shared bands

detected for each selective primer combination between Saccobolus versicolor

and S. verrucisporus species.

ToTAL N° SPECIES-SPECIFIC SHARED

PRIMER COMBINATION SPECIES OF BANDS BANDS BANDS

S. verrucisporus 32

M+G/E+ACG 10 28

S. versicolor 30

S. verrucisporus 26

M+C/E+AAG 7 18

S. versicolor 18

S. verrucisporus 71

M+T/E+ACG 10 49

S. versicolor 51

S. verrucisporus 69

M+A/E+ACG 5 52

S. versicolor 52

S. verrucisporus 52

M+C/E+ACG 11 46

S. versicolor 47

S. verrucisporus 72

M+A/E+AAG 11 60

S. versicolor 61

Degree of similarity and cluster analysis

The degree of similarity among species was low. Between S. citrinus and the

group formed by S. verrucisporus and S. versicolor (S = 0.57) and between these

cluster and S. pseudodepauperatus (S = 0.56). The group formed by the isolates

identified as S. saccoboloides clustered with the other three species with the

lowest coefficient of similarity (S = 0.53). The phenogram obtained by means

of UPGMA method (Fic. 1) showed that all the strains were grouped into

four clusters. Little distortion occurred while constructing this phenogram,

as implied by the value of the correlation cophenetic coefficients (r _ 0.9846).

AFLPs in Saccobolus ... 361

nan

“2

SMOdSion.uen “Ss

AOJOIISIBA “*S

C1 87

G1 $24

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C4 $26

cs $15

G2 $17

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wo o aA ao wa

snjesadnedapopnasd ‘gs = snuujio ‘Ss

076 0.88 Oss

Coefficient

FiGuRE 1. UPGMA phenogram showing relationships of Saccobolus species on the basis of simple

matching (SM) association coefficient estimated from AFLP loci.

The main group (Fic. 1, group G1) comprised two subgroups (SG1 and SG2).

Subgroup SG1 is formed by two sets (C1 and C2). The set C1 included all isolates

of S. verrucisporus and S. versicolor and set C2, the three isolates identified as

S. citrinus. The other Subgroup (SG2) comprised two sets, C3 and C4, formed

by all isolates of S. pseudodepauperatus.

A second group (Fic. 1, G2) included two sets, C5 and C6, that included all

isolates of S. saccoboloides.

Grouping analysis revealed that 14 isolates identified as S. verrucisporus

and as S. versicolor clustered in one nucleus (Fic. 1, C1) with an origin of

ramification in 0.92.

The set Cl is formed by two subsets (SC1, SC2) and one isolate S3

identified as S. verrucisporus separated from the rest. Only two pairs of isolates

presented 100% of similarity (association coefficient 1.0), and they belonged to

S. verrucisporus.

The ordination of isolates through the principal coordinated method let us

distinguish four groups (Fic. 2, groups 1-4) in three-dimensional dispersion

and showed nearly the same relations between isolates as the phenogram

(Fic. 1). The first group included all isolates of S. saccoboloides, very closely

attached in the three axes. The second set showed S. pseudodepauperatus

isolates differentiated in axis 1 but very closely in the other two axes. The third

362 ... Ramos & al.

°@ S. versicolor

group 1 Ges ;

. verrucisporus

S. saccoboloides 4

group 4

group 2

S. pseudodepauperatus

FiGuRE 2. Three-dimensional graphic showing relationships of Saccobolus species obtained with

Principal Coordinates ordination technique based on simple matching (SM) association coefficient

estimated from AFLP loci.

comprised the three isolates of S. citrinus. The fourth and last group included

the fourteen isolates identified as S. verrucisporus and as S. versicolor joined

closely together in the three axes, thus revealing a high degree of similarity.

Discussion

There are many traits that are used in the traditional and modern mycology

that contribute to taxonomical studies of fungi. These include morphological,

anatomical, biochemical, and DNA based methods. However, fungal species

descriptions are based mainly on morphological characters (Brasier 1997).

Although identification of Saccobolus species was possible, coinciding

morphological, cytological, and developmental characters make it difficult to

achieve a clear delimitation. Previous isoenzymatic studies (Ramos et al. 2000)

showed a low intraspecific variability and characterized the Saccobolus species

by some isoenzymatic bands.

In this study a high number of AFLP bands were analyzed, which confirmed

earlier results. The phenogram and ordination analysis both showed the same

four clearly separated clusters, three of them corresponding to each of the three

species identified as S. citrinus, S. saccoboloides, and S. pseudodepauperatus.

Isolates identified as S. versicolor using morphological characters shared almost

97.7% of AFLP bands with S. verrucisporus in all primer combinations tested

and joined together in the three axes of the ordination graphic and clustered

AFLPs in Saccobolus ... 363

very closely in the phenogram, suggesting that isolates previously identified

as S. versicolor could represent an intraspecific variant of S. verrucisporus.

A higher number of isolates will need to be assessed in order to resolve this

relationship.

Previous morphological and isoenzymatic studies in Saccobolus (Ranalli

& Mercuri 1995, Ramos et al. 2000) indicated low intraspecific variability.

A high number of AFLP bands obtained with the six primers allowed us to

detect intraspecific variability.

