IV ... MYCOTAXON 118

MY COTAXON

VOLUME ONE HUNDRED EIGHTEEN — TABLE OF CONTENTS

COVER SECTION

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OTP TICE LOT Rose B ihe Soy inn send de Ole Se al, Jb etly vin, Syd Rea xi

RESEARCH ARTICLES

New records of Geastrum from Japanese sand dunes

Taiga Kasuya, Kentaro Hosaka, Haruo Sakamoto,

Akitomo Uchida, Tamotsu Hoshino & Makoto Kakishima

Discrimination of Gigaspora species by PCR specific primers and

phylogenetic analysis Gladstone Alves da Silva, Erica Lumini,

Valeria Bianciotto, Paola Bonfante & Leonor Costa Maia

Contribution to the taxonomy of Bovista in Mexico

Silvia Bautista- Hernandez, Teofilo Herrera,

Elvira Aguirre-Acosta & Martin Esqueda

Subulicystidium curvisporum sp. nov. (Hydnodontaceae, Basidiomycota)

from the Patagonian Andes

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

New records of smut fungi. 4. Microbotryum coronariae comb. nov.

Cvetomir M. Denchev & Teodor T. Denchev

Lentinus giganteus revisited: new collections from Sri Lanka and Thailand

Samantha C. Karunarathna, Zhu L. Yang, Olivier Raspé, Thida W. Ko Ko,

Else C. Vellinga, Rui-Lin Zhao, A.H. Bahkali, Ekachai Chukeatirote,

Jerome Degreef, Philippe Callac & Kevin D. Hyde

Diversispora clara (Glomeromycetes)— a new species from saline dunes

in the Natural Park Cabo de Gata (Spain)

Beatriz Estrada, Javier Palenzuela, José-Miguel Barea,

Juan Manuel Ruiz-Lozano, Gladstone Alves da Silva & Fritz Oehl

Asterophora salvaterrensis (Basidiomycota, Agaricales), a new species

from Galicia (Spain) Jaime B. Blanco-Dios

Amicodisca castaneae sp. nov. (Hyaloscyphaceae, Helotiales) on Japanese

chestnut bur Jae-Gu Han, Tsuyoshi Hosoya & Hyeon-Dong Shin

Pseudocercospora epidendri sp. nov. on the neotropical orchid,

Epidendrum secundum from Brazil

Meiriele da Silva & Olinto Liparini Pereira

Eremiomyces magnisporus (Pezizales), a new species from central Spain

Pablo Alvarado, Gabriel Moreno, José L. Manjén & Miguel A. Sanz

103

OCTOBER-DECEMBER 2011... V

Peziza paludicola, the correct binomial for P. udicola, nom. inval.

Gabriele Cacialli, Angela Lantieri & Gianfranco Medardi 113

Two new species of Corynespora from northeastern Uttar Pradesh, India

Raghvendra Singh & Kamal 123

New lichenicolous fungi records for Kyrgyzstan, Uzbekistan, and Ukraine

Olga Nadyeina & Mehmet Gokhan Halici 131

New records of Agaricales from Atlantic Forest fragments of Pernambuco,

Northeast Brazil

Felipe Wartchow, Leonor C. Maia & M. Auxiliadora Q. Cavalcanti 137

Tremelloscypha gelatinosa (Sebacinales) from tropical deciduous

Gymnopodium forests in southern Mexico

Victor M. Bandala, Leticia Montoya & Rafael Villegas 147

New records of polypores from southern Florida

J. Vlasak, J. Kout, J. Vlasak Jr., & L. Ryvarden 159

Paraphysoderma sedebokerense, gen. et sp. nov., an aplanosporic relative of

Physoderma (Blastocladiomycota)

Timothy Y. James, Yoram Hoffman, Aliza Zarka & Sammy Boussiba 177

Microcyclospora rumicis, a new species on Rumex crispus from Iran

Mahdi Arzanlou & Mounes Bakhshi 181

Mycena guldeniana - a new alpine species from Norway

Arne Aronsen & Brian A. Perry 187

Neolecta vitellina, first record from Romania, with notes on habitat

and phenology Vasilica Claudiu Chinan & David Hewitt 197

Epitypification, morphology, and phylogeny of Tothia fuscella

Haixia Wu, Walter M. Jaklitsch, Hermann Voglmayr & Kevin D. Hyde 203

Exserohilum neoregeliae sp. nov., a new pathogen of Neoregelia carolinae

Takuya Sakoda & Takao Tsukiboshi 213

Species of Rhytismataceae on Camellia spp. from the Chinese mainland

Jiang-Lin Chen, Ying-Ren Lin, Cheng-Lin Hou & Shi-Juan Wang 219

A new species of Coccomyces (Rhytismatales, Ascomycota)

from Mt Huangshan, China Guo-Jun Jia, Ying-Ren Lin & Cheng-Lin Hou 231

New records of poaceous rusts from Pakistan

A. Ishaq, N.S. Afshan & A.N. Khalid 237

New records of smut fungi. 5

Cvetomir M. Denchev, Teodor T. Denchev & Brian M. Spooner 245

A new species of Marasmius from northern Argentina

Bernardo Lechner & Leandro Papinutti 251

Arrasia rostrata (Basidiomycota), a new corticioid genus and species from Italy

Annarosa Bernicchia, Sergio P. Gorjén & Karen K. Nakasone 257

Cortinarius mikedavisii sp. nov. from northern California

Dimitar Bojantchev 265

VI ... MYCOTAXON 118

Observations on gasteroid Agaricomycetes from the

Brazilian Amazon rainforest Larissa Trierveiler-Pereira, Allyne

Christina Gomes-Silva & Iuri Goulart Baseia

Septobasidium saurauiae sp. nov. (Septobasidiaceae) and

S. pseudopedicellatum new to China Suzhen Chen & Lin Guo

Hallenbergia (Agaricomycetes), a new corticioid genus

G.S. Dhingra & Priyanka

Cylindrochytridium johnstonii is a member of the Cladochytriales

Rebecca A. Steiger, D. Rabern Simmons & Joyce E. Longcore

A contribution to the study of smut fungi of Israel

Kyrylo G. Savchenko, Vasyl P. Heluta, Solomon P. Wasser & Eviatar Nevo

Species of Rhytismataceae on Lithocarpus spp. from Mt Huangshan, China

Qian Zheng, Ying-Ren Lin, Sheng-Ming Yu & Li Chen

Aschersonia conica sp. nov. (Clavicipitaceae) from Hainan Province, China

Jun-Zhi Qiu, Yu-Bin Su, Chong-Shuang Weng & Xiong Guan

273

283

289

293

303

SEL

325

Clarkeinda trachodes (Agaricales, Basidiomycetes), first record from Bangladesh

Md. Iqbal Hosen & Z.W. Ge

Glomus cubense sp. nov., an arbuscular mycorrhizal fungus from Cuba

Y. Rodriguez, Y. Dalpé, S. Séguin, K. Fernandez, F Fernandez & R.A. Rivera

Two new species of Exserticlava and Spiropes on decaying wood

from Guangdong, China Shou-Cai Ren, Jian Ma & Xiu-Guo Zhang

Discovery of Geastrum xerophilum from the Neotropics

Bianca Denise Barbosa da Silva,

Julieth de Oliveira Sousa & Iuri Goulart Baseia

A preliminary ITS phylogeny of Melanoleuca (Agaricales),

with special reference to European taxa

Alfredo Vizzini, Roberto Para, Roberto Fontenla,

Stefano Ghignone & Enrico Ercole

Geastrum species from the Amazon Forest, Brazil Anileide Gomes Leite,

Hannah Kathren de Assis, Bianca Denise Barbosa da Silva,

Helen Maria Pontes Sotao & Iuri Goulart Baseia

Hypoderma siculum sp. nov. from Italy Angela Lantieri, Peter R. Johnston,

Duckchul Park, Henrik Lantz & Gianfranco Medardi

Tuber sinoalbidum and T: polyspermum — new species from China

Li Fan, Cheng-Lin Hou & Jin-Zhong Cao

Hymenochaete in China. 2. A new species and three new records

from Yunnan Province Shuang-Hui He & Hai-Jiao Li

Lichenological notes 3: Sarcogyne plicata in California

Kerry Knudsen & Jana Kocourkova

Homolaphlyctis polyrhiza gen. et sp. nov., a species in the Rhizophydiales

(Chytridiomycetes) with multiple rhizoidal axes

Joyce E. Longcore, Peter M. Letcher & Timothy Y. James

337

349

355

361

383

393

403

411

423

433

OcCTOBER-DECEMBER 2011... VII

Eutypella phaeospora, a new species on Chenopodiaceae

Jacques Fournier & Christian Lechat 441

New combinations in Lactifluus. 1. L. subgenera Edules, Lactariopsis,

and Russulopsis A. Verbeken, J. Nuytinck & B. Buyck 447

Validation of combinations with basionyms published by Fries in 1861

Scott A. Redhead, Joseph F. Ammirati, Lorelei L. Norvell,

Alfredo Vizzini & Marco Contu 455

BOOK REVIEWS AND NOTICES Else C. Vellinga (EpiToR) 459

NOMENCLATURE

Nomenclatural novelties proposed in volume 118 463

PUBLICATION DATE FOR VOLUME ONE HUNDRED SEVENTEEN

MYCOTAXON for JuLY-SEPTEMBER, VOLUME 117 (I-x + 1-519)

was issued on November 22, 2011

vul ... MYCOTAXON 118

ERRATA FROM PREVIOUS VOLUMES

VOLUME 114

p- 429, line 4

VOLUME 117

p-20, lines 34-35

p-21, lines 17-19

p-27, line 20

p-27, line 42

for: DANILo. B

read: DANILO B.

for: smooth under the light microscope, but with bacillate

ornamentation under SEM (Figs.1 c-d, 2c).

read: smooth (Fig. 2b).

for: yellowish to yellowish brown in KOH, smooth, subfusiform to

ellipsoid, slightly thick-walled (up to 0.7 um) (Fig. 2c).

read: yellowish to yellowish brown in KOH, subfusiform to ellipsoid,

slightly thick-walled (up to 0.7 um), smooth under the light

microscope, but with bacillate ornamentation under SEM

(Figs.1c-d, 2c).

for: Albee-Scott S. 2007.

read: Albee-Scott S. 2007.

for: Mycological Progress (Online First).

read: Mycological Progress 10: 389-398.

p.32, between Arthrobotrys latispora heading and Latin diagnosis:

add: MycoBANK MB 560938 (assigned post publication)

p-368, line 10" from bottom

for: MycoBaANnK 512114

read: MYCOBANK MB 560939 (assigned post publication)

OCTOBER-DECEMBER 2011 ...

REVIEWERS — VOLUME ONE HUNDRED EIGHTEEN

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.

Vladimir Antonin

Boris Assyov

Dominik Begerow

Konstanze Bensch

Janusz Blaszkowski

Uwe Braun

Harold H. Burdsall, Jr.

Marina Capelari

Rafael F. Castafieda

Michael A. Castellano

Wen-Hsin Chung

Johannes C. Coetzee

Marco Contu

Vagner Gularte Cortez

Yu-Cheng Dai

Dennis Desjardin

Admir Jose Giachini

Sergio Pérez Gorjon

Bruno Tomio Goto

M. Gokhan Halici

Ian Robert Hall

Nils Hallenberg

Donald E. Hemmes

Cheng-Lin Hou

Seppo Huhtinen

Alfredo Justo Fernandez

Masoomeh

Ghobad-Nejhad

Admir Giachini

Alina G. Greslebin

Nils Hallenberg

Shuanghui He

Peter R. Johnston

Sergey Karpov

Taiga Kasuya

Ryan M. Kepler

Kerry Knudsen

Hanns Kreisel

Thomas Lzssge

D. Jean Lodge

Cristiano Losi

Guo-Zhong Lit

W. Wallace Martin

Eric H.C. McKenzie

John McNeill

Jamjan Meeboon

André August Remi de

Meijer

Jurgen Miersch

David W. Minter

Gabriel Moreno

Abdul Rehman Niazi

Lorelei L. Norvell

Fritz Oehl

Clark Ovrebo

Giovanni Pacioni

Omar Paino Perdomo

Shaun R. Pennycook

Sergio Pérez-Ortega

Donald H. Pfister

Martha J. Powell

Francois Rappaz

Scott A. Redhead

Jack D. Rogers

Ivan Sanchez-Castro

Conrad L. Schoch

Roger Graham Shivas

B.M. Sharma

Ewald Sieverding

Matthew E. Smith

Renata Gomes de Souza

Viacheslav Spirin

Sharon E. Standridge

Marcelo Aloisio

Sulzbacher

Haruki Takahashi

Zdenko Tkaléec

Kalman Vanky

Else C. Vellinga

Alfredo Vizzini

Michael Weif

Tim Wheeler

Merlin White

Meng Zhang

Tian-Yu Zhang

Wen-Ying Zhuang

x ... MYCOTAXON 118

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OCTOBER-DECEMBER 2011... XI

FROM THE EDITOR-IN-CHIEF

MycoTAxon 118 — Welcome to the (almost) on schedule October-December 2011

volume! Granted, you are reading this in 2012, but we have great hopes that the

hardcopies will be mailed in the first week of the first month of our new year. The year

2011 was the first where virtually all subscribers received their volumes entirely on-line.

Where once we believed that online publication would be speedier, we now understand

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A short while ago we discovered that INGENTACONNECT was citing the first day

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With warm best wishes for the New Year,

Lorelei Norvell, Mycotaxon Editor-in-Chief

December 30, 2011

MYCOTAXON

http://dx.doi.org/10.5248/118.1

Volume 118, pp. 1-15 October-December 2011

New records of Geastrum from Japanese sand dunes

TAIGA KAsuyA', KENTARO HoSAKA?, HARUO SAKAMOTO?,

AKITOMO UCHIDA‘, TAMOTSU HOSHINO®* & MAKOTO KAKISHIMA!

‘Laboratory of Plant Parasitic Mycology, Graduate School of Life and Environmental Sciences,

University of Tsukuba, 1-1-1, Ten-nodai, Tsukuba, Ibaraki 305-8572, Japan

*Department of Botany, National Museum of Nature and Science,

4-1-1, Amakubo, Tsukuba, Ibaraki 305-0005, Japan

°443-6, Nakaarai, Ageo, Saitama 362-0052, Japan

‘Shiretoko Museum, 49-2, Hon-machi, Shari, Hokkaido 099-4113, Japan

National Institute of Advanced Industrial Science and Technology (AIST),

2-17-2-1, Tsukisamu-higashi, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan

°Graduate School of Life Science, Hokkaido University,

North 10 West 8, Kita-ku, Sapporo, Hokkaido 060-0810, Japan

* CORRESPONDENCE TO: tkasuya@sakura.cc.tsukuba.ac.jp

ABSTRACT— Basidiomata of three earthstars —Geastrum campestre, G. corollinum,

G. hungaricum— were collected from sand dunes of the Japanese coasts. Those arenicolous

fungi are newly recorded for Japanese mycobiota. Descriptions, comments and illustrations

of basidiomata of the three fungi are provided.

Key worps— biogeography, coastal environment, gasteromycetes, Geastrales, taxonomy

Introduction

The genus Geastrum Pers. belongs to Geastrales, Phallomycetidae (Hosaka et

al. 2006) and includes currently about 50 taxa (Kirk et al. 2008). Most species of

the genus are known as saprobes and are distributed throughout all continents

except Antarctica. Several authors have systematically revised Geastrum

(Stanék 1958, Ponce de Leon 1968, Dorfelt & Miller-Uri 1984, Dorfelt &

Heklau 1987, Sunhede 1989). Several species have been recorded from multiple

continents, although their taxonomic identities are doubtful because there are

only a few molecular phylogenetic studies. Our preliminary morphological

and phylogeographical analyses of the globally distributed “G. triplex Jungh.”

indicate that the name represents an aggregation of multiple species (Kasuya

et al. 2011a). This suggests the possible existence of many cryptic species in the

genus.

2 ... Kasuya & al.

Although 18 Geastrum species have hitherto been recognized from Japan

(Imai 1936, Kawamura 1954, Ito 1959, Yoshimi & Hongo 1989, Sakamoto &

Kasuya 2008, Kasuya et al. 2009, 2011b), comprehensive taxonomical and

biogeographical studies of the genus have not yet been conducted. Therefore,

to clarify the geographical distribution, morphological variations, and

phylogenetic positions of Japanese Geastrum, we conducted morphological

and molecular phylogenetic analysis of the genus based on the materials from

multiple localities including Japan and other continents (Kasuya et al. 201 1a).

Several Geastrum species favor semiarid to arid environments, e.g., well-

drained sandy soils of coasts, deserts, and inland steppes. During our recent

floristic and taxonomic investigations of Japanese Geastrum (Sakamoto &

Kasuya 2008, Kasuya et al. 2009, 2011b), we recorded G. kotlabae V.J. Stanék,

G. minimum Schwein., and G. quadrifidum Pers. from coastal sand dunes of

Hokkaido and Honshu.

Some noteworthy Geastrum collections obtained by further fieldwork

conducted in sand dunes along the Japanese coasts include G. campestre,

G. corollinum, and G. hungaricum, which represent new distributional records

for Japan. We describe the Japanese collections of these three species with the

aid of illustrations showing morphological characters. We also compare them

with related taxa and discuss the biogeographical and ecological features of the

species.

Materials & methods

Fieldwork was carried out from September 2003 to October 2010 at three sites in

coastal sand dunes of Hokkaido and Honshu, Japan: (1) Minehama, Shari, Hokkaido

(43°55'34"N, 144°46'18"E); (2) Hitachi Kaihin Park, Hitachinaka, Ibaraki (36°23'54"N,

140°36'24"E); and (3) Fukude, Iwata, Shizuoka (34°39'51"N, 137°53'22"E). These sites

well preserve coastal vegetation dominated by poaceous and cyperaceous plants. For

morphological comparisons, we examined one additional G. corollinum specimen

collected from an inland area of Honshu.

The examined specimens are deposited in the mycological herbarium of the National

Museum of Nature and Science, Tsukuba, Ibaraki, Japan (TNS). Macroscopic characters

were described from dried and fresh material. For light microscopic observations,

freehand sections of the specimens were mounted in water, 3% (w/v) KOH, and 30%

ethanol solution on glass slides. Forty randomly selected basidiospores were measured

under a light microscope at 1000x magnification. Length measurements excluded the

apiculus; average dimensions + standard deviations are shown in brackets. The surface

features of basidiospores were also observed by scanning electron microscopy (SEM).

For SEM, a small portion of the gleba was dusted onto double-sided adhesive tape on a

specimen holder, coated with platinum-palladium using an E-1030 Ion Sputter Coater

(Hitachi, Tokyo, Japan), and examined with a S-4200 SEM (Hitachi, Tokyo, Japan)

operating at 20 kV.

Geastrum species new to Japan... 3

FIGURE 1. Geastrum campestre: macrocharacters (TNS-F-38710). A: Detailed structure of

expanded basidiomata. Note plicate peristome (black arrow) and short stalk (white arrow). B: The

mycelial layer of expanded basidiomata. C: Expanded and an unexpanded (arrow) basidiomata.

Scale bars = 20 mm.

Taxonomy

Geastrum campestre Morgan, Amer. Nat. 21: 1027, 1887, as “Geaster

campester’. Figs. 1-2

JAPANESE NAME: Hamabe-no-hida-tsuchigaki (coastal dune’s plicate earthstar, newly

named here).

UNEXPANDED BASIDIOMATA hypogeous to subhypogeous, depressed globose,

globose to subglobose when young, ca. 5-12 mm diam., surface encrusted

4 ... Kasuya & al.

with sand, whitish to slightly cineraceous. EXPANDED BASIDIOMATA 8-32 mm

across when dried, 15-37 mm across when wetted, exoperidium splitting into

6-8 rays, weakly to strongly hygroscopic, rays arched with straight to slightly

recurved tips in fresh state or when wetted, then partly or entirely covering

the endoperidial body when old or dried. MyceLiaL LAYER thin, whitish,

encrusted with sand and plant debris, persistent, attached to the fibrous layer for

a long time and without forming basal remnants attached to the basidiomata.

FIBROUS LAYER outer side almost completely covered with the mycelial layer,

whitish to grey. PSEUDOPARENCHYMATOUS LAYER persistent, pale beige in fresh

state, later becoming beige, reddish brown to dark brown. ENDOPERIDIAL BODY

stalked, depressed globose, globose to subglobose, 3-11 mm diam., with or

without apophysis. ENDOPERIDIUM pale brown, greyish brown to dark brown,

finely pruinate or almost smooth when old, but usually distinctly pruinose

or tomentose with whitish to beige crystalline material in fresh basidiomata,

with an indistinct circular area surrounding the peristome. PERISTOME

strongly plicate with 13-22 folds, almost concolorous or somewhat darker than

endoperidium, broadly conical to mammiform, distinctly delimited, 1-2.5 mm

long. STALK short but distinct, 0.5-1 mm long. COLUMELLA globose, ovoid to

club-shaped in mature state, persistent, distinct. MATURE GLEBA olivaceous

brown to brown.

MYCELIAL LAYER consisting of dimorphic hyphae: (1) 2-5 um thick, hyaline,

thick-walled, rarely branched; (II) 1.5-3.5 um thick, hyaline, thin-walled, with

clamp-connections, rarely branched. FiBRouUSs LAYER consisting of 2.5-6.5

um thick, hyaline, thick-walled hyphae. PsEUDOPARENCHYMATOUS LAYER

consisting of hyaline, yellowish brown to pale brown, thick-walled, almost

bladder-like but variously shaped cells. ENDOPERIDIUM consisting of dimorphic

hyphae: (I) 3.5-4.5 um thick, yellowish brown, brown to dark brown, thick-

walled, rarely dichotomously branched; (II) 2-4.5 um thick, hyaline to pale

yellowish brown, thin-walled, branched. HyPHAE OF THE COLUMELLA 2-6.5

um thick, hyaline, pale yellowish brown to pale brown, thick-walled, rarely

dichotomously branched. HyPHAE OF THE CAPILLITIUM hyaline to yellowish

brown, thick-walled, 2-8 um thick, tapered gradually towards subacute tips,

dichotomously branched, surface usually encrusted with numerous amorphous

remnants or crystalline materials but sometimes almost smooth. BAsrp1A

not seen. Basip1osporREs globose, densely verrucose, thick-walled, yellowish

brown, 4.1-[4.9+0.4]-5.7 um diam. excluding ornaments, 5-[5.5+0.3]-6.2 um

diam. including ornaments, verrucae conical to columnar-like, < 0.8 um high,

with flat, rounded to subacute apexes, basal apiculus prominent.

HABITAT AND DISTRIBUTION: Uncommon in Japan, where it is gregarious

on sand in coastal dunes in a warm-temperate zone near Carex kobomugi,

C. pumila, and Elymus mollis. Several fresh basidiomata were collected in

Geastrum species new to Japan... 5

FIGURE 2. Geastrum campestre: SEM micrographs (TNS-F-38710). A: Hypha of the capillitium

encrusted with amorphous remnants and crystalline materials. B: A basidiospore covered with

dense verrucae. C: Basidiospores with prominent apiculus. Scale bars: A = 3 um; B = 1.5um;

C=2 um.

September. Known from Japan (Ibaraki, new record), Europe (Sunhede 1989),

South Africa (Smith 1935), North America (Long & Stouffer 1948), Central

America (Esqueda et al. 2003), Hawaii (Gilbertson et al. 2001), and Australia

(Cunningham 1944).

SPECIMENS EXAMINED: JAPAN. IBARAKI PREFECTURE: Hitachinaka-shi, Nagasuna,

Hitachi Kaihin Park: September 26, 2004, H. Sakamoto, TNS-F-38710.

6 ... Kasuya & al.

COMMENTS: Geastrumcampestre is well characterized by hygroscopic exoperidial

rays, a mycelial layer encrusted with sand or debris, a stalked endoperidial body,

and a distinctly delimited, plicate peristome. The morphology of the Japanese

specimens agrees well with previous descriptions of G. campestre (Cunningham

1944, Long & Stouffer 1948, Sunhede 1989). However, the expanded Japanese

basidiomata are usually smaller (< 37 mm wide when wet) than European

specimens (< 65 mm wide, Sunhede 1989).

Geastrum kotlabae and G. pouzarii V.J. Stanék are morphologically and

ecologically very similar to G. campestre. All three species have hygroscopic

exoperidial rays and plicate peristomes and share similar habitats such as well-

drained, sandy soil. However, the endoperidial body of G. kotlabae is completely

sessile (Sunhede 1989, Sakamoto & Kasuya 2008). Basidiospores of G. pouzarii

are larger (5.5-7 um diam. including ornaments, Sunhede 1989) than those of

G. campestre. Moreover, the mycelial layer of both G. kotlabae and G. pouzarii

easily peels off at basidiome expansion (Sunhede 1989, Esqueda et al. 2003)

whereas that of G. campestre remains attached to the fibrous layer for a long

time. Geastrum berkeleyi Massee, which also produces stalked endoperidial

bodies and plicate peristomes, clearly differs from G. campestre by its non-

hygroscopic, much larger basidiomata (Kasuya et al. 2009).

Japanese material of G. campestre has been collected from sand dunes along

the Pacific Ocean coast in the warm-temperate area of Eastern Honshu. The

type specimen of G. campestre was collected from Nebraska, U.S.A., where it

grows on dry open grasslands or on litter under conifers or deciduous trees

(Long & Stouffer 1948). European and Australian specimens have been obtained

from well-drained, sandy soil near coasts (Cunningham 1944, Sunhede 1989),

and the species has been recorded from tropical deciduous forests in Mexico

(Esqueda et al. 2003) and dry mountain forests in Hawaii (Gilbertson et al.

2001). These facts suggest that G. campestre prefers dry, well-drained open land

but can grow in a variety of environments.

Geastrum campestre is known from all continents except Antarctica and its

habitats are diverse. As several morphological variations have been reported

(Sunhede 1989), G. campestre as currently defined may be a species complex.

Geastrum corollinum (Batsch) Hollds, Gasterom. Ung.: 65, 1904, as “Geaster

corollinus”. Fics. 3-4

JAPANESE NAME: Chijire-tsuchigaki (Curled earthstar, newly named here).

UNEXPANDED BASIDIOMATA usually epigeous, rarely subhypogeous,

subglobose, ovoid to onion-shaped with an umbo, 6.5-13 mm wide, 14-17.5mm

high including apical umbo, surface almost smooth to minutely wrinkled and

not encrusted with sand or plant debris, ochraceous to pale brown. EXPANDED

BASIDIOMATA 10-24 mm across when dried, 16-28 mm across when wetted,

Geastrum species new to Japan... 7

FIGURE 3. Geastrum corollinum: macrocharacters (TNS—F-38711). A: Two expanded basidiomata

with distinctly delimited, fibrillose peristomes. B: Two unexpanded basidiomata with almost

smooth surfaces. Note apical umbos (arrows). C: Expanded basidiomata showing hygroscopic

exoperidial rays. Scale bars: A = 10 mm; B-C = 15 mm.

exoperidium splitting into 5-10 rays, hygroscopic, partly or entirely covering

the endoperidial body and thus curled when dried, tips of the rays sometimes

become recurved downwards in fresh state. MYCELIAL LAYER thin, yellowish

8 ... Kasuya & al.

white, pale ochraceous to pale brown, smooth, without sand or plant debris,

visible when unexpanded, but soon disappeared when basidiome expand.

FIBROUS LAYER firm, persistently attached to the pseudoparenchymatous

layer, outer side ochraceous to pale brown. PSEUDOPARENCHYMATOUS LAYER

persistent, thin (>0.5 mm thick), pale beige in fresh state, later becoming

ochraceous, reddish brown to dark brown, coriaceous when wetted or young,

very hard when dried or old. ENDOPERIDIAL BoDy sessile, depressed globose,

globose to subglobose, 6-11.5 mm diam., without apophysis. ENDOPERIDIUM

pale brown to greyish brown, almost smooth when old, but usually pruinate with

whitish crystalline material in fresh or young basidiomata, with a circular area

surrounding the peristome. PERISTOME surface fibrillose, almost concolorous

or somewhat darker than endoperidium, broadly conical to mammiform,

distinctly delimited, 1-3.5 mm long. COLUMELLA cylindrical to club-shaped,

slender, sometimes indistinct. MATURE GLEBA Olivaceous brown to brown.

MYCELIAL LAYER consisting of dimorphic hyphae: (I) 2-4.5 um thick,

hyaline, thick-walled, rarely branched; (II) 2-3.5 um thick, hyaline, thin-

walled, with clamp-connections, branched. FiBRoUS LAYER consisting of

3.5-7 um thick, hyaline to pale yellowish brown, thick-walled hyphae.

PSEUDOPARENCHYMATOUS LAYER consisting of thick-walled, hyaline, yellowish

brown to reddish brown, almost bladder-like but variously shaped cells.

ENDOPERIDIUM consisting of dimorphic hyphae: (I) 3.5-6 um thick, hyaline

to pale yellowish brown, thick-walled, rarely dichotomously branched; (II)

1.5-4.5 um thick, hyaline to pale yellowish brown, thin-walled, branched.

HYPHAE OF THE COLUMELLA 2-7 um thick, thick-walled, surface smooth,

hyaline to pale yellowish brown. HYPHAE OF THE CAPILLITIUM 2-6.5 um thick,

thick-walled, pale yellowish brown to yellowish brown, curved, tapered gradually

towards subacute tips, rarely dichotomously branched, surface almost smooth

or sometimes encrusted with amorphous remnants and crystalline materials.

BASIDIA not seen. BASIDIOSPORES globose, densely verrucose, thick-walled,

yellowish brown to olivaceous brown, 3.5-[4+0.3]-4.5 um diam. excluding

ornaments, 4-[4.7+0.4]-5.5 um diam. including ornaments, verrucae conical

to columnar-like, < 0.7 um high, with flat, rounded to subacute apexes, basal

apiculus prominent.

HABITAT AND DISTRIBUTION: Uncommon in Japan, solitary to gregarious

on sand near Carex kobomugi and C. pumila in coastal dunes or on the ground

in broad-leaved forests in a warm-temperate area. Fresh and old basidiomata

were collected in July and December. Known from Japan (Shizuoka and Osaka,

new record), China (Zhou et al. 2007), Europe (Hollés 1904, Sunhede 1989),

South Africa (Bottomley 1948; Coetzee et al. 1997), North America (Coker &

Couch 1928), Central America (Esqueda et al. 2003) and Hawaii (Gilbertson

et al. 2001).

Geastrum species new to Japan... 9

FiGuRE 4. Geastrum corollinum: SEM micrographs (TNS-F-38711). A: Hypha of the capillitium

encrusted with amorphous remnants and crystalline materials. B: A basidiospore with apiculus

(arrow). C: Basidiospores with dense verrucae. Scale bars = 2 um.

SPECIMENS EXAMINED: JAPAN. SHIZUOKA PREFECTURE: Iwata-shi, Fukude: December

3, 2003, I. Asai, TNS-F-38711. OSAKA PREFECTURE: Katano-shi, Kaigake-no-michi:

July 31, 2011, Y. Kotera, TNS-F-550009.

CoMMENTs: Geastrum corollinum is diagnosed by hygroscopic exoperidial

rays, a smooth mycelial layer not encrusted with sand or debris, and a distinctly

delimited, fibrillose peristome. The morphology of the Japanese specimens

10 ... Kasuya & al.

agrees well with previous descriptions of G. corollinum (Hollés 1904, Dissing &

Lange 1962, Sunhede 1989). However, the pseudoparenchymatous layer of the

exoperidium of the Japanese material is thinner (< 0.5 mm thick) than that of

European material (0.5-1.5 mm thick, Sunhede 1989).

Geastrum floriforme Vittad. and G. hungaricum are morphologically similar

to G. corollinum. However, G. floriforme basidiomata are hypogeous when

unexpanded and have indistinctly delimited peristomes and the mycelial layer

is encrusted with sand or plant debris (Sunhede 1989). For comparison with

G. hungaricum, see description and comments below. Geastrum arenarium

Lloyd, which resembles G. corollinum macroscopically, differs in its mycelial

layer strongly encrusted with sand or plant debris (Sunhede 1989) and smaller

basidiospores of G. arenarium (3.5-4.5 um diam. including ornaments,

Sunhede 1986).

Japanese specimens of G. corollinum have been collected from coastal

sand dunes along the Pacific Ocean and under broad-leaved forests of warm-

temperate areas of Central Honshu. In Europe, the type locality of G. corollinum,

it has been obtained from various habitats, under deciduous trees (Dissing &

Lange 1962, Sunhede 1989), under Juniperus communis (Sunhede 1989), and

in sandy grasslands (D6rfelt et al. 1979) and coastal dunes (Eyndhoven 1937).

Chinese specimens have been collected from grasslands, grazed fields and under

Cryptomeria japonica or several salicaceous trees (Zhou et al. 2007). Geastrum

corollinum has also been recorded from tropical deciduous forests in Mexico

(Esqueda et al. 2003) and dry, coastal to montane forests in Hawaii (Gilbertson

et al. 2001). Ecologically, these facts suggest that the present fungus adapts to

varied environments. In Japan, further fieldwork is needed to clarify whether

the distribution of G. corollinum is limited to coastal sand dunes or not.

Geastrum corollinum has been recorded from all continents except

Antarctica. Given the variable morphological characters (Dissing & Lange

1962, Sunhede 1989) and diverse habitats, G. corollinum probably comprises a

species complex.

Geastrum hungaricum Hollds, Math. Természettud. Ertes. 19(5): 506, 1901,

as “Geaster hungaricus”. Fics. 5-6

JAPANESE NAME: Arechi-no-himetsuchiguri (dry wastelands earthstar, newly named

here).

UNEXPANDED BASIDIOMATA hypogeous to subhypogeous, depressed globose,

subglobose to ovoid, ca. 3-10 mm diam., surface encrusted with sand and plant

debris, whitish to slightly cineraceous. EXPANDED BASIDIOMATA 5-12 mm

across when dried, 13-17 mm across when wetted, exoperidium splitting into

6-11 rays, strongly hygroscopic, partly or entirely covering the endoperidial

body when dried, tips of the rays sometimes become curved downwards in

Geastrum species new to Japan... 11

FIGURE 5. Geastrum hungaricum: macrocharacters. A: An expanded basidioma (TNS-F-38713)

with distinctly delimited, fibrillose peristome. B: Expanded basidiomata (TNS-F-38712) showing

strongly hygroscopic exoperidial rays. Note basal remnants of the mycelial layer attached to

basidiomata with sand and plant debris (arrows). C: An expanded basidioma (TNS-F-38713) in

the natural habitat. Scale bars: A, C= 5 mm; B = 10 mm.

fresh state. MYCELIAL LAYER thin, whitish, with sand and plant debris, easily

peeling off and usually forming basal remnants attached to the basidiomata.

FIBROUS LAYER firmly, persistently attached to the pseudoparenchymatous

layer, outer side white. PSEUDOPARENCHYMATOUS LAYER persistent, cream

to pale beige in fresh state, later becoming pale brown, reddish brown to

12 ... Kasuya & al.

dark brown. ENDOPERIDIAL Bopy sessile, globose, subglobose to ovoid, 2-9

mm diam., without apophysis. ENDOPERIDIUM pale brown to greyish brown,

almost smooth when old, but usually pruinate with whitish to beige crystalline

material in fresh basidiomata, with a circular area surrounding the peristome.

PERISTOME surface fibrillose, almost concolorous or somewhat darker than

endoperidium, discoid to broadly conical, distinctly delimited, 0.5-1.5 mm

long. COLUMELLA cylindrical, slender, sometimes indistinct. MATURE GLEBA

olivaceous brown to brown.

MYCELIAL LAYER consisting of dimorphic hyphae: (I) 2-5.5 um thick,

hyaline, thick-walled, rarely branched; (II) 2-5 um thick, hyaline, thin-

walled, rarely with clamp-connections, sparsely branched. FIBROUS LAYER

consisting of 3-7.5 um thick, hyaline to pale yellowish brown, thick-walled

hyphae. PSEUDOPARENCHYMATOUS LAYER consisting of thick-walled, hyaline

to pale brown, almost bladder-like but variously shaped cells. ENDOPERIDIUM

consisting of 2.5-5 um thick, hyaline, pale yellowish brown to pale brown,

thick-walled hyphae encrusted with amorphous crystalline materials. HYPHAE

OF THE COLUMELLA 1-6 um thick, thick-walled, surface smooth, pale

yellowish brown. HYPHAE OF THE CAPILLITIUM 1-8 um thick, thick-walled,

pale yellowish brown to yellowish brown, tapered gradually towards subacute

tips, sometimes dichotomously branched, surface almost smooth or encrusted

with some amorphous remnants and crystalline materials. BastD1A not seen.

BasIDIospoREs globose, warty, thick-walled, olivaceous brown to dark brown,

4.3-[4.9+0.3]-5.3 um diam. excluding ornaments, 4.8-[5.4+0.3]-5.8 um

diam. including ornaments, warts radiating from the base of the apiculus and

connecting to adjacent verrucae thus showing wing-like, < 0.5 um high, with

rounded to subacute apexes, basal apiculus prominent.

HABITAT AND DISTRIBUTION: Uncommon in Japan, gregarious or solitary

on sand among mosses near Elymus mollis in coastal dunes in a cool-temperate

area. One fresh basidioma was collected in September; several overwintered,

old specimens were found in May. Known from Japan (Hokkaido, new record),

Mongolia (Dorfelt & Otto 1985), Russian Karachay-Cherkessia (Hollés 1904),

Hungary (Hollés 1904), Czech Republic (Stanék 1958), and Germany (Dorfelt

et al. 1979).

SPECIMENS EXAMINED: JAPAN. HOKKAIDO: Shari-gun, Shari-cho, Minehama: May 29,

2009, T. Hoshino and A. Uchida, TNS-F-38712; September 27, 2010, A. Uchida, TNS-

F-38713.

COMMENTS: Geastrum hungaricum, one of the smallest Geastrum species of

the genus, is well characterized by strongly hygroscopic exoperidial rays, an

endoperidium with a pruinose surface with whitish to beige crystalline material,

and a distinctly delimited, fibrillose peristome. The morphology of the Japanese

specimens agrees well with previous descriptions of G. hungaricum (Holl6s

Geastrum species new to Japan... 13

FIGURE 6. Geastrum hungaricum: SEM micrographs. A: Hypha of the capillitium sparsely

encrusted with amorphous remnants and crystalline materials (TNS-F-38712). B: A basidiospore

covered with dense verrucae (TNS-F-38712). C: A basidiospore with prominent apiculus

(TNS-F-38713). Note wing-like verrucae radiating from the base of the apiculus. Scale bars: A=3 um;

B-C = 1.5 um.

1904, Stanék 1958, Sunhede 1989), except that the basidiospores are slightly

narrower than in European materials (5-6 um diam. including ornaments,

Sunhede 1989).

14 ... Kasuya & al.

Two other small-sized earthstars, G. corollinum and G. floriforme, resemble

G. hungaricum. However, unexpanded basidiomata of G. corollinum are

epigeous, onion-shaped with an umbo, with brownish surfaces, and not

encrusted with sand or plant debris (Sunhede 1989; see also our description

of G. corollinum, above) and the expanded basidiomata are larger (< 23 mm

diam., Sunhede 1989). Geastrum floriforme clearly differs from G. hungaricum

by its indistinctly delimited peristome, smooth endoperidial surfaces, and

larger basidiospores (< 7 um diam., Sunhede 1989).

Japanese material of G. hungaricum was collected from a coastal sand dune

along the Sea of Okhotsk in the cool-temperate area of eastern Hokkaido, while

the type was collected from Hungary. In Europe it has been collected from

sandy, rocky or grazed ground (Hollds 1904, Stanék 1958, Sunhede 1989), and

Russian or Mongolian specimens were also found in semiarid areas (Hollds

1904, Dorfelt & Otto 1985), suggesting that G. hungaricum habitats are limited

to dry, sandy, stony or grazed ground in northern cool-temperate regions.

Acknowledgments

We thank Mr. Ikuo Asai (Saitama, Japan), Dr. Tsuyoshi Hosoya (National Museum of

Nature and Science, Japan) and Mr. Yuzo Kotera (Kyoto, Japan) for their collaborations

and suggestions to our study. The authors are also grateful to Dr. Don Hemmes

(University of Hawaii, U.S.A.) and to Dr. Vagner Gularte Cortez (Universidade Federal

do Parana, Brazil) for their critical reviews of the manuscript. Thanks are also owed to

Dr. Francisco D. Calonge (Madrid Botanical Garden, Spain) for his valuable comments

on this study. This research was financed in part by a JSPS grant-in-aid (No.222635) to

TK and a JSPS grant-in-aid for young scientists B (No. 21770096) to KH. This work is in

partial fulfillment of the requirements for the Ph.D. degree of TK.

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MY COTAXON

http://dx.doi.org/10.5248/118.17

Volume 118, pp. 17-26 October-December 2011

Discrimination of Gigaspora species by

PCR specific primers and phylogenetic analysis

GLADSTONE ALVES DA SILVA’, ERICA LUMINI”?, VALERIA BIANCIOTTO?,

PAOLA BONFANTE” & LEONOR CosTA MaIA?

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

Av. Prof. Nelson Chaves, s/n. 50670-420 Recife, PE, Brasil

*Dipartimento di Biologia Vegetale dell’ Universita and Istituto per la Protezione delle Piante,

(Sezione di Torino) del CNR - Viale P.A. Mattioli 25, 10125 Torino, Italy

*CORRESPONDENCE TO: gladstonesilva@yahoo.com

ABSTRACT — Species of arbuscular mycorrhizal fungi (AMF) are usually identified by

the morphological characteristics of their spores. However, considering the difficulties in

diagnosing their taxa, the construction of species-specific primers has been proposed as an

identification alternative. In this paper the problem of distinguishing different Gigaspora

species with slight morphological differences was solved using species-specific primers and

SSU and LSU rDNA sequence analyses of 18 AM fungal isolates comprising seven species.

Neighbor joining, maximum parsimony, and maximum likelihood analyses were performed

to evaluate the phylogenetic affiliation of the isolates, and a new reverse PCR primer (ALB1)

specific for Gigaspora albida was designed and tested with 11 Gigaspora isolates (four species).

The results confirmed misidentification of “G. albida’ FL 927 and ‘G. margarita’ BR 444 and

supported referring FL 927 to G. rosea and BR 444 to G. albida.

Key worps — phylogeny, ribosomal sequences, Glomeromycota

Introduction

There are approximately 220 species of arbuscular mycorrhizal fungi

(AMF) (Stockinger et al. 2010). Morphological differences in spore structure

are usually used to distinguish between individual species, but this requires

a great deal of experience (Morton 1993, Bentivenga & Morton 1994). More

practical methods are needed, so that AMF can be identified directly not

only in the rhizosphere, but also after host root colonization. DNA analytical

methods involving electrophoretic profiles (Wyss & Bonfante 1993), sequence

comparisons (Daniell et al. 2001, Husband et al. 2002, Mummey & Rillig 2007,

Stukenbrock & Rosendahl 2005), PCR-DGGE (Souza et al. 2004), species-

18 ... Silva & al.

specific primers (Gamper & Leuchtmann 2007, Geue & Hock 2004, Lanfranco

et al. 1995, 1999, 2001, Millner et al. 1998, 2001a, 2001b, Redecker 2000), and

DNA barcodes (Stockinger et al. 2010) are being developed that would allow

identification of individual AMF species in an ecological context. For example,

species-specific PCR primers could be used to discriminate morphologically

similar species or even to identify species within colonized roots. However, to

date only a small number of species-specific probes has been developed.

After Oehl et al. (2008) divided the Gigasporaceae into four families and

six genera, Morton & Msiska (2010) proposed dividing Gigasporaceae into just

three genera —Gigaspora, Racocetra, Scutellospora. Gigaspora has five species

(Bentivenga & Morton 1995) in which the morphological variation is low

(Bago et al. 1998, Souza et al. 2004, Lanfranco et al. 2001), and only diameter,

color and spore wall thickness of glomerospores have been used for species

identification (Bentivenga & Morton 1995). Species-specific primers designed

for G. margarita (Lanfranco et al. 1999) and G. rosea (Lanfranco et al. 2001)

have shown to be effective in solving such morphological conflicts. In this study

we show how a species-specific PCR primer constructed for G. albida helps

solve problems associated with discriminating Gigaspora species and evaluate

Gigaspora phylogenetically through SSU and LSU rDNA analyses.

Materials & methods

AM fungi

Eleven reference isolates (TABLE 1) representing Gigaspora propagated in pot cultures

were selected.

Intracellular bacteria

Presence of intracellular bacteria was assessed with the Live/Dead Bac-Light

bacterial viability kit (Molecular Probes, Inc., Eugene, OR, USA) following Bianciotto

et al. (1996).

DNA extraction

Ten to 50 spores were washed in distilled water, sonicated 3-4 times, crushed in

50-100 ul of lx REDTaq PCR Reaction Buffer (10 mM Tris-HCl pH 8.3, 50 mM KCI, 1.1

mM MgCl, and 0.01% gelatin) (Sigma-Aldrich, Milan, Italy), and centrifuged at 1000

RPM for 2 min. The supernatant containing the DNA was incubated at 95°C for 13 min.

After extraction, the DNA was stored at —20°C.

Design of PCR primers, amplification and sequencing

A new species-specific reverse PCR primer ALB1 (5’-CCCAACTAAAAATACTTCAGTC-

3’) was designed for G. albida based on the GenBank ITS sequence AJ239118 and

combined with GilTS1 (Lanfranco et al. 1999) to create a 385 bp fragment. For

G. margarita and G. rosea, combinations of primers GiITS1+GiITS2 (Lanfranco et al.

1999) and GiITS1-GiR3 (Lanfranco et al. 2001) were used, respectively.

Discrimination of Gigaspora spp. ... 19

TABLE 1. Glomeromycota isolates used in PCR amplification tests with

species-specific primers and phylogenetic analysis.

SPECIES* ISOLATE ORIGIN Source?

Gigaspora albida N.C. Schenck & G.S. Sm. BR 601 Brazil INVAM

G. albida FL 927 USA UFPE

G.decipiens I.R. Hall & L.K. Abbott BEG 45 Australia BEG

G. gigantea (T.H. Nicolson & Gerd.) Gerd. & Trappe MN 414D USA INVAM

G. gigantea NC 150 USA INVAM

G. gigantea WV 932 USA INVAM

G. gigantea NC 199A USA INVAM

G. margarita W.N. Becker & I.R. Hall MAFF 52 Japan MAFE

G. margarita MAFF 54 Japan MAFF

G. margarita BEG 34 New Zealand = BEG/UNITO

G. margarita BR 444 Brazil INVAM

G. rosea T.H. Nicolson & N.C. Schenck UT 102 USA INVAM

G. rosea MAFF 62 Japan MAFF

G. rosea BRI51A Brazil INVAM

G. rosea DAOM 194757 NA NA

Scutellospora calospora (T.H. Nicolson & Gerd.) BEG 32 UK BEG

C. Walker & EE. Sanders

S. calospora HDAM-3 na na

Scutellospora sp. W 3485 UK C. Walker

* Species used in the test with species-specific primers (in bold). "INVAM: International Culture Collection

of Arbuscular and Vesicular-Arbuscular Mycorrhizal Fungi, USA; UFPE: Universidade Federal de

Pernambuco, Departamento de Micologia, Brasil; MAFF: Ministry of Agriculture, Forest and Fisheries,

Japan; BEG European Bank of Glomales, France; UNITO: Universita di Torino, Dipartimento di Biologia

Vegetale, Italy; na: information not available.

The 28G1 and 28G2 primers (Silva et al. 2006) were used to amplify the partial LSU

rDNA region. NS1, NS2, NS3, NS4, NS5 and NS8 were used to obtain the SSU rDNA

amplicons (White et al. 1990).

PCR reactions were carried out in a volume of 50 ul, containing 10 mM Tris-HCl

pH 8.3, 50 mM KCI, 1.1 mM MgCl, 0.01% gelatin, 200 uM each dNTPs, 1 uM of each

primer and 2 units of REDTaq™ DNA polymerase (SIGMA) (Sigma-Aldrich, Milan,

Italy). Cycling parameters were 45 s at 94°C, 1 min at 55°C, 1 min at 72°C for 40 cycles,

with a final elongation of 7 min at 72°C followed the last cycle. The amplicons were

visualized on 1.5% agarose gel with ethidium bromide. The amplified products for

SSU and LSU rDNA were purified with a QIAquick kit (Qiagen S.p.A., Milan, Italy)

following the manufacturer's instruction and sequenced (accession numbers are listed

in TABLE 1). Sequencing was made by GeneLab (Roma, Italy).

Sequence alignment and phylogenetic analysis

Prior to phylogenetic analysis, a BLASTn query of the National Center for

Biotechnology information databases demonstrated that sequences obtained from AMF

species were affiliated with the genus Gigaspora.

20 ... Silva & al.

Sequences of 15 Gigaspora and 11 Scutellospora isolates (some obtained from

GenBank) were selected to align SSU and LSU rDNA (TABLE 1) using Clustal X (Larkin

et al. 2007) and edited with BioEdit (Hall 1999).

For phylogenetic analyses and tree construction, neighbor joining (NJ), maximum

parsimony (MP), and maximum likelihood (ML) analysis with 1,000 bootstrap

replications were performed using PAUP4 (Phylogenetic Analysis Using Parsimony

vers. 4, Swofford 2003). NJ and ML analyses were performed using parameters obtained

from ModelTest 3.7 (Posada & Crandall 1998). Scutellospora calospora and Scutellospora

sp. were used as outgroup.

Results

Morphological observations of isolates FL 927 and BR 444

Isolates FL 927 (G. albida’) and BR 444 (‘G. margarita’) had morphological

characters similar to other Gigaspora species: spore wall thickness [9.6-(12.2)-

14.4 um and 12-(13.9)-16.8 um, respectively] and spore diameter [230-(282)-

360 um and 250-(320)-384 um, respectively]. However, the ‘G. margarita’

BR 444 spores were hyaline/white to yellow and lacked the characteristic green

(G. albida) or pink (G. rosea) coloration observed in reference isolates G. albida

BR 601 and G. rosea UT 102. All isolates of “G. margarita BR 444 and G. albida

BR 601 contained endobacteria, which were absent in all isolates of G. rosea

and ‘G. albida’ FL 927.

Testing species-specific primers for taxa of Gigaspora

Eleven Gigaspora isolates were screened for PCR amplification of a short

partial nuclear ITS rDNA fragment (385bp) with species-specific primers

(TABLE 1). The primers GilTS1+GiITS2 amplified only the G. margarita species,

excluding isolate “G. margarita BR 444, which did not amplify at all (Fic. 1A).

The combination of the species-specific primer constructed for G. rosea (GiR3)

with GiITS1 amplified not only the isolates of G. rosea, but also ‘G. albida’

FL 927 and showed a slight reaction to the DNA from spores of G. albida BR

601 and ‘G. margarita’ BR 444 (Fic. 1B), while the GiITS1+ALB1 primer pair

amplified only G. albida BR 601 and ‘G. margarita’ BR 444 (Fic. 1C). These

results support a misidentification for “G. albida FL 927 and ‘G. margarita’

BR 444.

Phylogenetic analysis of Gigaspora from SSU and LSU rDNA

In the SSU rDNA phylogenetic analyses (NJ, MP and ML), the genus

Gigaspora was supported as a monophyletic group with 100% of the bootstrap

value (Fic. 2). Three subclades were observed in the genus, the first grouping

‘G. margarita BR 444 with all isolates of G. albida and G. rosea. In the

second cluster just G. gigantea was present and in the last group all isolates of

G. margarita (except ‘G. margarita’ BR 444) appeared together with G. decipiens.

In the tree constructed with the LSU rDNA, Gigaspora was monophyletic

with high bootstrap support (Fic. 3). All isolates of G. margarita (except

Discrimination of Gigaspora spp... 21

Fic. 1. Agarose gels (1.5%) with PCR products of the Gigaspora spp. Different combinations of

primers were used: A) GilTS1-GiITS2; B) GilTS1-GiR3; C) GiITS1-ALB1. M = pUC 18 digested

with Hae III. NTC = no template control.

‘G. margarita’ BR 444) clustered together. Two isolates of Gigaspora gigantea

grouped in the same clade, Gigaspora albida BR 601 and ‘Gigaspora margarita’

BR 444 clustered together and Gigaspora rosea UT 102 was close to “Gigaspora

albida’ FL 927. Based on SSU and LSU rDNA analyses ‘Gigaspora margarita BR

444 has been misidentified.

Discussion

Due to the low morphological variation amongst Gigaspora species

(Bentivenga & Morton 1995), some papers have reported species misidenti-

fication (Bago et al. 1998, Souza et al. 2004, Lanfranco et al. 2001). These

studies also report the usefulness of molecular approach as an identification

tool. Based on sequences of SSU rDNA and isozyme analysis, Bago et al. (1998)

divided Gigaspora into three subgeneric groups: the first with G. margarita and

G. decipiens, the second with G. albida and G. rosea, and the third with G. gigantea.

In this paper these same groups were observed in the phylogeny generated by

22 ... Silva & al.

S. calospora AJ306443 BEG32

Scutellospora sp. AJ306437 W3485

G. albida Z14009 FL927

G. margarita AJ852603 BR444

65 G. rosea X58726 DAOM194757

5 G. albida AJ852599 BR601

G. albida AJ852600 FL927

52 G. rosea AJ852608 MAFF520062

68 50

| | G rosea AJ852607 BR151A

G. rosea AJ852606 UT102

G. gigantea AJ852601 MN414D

100) 78 G. gigantea AJ852602 NC150

100) 514

100| 52 | G gigantea EF014362 NC199A

G. gigantea Z14010 WV932

G. margarita AJ852604 MAFF520052

G. decipiens U96146 BEG45

hc G. margarita AJ852605 MAFF520054

8 G. margarita AM181144 BEG34

a G. margarita AM181143 BEG34

0.001

Fic. 2. Phylogenetic reconstruction of the Gigaspora obtained from SSU rDNA sequences (~1700

bp). The NJ and ML analyses were performed with HKY85 + I substitution model. Bootstrap values

(in %) are from neighbor joining (NJ), maximum parsimony (MP) and maximum likelihood (ML)

analyses (1000 bootstraps), respectively. Only topologies with bootstrap values of at least 50% are

shown. (Consistency Index = 0.88; Retention Index = 0.93).

SSU rDNA, and just “G. margarita BR 444 was out of the correct clade. In the

LSU rDNA tree, the isolate BR 444 also grouped together with G. albida BR

601, whereas the isolate FL 927 (identified as G. albida) was clustered with

G. rosea. Bago et al. (1998) observed that the isolates BR 444, identified initially

as G. margarita, and FL 927 (described as G. albida), should instead be ascribed

to G. albida and G. rosea, respectively. The PCR amplification tests with species-

specific primers in this study also indicated that these two isolates should be

reclassified as G. albida and G. rosea. Nevertheless recently Msiska & Morton

Discrimination of Gigaspora spp. ... 23

S. calospora AJ510231 BEG32

S. calospora EU346867 HDAM-3

G. albida AJ852008 FL927

G. rosea AJ852015 UT102

G. albida AJ852007 BR601

G. margarita AJ852012 BR444

G. gigantea AJ852009 MN414D

G. gigantea AJ852010 NC150

G. margarita AJ852013 MAFF520052

G. margarita AJ852014 MAFF520054

G. margarita AJ852011 BEG34

0.01

Fic. 3. Phylogenetic reconstruction of the Gigaspora obtained from LSU rDNA sequences (~ 450

bp). The NJ and ML analyses were performed with GTR substitution model. Bootstrap values (in

%) are from neighbor joining (NJ), maximum parsimony (MP) and maximum likelihood (ML)

analyses (1000 bootstraps), respectively. Only topologies with bootstrap values of at least 50% are

shown. (Consistency Index = 0.93; Retention Index = 0.93).