The AFLP technique permits differentiation of Saccobolus species by

providing a greater number of species-specific bands. This result coincides with

previous isozyme studies (Ramos et al. 2000), where ALP (alkaline phosphatase)

was the only system that showed a diagnostic electromorph for each species.

Moreover, AFLPs are better able to differentiate species and detect intraspecific

variability than isozymes because they allow extensive genome sampling and

provide a great number of DNA markers and therefore a higher number of

species-specific markers.

This technique could become a powerful tool for problematical genera such

as Saccobolus, in which overlapping characters make species identification

difficult and where a high intraspecific homogeneity does not allow detection

of variability using traditional taxonomic and isozyme methods.

Acknowledgments

This study was supported by a grant from Consejo Nacional de Investigaciones

Cientificas y Técnicas (CONICET) and Universidad de Buenos Aires, Argentina. We

thank, Dr. Lina Bettucci and Dr. Sandra Lupo for the critical review of the manuscript.

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MY COTAXON

Volume 116, pp. 365-379 April-June 2011

DOI: 10.5248/116.365

Glomeromycota: two new classes and a new order

FRITZ OEHL', GLADSTONE ALVES DA SILVA?, BRUNO TOMIO GOTO},

LEONOR CosTA MAIA?& EWALD SIEVERDING*

'Federal Research Institute Agroscope Reckenholz-Tanikon ART, Ecological Farming Systems,

Reckenholzstrasse 191, CH-8046 Ziirich, Switzerland

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

Av. Prof. Nelson Chaves s/n, Cidade Universitaria, 50670-420, Recife, PE, Brazil

*Departamento de Botanica, Ecologia e Zoologia, CB, Universidade Federal do Rio Grande

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

‘Institute for Plant Production and Agroecology in the Tropics and Subtropics,

University of Hohenheim, Garbenstrasse 13, D-70599 Stuttgart, Germany

*CORRESPONDENCE TO: fritz.oehl@art.admin.ch

ABSTRACT — Based on concomitant molecular analyses of the ribosomal gene and

morphological characteristics, we divide the phylum Glomeromycota into three classes:

Glomeromycetes, Archaeosporomycetes, and Paraglomeromycetes. Glomeromycetes are newly

organized in three orders: Glomerales and Diversisporales, both forming typical vesicular

arbuscular mycorrhiza with higher plants, and Gigasporales, forming arbuscular mycorrhiza

without vesicles in the roots but with extra-radical auxiliary cells. Within the phylum,

Archaeosporomycetes comprise exclusively bimorphic families and genera. The monogeneric

Paraglomeromycetes species form glomoid spores that typically germinate directly through

the spore wall instead through their subtending hyphae.

Key worps — evolution, Gigasporineae, Gigasporaceae, molecular phylogeny, rDNA

Introduction

In 1998, a new fungal class, the Glomeromycetes, was proposed, originally

within the Zygomycota, but subsequently transferred to its own phylum, the

Glomeromycota C. Walker & A. Schiissler (Schiifler et al. 2001). The phylum is

currently represented by 230 arbuscular mycorrhizal (AM) fungiand one fungus

living in symbiotic association with algae. The division of Glomeromycota into

three major clades indicated by the genetic studies of Schiifler et al. (2001)

has since been confirmed by other studies (e.g. Redecker & Raab 2006, Msiska

& Morton 2009, Oehl et al. 2011a). The first objective of the present study

was to confirm the phylogenetic findings and to elucidate the morphological

366 ... Oehl & al.

homologies within and the differences between these three major clades.

Consequently, two new classes within the Glomeromycota are described in this

paper. Our analyses moreover suggest a further division of orders within the

major group, the Glomeromycetes. We propose to establish a new order for the

sporogenous cell-forming AM fungi, which is supported by a series of unique

features in addition to the presence of sporogenous cells, such as spore wall

and germination characteristics and the presence of extra-radical auxiliary cells

instead of intra-radical vesicles (Gerdemann & Trappe 1974).

Material & methods

Morphological analyses

Morphological analyses were performed on type and non-type specimens as

described and summarized in a series of recent publications for species of Glomerales,

Diversisporales, Paraglomerales (Oehl & Sieverding 2004, Sieverding & Oehl 2006, Oehl

et al. 2006, 2008, 2010, 2011a,b, Palenzuela et al. 2008, 2010) and Archaeosporales (Spain

et al. 2006, Palenzuela et al. 2011). Specimens representing >85% of the known 230

AM fungal species were obtained from public herbaria (OSC, FH, HAC, PDD, Z+ZT,

DPP, URM) and collections (Embrapa Agrobiologia (Seropédica, Brazil), International

Culture Collection of (Vesicular-)Arbuscular Mycorrhizal Fungi (INVAM)); private

collections held by Sieverding, Oehl, Trappe, Blaszkowski, Goto, Herrera, and McGee

collection; and the Hall & Abbott (1979) photographic collection. Older (pre-1990)

specimens on microscopic slides were mounted in lactophenol; more recent specimens

were fixed with polyvinyl alcohol-lactic acid-glycerol (PVLG) or a mixture of PVLG

+ Melzer’s reagent, since 1990 the major fixing media (Brundrett et al. 1994). Newly

mounted spores and sporocarps from the collections or from pure cultures were fixed by

using the latter two media and sometimes also in a mixture of lactic acid to water at 1:1,

in Melzer’s reagent, and in water. When available, spores freshly isolated from soils or

bait cultures were also mounted and analyzed. Spore morphology terminology follows

Oehl et al. (2008, 2011a).

Phylogenetic analyses

Partial sequences of B-tubulin and rRNA (SSU and LSU) genes (obtained from

public data bases) were used to reconstruct independent phylogenetic analyses of the

Glomeromycota. The B-tubulin gene intron sequences were excluded, with only exon

regions analysed.

Recent studies (Morton & Msiska 2009) have simultaneously analysed the 6-tubulin

and rRNA gene sequences. However, as the nucleotide substitution rate in the LSU

rRNA clearly differs from that found in the SSU region and 6-tubulin gene, we used

different nucleotide substitution models in analysing these genes. Furthermore, the

rRNA gene fragment (LSU and SSU) sequenced in Glomeromycota is represented for

> 2400 nucleotide bases whereas there are 600 bases for the B-tubulin sequences, so

a comparison is not possible without weighting the data sets correspondingly. Fewer

than 50 AM fungal species have been sequenced for both 6-tubulin and rRNA genes.

We consider it problematic that some sequences were obtained from different isolates

Gigasporales, Archaeosporomycetes & Paraglomeromycetes (Glomeromycota) ... 367

of the same putative species. Some problems with misidentifications that have been

reported (see Bago et al. 1998, Lanfranco et al. 2001, Souza et al. 2004, Spain et al. 2006,

Sieverding & Oehl 2006) have been repeated in the molecular analyses of such isolates.

Thus, we used only partitioned analyses for the different genes and rRNA subunits

(6-tubulin, LSU and SSU rDNA) for this study. For consensus analyses, the LSU and

SSU trees were considered as only one tree because the sequences of these regions

cannot be considered independent data sets. A consensus tree was obtained just for the

organization of taxonomic order level or above.

The sequences (all obtained from the National Center for Biotechnology Information-

NCBI) were aligned using ClustalX (Larkin et al. 2007) and edited with BioEdit (Hall

1999).

Maximum parsimony (MP) and neighbor joining (NJ) analyses with 1000 bootstrap

replications were performed using the Phylogenetic Analysis Using Parsimony (PAUP)

program version 4 (Swofford 2003). Bayesian (two runs over 1 x 10° generations with

a burnin value of 2500) and maximum likelihood (1000 bootstrap) analyses were

executed, respectively, in MrBayes 3.1.2 (Ronquist & Huelsenbeck 2003) and PhyML

(Guindon & Gascuel 2003), launched from Topali 2.5. The nucleotide substitution

model was estimated using Topali 2.5 (Milne et al. 2004). Sequences from Neurospora

crassa Shear & B.O. Dodge, Boletus edulis Bull., and Rhizophydium sphaerotheca Zopf

were used as outgroups for Glomeromycota.

Results

General phylogenetics and relations to morphological features

Paraglomerales and Archaeosporales have low phylogenetic relationship to

Glomerales and Diversisporales (Fics. 1-3). Species of Archaeospora, Intraspora,

Ambispora, and Paraglomus form extra-radical mycelia and mycorrhizal

structures that stain only faintly or not at all in trypan blue (Spain & Miranda

1996, Spain 2003, Spain et al. 2006, Sieverding & Oehl 2006, Walker et al. 2007,

Palenzuela et al. 2011). We do not know the reasons for this behaviour. Vesicle

formation is rarely reported or might have been based on misinterpretations

in Archaeospora, Intraspora, and Paraglomus. In constrast, in the glomeralean

and diversisporalean species, fungal structures stain blue to deep blue with

trypan blue and typical vesicular arbuscular mycorrhiza formation is regularly

reported (Schtifler et al. 2001). Within the Glomerales and Diversisporales,

only Gigasporaceae, Scutellosporaceae, Racocetraceae, and Dentiscutataceae

apparently do not form intraradical vesicles, but their fungal structures also

stain blue to deep blue with trypan blue as in all other glomeralean and

diversisporalean families (Bentivenga & Morton 1995, Oehl et al. 2008, 2010).

Because such a feature is general and related to the mycorrhiza formation, and

as the morphological and physiological characteristics are congruent with the

higher phylogenetic clades, there is support for separating the Archaeosporales

and Paraglomerales from the Glomeromycetes and for establishing new fungal

classes for them in the Glomeromycota.

368 ... Oehl & al.