(2009), based on $-tubulin gene, reported that “G. margarita’ BR 444 did not

group with G. albida or G. rosea, but with G. decipiens.

Using PCR-DGGE analysis, Souza et al. (2004) already observed, as we

confirmed here, that the FL 927 isolate, originally identified as G. albida, should

be ascribed to G. rosea, indicating identification problems. Endobacteria are

usually observed in glomerospores of Gigaspora species, with the exception of

24 ... Silva & al.

G. rosea isolates. Bacteria were not found in “Gigaspora albida’ FL 927, further

substantiating the need for reclassification.

Van Tuinen et al. (1998) were the first to design specific primers for Gigaspora

species (G. rosea) that amplify a region of the LSU rDNA. Because these

authors tested cross-amplification only with species of other AMF genera, it is

not possible to establish whether the primers amplify other Gigaspora species.

Currently there are primers specific for two Gigaspora species (G. margarita

and G. rosea); nevertheless, we report that the reverse primer GiR3 (Lanfranco

et al. 2001) also amplifies G. albida isolates. As Lanfranco et al. (2001) did not

use G. albida and G. decipiens in their analyses with species-specific primers,

it was not possible to establish how specific the primers are to G. rosea and

G. margarita. Of the five known Gigaspora species (Bentivenga & Morton

1995), only G. decipiens was not used in this work.

In summary, our results show that different specific PCR primer pairs

and phylogenetic studies can help discriminate among Gigaspora species

with similar spore morphologies and support the utility of these primers and

phylogenetic analysis for solving problems in identifying groups of species in

Gigasporaceae with a low morphological variation.

Acknowledgements

We are grateful to Dr. Fritz Oehl and Dr. Renata Gomes de Souza for the valuable

comments on the manuscript. L.C. Maia acknowledges the Conselho Nacional de

Desenvolvimento Cientifico e Tecnolé6gico (CNPq) for a research grant. The authors

wish to thank Dr. J. Morton and Dr. M. Saito for providing fungal isolates. We are

grateful to Dr. David Bousfield for the critical reading of the manuscript and appreciate

the corrections by Shaun Pennycook, Nomenclatural Editor, and suggestions by Lorelei

L. Norvell, Editor-in-Chief. This research was funded by the Italian National Council

of Research (CNR), Cassa di Risparmio di Torino and Centro di Eccellenza per la

Biosensoristica Vegetale e Microbica.

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MYCOTAXON

http://dx.doi.org/10.5248/118.27

Volume 118, pp. 27-46 October-December 2011

Contribution to the taxonomy of Bovista in Mexico

SILVIA BAUTISTA- HERNANDEZ’ , TEOFILO HERRERA’,

ELVIRA AGUIRRE-ACOSTA* & MARTIN ESQUEDA?

‘Laboratorio de Micologia, Departamento de Botanica,

Escuela Nacional de Ciencias Biolégicas, IPN,

Prolongacion de Carpio y Plan de Ayala s/n, Distrito Federal 11340, México

*Departamento de Botanica, Instituto de Biologia, UNAM,

Apartado Postal 70-233, Distrito Federal 04510, México

°Centro de Investigacion en Alimentacion y Desarrollo, A.C.,

Apartado Postal 1735, Hermosillo, Sonora 83304, México

CORRESPONDENCE TO *: silbh_11@yahoo.com

ABSTRACT — Specimens of the genus Bovista from eight Mexican herbaria (CESUES, ENCB,

FCME, HEMIM, IBUG, IZTA, MEXU, XAL) were examined and studied, and fifteen taxa

were identified: Bovista aestivalis, B. brunnea, B. dermoxantha, B. dominicensis, B. fusca,

B. grandipora, B. graveolens, B. heterocapilla, B. leacoderma, B. oblongispora var. longispora,

B. oblongispora var. oblongispora, B. plumbea, B. pusilla, B. sempervirentium, and B. tomentosa.

The new information obtained indicates a wider distribution for B. aestivalis, B. dermoxantha,

B. fusca, B. graveolens, B. leucoderma, B. oblongispora var. longispora, B. pusilla, and

B. tomentosa. New records for the Mexican mycobiota are: B. dominicensis, B. grandipora,

B. heterocapilla, B. oblongispora var. oblongispora, and B. sempervirentium.

Key worps — Basidiomycota, taxonomy, gasteroid fungi

Introduction

The genus Bovista Pers. is characterized by globose to pyriform basidiomes

with/without a short pseudostipe and with/without mycelial cords (Pegler et

al. 1995) and a thin, fragile, scaly exoperidium. On maturing the basidiome

may form plates, areolae, verrucae, or scales microscopically composed of

hyphal elements, sphaerocysts, claviform cells (Kreisel 1967), or mycosclereids

(Calonge et al. 2005). The endoperidium is persistent, in consistency

papyraceous, pseudopapyraceous, or rigid, smooth and generally shiny or with

a metallic luster (Coker & Couch 1928), although this does not distinguish

Bovista from other related genera. According to Smith (1951), spores are

28 ... Bautista- Hernandez & al.

smooth or ornamented. Shape varies, from globose, subglobose to oblong, with

a pedicel, which may be short or long, straight or curved, corresponding to

the sterigma of the basidium. The subgleba, present only in some species, has

a filamentous, compact appearance and is generally reduced in size and not

well developed. There are three types of capillitium: Lycoperdon, Bovista, and

intermediate (Calonge 1998; Kriiger et al. 2001).

Bovista, traditionally placed in Lycoperdaceae but currently classified in

Agaricaceae (Kirk et al. 2008), is divided into two subgenera based on capillitial

type. Taxa characterized by a Lycoperdon or intermediate capillitium are placed

in subg. Globaria, while those with either a Bovista or heteromorphic (bovista-

+ intermediate) capillitium belong in subg. Bovista. These subgenera are further

divided into sections and series based on capillitium type, absence or presence

of capillitial pores, and presence or absence of a subgleba (Kreisel 1967).

In Mexico few researchers have addressed the taxonomy of Bovista (Herrera

1959); the genus is generally included in general gasteromycete treatments

(Salcedo & Herrera 1966, Guzman & Herrera 1969, 1973, Rodriguez & Herrera

1970, Urista et al. 1985, Esqueda et al. 1990, 1996, 1998, Pardavé 1991, Pérez-

Silva et al. 1994, Calonge et al. 2004a,b, Ochoa & Moreno 1996, 2006). Bovista

has been confused with Lycoperdon Pers., Disciseda Czern., Calvatia Fr.,

Vascellum F. Smarda, Bovistella Morgan, and Calbovista Morse ex M.T. Seidl,

owing to shared macro- and microscopic characteristics. There are, however,

differences that allow them to be quite easily separated. Confusion surrounding

the correct identification of species has prompted this revision of Bovista with

the aim of contributing to the knowledge of the genus in Mexico.

Materials & methods

Herbarium material was examined from CESUES (Centro de Estudios Superiores del

Estado de Sonora, Unidad Hermosillo), ENCB (Escuela Nacional de Ciencias Bioldgicas,

IPN), FCME (Facultad de Ciencias, UNAM), HEMIM (Herbario Micoldgico de Morelos,

UAEM), IBUG (Instituto de Botanica, Universidad de Guadalajara), IZTA (Facultad de

Estudios Superiores campus Iztacala, UNAM), MEXU (Instituto de Biologia, UNAM),

and XAL (Instituto de Ecologia, A.C., Unidad Xalapa).

Macroscopic characters were transcribed from specimen labels or recorded directly

from the basidiomes. For microscopic examination, temporary preparations were made

using 5% potassium hydroxide (KOH) for observing and measuring basidiospores and

capillitial elements with an optical microscope (OM). Lactophenol cotton blue was used

for observing the paracapillitium. Material was also prepared for observation using a

scanning electron microscope (SEM) in order to properly describe spore ornamentation

and perforations in the hyphal walls of the capillitium.

The keys of Kreisel (1967), Demoulin & Marriot (1981), Calonge (1992, 1998) and

Pegler et al. (1995) were mainly used for species identification.

Bovista in Mexico ... 29

Results

Of the 15 Bovista taxa identified and included in the taxonomic key,

9 represented subgenus Globaria (B. aestivalis, B. dermoxantha, B. dominicensis,

B. grandipora, B. heterocapilla, B. oblongispora var. longispora, B. oblongispora

var. oblongispora, B. pusilla, B. sempervirentium) and 6 subgenus Bovista

(B. brunnea, B. fusca, B. graveolens, B. leucoderma, B. plumbea, B. tomentosa).

Five taxa are new records for the Mexican mycobiota and are described below:

B. dominicensis, B. grandipora, B. heterocapilla, B. oblongispora var. oblongispora,

and B. sempervirentium.

Key to the taxa of Bovista recorded from Mexico

1. Basidiomes with intermediate or Lycoperdon capillitium ...................2-.. 2

li Basidiomes with Bovista type-cap itt Uenn: aces sovarshe ssaracahs seavacche svavarcshe averse oats 11

ZltiterMmediaterty pe capil MAUI ase Tasca se Yun, aaa Tan ahceelgn asa Tge aia Tao nis evans ae eahe eee 3

ZL COP CPAOM TY PO CAPU TT UTNE § oe eb cose ttn wen © epaeye H array W moasigd ¥ sneeteyt Hydttesiey’ Sgpllenen Heat i;

3. Basidiomes with ellipsoid spores, short pedicels .......... 0.0... cece eee eee eee +

BZ BASIGIOMES WIT ClODOSESPOL CS ec apa on at oil Pepe PR ag le net oP Piags Pani oR ceo BP ae 5

4 capil lrtnuir wat PepOres) <6. ac’ Wis wale Wis Mal. Wal els Rk ae B. oblongispora var. longispora

4uCapillitinm: pores abSentt ns nk salen assess B. oblongispora var. oblongispora

5:-Heteromorphic-capillitivi, 5 gu), dios. douhes Sdocbe > Sehe 4 aouchet aul tbe Mate ide 6

5 Intermediate capillitiam. 23 vena. cx veoh see steals shale seo teal ed a's OR Sal OH 7

Ge Spores wath pedicels <alWipttne . 25116525 65 a nn pel nye Selene ele ain B. pusilla

GL Spores gvithsped ice) sree lye (iit, Me ait Med wats Ried wake Weed eats Nin Wale Wd Oates B. heterocapilla

7. Echinulate spores with pedicels:< 34 dim o.<-cta. co lense Yooleiten Holes & B. dominicensis

TE OPOLes WAL STG h VETEUCAS enh Whe, Beals beth y Wades bea Res Wadd se body cite e be wited 8

8. Gaprllituimewithbrancnest . Petey Msctew 1A Ti BeenRe Mey eh Ries ty aetna givens tae 9

§) Capillitinm withoutbranghes, ©... 6... 5 ats <.-scayse test ea eye Sa ee eee ee 10

9. Capillitium with amphiseptal branches, numerous pores,

Sivine-avacHo atesappearanice. ts 4H. dl tal tah Mtoe wlaMals © B. grandipora

9°. Capillitium with subseptal branches, with pores ................ B. dermoxantha

10. Exoperidium verrucae made of sphaerocysts .................0000- B. aestivalis

10’ Exoperidium verrucae made of hyphal elements ............ B. sempervirentium

Lil, Basidiomes medivi:tolargey > So Ati es aie eet mene bE arene oH sn tye eaellgnen weal 12

MeBasidiomesismallteneciyiiny:s SeaMIil 29s fie 08s vel ook hae te noe ee GN od 13

12¢Spores subsloboseto.ovoids pedicels: str dighit-c. bate t Aste eter aloo, B. fusca

12° Spores globose to subglobose, pedicels curved..................04. B. graveolens

LC ail Witney athe pores. sre oie Peg Tors ves Tenants awes late peias Vownaset tae aie mner ate eee 14

13° Capillitium pores absent, endoperidium lead colored ............... B. plumbea

30 ... Bautista-Hernandez & al.

FAP SPOKestovoid. 7. iawn 2; hae tates nae nea hea ae eee eee SS B. tomentosa

14, Spores: Slobose te-SulpSloDose Me. oats aes ha a Ola ode ela ob ele cece Oe seer shes 15

15. Capillitium with thick walls, LH = 0.2-0.5............. 0. esse eee eae B. brunnea

15° Capillitium with moderately thin walls, LH = 0.5-0.7 ............. B. leucoderma

Taxa studied

Subgenus Globaria

Bovista aestivalis (Bonord.) Demoulin, Beih. Sydowia 8: 143, 1979. Figs 1-2

SPECIMENS EXAMINED: MEXICO. HipaLGo: MINERAL DEL CHICO MUNICIPALITY,

near Las Ventanas, El Chico National Park, October 5, 1980 G. Guzman 19099 (ENCB).

JALISCO: SAN DIEGO DE ALEJANDRIA MUNICIPALITY, San Diego de Alejandria-San

Julian Highway, 7.5 km from the town of San Diego de Alejandria, August 30, 2000 O.

Rodriguez 2332 (IBUG). EsTADO DE MExIco: Near Amecameca, August 31, 1969 I. Uribe

11 (ENCB); Near La Marquesa, Miguel Hidalgo National Park, July 5, 1963 E. Gonzdlez

12 (ENCB); ATIZAPAN DE ZARAGOZA MUNICIPALITY, Sayavedra Farm subdivision, June

18, 2000 A. Lopez-Villalobos 24 (XAL). Oaxaca: San Pedro Macuiltianguis, June 15,

1984 A. Gonzdlez (XAL).

ComMENTs — ‘this species is characterized by a compact subgleba, evident or

barely developed. Unlike Bovista dermoxantha, B. aestivalis has an intermediate

type of capillitium and under the OM the spores show an ornamentation that

is not very evident. Pegler et al. (1995) mention that it is a very polymorphic

species and thus taxonomically difficult to identify.

Bovista aestivalis has been cited for Baja California (Ochoa & Moreno 2006),

Chihuahua (Moreno et al. 2010), Estado de México, Oaxaca and Veracruz

(Calonge et al. 2004b). This study extends its distribution range to the states of

Hidalgo and Jalisco.

Bovista dermoxantha (Vittad.) De Toni, Syll. Fung. 7: 100, 1888. Figs 3-4

SPECIMENS EXAMINED: MEXICO. Baya CALIFORNIA: km 2, close to the turnoff from

Ensenada to the San Felipe highway, on the way to Hanson Lagoon, in the foothills of

the Juarez range, March 02, 1984 G. Guzman 24326-A (XAL). CHIHUAHUA: TEMOSACHI

MunicIPatity, Nabogame, July 31, 1987 J. E. Laferriére 624 (XAL); 65 km NW of

Janus, September 20, 1972 P. Huerta (ENCB). Hipaueo: San Gregorio, north side of

Xihuingo Hill, July 26, 1963 G. Guzmdn 3865 (ENCB). EstaDo DE MExiIco: Close to

the Guadalupe Dam, July 02, 1967 G. Guzman 5862 (ENCB). Sonora: km 12 on the

Santa Rosa-Yécora highway, August 6, 1989 A. Aparicio 4 (MEXU 22589). TLAXCALA:

HUAMANTLA MUNICIPALITY, 2 km south of La Malinche mountain, July 19, 1987 Mata

217 (XAL). VERACRUZ: Montepio, June 19, 1969 R. Singer & T: Herrera (MEXU 6897);

IZTACZOQUITLAN MUNICIPALITY, in the region of Metlac Gorge, August 09, 1995 P

Navarro 1042; September 13, 1996 P Navarro 1095 (XAL).

CoMMENTS — Pegler et al. (1995) mention that this species is close to B. aestivalis

but differs by its conspicuously ornamented spores and the Lycoperdon type

capillitium.

Bovista in Mexico ... 31

Fics. 1-6. Bovista aestivalis: 1. Spores. 2. Pitted capillitium. Bovista dermoxantha: 3. Spores.

4, Pitted capillitium. Bovista dominicensis: 5. Spores. 6. Spores ornamentation detail. Scale bars:

1-4,6=1um;5=5 um.

Bovista dermoxantha has been reported for the states of Baja California

(Calonge et al. 2004b; Ochoa & Moreno 2006), Chihuahua, Tlaxcala and

Veracruz (Calonge et al. 2004b). This study extends its distribution range to

Hidalgo, Estado de Mexico and Sonora.

Bovista dominicensis (Massee) Kreisel, Feddes Repert. 69: 202, 1964. Fics 5-6

Basidiome 12-37 mm, globose to flattened-globose, exoperidium forms

small pyramidal scales like spines, blackish-brown to greyish in color, formed

by sphaerocysts that are reddish-brown in KOH; endoperidium whitish, of

intertwined hyaline hyphae. Gleba white to olive green. Subgleba compact.

32 ... Bautista-Hernandez & al.

Lycoperdon type capillitium, hyphae thin walled, 4-5 um, light brown

in color, pores absent, barely branched, septa scarce. Paracapillitium absent.

Spores globose, 4-5 um, equinulate, hyaline. Pedicel 13-30(-34) x 1 um,

straight, hyaline.

ECOLOGY & DISTRIBUTION — Gregarious, humicolous in cloud forests,

Pinus-Quercus in transition with cloud forest and tropical vegetation with

columnar cacti, at an altitude of 1270-1550 m.

SPECIMENS EXAMINED: MEXICO. PugEBra: La Magdalena Farm, S of Teziutlan, October

14, 2004 E. Gandara 1161. SINALOA: El Fuerte road to stones with hieroglyphics, after

the El Fuerte River, August 24, 2004 G. Guzmdn 36091. VERACRUZ: Surroundings

of the CONECALLI-DIF social services building, km 2.5 Xalapa-Coatepec Old

Highway, September 7, 1993 D. Fernandez 38; X1co MUNICIPALITY, road from Xico to

Matlalapa, before arriving at Xico Viejo, September 3, 2000 D. Jarvio 677; BANDERILLA

Municipa.ity, La Martinica Hill, July 01, 2004 V. Ramirez-Cruz 94; La Martinica Hill,

August 21, 2006 G. Guzman 36569; COATEPEC MunIcIPALITY, Zoncuantla, royal road

from Xalapa to Coatepec near km 6 of the Xalapa-Coatepec Old Highway (residence),

September 18, 1998 G. Guzman 32468; September 20, 1998 G. Guzman 32482; September

29, 1998 G. Guzman 32666; December 30, 1998 G. Guzmdn 32771; June 24, 1999

G. Guzman 32995; June 27, 1999 G. Guzman 33022; June 01, 2000 G. Guzman 33495;

July 13, 2003 G. Guzman 35488; October 03, 2003 G. Guzmdn 35568; November 02,

2003 G. Guzman 35704; October 02, 2004 G. Guzman 36119; G. Guzman 36122; October

05, 2004 G. Guzmdn 36125; G. Guzman 36127; October 07, 2004 G. Guzman 36133;

G. Guzman 36277; July 10, 2005 G. Guzman 36228; July 26, 2005 G. Guzman 36311;

G. Guzman 36328; October 23, 2005 G. Guzman 36367; July 02, 2006 G. Guzman 36440;

September 15, 2006 G. Guzman 36678; XALAPA MUNICIPALITY, FJ. Clavijero Botanical

Garden, km 2.5 on the Old Highway to Coatepec, June 03, 1995 Garcia- Velazquez 779;

July 03, 1995 Garcia- Velazquez 780; July 10, 2003 FE. Escalona 196; September 30, 2004

E. Gandara 892; October 07, 2004 E. Gandara 932; V. Ramirez-Cruz 189 (XAL).

ComMENTS — Bovista dominicensis, is characterized microscopically by globose

and echinulate spores with long pedicels from 13 up to 34 um. Macroscopically

it can be confused with Lycoperdon because the basidiome is pyriform owing to

the presence of a pseudostipe.

Part of the material examined had been identified as B. longissima Kreisel

and thus reported as a new record for the Americas (Calonge et al. 2004b).

However, B. longissima has verrucose spores and pedicels < 38 um long and to

date it has been recorded only from Japan (Kreisel 1967). Spore ornamentation

places it in B. dominicensis, as it is the same as that in the SEM images shown in

the study by Calonge et al. (2005). Although those authors describe the spores

as verrucose, Kreisel (1967) described them with fine verrucae to delicate

spines. ‘The pedicels of the examined specimens were longer (< 34 um) than

those reported by Kreisel (10-21 um) and Calonge et al. (20 um).

Worldwide, B. dominicensis has been reported for Brazil (Kreisel 1967,

Homrich 1969 [as Lycoperdon dominicensis], Trierveiler et al. 2010), the United

States of America, the Dominican Republic (Kreisel 1967), Colombia (Guzman

Bovista in Mexico ... 33

et al. 2004), and Costa Rica (Calonge et al. 2005). This is the first report of

B. dominicensis for Mexico, where it has been recorded from the states of

Puebla, Sinaloa and Veracruz.

Bovista grandipora Trierv.-Per., Kreisel & Baseia, Mycotaxon 111: 415, 2010.

Fics 7-8

Basidiome 10-12 mm in diameter, globose. Exoperidium white, breaking

into small scales, furfuraceous in appearance, made up of short fragments of

hyphae, orange-red color in KOH. Endoperidium greyish brown to dark brown.

Gleba white to mustard yellow. Subgleba absent. Mycelial cord radicant.

Lycoperdon type capillitium, with amphiseptal septa, pores abundant, giving

a vacuolate appearance. Globose spores, 4—5 tm, verruculose, greenish-yellow.

Short pedicel, 0.5-1 tm, hyaline.

ECOLOGY & DISTRIBUTION —Terricolous, gregarious in the grass. Found in

open Juniperus deppeana forest and in tropical vegetation at altitudes of 2500

and 1500 m, respectively.

SPECIMENS EXAMINED: MEXICO. JALISCO: SAN PEDRO TESISTAN MUNICIPALITY,

highway to Azuayo, W side of the Chapala Lagoon, July 07, 1974 G. Guzman 11582-B

(ENCB); ZAPOPAN MUNICIPALITY, 2 km NW of La Venta, September 27, 1970 D. Garcia

452 (IBUG). PUEBLA: Santa Cruz, between San Martin Texmelucan and La Blanca,

August 25, 1956 T. Herrera (MEXU 901).

CoMMENTS — ‘The specimens examined coincide with the description of

Bovista grandipora given by Trierveiler-Pereira et al. (2010), Microscopically,

B. grandipora is characterized by a capillitium of the Lycoperdon-type with

numerous pores, 0.5-2.4 um, round to ellipsoid or irregularly shaped, and

globose, verruculose with short pedicel (0.5-1 um) basidiospores.

Worldwide, this species has been recorded for Brazil, Cuba, Dominican

Republic, India, Japan, Nepal, Puerto Rico and Spain (Trierveiler-Pereira et al.

2010). This is its first record for Mexico, specifically for the states of Jalisco and

Puebla.

Bovista heterocapilla Kreisel, Beih. Nova Hedwigia 25: 224, 1967. Figs 9-11

Basidiome globose, subglobose to turbinate. Endoperidium papyraceous,

smooth, brown in color, made of intertwined hyphae. Exoperidium is white,

ephemeral, persistent small spines (spinulose), blackish-brown, made up

of sphaerocysts. Subgleba absent or if present, compact, olive brown to dark

brown.

Bovista type or heteromorphic capillitium in the center of the gleba, hyphae

with pseudosepta frequent, true septa and pores absent. Spores 5-6 um,

verrucose, pedicellate. Pedicel 8-17 um, straight, hyaline.

ECOLOGY & DISTRIBUTION — Gregarious, terricolous in Abies, Abies-Quercus

and cloud forest at an altitude of 2330 to 2800 m.

34 ... Bautista-Hernandez & al.

SPECIMENS EXAMINED: MEXICO. Distrito FEDERAL: Contreras, Cerro Tarumba,

October 21, 1956 T: Herrera (MEXU 1426). Hipateco: El Chico National Park, Las

Ventanas, September, 1970 R. Ramos 105 (ENCB); El Chico National Park, October

25, 1975 R. Herndndez (MEXU 12871); Piedras Voladas, Real del Monte, June 13, 1982

E. Pérez-Silva & R. Hernandez (MEXU 17283); Pueblo Nuevo, October 08, 1975 E. Pérez-

Silva (MEXU 11312). Estapo DE MExico: Amecameca, September 18, 1963 A. Najera

(MEXU 2761); La Marquesa-Tenango Highway km 6, October 05, 1975 R. Lamothe &

E. Pérez-Silva (MEXU 10163); La Marquesa, October 21, 1973 R. Lamothe & E. Pérez-

Silva (MEXU 8442); Purificacién NE of Texcoco, September 05, 1976 G. Guzman 16477

(ENCB).

ComMENTsS — ‘This species is characterized by a Bovista type capillitium;

heteromorphic in some specimens, i.e., of the intermediate+Bovista type.

Although Kreisel (1967) mentions that it has the Lycoperdon type of capillitium,

the characteristics of the hyphae and the spores match those cited for

B. heterocapilla.

One species close to B. heterocapilla is B. albosquamosa Kreisel. These

are distinguished by the characteristics of the exoperidium and the spore

ornamentation. In B. heterocapilla the exoperidium is uniformly granular-

verrucose and the spores are verrucose, while in B. albosquamosa the

exoperidium is furfuraceous areolate and the spores are dotted or verruculose

(Kreisel 1967).

Bovista heterocapilla has been reported only for Jamaica (Kreisel, 1967). This

is the first record for Mexico.

Bovista oblongispora var. longispora (Kreisel) A. Ortega & Buendia, Cryptogamie,

Mycol. 6: 285, 1985. Fics 12-13

Basidiome 13-20 x 10-15 um, subglobose to slightly pyriform; when young,

the exoperidium is reddish in color, later it disintegrates, leaving reddish-brown

scaly remains, exposing an ochraceous endoperidium, shiny and papyraceous in

consistency. Gleba whitish, yellowish to dark chocolate brown, fleshy-alveolate

to powdery in consistency. Subgleba present.

Intermediate type capillitium, hyphae 4-5(-6) um, pores moderately

frequent, wall thin, septate, with dichotomous branching. Paracapillitium

absent. Spores 5-7 x 3-4 um, ellipsoid to cylindrical, verrucose; verrucae

irregularly distributed. Pedicel 1-2.5 um, occasionally up to 8 um.

ECOLOGY & DISTRIBUTION — Gregarious, terricolous in Quercus—Pinus

forest, cloud forest, pastures and coffee plantations at altitudes of 1250-2300 m.

SPECIMENS EXAMINED: MEXICO. PuEBLA: Close to the Ocochiguaya Gorge, E side

of Chignahuapan, July 12, 1975 Herrera, Hernandez, Cruz & Guzman (MEXU 25336).

SONORA: YECORA MUNICIPALITY, km 3.4 along the Yécora- Las Cabafias road, September

12, 1996 M. Esqueda, A. Armenta, A. Nufiez & R. Santos (CESUES 3181); September

13, 1996 (CESUES 3013); Onavas MunIcIPALITY, km 204.5 on the Hermosillo-Yécora

road, October 06, 1995 E. Pérez, T. Herrera, M. Esqueda, A. Armenta, A. Nunez,

R. Rodriguez & R. Santos (CESUES 2107). VERACRUZ: Highway to Fortin, 10 km from

Bovista in Mexico ... 35

Fics. 7-13. Bovista grandipora: 7. Spores. 8. Pitted capillitium. Bovista heterocapilla: 9. Spore under

SEM. 10. Spores under OM x1600. 11. Capillitium and spores under OM x400. Bovista oblongispora

var. longispora: 12. Spores. 13. Pitted capillitium. Scale bars = 1 um.