Boletus edulis DQ534675

Neurospora crassa AY046271

100 _» Paragiomus occultum AJ276081

A 100[* P occultum DQ322629 1

‘ 100 P. brasilianum AJ301862 ‘araglomeromycetes

' 1.00 P. laccatum AM295493

: Archaeospora trappei AM114274

+ 86 A. trappei Y17634

os Geosiphon pyriforme AJ276074

4.00 G. pyriforme X86686 Archaeosporomycetes

Ambispora fennica AM268195

A. callosa AB047308

A. appendicula AB047302

A. appendicula ABO15052

97 Glomus fulvum AM418543

G. versiforme AJ276088

Diversispora spurca AJ276077

D. celata AM713421

G. eburneum AM713429

100 Kuklospora colombiana Z14006

ue Acaulospora spinosa 214004

A. laevis Y17633

A. scrobiculata AJ306442

A. longula AJ306439

100 A. rugosa Z14005

100 Pacispora scintillans AJ619944

100 P. scintillans AJ619948

1.00 Scutellospora aurigloba AJ276093

100 S. calospora AJ306445

Gigaspora gigantea AJ852602

100 G. rosea AJ852606

1.00 G albida AJ852599

Racocetra castanea AF038590

Dentiscutata cerradensis AB041345

Fuscutata heterogama AJ852609

F. heterogama AJ306434

aan Glomus viscosum Y17652

700 G. etunicatum AJ852598

1.00 G. lamellosum AJ276083

G. claroideum AJ276075

G. luteum AJ276089

Glomus sp AJ301857

G. hoi AF485889

G. africanum HM153416

4100 G. constrictum AJ534309

100 G. coronatum AJ276086

100 G. mosseae Z14007

1.00 G mosseae AY635833

G. caledonium Y17653

G. fragilistratum AJ276085

G. geosporum AJ245637

G. verruculosum AJ301858

G. indicum GU059541

G. iranicum HM153420

G. perpusilum FJ164236

G proliferum AF213462

G. sinuosumAJ133706

G. coremioides AJ249715

G. clarum AJ852597

G. manihotis Y17648

G. vesiculiferum L20824

G. fasciculatum Y17640

G. irregulare FJ009618

G. intraradices AJ852526

G. intraradices AY635831

0.71

sajesodsisianiq

100

v 100

1.00

sayesodsebin

°o

) io

= N

-|-------------- ep ee e eee >

1.00

sajaoAwosauo]s)

Sajesaulojy

0.1

Fic. 1. Phylogenetic reconstruction of the Glomeromycota obtained from partial SSU rDNA

sequences (~1800 bp). The NJ, ML and Bayesian analyses were performed with GTR+G+I

substitution model. Sequences are labeled with their database accession numbers. Support values

are from neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and

Bayesian analyses, respectively. Only topologies with bootstrap values of at least 50% are shown.

(Consistency Index = 0.47; Retention Index = 0.81).

Fic. 2 (right). Phylogenetic reconstruction of the Glomeromycota obtained from partial LSU

rDNA sequences (~600 bp). The NJ, ML and Bayesian analyses were performed with GTR + G

substitution model. Sequences are labeled with their database accession numbers. Support values

are from neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and

Bayesian analyses, respectively. Only topologies with bootstrap values of at least 50% are shown.

(Consistency Index = 0.42; Retention Index = 0.78).

1.00)

0.1

Gigasporales, Archaeosporomycetes & Paraglomeromycetes (Glomeromycota) ... 369

Boletus edulis AF336240

100

1.00

100

1.00

0.98

eeereneneenevnecanennenneaeneaeeeneneaeneeneaeeanvaneenaene

to)

‘

oon o

ONE

100

1.00

ees

Neurospora crassa AF286411

Glomus trimurales FJ461851

Diversispora celata DQ350453

1400 D. celata AY639233

ae Glomus eburneum EF067886

1.00 G. eburneum EF067888

G. versiforme AY 842573

G. versiforme AM947664

G. insculptum FJ461838

G. aurantium EF581864

G. aurantium EF581861

Scutellospora calospora AJ510231

Fuscutata heterogama AY900503

F. heterogama AY900500

Dentiscutata nigra AY900498

D. nigra AY900494

Gigaspora rosea Y12075

G. margarita AF396782

G. margarita AF396783

G. gigantea AY900506

G. gigantea AY900504

Racocetra gregaria AJ510232

R. verrucosa AY900508

R. castanea Y12076

Cetraspora pellucida AY639313

r Am pellucida AF396784

100 fr Pacispora scintillans FM876831

100 ' P scintillans FM876832

1.00 Kuklospora colombiana FJ461804

K. kentinensis FM876822

K. kentinensis FM876830

Acaulospora longula AJ510228

A. paulinae AY639328

A. paulinae AY639263

A. laevis AJ510229

A. lacunosa AJ510230

Glomus diaphanum AJ972461

G. diaphanum AJ9372460

G. sinuosum FJ461846

G. intraradices AY842577

G. intraradices AF396797

G. proliferum FM992402

G. proliferum FM992398

G manihotis AM158947

G. clarum AJ510242

G. clarum AJ510243

Glomus coronatum AF 145740

G. fragilistratum AF 145747

G. coronatum AF 145739

G. mosseae AF 145735

G. mosseae AY639273

G. caledonium AJ510239

G. caledonium AJ628059

G. geosporum AF 145743

G. geosporum AF 145742

G. deserticola AJ746249

G. constrictum FJ461827

G. xanthium AJ849467

Glomus luteum FM876809

8 G. claroideum AF235007

G. claroideum AJ271929

G. lamellosum AY541863

G. etunicatum AJ623310

G. etunicatum AF 145749

G. drummondii AJ972466

G. walkeri AJ972468

Geosiphon pyriforme AM183920

Ambispora gerdemannii AJ271712

A. fennica AM268202

Paraglomus occultum DQ273827

P. laccatum AM295494

saJaDAWOsBWIOJH

Archaeosporomycetes

Paraglomeromycetes

370 ... Oehl & al.