Huatusco, July 20, 1983 S. Chacon 1189 (XAL); highway to Fortin, Tepampa turnoff, 10

km from Huatusco, July 27, 1983 G. Guzman 23527 (XAL); COATEPEC MUNICIPALITY,

Zoncuantla, close to the road to Coatepec, August 15, 1994 G. Guzman 30930 (XAL).

Comments — For the identification of this species the criteria of Ortega &

Buendia (1985) were applied. They propose that B. longispora and B. oblongispora

36 ... Bautista-Hernandez & al.

are varieties of a single species. They argue that the difference between the two

lies in the respective presence or absence of pores in the capillitium.

In his monograph, Kreisel (1967) treats the two taxa as separate species,

stating that B. longispora has a Lycoperdon type capillitium with pores and

compact subgleba, while B. oblongispora has an intermediate type capillitium,

no pores, and no subgleba.

Bovista oblongispora var. longispora has previously been reported from

Mexico as B. longispora (Calonge et al. 2004b; Esqueda et al. 1996; 1998 and

Pérez-Silva et al. 1994).

Bovista oblongispora (Lloyd) Bottomley, Bothalia 4(3): 580, 1948

var. oblongispora Fics 14-15

Basidiome globose to subglobose 17-18 mm in diameter. Exoperidium

formed by small needles separated by approximately 0.5 mm, comprised of

hyphae and sphaerocysts; endoperidium brown in color to dark chocolate

brown, membranous. Gleba powdery, yellowish-brown to dark brown in color.

Subgleba yellowish-brown, compact.

Intermediate type capillitium, hyphae 6-7(-8) um, pores absent, wall thin,

dichotomous branching. Paracapillitium absent. Basidiospores 5-7 x 2.5-3

um, oblong to ellipsoid, verrucose, with a regular longitudinal arrangement.

Pedicel 0.5 um, hyaline.

ECOLOGY & DISTRIBUTION — Solitary, terricolous in cloud forest at an

altitude of 1300 m.

SPECIMEN EXAMINED: MEXICO. VERACRUZ: XALAPA MUNICIPALITY, km 2.5 on the

Xalapa-Coatepec Old Highway, Francisco Javier Clavijero Botanical Garden, May 14,

2005 Gandara 1268 (XAL).

ComMENTs — ‘his variety can be distinguished from B. oblongispora var.

longispora by the absence of pores and the generally predominant ellipsoid

shape of the spores. This is the first record for Mexico.

Bovista pusilla (Batsch) Pers., Syn. Meth. Fung. 1: 138, 1801. Fics 16-17

SPECIMENS EXAMINED: MEXICO. JALISCO: ZAPOTLAN EL GRANDE MUNICIPALITY, the

foothills of Nevado de Colima, Las Viboras track, Floripondio, June 27, 1999 Montoya-

Fuentes 14 (IBUG). Estapo DE MExico: La Marquesa, October 18, 1976 A. Gonzdlez

(FCME 496); NauUCALPAN MuNICcIPALITY, Los Remedios, October 7, 1956 T. Herrera &

O. Sanchez (MEXU 1416). SonorA: YECORA MunIcIPALiTy, km 258 on the highway

from Hermosillo to Yécora, October 5, 1995 E. Pérez-Silva, T. Herrera, M. Esqueda,

A. Armenta, A. Nufiez, R. Rodriguez & R. Santos (CESUES 2073).

Comments — For B. pusilla, we do not follow the species concept of Kreisel

(1967), who characterizes this species by small basidiomes with a Lycoperdon-

type capillitium and non-pedicellate spores, characters not seen in the recently

examined material. Larsson et al. (2009) amply discuss the taxonomic,

Bovista in Mexico ... 37

Fics. 14-19. Bovista oblongispora var. oblongispora: 14. Spores under SEM. 15. Spores under OM

1000. Bovista pusilla: 16. Spores under SEM. 17. Spores under OM x1000. Bovista sempervirentium:

18. Spores. 19. Pitted capillitium. Scale bars = 1 um.

ecological, and phylogenetic aspects of B. pusilla that were considered in the

determination of the studied materials: finely warted spores of 4.5-5.5 um,

pedicels of 3-9 um, heteromorphic capillitium, a temperate distribution, and

an association with mosses. These characteristics totally match the studied

material.

Following Kreisel’s (1967) concept, B. pusilla has been cited from seven

Mexican federative entities (Guzman & Herrera 1969; 1973; Rodriguez &

Herrera 1970; Urista et al. 1985; Esqueda et al. 1990; 1996; Pérez-Silva et al.

1994); but following the criteria of Larsson et al. (2009), this species is known

only from the states of Mexico, Jalisco, and Sonora.

38 ... Bautista-Hernandez & al.

Worldwide, B. pusilla sensu Kreisel (1967) is widely distributed in Europe

and America and sensu Larsson et al. (2009) is registered from the northern

Europe region.

Bovista sempervirentium Kreisel, Beih. Nova Hedwigia 25: 107, 1967. Fics 18-19

Basidiome 10-20 mm in diameter, globose. Exoperidium formed by

small furfuraceous verrucae, comprised of hyphal elements. Endoperidium

pseudopapyraceous, ochre-brown in color. Gleba is olive brown. Subgleba

compact.

Intermediate type capillitium, hyphae 4-5 um, with small pores, without

septa or pseudosepta. Paracapillitium absent. Spores 3.5-5 um, globose,

verruculose, with a short pedicel.

ECOLOGY & DISTRIBUTION — Solitary, terricolous in Abies and Pinus forest,

although Kreisel (1967) reported it for a Quercus virginiana forest.

SPECIMENS EXAMINED: MEXICO. DistRITO FEDERAL: Los Leones Desert, July 02, 1950

T. Herrera (MEXU 1438). Estapo DE MExico: El Carmen Hill, Chimalpa, October 28,

1956 T. Herrera & O. Sanchez (MEXU 1430).

ComMENTS — The examined specimens match B. sempervirentium as described

by Kreisel (1967): they have an intermediate type capillitium with small pores

but without septa or pseudosepta, verruculose spores with a short pedicel, an

exoperidium composed of hyphal elements, and a subgleba.

The species has been reported only for the United States of America (Kreisel

1967), thus this is the first record for Mexico, specifically for the Distrito Federal

and the Estado de México.

Subgenus Bovista

Bovista brunnea Berk., Fl. Nov.-Zel. 2: 189, 1855. Fics 20-21

SPECIMENS EXAMINED: MEXICO. DistRITO FEDERAL: Los Leones Desert, July 10, 1949

M. Ruiz-Oronoz e& T. Herrera (MEXU 1418). VERACRUZ: Las ViGAsS MUNICIPALITY,

Surroundings of El Volcancillo, SW of Encino Gacho, Xalapa-Perote road, November

30, 1991 F. Tapia 922 (XAL).

ComMENTS — ‘This species is characterized by Bovista type capillitium with

pores and no septa and a hyphal-vesiculate exostratum. An exoperidium was

not observed in the examined material; only some amorphous structures could

be seen, possibly owing to the weathering of the specimen. Bovista brunnea

is placed in the Globisporae series, which also includes B. echinella Pat.,

B. verrucosa (G.H. Cunn.) G.H. Cunn., and B. leucoderma. Bovista echinella is

distinguished by a hyphal exostratum and a thin papyraceous endoperidium,

B. verrucosa has a distinctive mycelial cord, and B. leucoderma has a white

exoperidium that forms small plates on the endoperidium and a capillitium

with infrequent septa.

Bovista in Mexico ... 39

Fics S. 20-23. Bovista brunnea: 20. Spores under OM x500. 21. Capillitium and spores x600.

Bovista fusca: 22. Spores. 23. Capillittum under OM x100. Scale bars = 1 um.

Herrera (1959) mistakenly cited B. brunnea for the Estado de México but

later redetermined the collection as B. dealbata [= B. leucoderma] (Herrera

1964, Guzman & Herrera 1969), and thus the B. brunnea record for the state

must be discarded. The only authentic previous record from Mexico is from

Veracruz (Calonge et al. 2004b).

Bovista fusca Lév., Ann. Sci. Nat., Bot., sér. 3, 5: 303, 1846. Figs 22-23

REPRESENTATIVE SPECIMENS EXAMINED (total collections = 103): MEXICO.

DIsTRITO FEDERAL: Monte Alegre, Ajusco Mountains, September 20, 1980 E. Aguirre

(MEXU 17044). DURANGO: PUEBLO NuEVo MuniciPa.ity, Las Bayas Property, Los

Otates stream, July 16, 2009 S. Bautista & E. Aguirre (MEXU 26301). GUERRERO:

CHICHIHUALCO MuNICIPALITY, km 8.5 between El Carrizal and Atoyac, M. Robledo 61

July 12, 1980, (FCME 986). H1ipAeo: Piedras Voladas, Real del Monte, June 13, 1982 E.

Pérez-Silva & R. Hernandez (MEXU 17282). JALISCO: CIUDAD GUZMAN MUNICIPALITY,

Nevado de Colima Mountain foothills, El Floripondio, September 25, 1992 V. Chaparro

3 (IBUG, MEXU 25378). EsTADO DE MEXxICco: JALATLACO MUNICIPALITY, km 13.5

on the Jalatlaco-El Ajusco Highway, August 4, 1986 A. Calderén & E. Aguirre (MEXU

21594). MICHOACAN: Mariposa Monarca Sanctuary, near Angangueo, February 5, 1981

S. Acosta 584 (ENCB). MoRELOs: Zempoala Lagoons, July 27, 1985 A. Calderén (MEXU

19857). OAXACA: 3 km SE of Plan de Guadalupe, La Cafiada, June 13, 1991 Pérez-Silva et

AO ... Bautista-Hernandez & al.

al. (MEXU 22788). PUEBLA: CHIGNAHUAPAN MUNICIPALITY, Apizaco-Chignahuapan

Highway, Cieneguilla Larga, July 13, 2000, F. Tapia 2050 (XAL). TLAxcaLa: San

Salvador, November 13, 1983 C. Martinez (IZTA 2540). VERACRUZ: RAFAEL LUCIO

MuNICcIPALITY, Santa Barbara Farm, 10 km NE of the Xalapa-La Joya Highway, July 8,

1998 F. Ramirez-Guillén 100 (XAL).

ComMENTS — This species is characterized macroscopically by a papyraceous

shiny peridium, especially in mature specimens; on maturing the basidiomes

separate easily from the substrate. Macroscopically it can be confused with

B. graveolens, however the difference lies mainly in the shape of the pedicels.

Some authors have confused B. fusca with B. nigrescens Pers. (Calonge et al.

2004b), although that species occurs only in Europe; Calonge et al. (2005)

verified that the XAL specimens earlier identified as B. nigrescens represent

instead B. fusca.

Observed in some of the examined specimens were clamp connections on

brown dichotomously branching hyphae with thin walls and thin-walled septate

ochraceous hyphae with projections shaped like small tubercles interspersed in

the capillitium. Additionally, the pedicel surface appears smooth under the OM

but rough and wrinkly with the SEM. It is notable that these details have not

been reported in previous studies, although these are not diagnostic, with the

exception of the pedicel, which is unique to B. fusca.

The taxonomy of B. fusca is complex and not very clear, because the material

examined, particularly in the collections of young specimens, was easily

confused with Lycoperdon basidiomes, given that they have a rigid granulose-

squamulose peridium. As mentioned previously, B. fusca specimens are easily

confused with other taxa precisely because our knowledge of this species is so

scarce. Herrera (1959) described Mexican material as a new species, B. ruizii,

which Kreisel (1967) synonymized with B. fusca.

The widely distributed B. fusca is very common in Méxicos temperate forests

and has been recorded for Aguascalientes (Pardavé 1991 [as B. ruizii]), Distrito

Federal, Estado de México (Kreisel 1967), and Chihuahua (Moreno et al. 2010).

This study extends its range to the states of Durango, Guerrero, Hidalgo, Jalisco,

Michoacan, Morelos, Oaxaca, Puebla, Tlaxcala and Veracruz.

Bovista graveolens Schwalb, Lotos N. F. 13: 53, 1893. Fics 24-25

SPECIMENS EXAMINED: MEXICO. Estapo pe México: La Marquesa-Tenango

Highway, August 18, 1979 E. Pérez-Silva (MEXU 16052); ZINACANTEPEC MUNICIPALITY,

km 17 Zultepec-Nevado de Toluca Highway near the town of Raices, June 18, 1993 A.

Castellanos (IZTA 2536). MICHOACAN: ZINAPECUARO MUNICIPALITY, Los Azufres

long lagoon under forest protection, July 25, 1987 Pompa-Gonzdlez 14 (FCME 13906).

TLAXCALA: 2 km south of the La Malinche hostel, La Malinche Hill, July 19, 1987 G.

Mata 200 (XAL). VERACRUZ: La Joya, El Rodeo, February 18, 1985 S. Chacén 3387 B

(XAL); RAFAEL Lucio MunicIPALity, La Joya in front of the “La cabafia del chivo”

restaurant, July 8, 1987 F. Ramirez-Guillén 102 (XAL); X1co MUNICIPALITY, eastern

Bovista in Mexico ... 41

Fics. 24-28. Bovista graveolens: 24. Spores under SEM. 25. Spores under OM x1000. Bovista

leucoderma: 26. Spores. 27. Pitted capillitium. Bovista plumbea: 28. Spore. Scale bars = 1 um.

region of the Cofre de Perote mountain, Los Gallos, 15 km N of the El Rosario sugar

refinery, May 15, 1995 J. Rico 756 (XAL).

COMMENTS — Bovista graveolens is characterized microscopically by the

curved C-shaped pedicels on the majority of its spores that sometimes occur

at the end of a widening corresponding to the basidium wall, although there

are also spores with straight pedicels. The capillitium is very branched and the

secondary hyphae are very long, compared with those of B. fusca. Although this

species is easily confused with B. fusca, because its basidiomes are very similar,

the differences become markedly evident under the microscope.

Bovista graveolens was known only for the state of Tlaxcala (Calonge et al.

2004b), but the present study broadens its distribution to include Estado de

México, Michoacan, and Veracruz.

Bovista leucoderma Kreisel, Feddes Repert. 69: 203, 1964. Fics 26-27

SPECIMENS EXAMINED: MEXICO. Distrito FEDERAL: S. Gonzdlez 97 (ENCB).

H1patGo: Loma Ancha, Marqués Dam, San Bartolo Socalpa, November 06, 1974

42 ... Bautista-Hernandez & al.

R. Cruz (ENCB); OMITLAN DE JUAREZ MUNICIPALITY, Vicente Guerrero Ranch, ahead

of the Real del Monte Mexico-Tampico Highway, short route km 115, July 05, 1986

A. Calderon et al. (MEXU 20142); EPAzoYUCAN PENAS LARGAS MUNICIPALITY, August

03, 1975 M. Medina & I. Garcia 1102 (ENCB); ZEMPOALA MUNICIPALITY, Tepa,

November 20, 1976 J. Baca del Moral (ENCB); San Miguel Regla, August 16, 1980 E.

Pérez-Silva (MEXU 17180). Estapo DE MExico: La Palma Hill, Salazar, October 20,

1957 M. Ruiz-Oronoz (MEXU 141); Between Chalco and Amecameca, September 05,

1965 J. Alvarez (MEXU 19864); km 39 Xochimilco-Oaxtepec Highway, August 24,

1980 E. Fuentes (MEXU 17098); San Pablo Teacalco, NE of Los Reyes, Mexico-Pachuca

Highway, June 30, 1968 J.L. Magaria 38-B (ENCB); Tlamacas, foothills of Popocatépetl,

August 31, 1958 E. Pérez-Silva (MEXU 889); Salazar, October 01, 1961 Herrera,

Zenteno & Riba (MEXU 7805); October 19, 1969 Herrera, Ulloa & Vazquez (MEXU

6772); Purificacién, NE of Texcoco, September 05, 1976 G. Guzman 16482 (ENCB);

TLALNEPANTLA MUNICIPALITY, Guadalupe Lake, September 15, 1968 F. Martinez 61

(ENCB). PuEBLa: Tepeyahualco, May, 1999 L. Mohedano 3 (XAL). VERACRUZ: Foothills

of the Cofre de Perote mountain, October 30, 1982 L. Guzmdn-Davalos 724 (ENCB).

ComMENTS — Bovista leucoderma is mainly characterized by basidiomes with

a white exoperidium that forms small plates in mature specimens. The Bovista

type capillitium contains pores, pseudosepta and true septa.

This species has been cited for the states of Baja California (Ochoa 1993),

Distrito Federal, Hidalgo, Puebla (Guzman & Herrera 1973), and Estado de

México (Kreisel 1967; Guzman & Herrera 1973). Here it is reported for the first

time for the state of Veracruz.

Bovista plumbea Pers., Ann. Bot. (Usteri) 15: 4, 1795. Fic 28

SPECIMENS EXAMINED: MEXICO. Baja CALIFORNIA: Dofia Petra Canyon, N of

Ensenada, January, 1971 S. Guzman del Proo & S. de la Campa (ENCB); El Junco, 15

km NE of Ensenada, February, 1983 N. Ayala 46-B (XAL); Vallecitos, San Telmo to

Observatory of the UNAM road, San Pedro Martir mountains, February 25, 1984 G.

Guzman 24277-A (XAL).

ComMENTs — This species is easily recognized by its white exoperidium which,

when removed, reveals the lead grey endoperidium.

In México, B. plumbea has been recorded only for Baja California (Ochoa

& Moreno 1996, Ochoa et al. 2000); a previous report for Veracruz is

rejected (Calonge et al. 2004b; Ochoa & Moreno 2006) as the cited specimen

(L. Montoya-Bello 698 XAL) has been redetermined to B. fusca. Kreisel (1967)

mentions that B. plumbea is distributed in northern New Zealand and over a

vast region in Europe, Asia, and North America.

Bovista tomentosa (Vittad.) De Toni, Syll. Fung. 7: 97, 1888. Fics 29-32

SPECIMENS EXAMINED: MEXICO. Baja CALIFORNIA: Turnoff at km 55 on the

Ensenada-San Felipe Highway, km 15 road to the Hanson Lagoon, Juarez mountains,

March 2, 1984 Ayala 365; G. Guzmdn 24328-A (XAL). HiDALGo: Surroundings of

San Gregorio, N side of Xihuingo Hill, July 26, 1963 G. Guzman 3875 (ENCB; MEXU

5548); Highway from Pachuca to Venados, turnoff to El Chico, Barranca Oriental, June

Bovista in Mexico ... 43

Fics. 29-32: Bovista tomentosa: 29-31. Spores. 32. Pitted capillitium. Scale bars: 29 = 5 um;

30-32 = 1 um.

24, 1973 G. Guzman 10874 (ENCB). Estapo DE MExico: Satélite City, June 29, 1982

E. Pérez-Silva (MEXU 22489); ZUMPANGO MUNICIPALITY, San Pablo Teacalco, June 30,

1968 E. Quezada-Guzmdn 10 (ENCB).

COMMENTS — Specimens from the XAL herbarium identified as B. minor

Morgan and B. tomentosa by Calonge et al. (2004b) were compared but no

significant microscopic differences were found. Examination of an herbarium

specimen from Belgium identified as B. tomentosa by Demoulin revealed

that the XAL Mexican specimens cited above coincide both macro- and

microscopically with the Belgian material.

Kreisel (1967) placed B. tomentosa (European) and B. minor (North

American) in subg. Globaria sect. Ovisporae because of their ovoid spores and

the Bovista type capillitium with pores. Microscopically, these two species are

very similar, and Kreisel differentiated them based on their depth in the soil: B.

tomentosa has one third of its basidiome immersed in the substrate and grows in

arid dry open regions, while B. minor produces subhypogeous basidiomes with

only the upper part above the soil and grows in humid regions in the shade. We

believe that the relationship between basidiome and substrate is an ecological

character lacking in taxonomic value. Because of the macro- and microscopic

similarities between the two species, we suggest that they are conspecific, with

B. tomentosa as the prior name.

44 ... Bautista-Hernandez & al.

Bovista tomentosa is characterized by the presence of mainly ovoid verrucose

pedicellate spores and a Bovista type capillitium with pores. In some specimens

both the primary and secondary hyphae have pores, while in others only the

secondary hyphae have them.

Bovista tomentosa has been cited for the state of Baja California (Calonge et

al. 2004b). This study extends its distribution range to Estado de México and

Hidalgo.

Acknowledgements

We wish to express our gratitude to Dr. H. Kreisel and Dr. G. Moreno for reviewing

the manuscript and their useful comments. The authors are grateful to the Posgrado

en Ciencias Biolégicas at UNAM, to CONACYT and to PAPIIT IN-218008 and IN-

207311 for the funding awarded to carry out this project; to Berenit Mendoza for taking

the SEM photographs; to the curators of the herbaria: E. Aguirre (MEXU), J. Cifuentes

(FCME), M.L. Coronado (CESUES), I. Frutis (IZTA), G. Guzman (XAL), L. Guzman-

Davalos (IBUG), R. Valenzuela (ENCB) for the loan of material; and to Aldo Gutiérrez

(CIAD) for preparing the plates and formatting the text. Bianca Delfosse translated the

text from the Spanish original.

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MY COTAXON

http://dx.doi.org/10.5248/118.47

Volume 118, pp. 47-52 October-December 2011

Subulicystidium curvisporum sp. nov. (Hymenochaetales,

Basidiomycota) from the Patagonian Andes

SERGIO P. GoRjJON™?, ALINA G. GRESLEBIN’” & MARIO RAJCHENBERG’”

'Centro de Investigacion y Extension Forestal Andino Patagonico,

Area de Proteccién. CC 14, 9200 Esquel, Chubut, Argentina

?Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET) Argentina

*CORRESPONDENCE TO: spgorjon@usal.es

ABSTRACT — Subulicystidium curvisporum is described as a new species from the Patagonian

Andes forests. It differs from its closest related species, S. longisporum and S. perlongisporum,

in considerably longer and characteristically curved basidiospores. A key to all known

Subulicystidium species is included, and the new species is compared with other species in

the genus.

Key worps — Argentina, corticioid fungi, Nothofagus dombeyi, Patagonia, taxonomy

Introduction

The genus Subulicystidium Parmasto comprises 9 described species (Duhem

& Michel 2001, Parmasto et al. 2004). Subulicystidium is an easily distinguished

genus characterized above all by the distinctive cystidia encrusted with

rectangular crystals arranged crosswise to the main axis (“two rows of “ribbon-

like” structures,’ Julich 1975), the presence of repetobasidia, and by the

cylindrical to acicular basidiospores sharing a more or less vermiform shape.

Molecular studies by Hibbett & Binder (2002) and Larsson et al. (2004) show

that Subulicystidium is closely related to Tubulicium vermiferum (Bourdot)

Oberw. ex Jiilich, and both are included in the trechisporoid clade next to other

Trechispora P. Karst. species, even if they do not share the same morphological

and ecological characters.

As a result of collecting trips in the Patagonian Andes forests of southern

Argentina, we have found growing on Nothofagus dombeyi (Mirb.) Oerst.

(Nothofagaceae)aSubulicystidiumspecieswithlongacicularandcharacteristically

curved basidiospores, distinct from previously described species, which we

propose as new below. Some species in Tulasnella J. Schrét. have spirally curved

48 ... Gorjén, Greslebin & Rajchenberg

basidiospores that often produce secondary spores by replication, but basidia

are differentiated by the large globose to clavate sterigmata. We considered the

possibility that a Tulasnella species inhabited the same substrata where the

Subulicystidium grew, but we have not found any trace of a tulasnelloid fungus.

Tulasnella helicospora Raunk., although somewhat inconspicuous, clearly differs

in the grayish pruinose basidiome, more variably shaped wider basidiospores

with homogeneous oily contents that germinate secondary spores, and typical

basidia with obclavate distally tapered sterigmata.

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

cyanophily of basidiospores. The term Q expresses the ratio Length/Width of the

basidiospore size. Line drawings were made with a camera lucida attachment. Specimens

are deposited in the herbarium of the “Centro de Investigacién y Extension Forestal

Andino-Patagonico” (CIEFAP, Esquel, Argentina), BAFC, and SALA.

Taxonomy

Subulicystidium curvisporum Gorjon, Gresl. & Rajchenb., sp. nov. PLATES 1,2

MycoBank MB 561691

Ab Subulicistidium longisporum et Subulicistidium perlongisporum differt basidiosporae

longioribus et recurvis.

Ho.otypus: Argentina, Chubut, Los Alerces National Park, Alerzal milenario

(42°36'34"S 71°53'28"W), 530 m a.s.l., on decayed wood of Nothofagus dombeyi, 3 May

2010, leg. S.P.Gorjon coll. 2689. Holotype, BAFC; isotypes, SALA and herbarium of

Centro de Investigacién y Extensién Forestal Andino-Patagonico.

ETyMOLoGy: curvisporum, Latin, referring to the curved shape of the basidiospores.

BASIDIOMATA annual, fragile, resupinate, effused, arachnoid, smooth and finely

velutinous due to the protruding cystidia, white to light grey, margin abrupt.

HYPHAL SYSTEM monomitic, generative hyphae with clamps, thin- to thick-

walled, 3-4 um wide, smooth or encrusted with amorphous hyaline material.

CYSTIDIA numerous, subulate, 70-80(-100+) x 3-4 um, projecting 60-80 um

above the basidia, encrusted over all the length, except in the smooth apical

part, with rectangular crystals circularly to spirally arranged along the cystidial

wall. BAsIDIA urniform, basally clamped, some of them repetitive, thin-walled,

12-15 x 5-7 um, with 4 stout sterigmata. BASIDIOSPORES long acicular, spirally

curved, with a vermiform shape, smooth, thin-walled, (25-)27-35(-38) x

(1.8-)2-2.5(-2.8) um, inamyloid, nondextrinoid, acyanophilous, guttulate

(Plates 1, 2).

DISTRIBUTION AND EcoLocy —Subulicystidium curvisporum is only

known from the Patagonian Andes forest of Argentina (Valdivian rainforest).

Subulicystidium curvisporum sp. nov. (Argentina) ... 49

PiatE 1. Subulicystidium curvisporum.

Hymenial elements— a) basidiospores, b) basidia, c) cystidia, d) generative hyphae.

coll. S.P.Gorjén 2689, holotype.

50 ... Gorjén, Greslebin & Rajchenberg

PLATE 2. Subulicystidium curvisporum. Basidiospores,

coll. S.P.Gorjén 2689, holotype.

Subulicystidium curvisporum sp. nov. (Argentina) ... 51

It grows on decayed wood of Nothofagus dombeyi, an endemic southern

South American perennial hardwood, in cold rainforests mixed with Fitzroya

cupressoides (Molina) I.M. Johnst. (Cupressaceae), Luma apiculata (DC.) Burret

(Myrtaceae), and Chusquea culeou E. Desv. (Poaceae).

ADDITIONAL SPECIMENS EXAMINED — Subulicystidium curvisporum. ARGENTINA.

Cuusvut: Los Alerces National Park, Alerzal milenario, on decayed wood of N. dombeyi,

3 Mar 2011, leg. S.P.Gorjon, SPG 3023.

Subulicystidium longisporum. ARGENTINA. TIERRA DEL FUEGO: Depto. Ushuaia,

Estancia El Valdez, on dead wood of Nothofagus betuloides and N. pumilio, 4 Mar 1996,

leg. A. Greslebin, AG 340, 382; Monte El Martial, on N. pumilio, 28 Mar 1998, leg.

M. Rajchenberg, MR 11564. CHILE. X REGIon: Puyehue, Entre Lagos, close to Gol-Gol

river bridge, on decayed wood of N. dombeyi, 21 Feb 2010, leg. N. Hallenberg & S.P.

Gorjon, SPG 2600.

Tulasnella helicospora. ARGENTINA. Rio NEGRO: Mallin Ahogado, El Guadal, on

bark of fallen trunk of Austrocedrus chilensis (D. Don) Pic.Serm. & Bizarri (Cupressaceae),

5 Sep 1995, leg. M. Rajchenberg, MR 11015.

ComMENTS — Subulicystidium curvisporum belongs to the group of

Subulicystidium species with long filiform to acicular basidiospores with a

Q > 4. Among them, Subulicystidium longisporum (Pat.) Parmasto, a common

globally distributed species, has the shortest basidiospores with a Q = 4-7. The

closest related species seems to be Subulicystidium perlongisporum Boidin &

Gilles (known from Réunion Island, Tropical Africa, Japan, France, and the

Caucasus) with smaller and straight basidiospores (Boidin & Gilles 1988).

Subulicystidium cochleum Punugu, known from St. Lucia in the Lesser Antilles,

has smaller basidiospores with the distal end slightly folded back or twisted

and curved; S. cochleum differs also in the cystidial ornamentation composed

of long acicular crystals arranged in a cylinder and restricted to the middle part

(Punugu et al. 1980). The remaining species —S. allantosporum Boidin & Gilles,

S. brachysporum (P.H.B. Talbot & V.C. Green) Jiilich, S. meridense Oberw.,

S. naviculatum Oberw., S. nikau (G. Cunn.) Jilich, S. obtusisporum Duhem & H.

Michel— have differently shaped basidiospores, usually reniform or cylindrical

to fusiform and with a Q<4. Limits between some Subulicystidium species are

unclear, and spore size frequently overlaps. As Liberta (1980) already stated, it

is likely that some species belong to a single species complex.

Key to Subulicystidium

bg, Basidiospores were mals Crt. ra.87. a8, OE LAN Oe lee OR Ao OR Oe OR al gt ee alas 2.

1b. Basidiospores fusiform, cylindrical to reniform, Q <4 ................0. 0. eee 5

2a. Basidiospores (9-)12-16(-18) x (1.8-)2-3 um, Q = 4-7 .......... S. longisporum

2beBasidiospores longer es >O8 x31 OR vgs eta Aye et RG NO Ree, AS Rn REA 3

3a. Basidiospores spirally curved, (25-)27-35(-38) um long ......... S. curvisporum

3b. Basidiospores straight or only slightly curved, shorter ....................000. 4

52 ... Gorjon, Greslebin & Rajchenberg

4a. Basidiospores 20-27 x 2-3 um, cystidial encrustation restricted to the middle part

OP ENE CY StI IN Bassa octet tie Nee ete sible sary alle dter tls bap y either tags alla S. cochleum

4b. Basidiospores 16-25 x 1.5-2.3 um, cystidia encrusted over the entire length

SER ae aa UN eta, ge Aha mie A E airet eN Giowl oe el S. perlongisporum

Hake BasicOspores 4a 5 ih WA S:ion A eh MO aM Oo! Ml LAAN Ula! Met MOL CBE So} 6

bb; Basidiospores ups tO Ag Wale... 2 liste. -a tists, ea tidteer att ber 3 ti ber gLite A Bea nes 7

6a. Basidiospores navicular, 10-12 x 4.5-5 um ........ eee eee eee S. naviculatum

6b. Basidiospores phaseoliform, 7-9.2 x 4-5 UM .... 2... eee eee eee eee S. nikau

7a. Basidiospores :(2;5=)3—4-ymk Wide... 5 ios ete kb ec Ed eee hd eee tie gene to 8

7beBasidigsporesO=2. 5a )auim Wide 37 eia.ct fist det we) eee Bh ee 9

8a. Basidiospores shorth cylindrical, 6-8 x 3-3.6 um.............. S. allantosporum

8b. Basidiospores cylindrical to fusoid, slightly curved,

(9-)11-13(-15) x (2.5-)3-4 um oo. eee eee ee S. obtusisporum

9a. Basidiospores cylindrical, 6.5-8.5 x 2.2-2.5 um .............. 000 S. meridense

9b. Basidiospores cylindrical to fusiform or banana-shape,

O.5e725-10 X22 (HS. See eh a uh 3 og ok gh aig S. brachysporum

Acknowledgments

Drs. Harold H. Burdsall, Jr. and Nils Hallenberg acted as pre-submission reviewers.

The Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET, Argentina)

supported this research through PIP 80101000. Sergio Pérez Gorjén is a postdoctoral

research fellow of the Agencia Espafiola de Cooperacién Internacional (MAEC-

AECID).

Literature cited

Boidin J, Gilles G. 1988. Basidiomycétes Aphyllophorales de Vile de la Réunion XII - le genre

Subulicystidium Parmasto. Bulletin de la Société Mycologique de France 104(3): 191-198.

Duhem B, Michel H. 2001. Contribution a la connaissance du genre Subulicystidium Parmasto

1968 (Basidiomycota, Xenasmatales). Cryptogamie, Mycologie 22(3): 163-173.

Hibbett DS, Binder M. 2002. Evolution of complex fruiting-body morphologies in homo-

basidiomycetes. Proceedings of the Royal Society of London 269: 1963-1969.

http://dx.doi.org/ 10.1098/rspb.2002.2123

Julich W. 1975. Studien an Cystidien. I. Subulicystidium Parm. Persoonia 8: 187-190.

Larsson KH, Larsson E, Koljalg U. 2004. High phylogenetic diversity among corticioid

Homobasidiomycetes. Mycological Research 108(9): 983-1002.

http://dx.doi.org/10.1017/S0953756204000851

Liberta AE. 1980. Notes on the genus Subulicystidium. Mycotaxon 10: 409-412.

Parmasto E, Nilsson RH, Larsson KH. 2004. Cortbase version 2. Extensive updates of a

nomenclatural database for corticioid fungi (Hymenomycetes). Phyloinformatics 1: 5.

Punugu A, Dunn MT, Welden AL. 1980. The peniophoroid fungi of the West Indies. Mycotaxon

10(2): 428-454.

MYCOTAXON

http://dx.doi.org/10.5248/118.53

Volume 118, pp. 53-56 October-December 2011

New records of smut fungi. 4. Microbotryum coronariae comb. nov.

CVvETOMIR M. DENCHEV & TEODOR T. DENCHEV

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

2 Gagarin St., 1113 Sofia, Bulgaria

* CORRESPONDENCE TO: cmdenchev@yahoo.co.uk

AxsstRAcT — For Ustilago coronariae on Lychnis flos-cuculi, a new combination in

Microbotryum, M. coronariae, is proposed. It is reported as new to Bulgaria.

Key worps — Microbotryaceae, taxonomy

Introduction

A new combination in Microbotryum, M. coronariae, is proposed for Ustilago

coronariae on Lychnis flos-cuculi.

Material & methods

Dried specimens from SOMF and (on loan) BP, BPI, COLO, DAOM, FH, H, ILL,

ILLS, ISC, KRAM, M, KSC, MICH, NY, and W were examined under light (LM) and

scanning electron (SEM) microscopes. For LM observations, spores were mounted

in lactophenol solution on glass slides, gently heated to boiling point to rehydrate the

spores, and then cooled. Spore measurements are given in the form: min-max (extreme

values, if necessary) [mean + 1 standard deviation]. The total number of spores (n) from

all collections (x) measured are given in the form ‘(n/x). For SEM, spores were attached

to specimen holders by double-sided adhesive tape and coated with gold with an ion

sputter. The surface structure of spores was observed at 10 kV and photographed with a

JEOL SM-6390 scanning electron microscope.

Taxonomy

Liro (1924) described Ustilago coronariae as a species morphologically

identical with U. dianthorum Liro, U. lychnidis-dioicae Liro, and U. stellariae

(Sowerby) Liro but with a narrow specialization on Coronaria flos-cuculi (L.)

A. Braun (currently Lychnis flos-cuculi). Using artificial infections with Ustilago

coronariae, Liro (1924: 319) demonstrated that the following caryophyllaceous

plants were not hosts: Agrostemma githago L., Dianthus deltoides L., Lychnis

54 ... Denchev & Denchev

chalcedonica L., Silene dioica (L.) Clairv. (as Melandrium rubrum), Silene latifolia

subsp. alba (Mill.) Greuter & Burdet (as Melandrium album), Silene nutans L.,

Silene vulgaris (Moench) Garcke (as S. inflata), Stellaria graminea L., Stellaria

holostea L., and Viscaria vulgaris Bernh. Later, Ustilago coronariae was reduced

to a synonym of Ustilago violacea and, after its transfer to Microbotryum, to

M. violaceum s. lat. (cfr Vanky 1994: 156). Based on physiological (Liro 1924)

and molecular phylogenetic (Le Gac et al. 2007, Refrégier et al. 2008, Devier

et al. 2010) inferences, we propose that Ustilago coronariae be transferred to

Microbotryum as an independent species.

Microbotryum coronariae (Liro) Denchev & T. Denchev, comb. nov. FIGs 1-2

MycoBank MB 561573

= Ustilago coronariae Liro, Annales Academiae Scientiarum Fennicae, Ser. A

17(1): 38, 1924. — Lectotype on Coronaria flos-cuculi (design. by Lindeberg

1959: 142), Finland, Regio aboénsis, Turku [Abo], 11 July 1916, J.I. Liro &

E. Kitunen; isolectotypes in Liro, Mycoth. fennica, no. 347.

SorI in anthers. SPORE MASS powdery, dark livid, dark purple or dark

vinaceous (based on the Rayner (1970) colour chart). Spores mainly globose

or subglobose, sometimes broadly ellipsoidal or ovoid, 6.5-10(-11) x 5.5-9.5

[7.7 + 0.7 x 7.2 + 0.6] um (n/,,=1750); spore wall reticulate, 6-8 meshes per

spore diameter, meshes irregularly polygonal, (0.5-)0.7-1.4(-1.6) um long; in

SEM interspaces smooth.

SPECIMENS EXAMINED — On Lychnis flos-cuculi L.: BULGARIA, Mt. Vitosha, near

Chouypetlyovo, 1250 m, 24 July 1992, C.M. Denchev (SOMF 29192); EstrontA, island

Saaremaa, 1899, T. Vestergren (as U. violacea, DAOM 215227); FAEROE ISLANDS, Kals6,

5 August 1897, J. Hartz & C. Ostenfeld (as U. coronariae, H); FINLAND, Ab, isolectotype,

Mycoth. fennica, no. 347 (H); Ab, Turku, Katariinan laakso, 11 July 1916, E. Kitunen

& J.I. Liro (as U. coronariae, H); N, Vantaa, Tikkurila, 21 June 1919, A. Rainio (as U.

violacea, H); Satakunta, Kankaanpaa, Kunintaanlahde, 8 August 1935, M. Laurila (as U.

coronariae, H); Satakunta, Ylane, Elijarvi, Vastalahti, 60°54’ N, 17 July 1950, L.E. Kari (as

U. violacea, Fungi exs. fennici, no. 710, MICH, NY, W 8496); Tb, Jyvaskyla, Vuoritsalo, 7

July 1916, K. Linkola (as U. coronariae, H); FRANCE, 30 May 1880, N. Patouillard (as U.

antherarum, FH); GERMANY, Bayern, Bez. Kelheim, Ulrain, 12 June 1940, E. Eichhorn

(as U. coronariae, M); Nordrhein-Westfalen, Kr. Siegen, Freudenberg, 17 June 1934, A.

Ludwig (as U. coronariae, BPI 159712, FH); Brandenburg, Berlin, Rahnsdorf, 16 June

1935, Fahrendorff (as U. coronariae, M, W 8999); Genshagen s. Berlin, 23 June 1899, P.

Sydow (as U. violacea, Sydow, Ustilag., no. 214, FH, KSC, M, NY); Mellen bei Zossen, 9

June 1905, P. Sydow (as U. violacea, Sydow, Mycoth. german., no. 367, COLO F-6367, FH

1094144, ILL 17344, M, MICH; the host plant wrongly given as Viscaria vulgaris); Kr.

Teltow, Trebbin, April 1946, W. Lempke (as U. coronariae, BPI 159710); LATVIA, prov.

Vidzeme, Triedaine, 16 July 1932, A. Kirulis (as U. coronariae, BPI 159709, ILLS 23414);

NETHERLANDS, Wageningen, 26 June 1923, J.I. Liro (as U. violacea, H); POLAND,

Leknica (on the label as “Muskau, Lugknitz”; cfr Scholz & Scholz 1988: 231), June 1891,

P. Sydow (Sydow, Mycoth. march., no. 3222) (as U. violacea, BP 3033, NY); Dabroszyn

(formerly German Tamsel), 28 June 1925, P. Vogel (as U. coronariae, Sydow, Mycoth.

german., no. 2290, BPI 159708, COLO F-8290, FH 1095112, ISC 371037, MICH, W

922); “Warthewiesen bei Tamsel’, 28 June 1925, P. Vogel (as U. violacea f. sp. lychnidis-

Microbotryum coronariae comb. nov. ... 95

Fics 1-2. Spores of Microbotryum coronariae on Lychnis flos-cuculi in SEM (Isolectotype, H).

Scale bars: 1 = 10 um; 2 = 1 um.

flos-cuculi, Zillig, Ustilag. Europ., no. 59, M, W 10982); ditto, June 1926, P. Vogel (as

U. coronariae, Petrak, Mycoth. gener., no. 1077, BPI 159713); Kérnik prope Poznan,

June 1927, A. Wrdblewski (Wroblewski et Siemaszko, Fungi polon. selec. exs., no. 7,

KRAM-F 2964); Russta, Karelia, Kpocc, Iso-Keilak, 1 August 1896, Bergroth & J.I. Liro

(as U. coronariae, H); Karelia, Kpor, Jernema - Somba, 17 August 1899, J.I. Liro (as U.

coronariae, H); Karelia, Kpor, Prilug - Siftuga, 12 August 1899, J.I. Liro (as U. coronariae,

H); Karelia, Kpor, Tamitsa, 26 July 1899, J.I. Liro (as U. coronariae, H); Karelia, K],

Kankala, ad rivulum lacuum Valkialampi et Parijarui, 1 August 1935, M. Laurila

(as U. coronariae, H); Sieverskaja, prov. Petropolitanae, 6/18 June 1898, Tranzschel

(Jaczewski, Komarov & Tranzschel, Fungi rossiae exs., no. 207) (as U. violacea, FH, NY);

Mikhaylovskoe, Moskow Gubernia, 28 June 1917, FE Bucholtz (as U. violacea, FH, NY);

SWEDEN, prope Stockholm, 20 June 1883, E. Eriksson (as U. violacea, Eriksson, Fungi

paras. scand. exs., no. 153a, NY); Ostergétland, Gryt parish, Korsudden, 16 July 1957,

J.A. Nannfeldt, no. 14874 (as U. violacea, Fungi exs. suecici, no. 3017, W 13061); Ol,

Stara Rév., June 1900, G. Lagerheim (as U. violacea, FH, H); Ramsasa, 28 June 1929,

H. Christoffersson (as U. coronariae, H); SWITZERLAND, Canton de Vand, Emrions de

Leysiu, 11 July 1917, E. Mayor (as Ustilago coronariae, BPI 159 711).

DISTRIBUTION — On Caryophyllaceae: Lychnis flos-cuculi, Europe (Austria, Bulgaria,

Czech Republic, Estonia, the Faeroes, Finland, France, Germany, Latvia, Lithuania, the

Netherlands, Poland, Romania, Russia, Slovakia, Spain, Sweden, Switzerland, UK) (Liro

1924; Savulescu 1957; Lindeberg 1959; Ignatavicitité 1975, 2001; Mordue & Ainsworth

1984; Vanky 1985; Zogg 1986; Scholz & Scholz 1988; Karatygin & Azbukina 1989;

Almaraz 2002; Zwetko & Blanz 2004; Legon et al. 2005; etc.). The smut is reported here

as new to Bulgaria.

56 ... Denchev & Denchev

Acknowledgements

We gratefully acknowledge Dr Kalman Vanky (Herbarium Ustilaginales Vanky,

Tubingen, Germany) and Dr Roger G. Shivas (Agri-Science Queensland, Australia) for

critically reading the manuscript and serving as pre-submission reviewers, and curators

of the herbaria (listed in Material & methods) for loans of the cited specimens. The

financial support from the Bulgarian National Science Fund (grant no. DO 02-181/2008)

is gratefully acknowledged.

Literature cited

Almaraz T. 2002. Bases coroldgicas de flora micolégica Ibérica. Numeros 1766-1932, in F Pando &

J.C. Hernandez (eds), Cuadernos de trabajo de flora micoldgica Ibérica 17: 1-124.

Devier B, Aguileta G, Hood ME, Giraud T. 2010. Using phylogenies of pheromone receptor

genes in the Microbotryum violaceum species complex to investigate possible speciation by

hybridization. Mycologia 102: 689-696. http://dx.doi.org/10.3852/09-192

Ignataviciuté M. 1975. The smut fungi of the Baltic Region. Mintis, Vilnius. 278 pp. (In Russian)

Ignataviciuteé M. 2001. Ustilaginales of Lithuania. 1-199, in Mycota Lithuaniae, vol. 4. UAB

‘Valstieciy Laikrastis, Vilnius. (In Lithuanian)

Karatygin IV, Azbukina ZM. 1989. Ordo Ustilaginales 1. Familia Ustilaginaceae. Definitorium

Fungorum URSS. Nauka, Leningrad. 220 pp. (In Russian)

Le Gac M, Hood ME, Fournier E, Giraud T. 2007. Phylogenetic evidence of host-specific cryptic

species in the anther smut fungus. Evolution 61: 15-26.

http://dx.doi.org/10.1111/j.1558-5646.2007.00002.x

Legon NW, Henrici A, Roberts PJ, Spooner BM, Watling R. 2005. Checklist of the British & Irish

Basidiomycota. Royal Botanic Gardens, Kew. 517 pp.

Lindeberg B. 1959. Ustilaginales of Sweden (exclusive of the Cintractias on Caricoideae). Symbolae

Botanicae Upsalienses 16(2):1-175.

Liro JI. 1924. Die Ustilagineen Finnlands 1. Annales Academiae Scientiarum Fennicae, Ser. A 17(1):

1-636.

Mordue JEM, Ainsworth GC. 1984. Ustilaginales of the British Isles. Mycological Papers 154:

1-96.

Rayner RW. 1970. A mycological colour chart. CMI, Surrey and the British Mycological Society,

Kew.

Refrégier G, Le Gac M, Jabbour F, Widmer A, Hood ME, Yockteng R, Shykoff JA, Giraud T. 2008.

Cophylogeny of the anther smut fungi and their caryophyllaceous hosts: Prevalence of host

shifts and importance of delimiting parasite species. BMC Evolutionary Biology 8: 100.

Savulescu T. 1957. Ustilaginalele din Republica Populara Romina. Vols 1-2. Editura Academiei R.P.

Romine, Bucuresti. 1168 pp.

Scholz H, Scholz I. 1988. Die Brandpilze Deutschlands (Ustilaginales). Englera 8: 1-691.

http://dx.doi.org/10.2307/3776736

Vanky K. 1985. Carpathian Ustilaginales. Symbolae Botanicae Upsalienses 24(2): 1-309.

Vanky K. 1994. European smut fungi. Gustav Fischer Verlag, Stuttgart, Jena, New York. 570 pp.

Zogg H. 1986 [“1985”]. Die Brandpilze Mitteleuropas unter besonderer Beriicksichtigung der

Schweiz. Cryptogamica Helvetica 16: 1-277.

Zwetko P, Blanz P. 2004. Die Brandpilze Osterreichs. Doassansiales, Entorrhizales, Entylomatales,

Georgefischeriales, Microbotryales, Tilletiales, Urocystales, Ustilaginales. I-IV, 1-241 + CD, in F

Ehrendorfer (ed.), Catalogus Florae Austriae, vol. 3(3), Biosystematics and Ecology Series, vol.

21. Verlag der Osterreichischen Akademie der Wissenschaften, Wien.

MY COTAXON

http://dx.doi.org/10.5248/118.57

Volume 118, pp. 57-71 October-December 2011

Lentinus giganteus revisited:

new collections from Sri Lanka and Thailand

SAMANTHA C. KARUNARATHNA®”°, ZHU L. YANG?’, OLIVIER RASPE’,

THIDA W. Ko Ko’, ELSE C. VELLINGA®, RUI-LIN ZHAO‘, A.H. BAHKALY’,

EKACHAI CHUKEATIROTE’, JEROME DEGREEF*, PHILIPPE CALLAC®

& KEVIN D. HYDE?” ”?

'School of Science, Mae Fah Luang University,

333 Mool, Tasud, Muang, Chiang Rai 57100, Thailand

?International Fungal Research & Development Centre, Research Institute of Resource Insects,

Chinese Academy of Forestry, Kunming 650034, China

°Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany,

Chinese Academy of Sciences, Kunming 650204, China

‘Department of Cryptogamy, National Botanic Garden of Belgium,

Domein van Bouchout, Nieuwelaan 38, 1860 Meise, Belgium

‘Department of Plant and Microbial Biology, University of California,

Berkeley CA 94720-3102, U.S.A.

°Key Laboratory of Forest Disaster Warning and Control in Yunnan Province,

Faculty of Conservation Biology, Southwest Forestry University, Kunming 650224, China

“Botany and Microbiology Department, College of Science, King Saud University,

Riyadh, Saudi Arabia

8INRA, MYCSA (Mycologie et sécurité des aliments),

BP 81, 33883 Villenave d’Ornon cedex, France

*Mushroom Research Foundation,

128 M.3 Ban Pa Deng T. Pa Pae, A. Mae Taeng, Chiang Mai 50150, Thailand

“CORRESPONDENCE TO: fungiamanita@gmail.com

ABSTRACT— A new collection of Lentinus giganteus from Sri Lanka, where it was originally

described, is used to epitypify the species after comparison with the type protologue and

drawings held in Peradeniya, Sri Lanka; a full description and illustrations are provided.

Additional collections were made at three sites in northern Thailand. Phylogenetic ITS-

1-5.8S-ITS2 rDNA sequence analyses using maximum likelihood, maximum parsimony

and Bayesian inference all support the transfer of L. giganteus to Pleurotus. Although the

collections from Thailand differ slightly morphologically and phylogenetically from

P. giganteus sensu stricto, these differences do not yet merit specific status. Instead, P giganteus

is maintained as one widely variable species represented by relatively large fruiting bodies.

Saprobic on buried well-rotted wood in forests, P. giganteus is widely consumed in Sri Lanka

and might be profitably cultivated in Thailand.

Key worps— edible fungi, morphology, new records, new combination, taxonomy

58 ... Karunarathna & al.

Introduction

Research on macrofungi of the Mushroom Research Centre and surrounding

areas in northern Thailand has documented a remarkable biodiversity and

produced many new species and new records (Le et al. 2007a,b; Sanmee

et al. 2008; Kerekes & Desjardin 2009; Wannathes et al. 2009a,b; Zhao et al.

2010). Recently we have focused on potentially cultivatable and edible genera

(Karunarathna et al. 2011). This paper presents the first report of Lentinus

giganteus from Thailand.

Lentinus Fr. is a cosmopolitan genus with an estimated 40 species (Kirk et al.

2008) distributed across a wide temperature range, being abundant in the tropics

and often found in temperate regions (Pegler 1983). Lentinus species, normally

wood decaying basidiomycetes, are characterized by decurrent lamellae,

dimitic tissues, and hyaline ellipsoid to cylindrical basidiospores. Species in

subgenus Lentinus have hyphal pegs (Corner 1981, Pegler 1983). Generally

the xeromorphic long-lived basidiomes are tough and firm when dry, but in

Thailand they fruit only at the beginning of the rainy season (Sysouphanthong

et al. 2010, Karunarathna et al. 2011). Traditionally, Lentinus was placed in the

agaric family Tricholomataceae based on the lamellate hymenophore and white

spore print (e.g., Miller 1973).

Previous researchers (e.g., Redhead & Ginns 1985; Hibbett & Vilgalys

1991, 1993; Hibbett & Thorn 1994; Hibbett et al. 1993) noted the existence

of a Lentinus—Pleurotus-Panus complex and showed that Lentinus sensu

Pegler is polyphyletic, with the monophyletic Lentinus subg. Lentinus sensu

Pegler belonging to the Polyporales (Hibbett 1991, Kruger & Gargas 2004).

Panus Fr., Pleurotus (Fr.) P. Kumm., and Neolentinus Redhead & Ginns are

also monophyletic, with Pleurotus belonging to the Agaricales (Fleming 1994,

Hibbett et al. 1994). Corner (1981), Kithner (1980), Pegler (1975, 1983) and

Singer (1986) all classified these genera differently.

Lentinus giganteus, originally described from Sri Lanka (Berkeley 1847),

bears many structures that are atypical of Lentinus, and its taxonomic position

has long been unresolved (Pegler 1983). Corner (1981: 54) suggested it as type

(and only species) of his new subgenus, Panus subg. Gigantopanus Corner.

Pegler (1983: 168) transferred this subgenus (as a section) to Lentinus: L. sect.

Gigantopanus (Corner) Pegler. Even though the largest spores become oblong-

ellipsoid, mature spores are not cylindrical but somewhat broadly ellipsoid. The

oil guttule present inside the spore is characteristic of Lentinus. The generative

hyphae are narrower than the skeletal hyphae. The lamellar edge is distinct with

a broad, sterile layer of lecythiform cheilocystidia similar to those observed

in some Pleurotus species. The lamellae are wide and well separated, and the

development process is metavelangiocarpic. In many ways this species might

be more properly positioned in Pleurotus, rather than in Lentinus. However,

Pleurotus giganteus comb. nov. (Sri Lanka, Thailand) ... 59

the skeletal hyphae of the dimitic hyphal system dominate to produce a

tough basidiome and the radiate construction of the hymenophoral trama

differentiates with the descending trama observed in Pleurotus.

Lentinus giganteus, referred to as “uru paha” in Sri Lanka, is one of the largest

edible mushrooms and —as noted in Buddhist literature— has been treated as

a special food since ancient times (Udugama & Wickramaratna 1991; Berkeley

1847).

Objectives of the present study were to redescribe L. giganteus from fresh

collections from the type locality, epitypify L. giganteus with a fresh Sri Lankan

collection, decide on the taxonomic position of L. giganteus to determine whether

it is related to Lentinus or Pleurotus using ITS-1-5.8 S-ITS2 rDNA sequence

data, and introduce the species to Thailand as a new edible mushroom.

Materials & methods

Lentinus giganteus samples were collected in northern Thailand and Sri Lanka

between June 2008 and October 2010 and processed as in Karunarathna et al. (2011).

Ethnomycological survey

Thirty locals including mushroom sellers in markets were surveyed through a general

questionnaire in Chiang Mai and Chiang Rai provinces, Thailand.

Morphological examination

Macro-morphological characters were described from fresh material and documented

by photographs. Colour designations (e.g., 4B5) follow Kornerup & Wanscher (1978),

while the colour names (e.g., grayish yellow) follow Ridgway (1912). Specimens were

dried, placed in separate plastic bags, and deposited in the Herbarium of Mae Fah

Luang University (MFLU). For microscopical examination, sections were cut with

a razor blade from dried specimens, mounted on slides in 5% KOH and Congo red,

observed, measured, and illustrated using a Zeiss Axioskop 40 compound microscope.

Basidiospore measurement abbreviations: n = number of spores measured; L_ = mean

spore length; W_, = mean spore width; Q = length/width ratio (L/W) of a spore in side

view; Q. = average Q of all spores measured.

Molecular & phylogenetic methods

DNA EXTRACTION— Genomic DNA was extracted from dried mushroom samples

with cetyl trimethylammonium bromide (CTAB) according to Doyle & Doyle (1990)

with some modifications. DNA concentrations were estimated visually in agarose gel by

comparing band intensity with a 1000 bp DNA ladder (Transgen Biotech).