Neurospora crassa M13630

Rhizophydium sphaerotheca AY944848

Archaeospora trappei FJ174311

Ambispora leptoticha FJ174312 Archaeosporomycetes

Glomus deserticola FJ174295

G diaphanum FJ174300

50 G sinuosum Fj174298

90 G clarum FJ174296

1.00 G manihotis FJ174297

G intraradices FJ174299

G proliferum AJ717321

58 G mosseae FJ174292

72 G coronatum FJ174290

0.99 G. geosporum FJ174291

G. fragilistratum FJ174293

G geosporum AJ717323

ae ine G luteum FJ174282

_ 100 G etunicatum FJ174280

87 1.00 |p G claroideum FJ174283

G etunicatum FJ174279

Fuscutata heterogama FJ174315

Gigaspora rosea FJ174274

G albida FJ174275

100 G. gigantea FJ174276

1.00] | | G decipiens FJ174277

G margarita FJ174278

Scutellospora calospora FJ174268

Racocetra persica FJ174272

R. verrucosa FJ174270

R. gregaria FJ174271

Cetraspora pellucida FJ174269

R. fulgida FJ174273

R. castanea AJ717326

100 — Glomus versiforme FJ174285

700 G. trimurales FJ174289

1.00 Diversispora spurca FJ174287

53 G eburneum FJ174288

Kuklospora colombiana FJ174310

97 Acaulospora koskei FJ174309

100 A. morrowiae FJ174302

a A. dilatata FJ174301

A. mellea FJ174303

K. kentinensis FJ174306

A. foveata FJ174304

A. lacunosa FJ174305

A. scrobiculata FJ174307

9 A. laevis FJ174308

00 Paraglomus occultum FJ174313

96 P. brasilianum FJ174314

Seei

Sa]es9WIO|5

0.87

sa}20AWOaWI0]5

sajesodsebig

eeenenenereneceeneenaecae

sajesodsisiaaig

3 Paraglomeromycetes

0.1

Fic. 3. Phylogenetic reconstruction of the Glomeromycota obtained from partial B-tubulin

sequences (~600 bp). The NJ, ML and Bayesian analyses were performed with GTR + G + I

substitution model. Sequences are labeled with their database accession numbers. Support values

are from neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and

Bayesian analyses, respectively. Only topologies with bootstrap values of at least 50% are shown.

(Consistency Index = 0.37; Retention Index = 0.70).

Archaeosporaceae and Ambisporaceae have two different morphs

(acaulosporoid & glomoid, or entrophosporoid & glomoid) and form mycorrhiza

with plants (Spain et al. 2006; Sieverding & Oehl 2006). Geosiphonaceae occur,

as far as known, only in association with algae and were until now found with

mosses, connected to rhizoid structures. Hence, species of Archaeosporales

have spores of two different morphs of other AM fungal classes (or order)

or are formed by two different groups of organisms. Genetical analyses (see

Fics. 1-3) support the establishment of a new class for these organisms.

Paraglomerales species form glomoid spores with one spore wall as known

for species in the Glomerales. However, spores germinate directly through the

Gigasporales, Archaeosporomycetes & Paraglomeromycetes (Glomeromycota) ... 371

spore wall (Spain & Miranda 1996, Oehl et al. 2011a), and their extra-radical

mycelia and mycorrhizal structures stain only faintly or not at all in trypan

blue. Phylogenetic analyses clearly indicate that they are quite distant to species

of all other orders in Glomeromycota. Hence, we place Paraglomerales in a new

fungal class.

The Glomeromycetes class splits into two major phylogenetic clades:

/glomerales and /diversisporales sensu Schiifsler et al. (2001) (Fics. 1-3). Spores

in these two clades also clearly differ morphologically. Species of Glomerales

were recently divided in two families (Glomeraceae and Claroideoglomeraceae)

according to spore morphology and phylogenetic analyses (Schiifler & Walker

2010, Oehl et al. 201 1a). The Diversisporales contain species with highly diverse

types of spore formation: ACAULOSPOROID & KUKLOSPOROID (with all spores

having three walls and forming on or in the subtending hypha of a sporiferous

saccule), GIGASPOROID & SCUTELLOSPOROID (spores forming on sporogenous

cells without or with germ shield formation), PACISPOROID (spores with

two walls, forming terminally on subtending hyphae, and germinating from

the inner wall), DIVERSISPOROID (spores forming terminally on subtending

hypha and having one wall that is not entirely continuous with the hyphal

mycelia wall; see Oehl et al. 2011a), and ENTROPHOSPOROID & OTOSPOROID

(spores with two walls and forming in or on hyphae of sporiferous saccules).

The Diversisporales currently comprises eight families: Acaulosporaceae,

Dentiscutataceae, Diversisporaceae, Entrophosporaceae, _Gigasporaceae,

Racocetraceae, Pacisporaceae, Scutellosporaceae.

Relationships between the families with gigasporoid & scutellosporoid

(sensu lato) spore formation and other glomeromycete families are not fully

understood. The SSU and LSU rDNA sequence analyses place these families

into Diversisporales, while the $-tubulin analyses put this group out of

Diversisporales.

Important morphological features for differentiating species within

the Glomeromycetes are intraradical vesicle and extra-radical auxiliary cell

formation, while species of the Gigasporineae form arbuscular mycorrhizas with

extraradical auxiliary cells instead of vesicles in roots. All other representatives

of the Glomerales and Diversisporales (excluding Gigasporineae) generally form

vesicular arbuscular mycorrhizas. Because genetic analyses also clearly indicate

sporogeneous cell forming species in a clade separate from the Diversisporales

and Glomerales, we suggest including sporogeneous cell forming species in a

new order.

Re-organization of classes in the Glomeromycota

Due to the molecular phylogenetic analyses and differences in the type

of the arbuscular mycorrhizal structures and spore formation, we divide the

Glomeromycota into three fungal classes, formally propose the new order

372 ... Oehl & al.

Gigasporales, and emend existing orders and families where necessary. An

overview of the new taxonomic organization of the Glomeromycota is given in

FIG. 4.

Glomeromycetes Caval.-Sm., emend. Oehl, G.A. Silva, B.T. Goto & Sieverd.

EMENDED DESCRIPTION: Glomerospores formed terminally, subterminally or

intercalary in hyphae, either in soils or sometimes in roots, either singly, in

spore clusters or multiple-spored loose to compact sporocarps, on subtending

hyphae (sH), on sporogenous cells, or laterally on or intrahyphally in the stalk

of sporiferous saccules, forming arbuscular or vesicular-arbuscular mycorrhiza,

with mycorrhizal structures that stain blue to dark blue in trypan blue.

Glomerales J.B. Morton & Benny, emend. Oehl, G.A. Silva, B.T. Goto & Sieverd.

EMENDED DESCRIPTION: Spores formed terminally on or intercalary in

hyphae, either in soils or sometimes in roots, either singly, in spore clusters or

multiple-spored loose to compact sporocarps, with a single mono-to-multiple

layered spore wall. Wall of sH conspicuously continuous with the spore wall

and colored the same as or slightly lighter than it or hyaline to subhyaline;

sH funnel-shaped, cylindrical or constricted; forming typical vesicular-

arbuscular mycorrhiza, with mycorrhizal structures that stain blue to dark blue

in trypan blue.

TYPE SPECIES: Glomus macrocarpum Tul. & C. Tul. (Clements & Shear 1931)

FAMILIES INCLUDED: Glomeraceae Piroz. & Dalpé, Claroideoglomeraceae C. Walker &

A. Schiissler

GENERA INCLUDED: Funneliformis C. Walker & A. Schiissler, Glomus Tul. & C. Tul.,

Claroideoglomus C. Walker & A. Schiissler, Septoglomus Sieverd. et al., Simiglomus

Sieverd. et al., Viscospora Sieverd. et al.

Diversisporales C. Walker & A. Schiissler, emend. Oehl, G.A. Silva & Sieverd.

EMENDED DESCRIPTION: Spore formation acaulosporoid, kuklosporoid,

entrophosporoid, otosporoid, or diversisporoid; acaulosporoid and

kuklosporoid spores with three walls: multiple layered outer wall, and hyaline

middle and inner walls; otosporoid & entrophosporoid spores with two walls:

multiple layered outer wall and hyaline inner wall; diversisporoid spores with

subtending hyphae (sH) with a conspicuous color change distant to the septum

most proximal to the spore base; spores with 1-3 wall layers (swL1-3), pore

rarely open. Forming typical vesicular-arbuscular mycorrhizae, formation of

auxiliary cells in root external mycelium unknown.

TYPE SPECIES: Diversispora spurca (C.M. Pfeiff. et al.) C. Walker & A. Schiissler

FAMILIES INCLUDED: Acaulosporaceae J.B. Morton & Benny, Diversisporaceae C. Walker &

A. Schiissler, Entrophosporaceae Oehl & Sieverd., Pacisporaceae C. Walker et al.

GENERA INCLUDED: Acaulospora Gerd. & Trappe, Diversispora C. Walker & A. Schiissler,

Entrophospora R.N. Ames & R.W. Schneid., Redeckera C. Walker & A. Schiissler,

Kuklospora Oehl & Sieverd., Otospora Oehl et al., Pacispora Sieverd. & Oehl

Gigasporales, Archaeosporomycetes & Paraglomeromycetes (Glomeromycota) ... 373

B. edulis DQ534675

N. crassa AY046271

Fic. 4. Cladogram generated by a consensus between the trees obtained from rRNA and $-tubulin

genes.

Gigasporales Sieverd., G.A. Silva, B.T. Goto & Oehl, ord. nov.

MycoBank MB 519688

Sporae terminaliter efformatae anguste adiacetae ad cellulas sporogeneas formans

structuras mycorrhizarum arbuscularum et celulas auxiliares; formatio vesiculis ignota.

KEY CHARACTERS: Spore formation gigasporoid and scutellosporoid, ice.

terminally on sporogenous cells, with one to four spore walls; germination

from germ warts positioned on the inner surface of single-walled spores or in

bi- to multiple-walled spores from germ tube initiations positioned on discrete

germination structures (germ shields); arbuscular mycorrhizae and extra-

radical auxilliary cells; intra-radical vesicle formation unknown.