PCR AMPLIFICATION & SEQUENCING—PCR reactions were performed in a 50 ul

volume (0.625 mM primers (White et al. 1990: ITS4: 5’TCCTCCGCTTATTGATATGC3’;

ITS5: 5°GGAAGTAAAAGTCGTAACAAGG3 ’); 10-20 ng DNA template, 10x buffer, 0.2 mM

dNTPs, 1.5 units Taq and sterile water. Thermal cycles: 3 m at 94 °C, 30-35 cycles at 93

°C for 30 s, 1 m at 55 °C, 1 m at 72 °C, final extension 10 m at 72 °C. PCR products were

verified by 1% agarose electrophoresis gels stained with ethidium bromide in 1xTris-

boric acid EDTA buffer. ITS 4/ITS 5 were used to sequence both DNA strands (White et

al. 1990) at the National Botanic Gardens of Belgium.

60 ... Karunarathna & al.

TABLE 1. Pleurotus giganteus and other taxa sequenced for phylogenetic analyses.

Taxa COUNTRY OF ORIGIN GENBANK accession numbers (ITS)

Lentinus squarrosulus Japan AB478883 (Sotome et al. 2009)

Panus sp. China HM245784 (unpublished)

P. australis New Zealand AY315764 (Zervakis et al. 2004)

P. cornucopiae Austria AY450341 (unpublished)

P. cystidiosus India AY315810 (Zervakis et al. 2004)

P. euosmus China EU424298 (unpublished)

P. giganteus ; :

(MELU 08 1370) Thailand (new sequence, from dried sample)

P. giganteus : ,

(MELU 08 1371) Thailand (new sequence, from dried sample)

P. giganteus . :

(MELU10 0141) Thailand (new sequence, from dried sample)

P. giganteus : ;

(MELU 11 0018, epitype) Sri Lanka (new sequence, from dried sample)

Pleurotus giganteus ; :

fastLebdinusieanteid Thailand DQ334857 (unpublished)

P. giganteus China HM245788 (unpublished)

[as Panus giganteus]

P. giganteus China HM245786 (unpublished)

[as Panus giganteus]

P. giganteus China ;

Lea gbantnsl HM245780 (unpublished)

P. fuscosquamulosus India AY315789 (Zervakis et al. 2004)

P. smithii Mexico AY315779 (Zervakis et al. 2004)

SEQUENCE ALIGNMENT & PHYLOGENETIC ANALYSIS— Taxa sequenced and GenBank

accession numbers are listed in TaBLE 1. Sequences for each strain were aligned

using Clustal X (Thompson et al. 1997). Alignments were manually adjusted to allow

maximum sequence similarity. Gaps were treated as missing data. Phylogenetic analysis

were performed using PAUP* 4.0b10 (Swofford 1998). Ambiguously aligned regions

were excluded from all analyses. Trees were inferred using the heuristic search option

with TBR branch swapping and 1000 random sequence additions. Maxtrees were

unlimited, branches of zero length were collapsed and all multiple parsimonious trees

were saved. Clade stability of the trees resulting from the parsimony analyses were

assessed by bootstrap analysis with 1000 replicates, each with 10 replicates of random

stepwise addition of taxa (Felsenstein 1985). Trees were figured in TreeView.

Results

The ITS sequence dataset comprised 16 sequences (TABLE 1) representing

three Panus giganteus collections and one Lentinus giganteus collection from

GenBank, three L. giganteus collections from northern Thailand, and one

collection from Sri Lanka. Outgroup taxa were Lentinus squarrosulus and Panus

sp. Of 732 total characters, 363 are constant, 112 are parsimony-uninformative,

and 257 are parsimony-informative. The MP tree was produced after 238,115

Pleurotus giganteus comb. nov. (Sri Lanka, Thailand) ... 61

AB478883 L. squarrosulus

AB509648 Panus sp.

AY315764 Pl. australis

100

0 AY315779 Pl. smithii Pleurotus subgenus

$9 Coremiopleurotus clade

99 AY315810 Pl. cystidiosus

AY315789 Pl. fuscosquamulosus

MFLUII 0018 PI. giganteus } Sri Lankan Giganteus forms a new clade

100

HM245788 Pl. giganteus*

86 68

79 HM245780 PI. giganteus* Giganteus clade I

(Chinese taxa belong to this clade)

96 HM245786 PI. giganteus*

MFLU10 0141 Pl. giganteus

89 MFLU08 1371 PI. giganteus Giganteus clade Il

96 Be MFLUO8 1370 Pl. giganteus (Thai specimens belong to this clade)

DQ334857 Pl. giganteus**

100 EU424298 Pl. euosmus

AY450341 Pl. cornucopiae

PLaTE 1. Maximum parsimony phylogram showing phylogenetic relationships among Pleurotus

giganteus from Thailand (MFLUO8 1370, MFLUO8 1371, MFLU10 0141) and epitype from Sri Lanka

(MFLU11 0018) with some selected Lentinus, Panus and Pleurotus species based on ITS-1-5.8 S-ITS2

rDNA sequences. Data were analysed with random addition sequence, unweighted parsimony and

gaps were treated as missing data. Values above the branches are parsimony bootstrap (2 50%). The

tree is rooted with Lentinus squarrosulus (AB478883) and Panus sp. (AB509648). Pl = Pleurotus;

** = as Lentinus giganteus in GenBank, * = as Panus giganteus in GenBank.

rearrangements; the best MP tree found (scored at 645) was chosen to represent

the phylogenetic position of L. giganteus (PLATE 1).

The three L. giganteus samples from Thailand (MFLU08 1370, MFLUO8 1371,

MFLU1O 0141) and Chinese collections from GenBank form one clade (PLATE 1)

that is sister (86% bootstrap support) to the only L. giganteus collection from

Sri Lanka. Both Sri Lankan and Thai collections are closely related to Pleurotus

cornucopiae and P. euosmus (89% bootstrap support). The L. giganteus clade,

P. cornucopiae, and P. euosmus cluster with Pleurotus subg. Coremiopleurotus

(100% bootstrap support). Coremiopleurotus comprises P. australis, P. smithii,

P. cystidiosus, and P. fuscosquamulosus.

These results support the view that L. giganteus is related to Pleurotus and not

Panus or Lentinus. Since the morphological data also supports the molecular

62 ... Karunarathna & al.

conclusions, we transfer L. giganteus to Pleurotus and epitypify the species

based on the collection from the holotype locality to enable future molecular

studies.

Taxonomy

Pleurotus giganteus (Berk.) Karunarathna & K.D. Hyde, comb. nov. PLATES 2-4

MycoBank MB 561087

= Lentinus giganteus Berk., Lond. Journ. Bot. 6: 493[bis], pl.17/18 f.2 (1847)

= Pocillaria gigantea (Berk.) Kuntze, Revis. Gen. Pl. 2: 866 (1891)

= Velolentinus giganteus (Berk.) Overeem, Bull. Jard. Bot. Buitenz, 3 sér., 9: 12 (1927)

= Panus giganteus (Berk.) Corner, Beih. Nova Hedwigia 69: 69 (1981)

Type: Sri Lanka, Central Prov., Hautane Range, on ground, July 1844, by Gardner, No 58

(holotype K; PLaTe 2A-C); Central Prov., Kandy Distr., Deliwala village, 7°14'43.55"N

80°33'51.40"E, elevation 1050 m, rainforest dominated by Swietenia spp. and Artocarpus

heterophyllus, 5 June 2009 (MFLU11 0018, epitype designated here [PLATE 4A]).

PitEus 60-310 mm in diameter, strongly convex to applanate becoming

slightly depressed in the centre, dark brown (7F5), towards the margin light

brown camel (6D4), grayish orange (5B4) at the marginal area, at centre

fibrillose-scaly, surface initially uniformly dark, fuscous brown, fuliginous or

black, then fading with age to pale ochraceous or yellowish brown (E8), with a

darker centre although sometimes remaining dark, dry, distrupted into small,

indefinite, radial, innate squamules, overlain by scanty, pale grey or blackish,

verrucose-floccose, concentrically arrange remnants of the veil; margin strongly

involute then straight, thin, slightly sulcate-striate. LAMELLAE moderately

crowded with lamellulae of five lengths, decurrent, slightly interveined and

anastomosing over the stipe apex, 2-3 mm broad, white to cream (3A2); edge

entire, pale ochraceous or yellowish brown (E8). STIPE up to 50-200 mm long,

7-10 mm broad at the apex, 10-15 mm at the base, fusiform, with radicating

base, solid, with surface concolorous with the pileus, paler at the apex, finely

tomentose with indefinite zones of paler velar remnants in the early stages; veil

thin, floccose, pale to dark brown (6F6), soon reduced to floccose remnants but

never forming an annulus on the stipe. CONTEXT 5-10 mm thick at the disk,

submembranous at the pileal margin, white in pileus and stipe, fleshy-spongy,

consisting of a dimitic hyphal system with skeletal hyphae.

GENERATIVE HYPHAE (PLATE 3e) 4-6 um in diameter, inflating with a

thick or slightly thickened wall, more or less radially parallel but frequently

branching and with large clamp connections. SKELETAL HYPHAE (PLATE 3d)

6-8 um in diameter, hyaline of intercalary or terminal origin, becoming very

thick-walled with a narrow lumen, tending to taper apically, occasionally with

a limited lateral branch. BAsIDIOSPORES (PLATE 3a) 7-9 x 6-7 um [n = 40, L_=

8.30 um, W_ = 6.36 um, Q= 1.18-1.46, Q_ = 1.33] broadly ellipsoid to ellipsoid,

white in mass, smooth, with one large oil drop or multiguttulate, inamyloid,

Pleurotus giganteus comb. nov. (Sri Lanka, Thailand) ... 63

PLATE 2. Lentinus giganteus: watercolour illustrations by Mr. William De Silva conserved at

Fungal Herbarium, Horticultural Crop Research and Development Institute (HORDI), Sri Lanka

[scanned by and reproduced with permission of Mrs. Srimathie Udugama, Director, Fungal

Herbarium, Horticultural Crop Research and Development Institute (HORDI), Sri Lanka]. A-C:

Sri Lanka, Hautane Range, on ground, July 1844, Gardner n.58 (holotype; Berkeley 1847); A:

habit of young basidiocarps; B. cap surface, upper view; C: side view of mature basidiocarp. D: Sri

Lanka, Peradeniya, July 1868, Thwaites n.688 (Berkeley & Broome 1873, as L. stenophyllus); habit

of basidiocarp.

thin-walled. The large spores are not cylindrical but rather broadly ellipsoid,

although the largest spores become oblong ellipsoid. Basip1A (PLATE 3c) 25-40

x 8-10 um, elongate, clavate, bearing 4 sterigmata. LAMELLA EDGE sterile with

a broad layer of CHEILOCYSTIDIA (PLATE 3b) 15-30 x 6-10 um, more or less

64 ... Karunarathna & al.

PLaTE 3. Pleurotus giganteus (MFLUO8 1370). a: Spores; b: Cheilocystidia; c: Basidia; d: Skeletal

hyphae; e: Generative hyphae; Scale bars: a~c = 20 um; d, e = 10 um.

PiaTE 4 (to right). A. Basidiocarp of Pleurotus giganteus from Sri Lanka (MFLU11 0018, epitype);

B-E: Basidiocarps of P. giganteus from northern Thailand (B: MFLU10 0141; C: MFLUO8 1371;

D, E: MFLUO8 1370). Scale bars: A = 20 cm; B, C = 10 cm.

Pleurotus giganteus comb. nov. (Sri Lanka, Thailand) ... 65

66 ... Karunarathna & al.

lecythiform with a ventricose base and a small capitellum (3-4 um) subtended

by a narrow neck, hyaline, thin-walled.

ECOLOGY AND DISTRIBUTION— solitary on buried rotten wood in rain

forest. Widely distributed in Australia, China, Malay Peninsula, Sabah, Sri

Lanka, Vietnam (Pegler 1983), Thailand (this study).

ADDITIONAL MATERIAL EXAMINED: THAILAND, CHIANG Mal PRov., MAE TAENG

Distr., Ban Pha Deng, Mushroom Research Centre, 19°17.123'N 98°44.009’E, elevation

900 m, rainforest dominated by Castanopsis armata, Erythrina sp, and Dipterocarpus

sp., 8 July 2008, Ruilin Zhao (MFLU08 1370); 21 June 2008, Samantha C. Karunarathna

(MFLU10 0138); 27 June 2008, Samantha C. Karunarathna (MFLU08 1371); 6 July 2008,

Samantha C. Karunarathna (MFLU08 1382); 22 July 2008, Samantha C. Karunarathna

(MFLU10 0137); 10 July 2010, Olivier Raspe (MFLU10 0153); Doi Suthep-Pui National

Park, Sangasabhasri Lane to Huai Kok Ma village, 18°48.62'N 98°54.60’E, elevation 1145

m, rainforest dominated by Castanopsis spp., Lithocarpus polistachyus and other trees,

9 June 2008, Samantha C. Karunarathna (MFLU10 0136); CHIANG Ral PRov., Highway

No.110 to Mae Sai, Doi Tung, 20°17'37"N 99°48'56"E, elevation 950 m. 15 July 2009,

Samantha C. Karunarathna (MFLU10 0140); 15 July 2009, Samantha C. Karunarathna

(MFLU10 0141); 8 August 2009, Samantha C. Karunarathna (MFLU10 0142);

8 August 2009, Samantha C. Karunarathna (MFLU10 0143); 16 July 2010, Samantha C.

Karunarathna (MFLU10 0154).

Discusston— Molecular evidence indicates that Lentinus giganteus is better

placed in Pleurotus. This is supported by morphology as (1) the lamella-edge is

well defined with a broad, sterile layer of differentiated cheilocystidia, similar

to those found in some Pleurotus species; (2) basidiomes are soft in texture with

short life span, similar to Pleurotus species; (3) the lamellae are broad and well

spaced; and (4) development is metavelangiocarpic. Although Corner (1981)

established L. subg. Gigantopanus for L. giganteus, our limited data do not

support this division.

We collected 11 Pleurotus giganteus specimens from three sites in northern

Thailand and one specimen from the original site in Sri Lanka cited in the

protologue. The new Sri Lankan collection, identical to that of Corner’s (1981)

description and our observation of the holotype, is designated as epitype of

P. giganteus. Molecular data groups our collections from Thailand with high

bootstrap support in a single clade that clusters with sequences of P giganteus

from China, suggesting strain similarity. The Sri Lankan P. giganteus collection

forms a sister group with Thai and Chinese collections with 86% bootstrap

support. This suggests that the Chinese and ‘Thai collections might have

diverged from the Sri Lankan species due to geographical isolation. There are

also micro-morphological differences (TABLE 2), although more collections

are needed to confirm whether these are different taxa. Thus we maintain P

giganteus as a single widely variable species. Both Thai and Sri Lankan collections

are more closely related to Pleurotus than to Panus and Lentinus. Pleurotus

australis, P. smithii, P. cystidiosus, and P. fuscosquamulosus (all Pleurotus subg.

Pleurotus giganteus comb. nov. (Sri Lanka, Thailand) ... 67

TABLE 2. Pleurotus giganteus morphological comparisons

Cap (diam.) SPORES BASIDIA CHEILOCYSTIDIA

ISOLATE/COLLECTION STIPE (length) (mm) (mm) (um)

(both in mm)

MFLU08 1370 Cap: 100-110 7-9 x 25-40 x 15-30 x

Stipe: 65-70 6-7 8-10 6-10

MFLU10 0138 Cap: 90-110 6.5-8.5 x 26-38 x 14-30 x

Stipe: 80-110 6-7 7.5-9.5 6.5-11

MFLU08 1371 Cap: 165-170 6.5-8.5 x 25-39 x 13.5-31 x

Stipe: 65-70 6-7 7.5-9.5 6-11

MFLU08 1382 Cap: 45-50 6.5-8.5 x 25.5-41 x 15-31 x

Stipe: 65-70 6.5-7 8-10.5 6-10.5

MFLU10 0137 Cap: 105-110 6.5-8.5 x 24-41 x 15-31 x

Stipe: 50-60 6-7 8-10 6.5-10.5

MFLU10 0153 Cap: 140-150 6.5-8.5 x 25-41 x 14-30.5 x

Stipe: 190-200 6.5-7 8.5-10.5 6-10

MFLU10 0140 Cap: 50-70 6.5-8 x 25-41 x 15-31 x

Stipe: 130-150 6.5-7 8.5-10.5 6.5-11

MFLU10 0143 Cap: 200-220 6-8 x 25-41.5 x 15-30.5 x

Stipe: 70-80 6-7 8-10 6-10

MFLU10 0141 Cap: 70-100 6.5-8 x 25-39.5 x 14-30.5 x

Stipe: 150-180 6.5-7.5 8.5-10 6.5-10.5

MFLU10 0142 Cap: 100-180 6.5-8.5 x 24-40 x 14-31 x

Stipe: 120-130 6.5-7 8.5-10 6.5-10.5

MFLU10 0154 Cap: 150 6.5-8.5 x 23.5-41 x 15.5-31 x

Stipe: 75 6.5-7 8-10 6.5-10

MFLU10 0136 Cap: 130 6.5-8.5 x 24-41 x 15-31 x

Stipe: 110 6.5-7 8-9.5 6.5-10.5

MFLU11 0018 Cap: 60-310 6-8 (-9) x 29-52 x 23-35(-38) x

(epitype) Stipe: 50-190 4.3-5.2 8-9.5 6-10

K Cap: 50-300 6-8 X 30-50 x 25-35 x

(holotype) Stipe: 50-180 4.5-5.2 8-9 6-10

Coremiopleurotus) group with all sequenced P giganteus collections with 100%

bootstrap support.

Pleurotus giganteus is closely related to P subg. Coremiopleurotus, a group

of edible species with high commercial value (Zervakis et al. 2004). The

relationship suggests that P giganteus is likely to be a good edible species,

confirmed by its present consumption in Sri Lanka (Pegler 1983; Udugama &

Wickramaratna 1991). Pleurotus giganteus is also considered edible in China

(Dai et al. 2010).

The most commonly and easily cultivated mushrooms in Thailand and other

southeast Asian countries are oyster mushrooms (Pleurotus ostreatus (Jacq.)

P. Kumm.), ear mushrooms (Auricularia polytricha (Mont.) Sacc.), and straw

mushrooms (Volvariella volvacea (Bull.) Singer). Other mushroom species in

68 ... Karunarathna & al.

Lentinula, Lentinus, Ganoderma, and Macrocybe (Hanko 2001, Karunarathna

et al. 2011, Boa 2007) and Agrocybe can also be cultivated successfully but

require more attention and knowledge (Boa 2007).

Wild mushrooms are one of the higher valued non-timber forest products

in northern Thailand (Sysouphanthong et al. 2010, Karunarathna et al. 2011).

They provide locals with seasonal food, medicine, and an alternative income

while maintaining forest health (Sysouphanthong et al. 2010). The richness

of wild mushrooms is also one bioindicator of ecosystem health (Dai & Yang

2009; Du et al. 2011a,b; Sysouphanthong et al. 2010; Egli 2010). Cultivated,

non-mycorrhizal mushrooms (e.g., Lentinula edodes (Berk.) Pegler, Pleurotus

sajor-caju (Fr.) Singer, Flammulina velutipes (Curtis) Singer, and species of

Agaricus, Auricularia, Pleurotus, Agrocybe, and Volvariella) are available year

around in northern Thai markets. However, edible wild mushrooms can be

found only in the wet season from June to September (Sysouphanthong et al.

2010, Karunarathna et al. 2011). As Pleurotus species form a heterogeneous

commercially valuable group of edibles, it is therefore desirable to try to introduce

members of this genus as commercial species. Although there are many studies

on cultivated and wild edible mushrooms and their nutritional value in the

northern hemisphere (Aletor 1995; Latiff et al. 1996; Manzi et al. 1999, 2001;

Dermirbas 2000), there is little information available concerning the taxonomy,

biodiversity, and economic potential of Pleurotus species in the tropics. Scientific

information on wild mushrooms is essential for the introduction of new species

for the table (Karunarathna et al. 2011, Sysouphanthong et al. 2010).

Based on the survey of 30 locals through a questionnaire in Chiang Mai and

Chiang Rai provinces, we were unable to obtain a clear idea as to whether or

not they regard P. giganteus as an edible mushroom. Pleurotus giganteus is not

sold in Thai markets during wet season, and most inhabitants of Chiang Mai

and Chiang Rai provinces do not consume this wild edible mushroom. Even

though P giganteus has a very good taste (Udugama & Wickramaratna 1991), it

is not yet cultivated in Thailand as a commercial mushroom.

Acknowledgements

Weare grateful to Jian Kui Liu, Naritsada Thongklang and Phongeun Sysouphanthong

for their help in collecting and suggestions, Pheng Phengsintham and Nilam Wulandari

are gratefully acknowledged for their valuable discussions. We wish to acknowledge

the help with field work provided by Kobeke Van de Putte, Nalin Wijewardane, Putrak

Chomnuti, Rungtiva Pookamsak, Saowanee Wikee, Stefan D. Baros, Joshua Mark

Birkebak, Anjel Creig, Don Nelson, Jie Chen, Michael Pilkington. Kanjana Niraphai

(MFLU) is thanked for her assistance in the herbarium. A special thank goes to Prof.

Nimal Adikaram for providing his laboratory facilities and his valuable suggestions. The

comments by the two reviewers, Dr. Eric McKenzie and Dr. Yu Chen Dai, are gratefully

acknowledged. This study was financially supported by the project “Value added products

Pleurotus giganteus comb. nov. (Sri Lanka, Thailand) ... 69

from basidiomycetes: Putting Thailand's biodiversity to use” (BRN049/2553). The Global

Research Network for Fungal Biology and King Saud University are also thanked for

supporting this research. Study in China was supported by a grant from the Ministry of

Science and Technology of the People’s Republic of China (2008FY 110300).

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http://dx.doi.org/10.1007/s13225-010-0050-4

MY COTAXON

http://dx.doi.org/10.5248/118.73

Volume 118, pp. 73-81 October-December 2011

Diversispora clara (Glomeromycetes)— a new species

from saline dunes in the Natural Park Cabo de Gata (Spain)

BEATRIZ ESTRADA’, JAVIER PALENZUELA’, JOSE-MIGUEL BAREA’,

JUAN MANUEL Ru1z-LOZANO', GLADSTONE ALVES DA SILVA? & FRITZ OEHL?

‘Departamento de Microbiologia del Suelo y Sistemas Simbioticos, Estacién Experimental del

Zaidin, CSIC, Profesor Albareda 1, 18008 Granada, Spain

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

Av. Prof. Nelson Chaves s/n, Cidade Universitaria, 50670-420, Recife, PE, Brazil

*Federal Research Institute Agroscope Reckenholz-Tdanikon ART,

Organic Farming Systems, Reckenholzstrasse 191, CH-8046 Ziirich, Switzerland

*CORRESPONDENCE TO: fritz.oehl@art.admin.ch

ABSTRACT — A new species of Diversispora (Glomeromycetes) was found in saline sand dunes

of the Natural Park Cabo de Gata (Almeria, Andalucia, Southern Spain) in the rhizosphere

of Asteriscus maritimus, a plant species especially adapted to saline environments. The new

fungal species forms brilliant white spores that are 79-130 x 75-125 um and have one wall

consisting of three layers. The subtending hyphae are, as typical for many Diversispora spp.,

thin-walled, hyaline, and cylindrical (or rarely constricted) and flexible and fragile below the

septa separating the spore and hyphal contents. The septa form regularly at the spore bases

or, less frequently, in subtending hyphae at short distances from the spore base. Phylogenetic

analyses of the ITS and partial 28S ribosomal gene confirm that D. clara forms a monophyletic,

independent clade within Diversispora.

Key worps — Glomus, Glomeromycota, Europe, extreme environments, rDNA

Introduction

During recent studies on arbuscular mycorrhiza (AM) fungal diversity

in sand dune systems of the Cabo de Gata Natural Park in Almeria (Spain),

a brilliant-white new glomeromycotean fungus was recovered from the

rhizosphere of Asteriscus maritimus, a plant species characteristic of saline

Mediterranean environments with elevated soil electrical conductivity. The

new fungus formed spores in AM fungal bait cultures predominantly in the

Asteriscus maritimus rhizosphere. The aims of the present study were to analyze

this particular fungus applying combined morphological and molecular tools

and to describe its characteristics.

7A ... Estrada & al.

Material & methods

Soil and plant sampling

Soil samples were taken in February 2010 from the rhizosphere of ten Asteriscus

maritimus plants growing in a natural sand dune system of the Natural Park Cabo de Gata

in Almeria (Andalucia, Spain). The site is located at 36°44’41"N 02°07'26"E. The samples,

air-dried in the laboratory, were used to analyze selected chemical soil parameters (pH,

soil organic carbon, total nitrogen, soil electrical conductivity) and spore populations.

The ten plants and the surrounding rhizosphere soil were also extracted and used as

bait cultures for propagation of AM fungal communities indigenous to the natural sand

dune system. At the site, the soil was sandy and with pH (H,O) 8.2, organic carbon 15.3

g kg", total nitrogen 1.9 g kg’, available P 27.0 mg kg", and soil electrical conductivity

1.5 uS cm”.

AM fungal bait cultures

Bait cultures were established and maintained as described in Palenzuela et al. (2008,

2011) by transplanting 10 Asteriscus maritimus plants with their rhizosphere soils into

1 L pots and transferring them to the greenhouse at EEZ in Granada immediately after

sampling. The pots were irrigated three times per week and fertilized every four weeks

with Long-Aston nutrient solution (Hewitt 1966). Pure cultures of the new fungus were

initiated in a mixed-culture of three Allium porrum L. and three Hieracium pilosella

L. plantlets in 750 mL pots grown together at three locations in the pots. The plant

rhizosphere was inoculated with 20 spores per pot and pot location. A sterile mixture

of Terragreen (American aluminum oxide, Oil Dry US special, type III R; Lobbe

Umwelttechnik Iserlohn, Germany) and Loess (mixture 3:1; with pH-KCl 6.2; organic

carbon 0.3%; available P (Na-acetate) 2.6 mg kg”; available K (Na-acetate) 350 mg kg"!

was chosen as culture substrate (Oehl et al. 2002). So far, pure cultures of the new fungus

have not been obtained.

Morphological analyses

AM fungal spores were separated from the soil samples by a wet sieving process

(Sieverding 1991). The morphological spore characteristics and their subcellular

structures were described from a specimen mounted in: polyvinyl alcohol-lactic acid-

glycerol (PVLG; Koske and Tessier 1983); a mixture of PVLG and Melzer’s reagent

(Brundrett et al. 1994); a mixture of lactic acid to water at 1:1; Melzer’s reagent; and

water (Spain 1990). The spore structure terminology follows Oehl et al. (2003, 2005,

2011a) for species with glomoid or diversisporoid spore formation. Photographs (Fics

1-10) were taken with a Leica DFC 290 digital camera on a Leitz Laborlux S compound

microscope using Leica Application Suite Version V 2.5.0 R1 software. Specimens

mounted in PVLG and the PVLG+Melzer’s mixtures were deposited at the herbaria

Z+ZT (ETH Zurich, Switzerland), GDA-GDAC (University of Granada, Spain), and

URM (Federal University of Pernambuco, Recife).