TYPE SPECIES: Gigaspora gigantea (T.H. Nicolson & Gerd.) Gerd. & Trappe

FAMILIES INCLUDED: Dentiscutataceae F.A. Souza et al., Gigasporaceae J.B. Morton &

Benny, Racocetraceae Oehl et al., Scutellosporaceae Sieverd. et al.

GENERA INCLUDED: Cetraspora Oehl et al., Dentiscutata Sieverd. et al., Fuscutata Oehl

et al., Gigaspora Gerd. & Trappe, Orbispora Oehl et al., Quatunica F.A. Souza et al.,

Racocetra Oehl et al., Scutellospora C. Walker & FE. Sanders

374 ... Oehl & al.

Archaeosporomycetes Sieverd., G.A. Silva, B.T. Goto & Oehl, cl. nov.

MycoBank MB 519686

Simbiosis formans inter radices plantarum et fungos vel entre algae et fungi si quod inter

radices plantarum et fungos esporae dimorphae; rarum secundum morphum ignotum;

arbusculae vel arbuscular et vesiculae formans, estructurae mycorrhizarum non vel pallide

tinguntur cum trypan blue.

KEY CHARACTERS: Forms symbiosis between plant roots and fungi, or between

algae and fungi; if between plants and fungi, fungal spores with two morphs,

rarely only one morph known, and forming vesicular arbuscular or arbuscular

mycorrhiza which do not or only faintly stain with trypan blue.

Archaeosporales C. Walker & A. Schiissler, emend. Sieverd., G.A. Silva, B.T. Goto &

Oehl

EMENDED DESCRIPTION: Forms simbiosis between plant roots and fungi,

or between algae and fungi. If between plants and fungi: fungal species with

regularly bi-morphic spore formation: ambi-acaulosporoid & ambi-glomoid,

archaeo-acaulosporoid & glomoid, or intra-entrophosporoid & glomoid spore

formation; inner walls form de novo; forming arbuscular mycorrhiza; mycelia

and mycorrhizal structures do not or only faintly stain in ‘trypan blue:

TYPE SPECIES: Archaeospora trappei (R.N. Ames & Linderman) J.B. Morton & D. Redecker

FAMILIES INCLUDED: Ambisporaceae C. Walker et al., Archaeosporaceae J.B. Morton &

D. Redecker, Geosiphonaceae Engl. & E. Gilg

GENERA INCLUDED: Ambispora C. Walker et al. (= Appendicispora Spain et al.),

Archaeospora J.B. Morton & D. Redecker, Geosiphon F. Wettst., Intraspora Oehl &

Sieverd.

Paraglomeromycetes Oehl, G.A. Silva, B.T. Goto & Sieverd., cl. nov.

MycoBank MB 519687

Sporae glomoideae; germinatio ex tunica sporae; estructurae mycorrhizae non tinguntur

‘trypan blue’; vesiculae ignotae.

KEY CHARACTERS: Spores formed terminally on hyphae; germination directly

through spore wall; arbuscular mycorrhiza without or with only faint reaction

when exposed to trypan blue; vesicle formation unkown.

TYPE SPECIES: Paraglomus occultum (C. Walker) J.B. Morton & D. Redecker

FAMILY INCLUDED: Paraglomeraceae J.B. Morton & D. Redecker

GENUS INCLUDED: Paraglomus J.B. Morton & D. Redecker

Discussion

Glomeromycota phylum has three classes and five orders

When Schiifler et al. (2001) described the Glomeromycota, they stated that

the phylum has one class, the Glomeromycetes, established by Cavalier-Smith

(1998) for all arbuscular mycorrhizal fungi. Our current molecular analyses

(see Figs 1-3) as well as those by the authors of the new phylum and many

Gigasporales, Archaeosporomycetes & Paraglomeromycetes (Glomeromycota) ... 375

other researchers (e.g. Msiska & Morton 2009, Kriiger et al. 2009, Gamper et

al. 2009, Oehl et al. 2011a) have all consistently demonstrated that no matter

which genes are analysed, the Glomeromycota is divided into three major

monophyletic clades, with a large majority of the known species belonging to

the Glomeromycetes. We, as Cavalier-Smith (1998) did, think that all species

in this class form typical arbuscular mycorrhizal structures in roots. The

Glomeromycetes can be separated from the other two classes by the staining

features of root internal fungal structures. The other two glomeromycotan

classes (Archaeosporomycetes, Paraglomeromycetes) are new, but were

previously recognized as orders (Schiifler et al. 2001). The absent or weak

staining of the fungal structures within both new classes suggests a relationship

between them. However, the clear genetical differences and the apparent

association of two morphologically different organisms (bi-morphic status)

in the Archaeosporomycetes versus the mono-morphic status of members of

Paraglomeromycetes species separate these two classes.

Archaeosporales is the only order in Archaeosporomycetes. ‘The class

contains three clades: the algal symbiont-forming Geosiphonaceae and the

Ambisporaceae and Archaeosporaceae. Sieverding & Oehl (2006) and Spain et al.

(2006) revised the Archaeosporaceae (including the species of Ambisporaceae).

Genetically, Geosiphonaceae appear more closely related to Ambisporaceae than

Archaeosporaceae. However, we have not yet investigated or tried to clarify the

relationship of Geosiphonaceae with Ambisporaceae, leaving that for future

research.