Molecular analyses

Five spores isolated from the trap cultures were surface-sterilized with chloramine

T (2%) and streptomycin (0.02%) (Mosse 1962) and crushed with a sterile disposable

micropestle in 40 uL milli-Q water (Ferrol et al. 2004). PCRs of the crude extracts

Diversispora clara sp. nov. (Spain) ... 75

were obtained in an automated thermal cycler (Gene Amp PCR System 2400, Perkin-

Elmer, Foster City, California) with a pureTaq Ready-To-Go PCR Bead (Amersham

Biosciences Europe GmbH, Germany) following manufacturer's instructions with 0.4

uM concentration of each primer. A two-step PCR amplified the partial SSU, ITS1,

5.88, ITS2 and partial LSU rDNA ribosomal fragment using the SsUmAf/LSUmAr

and SSUmCf/LSUmBr primers consecutively (Kriger et al. 2009). The second PCR

products were separated electrophoretically on 1.2% agarose gels stained with Gel Red™

(Biotium Inc., Hayward, CA, U.S.A.) and viewed by UV illumination. The band of the

expected size was excised with a scalpel and the amplified DNA was isolated from the

gel with the QIAEX II Gel Extraction kit (QIAGEN, Valencia, CA, USA), cloned into the

PCR2.1 vector (Invitrogen, Carlsbard, CA, USA), and transformed into one shot TOP10

chemically competent Escherichia coli cells. After plasmid isolation from transformed

cells, cloned DNA fragments were sequenced with vector primers (White et al. 1990) in

both directions by Taq polymerase cycle sequencing on an automated DNA sequencer

(Perkin-Elmer ABI Prism 373). Sequence data were compared to gene libraries (EMBL

and GenBank) using BLAST (Altschul et al. 1990). The new sequences were deposited in

the EMBL database under the accession numbers FR873629-FR873633.

PHYLOGENETIC ANALYSES: The AM rDNA ITS1+5.8S +ITS2 fungal sequences were

aligned in ClustalX (Larkin et al. 2007) and edited with BioEdit (Hall 1999) to obtain

a final alignment with Acaulospora laevis Gerd. & Trappe and A. lacunosa J.B. Morton

as outgroups. Prior to phylogenetic analysis, the model of nucleotide substitution

was estimated using Topali 2.5 (Milne et al. 2004). Bayesian (two runs over 1 x 10°

generations with a burn in value of 2500) and maximum likelihood (1000 bootstrap)

analyses were performed in MrBayes 3.1.2 (Ronquist & Huelsenbeck 2003) and PhyML

(Guindon & Gascuel 2003) respectively, launched from Topali 2.5, using the GTR +

G model. Neighbor-joining (established with the model cited above) and maximum

parsimony analyses were performed using PAUP*4b10 (Swofford 2003) with 1000

bootstrap replications.

Results

Diversispora clara Oehl, B. Estrada, G.A. Silva & Palenz., sp. nov. Fries 1-10

MycoBank MB 561583

Sporae albae, 79-130 x 75-125 um, tunica tribus stratis, 4.3-8.4 um. Stratum exterior

hyalinum; stratum medium, album ad rarum ochreo-album, 2.8-5.4 um crassum; stratum

interius album. Hypha adhaerenta, 8-13 um in diametrum. Tunica hyphae 1.0-1.8 um

crassa. Strata medium interiusque septo porum occludentes. Strata exterior mediumque

flava colorantes Melzeri.

Type: Spain. Andalucia, Almeria, Cabo de Gata Natural Park, sand dune, from the

rhizosphere of Asteriscus maritimus (L.) Less. (Asteraceae), isolation date February 2011,

by B. Estrada (holotype, ZT Myc 3796 [permanent slide 20-2001]; isotypes, ZT Myc

3797 [permanent slides 20-2003 to 20-2012], GDA-GDAC [permanent slides 20-2013

to 20-2018], URM [permanent slides 20-2019, 20-2020]).

ErymMo_oey: clara (Latin = clear, bright, brilliant, light), referring to the brilliant white

spores.

76 ... Estrada & al.

=o . ee

Pee

Cand

Fics 1-10. Diversispora clara: 1-4. Uncrushed spores with a triple-layered spore wall (sw11-3), a

single, cylindrical subtending hypha (sh), and a septum (sp) arising directly or at some distance

from the base. Fics 5-6. Spores crushed to show triple layered spore wall (SwL1-3). Fics 7-8. Spore

wall structure in PVLG + Melzer’s reagent. The swL1 or sw12 (or both) stain light to dark yellow

in Melzer’s, while no such staining reaction is seen on SwL3. However, the yellow stain is usually

most noticeable in the spore cell contents. Fic. 9. Septum at spore base formed by sw12 (sw13 is

not visible in uncrushed spore). Fic. 10. Septum at spore base formed by sw13.

SPoRES (Fics 1-4) singly formed in rhizosphere soils, terminally on subtending

hyphae, globose (80-110 um diam.) to subglobose (79-130 x 75-125 um), one-

walled, brilliant to creamy white.

SPORE WALL 4.3-8.4 um thick, three-layered (Swil, swL2, SwL3; FIGs 5-7);

outer layer (Swi1) hyaline, 0.8-1.5 um thick, evanescent and thus often

absent in mature spores; second layer (sw12) bright to (rarely) creamy white,

laminated, 2.8-5.4 um thick (Fic. 7) in uncrushed spores (sometimes < 6.6 um

in crushed spores when pressure is applied on the cover slip); inner layer (SwL3)

Diversispora clara sp. nov. (Spain) ... 77

brilliant white, 0.7-1.5 um thick (usually tightly adherent to sw12 and difficult

to observe when < 1.0 um; see Fics 5-6); both swi1 and sw12 generally light to

dark yellow in Melzer’s reagent (Fics 7-8, with spore cell contents often a more

intense yellow than the surrounding cell wall layers (Fic. 8).

SUBTENDING HYPHAE (sh) generally singly on spores; brilliant white

(Fics 1-3), cylindrical or (rarely) slightly constricted at the spore base, 4.0-8.0

um broad tapering to 3.2-6.1 um within 100 um of spore base, although the

distance may appear shorter (4.0-10(-25) um) because the sh walls taper from

1.0-1.8 um thick to 0.6-1.2 um within the first 10 um from the base causing the

flexible fragile portion to break from the mature spore (Fics 3, 4, 9) at a point

where the septum separates the spore contents from the hyphal contents. Spore

pores at the spore bases or in the sh normally closed by a septum (Fics 1-2, 4)

arising from sw12 (Fic. 9), swi3 (Fic. 10), or both layers (not shown); pores

rarely open (Fic. 3).

DISTRIBUTION: Known only in the natural sand dune system of the Cabo de

Gata Natural Park in Andalucia in the rhizosphere of Asteriscus maritimus.

MOLECULAR ANALYSES: Phylogenies derived from ITS (Fic. 11) and 28S (data

not shown) rDNA analyses cluster the new fungus within the Diversisporaceae

in a well-separated clade adjacent to several other Diversispora species, Otospora

bareae, and Tricispora nevadensis (Oehl et al. 2011b).

Discussion

Our morphological analyses, in particular those of the subtending hyphae

and spore bases, clearly support the new fungus in Diversispora, with many

characters identical to those of other Diversispora species (e.g., D. spurca,

D. eburnea; see Oehl et al. 2011a,b) such as the thin-walled, hyaline, cylindrical

(or rarely constricted) subtending hyphae that appear flexible and fragile behind

the septum that closes the pore at the spore base or in short distance from the

base. In Diversispora species with pigmented spores, the subtending hyphae

regularly change color conspicuously, becoming hyaline to white behind the

septum (Oehl et al. 2011a). This color change, however, was not confirmed for

D. clara, since the new fungus generally does not form pigmented spores.

Molecular analyses confirm the species as new: in the phylogenetic tree,

D. clara clusters in a independent, monophyletic clade within a polyphyletic

Diversispora. ‘These sequence analyses also confirm unequivocally using

morphology to identify diversisporoid species within the Glomeromycota (Oehl

et al. 201 1a,c).

Including D. clara, there are now 14 Diversispora spp. known in the

Glomeromycetes (Oehl et al. 2011a,b). The 9 species that form significantly

pigmented, yellow brown to brown or orange to orange brown spores are

D. arenaria (Blaszk. et al.) Oehl et al., D. aurantia (Blaszk. et al.) C. Walker

& A. Schiissler, D. epigaea (B.A. Daniels & Trappe) C. Walker & A. Schiissler,

78 ... Estrada & al.

D. insculpta (Btaszk.) Oehl et al., D. przelewicensis (Blaszk.) Oehl et al.,

D. pustulata (Koske et al.) Oehl et al., D. tenera (P.A. Tandy) Oehl et al.,

D. trimurales (Koske & Halvorson) C. Walker & A. Schiissler, and D. versiformis

(P. Karst.) Oehl et al. (Oehl et al. 2011a,b). Diversispora celata C. Walker et

al. forms triple-layered, ochre to ivory to pinkish cream spores (Gamper et al.

2009). Only D. spurca (C.M. Pfeiff. et al.) C. Walker & A. Schiissler, D. eburnea

(L.J. Kenn. et al.) C. Walker & A. Schiissler, and D. gibbosa (Blaszk.) Blaszk. &

Kovacs form hyaline to subhyaline spores that are, however, never brilliant-

white as observed for D. clara. Moreover, D. spurca and D. eburnea have bi-

layered spore walls (Kennedy et al. 1999), D. gibbosa has a five-layered wall

(Blaszkowski 1997), and D. clara has a three-layered wall.

In the past, large-spored or sporocarpic AM fungi were described from sand

dune systems, such as Gigaspora, Scutellospora, Pacispora or Glomus species

that were generally easy to isolate and recognize from field samples (Koske &

Gemma 1995, Gemma et al. 1989, Blaszkowski 1994). Likewise in the current

Cabo de Gata Natural Park sand dune system, where Funneliformis coronatus

(Giovann.) C. Walker & A. Schtissler, E mosseae (T.H. Nicolson & Gerd.)

C. Walker & A. Schiissler, Scutellospora calospora (T.H. Nicolson & Gerd.)

C. Walker & EE. Sanders, Racocetra persica (Koske & C. Walker) Oehl et al.,

and Glomus macrocarpum Tul & C. Tul. were identified (Estrada, unpublished).

When sand dune AM fungal communities were maintained and reproduced

in bait cultures, small-spored Glomus spp. were also sometimes detected (e.g.

Blaszkowski et al. 2009a,b, 2010). It was supposed that species with small, quickly

degrading spores were difficult to recover or identify only from field samples.

This might also be true for D. clara, even though its laminated wall structure

is clearly persistent. It will be interesting to see whether future taxonomists

will be able to identify the new species directly from field samples, now that

the existence and morphology of this unique, brilliant-white, conspicuous but

small-spored species is known. Morphological spore and molecular root and

spore analyses will hopefully tell us more about the ecology and biogeography

of this fungus that is thus far known only from a single Asteriscus maritimus

rhizosphere in the Natural Park Cabo de Gata of Almeria in southern Spain.

Acknowledgments

This study has been supported by the Junta de Andalucia (Spain), project P06-CVI-

01876 and by the Swiss National Science Foundation (SNSF; Project 315230_130764/1).

We acknowledge the valuable comments on the manuscript and revisions of Dr. Bruno

Tomio Goto (Universidade Federal do Rio Grande do Norte, UFRN, Natal, Brazil), Dr.

Ivan Sanchez-Castro (INRA, Dijon, France) and PD Dr. Ewald Sieverding (University

of Hohenheim, Germany) and appreciate the corrections by Shaun Pennycook,

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

Diversispora clara sp. nov. (Spain) ... 79

Acaulospora laevis AJ242499

A. lacunosa AJ891113

100

Redeckera pulvinata AM418550

R. pulvinata AM418549

41.00

93} 99 _ R megalocarpa AM418552

400| 95 g P

as ee R. megalocarpa AM418551

99) R. fulva AM418547

tool rR. fulva AM418548

1.00 R. fulva AM418545

R. fulva AM418546

72 Diversispora spurca FN547641

100 71

100 76 D. spurca FN547644

100 0.93

1.00 D. spurca FN547637

99 D. aurantia AJ849468

ar D. aurantia FN547661

ari D. aurantia FN547655

a4 D. aurantia FN547657

1.00 es D. celata AM713403

88 D. celata AM713402

81 0.86

83 D. celata AM713404

0.09 72| | |r D. eburnea AM713409

a D. eburnea AM713408

O97 D. eburnea AM713416

D. eburnea AM713407

D. clara FR873632

D. clara FR873629

ue D. clara FR873633

9,96 D. clara FR873631

D. clara FR873630

D. versiformis FN547635

98 D. versiformis AY842567

a5 D. versiformis FN547674

1.00 D. versiformis AY842569

D. versiformis FN547666

ton Otospora bareae FR865444

ape O. bareae FR865445

1007 Entrophospora nevadensis FR865447

408 E. nevadensis FR865448

1.00. EF nevadensis FR865446

0.1

Fic. 11. Diversisporaceae. ITS rDNA-based phylogenetic tree rooted by Acaulospora laevis and

A. lacunosa. Sequences are labeled with database accession numbers. Support values are from

neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and bayesian

analyses. Diversispora clara sequences are in bold. Only topologies with = 50% bootstrap values are

shown. (Consistency Index = 0.72; Retention Index = 0.84).

80 ... Estrada & al.

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two new genera in the arbuscular mycorrhizal Glomeromycetes. J. Appl. Bot. Food Qual. 80:

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methods and applications. Academic Press, San Diego, California.

MY COTAXON

http://dx.doi.org/10.5248/118.83

Volume 118, pp. 83-88 October-December 2011

Asterophora salvaterrensis (Basidiomycota, Agaricales),

a new species from Galicia (Spain)

JAIME B. BLANCO-DIOsS

Centro de Formacion e Experimentacion Agroforestal de Lourizan,

Conselleria de Medio Rural, Xunta de Galicia, RO. Box 127.36080, Pontevedra,Spain

* CORRESPONDENCE TO: jbblancodios@gmail.com

ABSTRACT—Asterophora salvaterrensis, collected in Pinus pinaster forests in Galicia

(northwest Iberian Peninsula), is described as new and compared with other known

Asterophora species. The new species is distinguished by its zonate greenish brown to

brownish ochre pileus with whitish margin that blackens, decurrent obtuse furfuraceous grey

lamellae, furfuraceous brown to blackish stipe, black context, nondistinctive odor and taste,

and small basidiospores. A key to all known Asterophora species is provided.

Key worps—Lyophyllaceae, parasitic fungi, taxonomy

Introduction

Mycological studies conducted during the last decades in the forests

of maritime pine (Pinus pinaster Aiton) in the Mifo river basin of Galicia

(northwestern Spain) have produced interesting discoveries, such as the new

species, Sparassis miniensis Blanco-Dios & Zheng Wang (Blanco-Dios et al.

2006). In this territory, several samples of a remarkable species of Asterophora

Ditmar were recently collected on a russulaceous host (Russula sp.).

Species of Asterophora (Lyophyllaceae) are among the few agarics that grow

on basidiomata of other fungi (the most common hosts are russulaceous species,

mainly in Lactarius Pers. and Russula Pers.). The genus is further distinguished

by the production of chlamydospores. Asterophora is phylogenetically placed in

Lyophyllaceae (Matheny et al. 2006).

Today, only three well documented species of Asterophora are accepted:

A. lycoperdoides (Bull.) Ditmar, A. parasitica (Pers.) Singer, and A. mirabilis

(T.W. May) Redhead & Seifert (May & Fuhrer 1995; Redhead & Seifert 2001a,b;

Vizzini 2009). Our collections, which do not fit the morphological concept of

any of these taxa, are described here as a new species.

A key to the known species of Asterophora is provided.

84 ... Blanco-Dios

Materials & methods

Macro- and micromorphological descriptions are based on observations of both

fresh and dried specimens. L = number of lamellae; Q = quotient of length /width of

an individual spore and avQ is the average Q. All the micromorphological structures

were measured in Melzer’s or 3% KOH, except for the pileipellis and stipitipellis

hyphae, which were measured in 10% ammonia. Spore measurements do not include

the hilar appendage. The length of intercalary chlamydospores was measured from the

septum of one subtending hypha to the septum of the other; the length of terminal

chlamydospores was measured from basal septum to apex. All specimens, including

the holotype, are kept in LOU-Fungi herbarium (Centro de Investigacién Forestal de

Lourizan, Pontevedra, Spain).

Taxonomy

Asterophora salvaterrensis Blanco-Dios, sp. nov. PLATE 1, Fic. 1

MycoBank MB 519728

Pileus 0.5-4 mm latus, planus leviter depressus, non hygrophanus, haud traslucenter

striatus, zonatus, ab brunneo-viridulus ad brunneo-ochraceus, ad medium brunneus,

aliunde albidus, nigrescens. Lamellae decurrentes, dispersae, obtusatae, furfuraceae,

griseae. Stipes 1.5-10 mm longus, 0.5-1.5 mm latus, plenus, furfuraceus, brunneus vel

brunneo-nigrescens. Caro nigrescens, parce conspicua. Odor et sapor haud notabiles. Sporae

2.5-3.2(-4.5) x (1.2-)1.8-2.5 um, Q = 1.29-2, ellipsoidaeae vel oblongae, laeves. Basidia

tetrasporigera, fibulata. Acies lamellarum fertilis. Chlamydosporae (12-)13-17.5(-20.5) x

(7.3-)8.5-10.5(-11.5) um Q=1.26-2, ellipsoidaeae vel oblongae, laeves. Fibulae praesentes.

Ad basidiomata corruptas Russulae nigricantis.

Type: Spain: Pontevedra, Salvaterra de Mifio, Leirado. Collected by J.B.Blanco-Dios,

1-XII-2009 (Holotype: LOU-Fungi 19491).

EryMo.oey: salvaterrensis, from the municipality of Salvaterra de Mino (Galicia,

Spain).

PiLEus 0.5-4 mm diam., plano-convex to flattened with depressed centre,

margin first involute, then broadly wavy, not hygrophanous or translucently

striate, zonate, brown at the disc, greenish brown to brownish ochre elsewhere

with the margin whitish, blackening with age or handling. LAMELLAE < 1 mm

broad, decurrent, scattered (L = 8-10), thick, not forked, entirely furfuraceous

over a grey pale or grey colour, edge concolourous, even, obtuse, lamellulae

absent. Stipe 1.5-10 x 0.5-1.5 mm, central, even or a bit expanded above or

attenuate, filled, entirely furfuraceous over a brown or blackish brown surface.

CONTEXT very thin, black when exposed, finally brown to ochre. ODOR AND

TASTE nondistinctive. SPORE PRINT whitish.

BASIDIOSPORES 2.5-3.2(-4.5) x (1.2-)1.8-2.5 um, Q = 1.29-2, avQ = 1.47,

(n = 30), sparse, ellipsoid to oblong, smooth, thin-walled, not amyloid. Bastp1a

(10-)13-19 X 4-7 um, sparse, broadly clavate, 4-spored, sterigmata up to 2.5

um long, siderophilous. CHLAMYDOSPORES (12-)13-17.5(-20.5)x (7.3-)

8.5-10.5(-11.5) um, Q = 1.26-2, avQ = 1.56, numerous in the lamellar trama,

Asterophora salvaterrensis sp. nov. (Spain) ... 85

PLaTE. 1. Asterophora salvaterrensis (holotype). Habit.

Photos by Amancio Castro.

86 ... Blanco-Dios

0006

9000

Fic. 1. Asterophora salvaterrensis (holotype).

a. basidiospores; b.basidia; c. chlamydospores. Scale bars = 10 um.

in the pileal trama above the lamellae, and on the pileus surface, numerous also

in the host tissue, intercalary or terminal, smooth, ellipsoid to oblong, hyaline

then pale yellow, at length with an inner wall up to 0.5 um thick which excludes

a segment adjacent to each subtending hypha (the basal portion of terminal

chlamydospores is similarly excluded), enclosing contents which are granular

and with small and large, refractive droplets. CysTip1a not seen. PILEIPELLIS a

cutis, consisting of parallel, cylindrical hyphae, 1.5-10 um diam., thin walled,

radially arranged, with ochre intracellular pigment. STIPITIPELLIS a cutis,

composed of parallel, cylindrical hyphae, 1.5-9 um diam., thin walled. CLamp

CONNECTIONS present in all tissues.

ECOLOGY & DISTRIBUTION— On blackened and rotten Russula nigricans

basidiomata, growing together with Asterophora lycoperdoides and A. parasitica

in a Pinus pinaster forest. Known only froma single locality in Spain. November-

December.

COLLECTIONS EXAMINED: SPAIN: PONTEVEDRA: SALVATERRA DE MINO, Leirado,

29TNG4565, 110 m, forest of Pinus pinaster, on basidiomata of Russula nigricans,

1.X1I.2009, leg. J.B.Blanco-Dios, LOU-Fungi 19491 (holotypus); 24.X1.2009, leg.

J.B.Blanco-Dios, LOU-Fungi 19490.

OTHER COLLECTIONS EXAMINED: Asterophora lycoperdoides - SPAIN: A CORUNA:

Brion, Adoufe, 29TNH2645, 70 m., on basidiomata of Russula nigricans, 19.XII.1977,

leg. L. Cabo Rey, LOU-Fungi 0114; Santraco, bosque de la Condesa, 29TNH3747,

Asterophora salvaterrensis sp. nov. (Spain) ... 87

250 m., on basidiomata of Russula nigricans, 23.X.1975, leg. L. Freire, LOU-Fungi

0112. PONTEVEDRA: A EstTRADA, Arnois, 29TNH4834, 60 m., on basidiomata of

Russula nigricans, 23.X.1975, leg. L. Freire, LOU-Fungi 0116; ViILAGARCIA DE AROUSA,

29TNH1916, 20 m., on basidiomata of Russula sp, 18.X1.1987, leg.: E.Valdés-Bermejo,

LOU-Fungi 0113.

Asterophora parasitica - SPAIN: A CoruNa: Bridn, Adoufe, 29TNH2645, 70 m.,

on basidiomata of Russula nigricans, 12.X1.1981, leg. L. Cabo Rey, LOU-Fungi 0117;

CAMBRE, Cecebre, Fraga de Cecebre, 29TNH5794, 100 m., on basidiomata of Russula sp.,

17.X1.1990, leg. L. Freire, LOU-Fungi 4557; PUEBLA DO CARAMINAL, Cabio, 29TNH0514,

10 m., on basidiomata of Russula densifolia Secr. ex Gillet, 25.X1.1989, leg. M. Martinez

Campos, LOU-Fungi 6930; on basidiomata of Russula nigricans, 08.X11.1984, leg. L.

Freire & M. L. Castro, LOU-Fungi 0121. Lugo: MONDONEDO, Couboeira, 29TPJ3215,

100 m., on basidiomata of Russula sp., 3.X.1992, leg. S. Cabanela, LOU-Fungi 5844.

ComMENTS. Asterophora salvaterrensis is unique with respect to the other

known Asterophora species —A. lycoperdoides, A. parasitica, A. mirabilis— by

the following combination of features: (i) the zonate pileus which is brown

at the disc, greenish brown to brownish ochre towards margin and whitish

at the margin, (ii) the pileus-surface turning blackish with age or handling,

(iii) the decurrent lamellae, which are furfuraceous and grey-tinged, (iv) the

furfuraceous stipe-surface over a brown or blackish-brown cortical layer, (v)

the context blackening when exposed and lacking a distinct smell and taste.

From the micromorphological point of view the small, ellipsoid to oblong

basidiospores associated with large, smooth, ellipsoid to oblong chlamydospores

are typical and diagnostic. Some stellate chlamydospores (from basidiomata of

Asterophora lycoperdoides),13-16.5 x 12-15 um, were seen in the pileipellis,

lamellae, and stipitipellis surface.

Asterophora lycoperdoides differs by having stellate chlamydospores, formed

in the upper pileal trama. Additionally, this species frequently does not produce

basidiospores, has a globose to pulvinate pileus with a powdery surface, and

shows often only rudimentary lamellae. Asterophora parasitica also frequently

does not produce basidiospores, has a convex to umbonate pileus with a

typically silky surface and obtuse but comparatively well-formed lamellae.

Both of these species cover a similar geographic range spanning through North

America, Europe, North Africa, East Asia, and Papua New Guinea (Corner

1966, Horak 1980, Singer 1986). Asterophora mirabilis, known from Australia

(Victoria and Tasmania), is similar to A. parasitica, but differs in producing

stellate chlamydospores that are numerous in the lamellar trama and pileal

trama above the lamellae and sparse to absent in the upper pileal trama and on

the pileus surface (May & Fuhrer 1995).

From the taxonomic point of view, owing to the fact that it produces only

smooth chlamydospores, Asterophora salvaterrensis seems to be close to

Asterophora parasitica from which it is easily distinguished by greenish-brown

to greenish-ochre zonate pileus and much smaller basidiospores.

88 ... Blanco-Dios

Key to the known species of Asterophora

bar GC blamiydosparesistellatees £4chcchs dds Ad BR a Rg Ey 2

LBAE WamMiyAOS POLES: SIO ORD 5h fon he fe aie Beale eee 2 be ae RN LAR Nas ot 3

2a. Pileus surface powdery; chlamydospores formed in the upper pileal trama

ws tis oS Siig Be Seta he Mey: Ma Sees WE Ricci, WE pe Se Mipy 8 Ri se IS A. lycoperdoides

2b. Pileus smooth; chlamydospores formed in the lamellar trama

and pileal trama above the lamellae............... 0... eee eee eee A. mirabilis

3a. Pileus white or grey with a silky surface, not zonate;

basidiospores, 5-6, 52554: Wiis e505 Fs FE AR Whaat UT A. parasitica

3b. Pileus brown-greenish-ochre, zonate;

basidiospores 2:5=3r -* 1.8-2.5 aime sn, See gee cle! A. salvaterrensis

Acknowledgements

The author is grateful to Drs. Scott A. Redhead and Tom W. May for kindly sending

relevant literature. Drs. Alfredo Justo, Alfredo Vizzini, and Marco Contu are thanked

for critically reviewing an earlier draft of this paper. Amancio Castro is gratefully

acknowledged for providing the photographs. Finally I express my most sincere thanks

to the direction and members of the Centro de Investigacién Forestal de Lourizan

(Conselleria de Medio Rural, Xunta de Galicia) for curating the herbarium LOU-

Fungi.

Literature cited

Blanco-Dios JB, Wang Z, Binder M, Hibbett DS. 2006. A new Sparassis from Spain described using

morphological and molecular data. Myc. Res. 110: 1127-1134. http://dx.doi.org/10.1016/

j-mycres.2006.07.012.

Corner EJH. 1966. A monograph of cantharelloid fungi. Ann. Bot. Mem. 2: 1-255.

Horak E. 1980. New and remarkable hymenomycetes from tropical forests in Indonesia (Java) and

Australasia. Sydowia 33: 39-63.

Matheny PB, Curtis JM, Hofstetter V, Aime MC, Moncalvo J-M, Ge Z-W, Yang Z-L, Slot JC,

Ammirati JF, Baroni TJ, Bougher NL, Hughes KW, Lodge DJ, Kerrigan RW, Seidl MT, Aanen

DK, DeNitis M, Daniele GM, Desjardin DE, Kropp BR, Norvell LL, Parker A, Vellinga EC,

Vilgalys R, Hibbett DS. 2006. Major clades of Agaricales: a multilocus phylogenetic overview.

Mycologia 98(6): 982-995.

May TW, Fuhrer BA. 1995. Nyctalis mirabilis (Fungi, Agaricales), a new species from Australia.

Muelleria 8: 385-390.

Redhead SA, Seifert KA. 2001a. Asterophora Ditmar ex Link 1809 versus Nyctalis Fries 1825, and

the status of Ugola Adanson 1763. Taxon 50: 243-268. http://dx.doi.org/10.2307/1224526

Redhead SA, Seifert KA. 2001b. Proposal to conserve the name Agaricus lycoperdoides Bull.

(=Asterophora lycoperdoides (Bull.) Ditmar against Asterophora agaricoides Fr.: Fr. and

Asterophora lycoperdoides Fr.: Fr. Taxon 50: 279-280. http://dx.doi.org/10.2307/1224530

Singer R. 1986. The Agaricales in modern taxonomy. 4th ed. Koeltz Scientific Books. Koenigstein.

Vizzini A. 2009. Revisione del genere Asterophora ed introduzione allo studio delle Lyophylleae. 371-

376, in: J-C Maire et al. (eds). Compléments a la Flore des champignons supérieurs du Maroc de

G. Malencon et R. Bertault. Confédération Européenne de Mycologie Méditerranéenne, Nice.

MY COTAXON

http://dx.doi.org/10.5248/118.89

Volume 118, pp. 89-94 October-December 2011

Amicodisca castaneae sp. nov.