Within Paraglomeromycetes the single order Paraglomerales appears quite

homogenous and monophyletic. Paraglomeromycetes have glomoid spore

formation, and we recognize that interspecific differentiation from members

of the Glomeromycetes is difficult if one is not familiar with the species-specific

characters of the Paraglomus spp. Taxonomic separation of Paraglomeromycetes

from Glomeromycetes species has been supported previously only by molecular

analyses (Morton & Redecker 2001). However, we found that the root internal

mycelium of Paraglomus spp. have different staining features than glomoid

(Glomerales) and diversisporoid (Diversisporales) species, and Paraglomus spp.

obviously lack vesicle formation in the roots, distinguishing them from the

vesicular-arbuscular mycorrhiza forming Glomeromycetes (Oehl et al. 201 1a).

Remarkably, while all other glomoid spores germinate through the subtending

hyphae, we observed that Paraglomeromycetes species generally germinate

through the spore wall (Oehl et al. 201 1a), an observation needing confirmation

for all Paraglomus species.

Differentiation of orders in Glomeromycetes

The revised class Glomeromycetes is divided in three orders: Glomerales,

Diversisporales, and Gigasporales. Within Diversisporales, Acaulosporaceae have

376 ... Oehl & al.

particular hyphal attachments, unique spore wall composition, and germination

characteristics shared only with ancestral species of Scutellosporaceae (Oehl

et al. 2008, 2011b). For Diversisporaceae (respective former Glomus clade

C), no attempts or assumptions have been made until very recently (Oehl et

al. 2011a) to explain how its species might differ morphologically from the

Glomerales sensu Oehl et al. (e.g. Schiifler et al. 2001, Walker & Schiifler 2004,

Redecker et al. 2007). The spore base of the Diversisporaceae is typical and the

structural wall layer appears to have formed like an “endospore”. In the current

study, the morphological differences between glomoid and diversisporoid

spores were described, and the family Glomeraceae (Pirozynski & Dalpé

1989) was accordingly revised to exclude several former Glomus species from

the family and transferring them to Claroideoglomus or Viscospora in the

Claroideoglomeraceae, to Diversispora or Redeckera in the Diversisporaceae, or

to Paraglomus in the Paraglomeraceae based on molecular and morphological

analyses (Oehl et al. 2011a).

Gigasporales do not include taxa with vesicles and glomoid spore formation.

Species of this new order have unique hyphal attachments (sporogenous cells),

complex spores, and distinct germination characteristics (Oehl et al. 2008, Goto

et al. 2010, 2011). Oehl et al. (2008, 2011b) have recently revised the families,

genera and species in Gigasporales.

Unsolved problems in the Glomeromycetes

Recently, Pacisporaceae and Entrophosporaceae were included into the

Diversisporales (Walker & Schiifler 2004, Sieverding & Oehl 2006).

Pacisporaceae fall in two different positions in the SSU and LSU rRNA

analyses, and there are no sequences for B-tubulin gene to confirm the

phylogenetic relationship between this family and either the gigasporalean

or other diversisporalean groups. Since we do not know the real position for

Pacisporaceae, it was decided to maintain this family within the Diversisporales.

More studies are required to elucidate the evolutive pathways in Pacisporaceae

and confirm or correct the actual proposition.

Entrophosporaceae was revised by Sieverding & Oehl (2006). Recent

observations suggest a split of the species of Entrophosporaceae according to

their phylogenetic positions (e.g. see public data bases, Palenzuela et al. 2010).

However, further molecular analyses are needed to confirm our observations.

Sieverding & Oehl (2006) recently emended the genus Entrophospora and its

type species.

Last considerations

Taxonomic congruence can be used to determine the true phylogenetic

relationships in a group of organisms (Kitching et al. 1998). However this

Gigasporales, Archaeosporomycetes & Paraglomeromycetes (Glomeromycota) ... 377

should be obtained by analyses of independent data sets. When the study

involves genes, these should not be genetically linked. Thus, we cannot use LSU

and SSU rDNA as two independent data sets to obtain taxonomic congruence.

Diversisporales (sensu Schiifler et al. 2001) form a polyphyletic clade in the

6-tubulin analysis and a monophyletic group in the rRNA trees. However, the

consensus tree does not establish the former Diversisporales as a monophyletic

clade in a congruent phylogeny with the two independent data sets (rRNA and

6-tubulin genes). Thus, the supports obtained by SSU rDNA and 6-tubulin

genes sustain Gigasporales as a natural taxon. This new order was decribed here

to solve the incongruence in Diversisporales (sensu Schifsler et al. 2001) and

maintain a natural classification in the Glomeromycota in the higher taxonomic

levels (order and above).

Acknowledgments

We acknowledge the valuable comments and revisions of several experts on the

manuscript and appreciate the corrections by Shaun Pennycook, Nomenclatural Editor,

and suggestions by Lorelei L. Norvell, Editor-in-Chief. This study was supported by the

Swiss National Science Foundation (SNSE, Project 315230_130764/1), the Conselho

Nacional de Desenvolvimento Cientifico e Tecnolégico (CNPq-Brasil), and by the

Fundacao de Amparo a Ciéncia e Tecnologia do Estado de Pernambuco (FACEPE) and

the Universidade Federal de Pernambuco (UFPE) which provided grants to E Oehl as

‘visiting professor.

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