(Hyaloscyphaceae, Helotiales) on Japanese chestnut bur

JAE-Gu Han’, TsuyosHi Hosoya? & HYEON-DONG SHIN”

"Division of Environmental Science and Ecological Engineering,

College of Life Sciences and Biotechnology, Korea University,

Seoul 136-701, Korea

*Department of Botany, National Museum of Nature and Science,

4-1-1 Amakubo, Tsukuba, Ibaraki 305-0005, Japan

*CORRESPONDENCE TO: hdshin@korea.ac.kr

ABSTRACT— Amicodisca castaneae sp. nov. was collected on Japanese chestnut bur. It differs

from all known members of Amicodisca by having brownish apothecia and smaller asci and

ascospores. A key to the accepted Amicodisca species is given.

KEY worpDs— Castanea crenata, discomycetes, species nova, taxonomy

Introduction

Svréek (1987) established a monotypic genus Amicodisca Svréek, typified

by Amicodisca brdensis (Velen.) Svréek (= Dasyscyphus brdensis Velen.), which

is characterized by yellowish to olivaceous colored excipulum and hairs,

ascospores bearing lemon-yellow pigment dissolving in NH,OH or KOH

solutions, and fimbriate dehiscence of the ascus pore. Amicodisca viridicoma

(Peck) J.H. Haines, A. svrcekii Raitv. & Huhtinen, and A. groenlandica Raitv.

were later additionally described (Haines 1989; Raitviir 2001, 2003). Since

Huhtinen (1994) proposed A. virella (P. Karst.) Huhtinen (with its earlier

basionym) as the correct name for the type species, and Raitviir (2004) listed A.

viridicoma as a synonym of A. virella, there are three species now recognized.

In the course of extensive survey on the fungal biodiversity in Korea, an

interesting discomycete with small apothecia surrounded with yellowish hairs

was collected on fallen burs of Japanese chestnut (Castanea crenata Siebold

& Zucc.). Its gross morphology and microscopic features fit well with those

of Amicodisca, but it is clearly different from the known species in having

remarkably smaller asci and ascospores and by host preference. Here, we

describe and illustrate this fungus as new to science.

90 ... Han, Hosoya & Shin

Materials & methods

Fresh materials were primarily mounted in distilled water to confirm the natural

colors of their microstructures. Dried materials were revived in 3-10% aqueous KOH.

Amyloid reactions were tested by Melzer’s reagent (MLZ) or Lugol's solution (IKI)

without KOH pretreatment. Olympus BX50 microscope equipped with a drawing

tube (Olympus U-DA) was used for measurements and line drawings. Structures were

measured at 1000x; measurements are reported as follows: minimum-—maximum

(length) x minimum-maximum (width) [mean length + standard deviation x mean

width + standard deviation, Q (I/w ratio) = average + standard deviation]. All specimens

examined are deposited in the Herbarium of Korea University, Seoul, Korea (KUS) or

National Museum of Nature and Science, Tsukuba, Japan (TNS).

Taxonomy

Amicodisca castaneae J.G. Han, Hosoya & H.D. Shin, sp. nov. Fics. 1-2

MycoBAank 561134

Apothecia sessilia, bruneta, receptaculo dense sulphureo longipiloso. Pili cylindro-conique,

2-5-septati, tenuiter tunicatis, 63-107 x 2-3 um. Excipulum ectale ex textura angularis ad

prismatica compositur. Asci non uncinati, cinereae, 38-57 x 4-5.2 um. Sporae ellipsoidea

ad clavato-ellipsoideae, hyalinae ad cinereae, 4.8-7 x 1.4-2 um. Ab Amicodiscae svrcekii

ascosporis brevius et pili septatae differens.

Type: The inner surface of chestnut bur (Castanea crenata), Korea, Goesan, Songnisan

National Park, 36°44'42.54"N, 128°53'53.33”E, 320 ma.s.L, 5 Oct. 2007, J.G. Han & H.D.

Shin (KUS-F51917 Holotype; TNS-F32105 Isotype).

ErymMo.ocy: The specific epithet refers to the generic name of the host.

APOTHECIA superficial, scattered to gregarious, developed on well-developed

anchoring hyphae, broadly sessile; RECEPTACLE at first globose to cupulate,

then becoming discoid, externally covered by yellowish white hairs, turning

reddish-brown when dry; Disc up to 3 mm in diameter, greyish-brown to

chestnut when fresh, turning dark brown to dark grey when dry; Harrs

cylindric-conical, straight, gradually tapering toward the apex, hyaline to

yellowish, 2-5-septate, thin-walled, smooth, apex not sharply pointed, up to

3 um wide near the base, 63-107 um long; ECTAL EXCIPULUM yellow to light

brown, composed of thin-walled angular to rectangular cells, 7-12 x 4-7 um;

AscI arising from simple septa, cylindric-clavate, yellowish, 8-spored, apical

pore blued in MLZ and IKI without KOH pretreatment, 38-57 x 4-5.2 um

(45.2 + 4.19 x 4.5 + 0.41 um, n = 50); Ascosporss biseriate, ellipsoid to clavate

or ovoid, hyaline to light yellow, aseptate, eguttulate, occasionally containing a

central guttule, smooth, 4.8-7 x 1.4—2 um (5.7 + 0.69 x 1.6 + 0.18 um, Q = 3.6 +

0.62, n = 120); PARAPHysSES filiform, hyaline to yellowish, septate, slightly bent,

sometimes branched, slightly exceeding the asci, up to 1.5 um wide.

ADDITIONAL SPECIMENS EXAMINED - the inner surface and spines of chestnut

bur: KOREA, Wownju, Chiaksan National Park, Geumdae valley, 37°17'44.44"N,

128°01'10.34"E, 370 m as.l, 9 Sep. 2006, J.G. Han & H.D. Shin (KUS-F51377);

Amicodisca castaneae sp. nov. (Korea) ... 91

Fic. 1. Amicodisca castaneae (holotype KUS-F51917). A: Fruiting bodies on a fallen bur of Castanea

crenata, B: Sessile apothecia densely gregarious, C: An immature apothecium, globose to cupulate,

D: A mature apothecium, expanded. Scale bar: C-D = 1 mm.

92 ... Han, Hosoya & Shin

BORYEONG, Oseosan recreation forest, 36°26'31.52”N 126°40'3.2"E, 240 m a.s.l., 27 Oct.

2007, J.G. Han & H.D. Shin (KUS-F51993).

Discussion

Amicodisca is a small group of hyaloscyphaceous discomycetes characterized

by a lemon-yellow colored excipulum, hairs, and ascospores (Svréek 1987).

Although its dark globose ectal cells and translucent greyish hymenium

suggest a relationship to Dermateaceae, many authors (Svréek 1987; Haines

1989; Huhtinen 1994; Huhtinen & Less@e 2001; Raitviir 2003, 2004) have

preferred to assign the genus to the Hyaloscyphaceae based on the presence of

distinct hairs. Dematioscypha Svréek is similar to Amicodisca in having a dark

excipulum and cylindric hairs with tapering apices, but differentiate the genus

by its hyaline hairs with glassy apices, inamyloid ascal pore, and association

with a Haplographium anamorph. Superficially, it is also reminiscent of

Dennisiodiscus Svréek in sessile apothecia with greyish hymenium and bright

colored surrounding hairs, but differs in hairs encrusted by reddish brown

granules and growing on culms or leaves of monocotyledonous hosts.

Macroscopically, the present fungus is easily discriminated from other

species by relatively large apothecia (< 3 mm diam.) with brownish hymenium.

In contrast, all known species of Amicodisca produce greyish or olivaceous

apothecia and are usually smaller than 1 mm in diameter. Amicodisca castaneae

is microscopically closest to A. svrcekii and A. virella, but their larger ascospores

(8-11 and 17-25 um long, respectively) and slightly narrower paraphyses

(0.8 and 1 um broad, respectively) are different. Amicodisca groenlandica

strikingly differs in much larger asci (92-116 x 12-15 um) and ascospores

(20-27 x 3-5 um).

The specificity of A. castaneae to chestnut burs seems to be another notable

feature. Hitherto all known Amicodisca species have been collected on woody

substrates (Raitviir 2004), such as Alnus, Betula, Salix, Sambucus, and other

deciduous trees.

A key to the accepted species of the genus is provided below.

Key to the accepted species of Amicodisca

la. Fruiting bodies formed on chestnut bur; disc brownish ............ A. castaneae

1b. Fruiting bodies formed on woods; disc greyish or olivaceous ...............4. 2

2a (1b). Disc pale greenish-olivaceous when fresh, becoming dark green when dry;

ascilonGer, than-SO iii tes eie we, Me tem hein, Mita s eet aes tann pests A. groenlandica

2b. Disc greyish when fresh and dry; asci shorter than 80 um ................006. 3

3a (2b). Ascospores 8-11 x 1.8-2.5 UM ......... eee eee eee ee eee A. svrcekii

Sb: ASCosporesyl 7225 9CA— Git” Le Paice pela a elaine dle lant a aed gn dlanatend as A. virella

Amicodisca castaneae sp. nov. (Korea) ... 93

gq lee

Tee CAE

Fic. 2. Amicodisca castaneae (holotype KUS-F51917). A: 8-spored asci, note on the apical pore blued

in MLZ, B: Ellipsoid to ovoid ascospores with pale yellowish pigments, C: Filiform paraphyses, D:

Cylindric hairs with tapering apices, E: Brownish ectal excipulum composed of texture angularis to

prismatica. Scale bars: A and C-E = 20 um, B = 10 um.

94 ... Han, Hosoya & Shin

Acknowledgements

The authors are grateful to Dr. Seppo Huhtinen (Herbarium, University of Turku,

Finland) and Dr. Peter Johnston (Landcare Research, New Zealand) for kindly reading

the manuscript and suggesting appropriate revisions. This work was supported bya grant

from Regional Subgenebank Support Program of Rural Development Administration,

Republic of Korea.

Literature cited

Haines JH. 1989. Studies in the Hyaloscyphaceae V: Species described by C.H. Peck. Mycotaxon

$5:3:17=352,

Huhtinen S. 1994. Finnish records of discomycetes: type studies on some Karsten species. Karstenia

34: 5-12.

Huhtinen S, Leessae T. 2001. Amicodisca - en skivesvampeslegt med to smukke, mennesten ens

arter. Svampe 43: 43-47.

Raitviir A. 2001. Taxonomic notes on Dematioscypha and Amicodisca. Czech Mycol. 52: 289-294.

Raitviir A. 2003. New or forgotten Helotiales from Greenland 1. Dermateaceae and Hyaloscyphaceae.

Mycotaxon 87: 359-378.

Raitviir A. 2004. Revised synopsis of the Hyaloscyphaceae. Scripta Mycol. 20: 1-133.

Svréek M. 1987. New or less known discomycetes. XV. Czech Mycol. 41: 16-25.

MYCOTAXON

http://dx.doi.org/10.5248/118.95

Volume 118, pp. 95-101 October-December 2011

Pseudocercospora epidendri sp. nov. on the neotropical orchid,

Epidendrum secundum from Brazil

MEIRIELE DA SILVA & OLINTO LIPARINI PEREIRA*

Departamento de Fitopatologia, Universidade Federal de Vicosa

Vicosa, Minas Gerais, 36570-000, Brazil

* CORRESPONDENCE TO: oliparini@ufv.br

ABSTRACT — A new leaf spot disease was observed on the orchid species Epidendrum

secundum in “high altitude grasslands” of Araponga, Minas Gerais State, Brazil. Inoculation

tests on healthy plants confirmed the pathogenicity of this fungus. Pseudocercospora epidendri

sp. nov., the causal agent of the leaf spot disease of E. secundum, is described, illustrated, and

compared with allied Pseudocercospora species on hosts of the family Orchidaceae.

KEY worps — cercosporoid hyphomycetes, phytopathology, plant disease, tropical fungi

Introduction

Epidendrum secundum is an orchid species with epiphytical or terrestrial

habitat, characteristic of the Brazilian “high altitude grasslands” (PLATE 1)

and the Brazilian Cerrado (Caiafa & Silva 2005, Neto et al. 2007). In 2002,

exploratory research was begun with the goal of describing the phytopathogenic

mycodiversity associated with Orchidaceae in the state of Minas Gerais (Pereira

et al. 2002, Pereira & Barreto 2004, Silva & Pereira 2007, 2008, Silva et al. 2008,

Lopes et al. 2009, Pereira & Silva 2009). During an initial botanical survey in

the “high altitude grasslands” of Parque Estadual da Serra do Brigadeiro in

Araponga in Minas Gerais State, Brazil, samples of the orchid E. secundum

with leaf spots caused by a cercosporoid hyphomycete were collected (PLATE

2-3). Our morphological characterization showed that the leaf-spotting fungus

represented a new species of Pseudocercospora, which we describe, illustrate,

and discuss below.

Material & methods

Samples of infected E. secundum leaves were collected, photographed (SONY

DSC-H9 digital camera), dried in a plant press, and deposited at the herbarium of

Universidade Federal de Vicosa (VIC). Fungal samples were removed from fresh

96 ... Silva & Pereira

> ww

le \ -

Pes . \

PiaTeE 1. Terrestrial Epidendrum secundum in the “high altitude grasslands” of the Parque

Estadual da Serra do Brigadeiro, Araponga, State of Minas Gerais, Brazil.

leaf spots under an Olympus SZX7 stereomicroscope and mounted on glass slides

with lactophenol. Material was also hand sectioned for stromata observation and

measurements. Structures were observed, measured (30 of each structural type), and

illustrated under 400x magnification using an Olympus BX 50 light microscope fitted

with a drawing tube. To determine pathogenicity, the fungus was isolated directly onto

PDA, brought into pure culture, and grown at 27°C for 20 days. Culture disks taken

Pseudocercospora epidendri sp. nov. (Brazil) ... 97

. )

\\ aN

-- in nd

PLATE 2-3. Pseudocercospora epidendri. Detail of infected leaf, showing delimited and

irregular necrotic spots. 2. Adaxial leaf surface. 3. Abaxial leaf surface.

from colony borders were used to inoculate four healthy young and mature leaves of

three E. secundum plants. The inoculated plants were maintained in moist chambers for

three days and then transferred to a greenhouse at 25°C and 70-80% humidity. Healthy

leaves, on which only PDA plugs were placed, served as control. This assay was carried

in three replications.

98 ... Silva & Pereira

PiaTE 4-5. Pseudocercospora epidendri (VIC 30553, holotype). 4. Pigmented conidiophores

with inconspicuous conidiogenous cells. 5. Solitary, pluriseptate conidia, with inconspicuous

unthickened and not darkened scars. Bar: 10 um.

Taxonomy

Pseudocercospora epidendri Meir. Silva & O.L. Pereira, sp. nov. PLATE 4-5

MycoBAnk 519630

Differt a Pseudocercospora odontoglossi conidiphoris non ramosae et brevioribus, 10-57

x 2-6 um; conidiis brevioribus 11-86 x 1-3 um.

Ho.otyPe: BRAZIL, Minas Gerais, Araponga, Parque Estadual da Serra do Brigadeiro,

on leaves of Epidendrum secundum Jacq. (Orchidaceae), 08 Jan. 2008, O. L. Pereira (VIC

30553, holotype). Ex-type culture OLP 30553 (Universidade Federal de Vicosa).

EryMo_oey: referring to the host genus Epidendrum.

Pseudocercospora epidendri sp. nov. (Brazil) ... 99

PLATE 6. Necrotic symptoms on leaves of Epidendrum secundum 8 days after inoculation with

Pseudocercospora epidendri.

Leaf spots distinct, scattered over leaves, amphigenous, irregular, pale brown,

delimited by a black margin on abaxial and adaxial leaf surfaces, 3-12 mm

diam. Stromata well-developed, immersed, becoming erumpent, brown,

47.5-166 um wide and 57-213 um high. Caespituli commonly hypophyllous,

brown. Conidiophores fasciculate, brown, becoming paler towards apex,

unbranched, smooth, 0-2 septate, subcylindrical, straight to curved, arising

from cells of a well-developed stroma, 10-57 x 2-6 um. Conidiogenous cells

terminal, integrated, pale brown, smooth 3-4.5 x 2-3.5 um. Conidiogenous

loci inconspicuous, not darkened, unthickened. Conidia solitary, acicular

to obclavate, 11-86 x 1-3 um, 1-9 septate, straight to curved, pale brown,

smooth, guttulate, base rounded to long obconically truncate hila unthickened,

not darkened.

Cultures on PDA slow growing, 1.5-2.0cm after 30 days at 27°C, stromaticand

immersed in center, grayish aerial mycelium, black reverse, no sporulation.

ComMENTS — Dark sunken necrotic symptoms were detected 8 days after

artificial inoculation on all inoculated leaves, with structures formed 20-30

days after inoculation. Uninoculated control leaves, on which only PDA plugs

were placed, remained healthy. The same fungus was then re-isolated, from

the symptom in the inoculation assay (PLATE 6). Thirteen cercosporoid species

are known to occur on orchidaceous hosts: Cercospora angraeci Feuilleaub.

100 ... Silva & Pereira

TABLE 1. Pseudocercospora spp. known to occur on Orchidaceae.

CONIDIOPHORES CONIDIA CAESPITULI HOST GENERA

SPECIES (um) (um)

P. cypripedii 10-40 x 3-5 30-150 x mainly Cypripedium

3-5 epiphyllous

P. dendrobii 10-100 x 2-5 15-80 x Dendrobium

hypophyllous

2-4.5

P. peristeriae 10-60 x 2.5-6 40-100 x mainly Peristeria

2-5 hypophyllous

P. odontoglossi 50-200 x 3-5.5 35-100 x hypophyllous Cattleya, Cymbidium,

3-5 Dendrobium, Epidendrum,

Laelia, xLaeliocattleya,

Odontoglossum

P. epidendri 10-57 x 2-6 11-86 x mainly Epidendrum

1-3 hypophyllous

& Roum., C. cephalantherae Ondiej & Zaviel, C. epidendronis Bolick,

C. epipactidis C. Massal., C. eulophiae M.S. Patil, C. habenariicola Meeboon et al.,

Ramularia epipactidis U. Braun & Rogerson, Pseudocercospora cypripedii (Ellis

& Dearn.) U. Braun & Crous, P. dendrobii Goh & W.H. Hsieh [= P. dendrobii

(H.C. Burnett) U. Braun & Crous, nom. illegit.], P. odontoglossi (Prill. & Delacr.)

U. Braun, P. peristeriae (H.C. Burnett) U. Braun & Crous, Stenella orchidacearum

U. Braun & Sivap., and S. cyrtopodii Dorn.-Silva et al. (Chupp 1954, Hsieh &

Goh 1990, Crous & Braun 2003, Braun & Crous 2007, Dornelo-Silva et al. 2007,

Meeboon et al. 2007). Pseudocercospora epidendri is the fifth Pseudocercospora

species known to occur on the Orchidaceae (TABLE 1) and the second known

to occur on Epidendrum (P. odontoglossi was the first reported to occur on that

genus.) Pseudocercospora cypripedii and P. odontoglossi can be distinguished

from P. epidendri by their longer and wider conidia (Chupp 1954), while

P. dendrobii produces narrower conidia (Crous & Braun 2003) and P. peristeriae

is diagnosed by superficial hyphae with solitary conidiophores (Meeboon et

al. 2007). Almost all P. epidendri caespituli were hypophyllous but not formed

through stomata, which may indicate an adaptation for the high solar incidence

typical of Brazilian “high altitude grasslands”.

Acknowledgments

The authors wish to thank Uwe Braun (Institute of Biology, Martin-Luther-

University, Germany) and Jamjan Meeboon (Faculty of Agricultural Technology,

Chiang Mai Rajabhat University, Thailand) for reviewing the manuscript. This work is

part of an ongoing program of surveying and describing the phytopathogenic and the

endophytic mycodiversity associated to the family Orchidaceae in the state of Minas

Gerais supported by grants from Fundacao de Amparo a Pesquisa do Estado de Minas

Gerais - FAPEMIG (project 279/07) and the CNPq fellowship for O.L. Pereira.

Pseudocercospora epidendri sp. nov. (Brazil) ... 101

Literature cited

Braun U, Crous PW. 2007. The diversity of cercosporoid hyphomycetes — new species, combinations,

names and nomenclatural clarifications. Fungal Diversity 26: 55-72.

Caiafa NA, Silva AF. 2005. Composicao floristica e espectro biolégicode um campo de altitude no

Parque Estadual da Serra do Brigadeiro, Minas Gerais-Brazil. Rodrigésia 56: 163-173.

Chupp C.1954. A monograph of the fungus genus Cercospora. Ithaca, Published by the author.

Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs: 1. Names published in Cercospora

and Passalora. CBS Biodiversity Series 1.571 p.

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

the Brazilian cerrado. Mycologia 99: 753-764. http://dx.doi.org/10.3852/mycologia.99.5.753

Hsieh WH, Goh TK. 1990. Cercospora and similar fungi from Taiwan. Maw Chang Book Company,

Taipei. 376 p.

Lopes UP, Zambolim L, Pereira OL. 2009. First report of Lasiodiplodia theobromae causing leaf

blight on the orchid Catasetum fimbriatum in Brazil. Australasian Plant Disease Notes 4:

64-65.

Meeboon J, Hidayat I, Nakashima C, To-Anun C. 2007. Cercospora habenariicola sp. nov. and some

new records of cercosporoid fungi from Thailand. Mycotaxon 99: 117-121.

Neto LM, Alves RJV, Barros F, Fonzza RC. 2007. Orchidaceae do Parque Estadual de Ibitipoca, MG,

Brasil. Acta Botanica Brasilica 21: 93-101.

Pereira OL, Barreto RW. 2004. First report of Sphenospora kevorkianii (Raveneliaceae) on the orchid

Catasetum fimbriatum in Brazil. Plant Pathology 53: 256.

http://dx.doi.org/10.1111/j.0032-0862.2004.00969.x

Pereira OL, Silva M. 2009. Two new hosts, Epidendrum secundum and Epidendrum xanthinum, for

the orchid rust Sphenospora kevorkianii (Raveneliaceae) in Brazil. Australasian Plant Disease

Notes 4: 62-63.

Pereira OL, Cavallazzi JRP, Rollemberg CL, Kasuya MCM. 2002. Sphenospora kevorkianii, a rust

fungus (Uredinales: Raveneliaceae) on the orchid Pleurothallis mentigera. Brazilian Journal of

Microbiology 33: 155-156. http://dx.doi.org/10.1590/S1517-83822002000200011

Silva M, Pereira OL. 2007. First report of Guignardia endophylicola leaf blight on Cymbidium

(Orchidaceae) in Brazil. Australasian Plant Disease Notes 2: 31-32.

http://dx.doi.org/10.1071/DN07015

Silva M, Pereira OL. 2008. Black mildew disease of the neotropical orchid Epidendrum secundum

caused by Lembosia epidendri sp. nov. from Minas Gerais, Brazil. Mycotaxon 104: 385-390.

Silva M, Pereira OL, Braga IF, Lelis SM. 2008. Leafand pseudobulb diseases on Bifrenaria harrisoniae

(Orchidaceae) caused by Phyllosticta capitalensis in Brasil. Australasian Plant Disease Notes 3:

53-56. http://dx.doi.org/10.1071/DN08022

MYCOTAXON

http://dx.doi.org/10.5248/118.103

Volume 118, pp. 103-111 October-December 2011

Eremiomyces magnisporus (Pezizales), a new species

from central Spain

PABLO ALVARADO”, GABRIEL MORENO’, JOSE L. MANJON’ & MIGUEL A. SANZ

‘Departamento de Biologia Vegetal - Universidad de Alcala, 28871, Alcala de Henares, Spain

*CORRESPONDENCE TO: pablo.alvarado@gmail.com

ABSTRACT — A new Species of Eremiomyces is described from central Spain. Eremiomyces

magnisporus is proposed to accommodate a single collection found in July 2010 in a marl-

gypsicolous soil at Mount Gurugu, Alcala de Henares (Spain). The habitat and macro-/

micromorphology of the fresh material are described. Molecular analyses of ITS and nLSU

DNA support the new species as closely related to but distinct from Eremiomyces echinulatus

from the Kalahari desert.

Key worps — Kalahari truffles, Mediterranean, hypogeous fungi

Introduction

The genus Eremiomyces Trappe & Kagan-Zur must be considered as a

member of the Pezizaceae Dumortt. (Pezizales J. Schr6t.) since it was proposed

by Ferdman et al. (2005) to accommodate the molecularly deviant species

Choiromyces echinulatus Trappe & Marasas [= E. echinulatus (Trappe & Marasas)

Trappe & Kagan-Zur]. Eremiomyces echinulatus is a truffle of the African

Kalahari, collected in South Africa, Botswana, and Namibia (Trappe et al. 2008,

2010) and, until now, the only known species in this genus.

In the present work, a second Eremiomyces species is described from the

steppe-like semi-arid hills around Alcala de Henares, central Spain. These

low hills present a marl-gypsum soil covered by introduced Pinus halepensis

as well as indigenous xerophilous shrubs such as Macrochloa tenacissima,

a Mediterranean poaceous plant with an Iberian and northern African

distribution (Fics. 1-2). Other fungi previously collected in this region

include Battarrea phalloides (Dicks.) Pers., Dictyocephalos attenuatus (Peck)

Long & Plunkett, Galeropsis desertorum var. bispora (Vassilkov) G. Moreno

et al., Gastrosporium simplex Mattir., Tulostoma spp., and Phaeomyces ibericus

(G. Moreno & Esteve-Rav.) E. Horak.

104 ... Alvarado & al.

ert) Fu

Figs. 1. Habitat under Pinus halepensis with Macrochloa tenacissima. 2. Sample collection site.

Materials & methods

Morphology

The type material is preserved in the herbarium of the Dept. of Plant Biology,

University of Alcala de Madrid (AH). Microscopical observations were recorded

from tissues mounted in KOH 5%, lactophenol cotton blue, and congo red. Spore

measurements do not include superficial structures. Scanning electron microscopy

(SEM) images were taken at the University of Alcala de Henares, using a Zeiss DSM-950

instrument.

DNA extraction, PCR amplification, and DNA sequencing

DNA extraction, PCR amplification, and amplicon purification from type collection

AH 38981 was performed as described previously (Alvarado et al. 2010). PCR primers

ITS1F and ITS4 (Gardes & Bruns 1993), and ITS4 (White et al. 1990) for the ITS region,

and LR1 and LR7 (van Tuinen et al. 1998, Vilgalys & Hester 1990) for the nLSU rDNA

region were employed for PCR amplification and sequencing purposes. Sequences were

visually inspected for reading errors in MEGA4 (Tamura et al. 2007).

Sequence alignment and phylogenetic analysis

The obtained sequences were aligned with the closest matches from BLAST searches

in the public databases. ITS and nLSU sequences came mainly from Lzessoe & Hansen

(2007) and Perry et al. (2007). Sequences were aligned in MEGA4.0 software using

its ClustalW application. The final alignment was assembled manually. Neighbour-

joining phylogenetic inference was performed in MEGA4 (pairwise deletion of gaps,

2000 bootstrap replicates). The aligned loci were loaded in PAUP* 4.0b10 (Swofford

2001) and a maximum parsimony phylogenetic tree reconstruction was performed

(2000 bootstrap replicates, TBR swapping algorithm, 50 sequence additions per

replicate, MULTrees not in effect). Aligned loci were also subjected to MrModeltest 2.3

(Nylander 2004) in PAUP* 4.0b10 to search for the evolutionary models that best fit

the data. These models were simulated during the analysis of each locus in MrBayes 3.1

(Ronquist & Huelsenbeck 2003). A Bayesian maximum likelihood (BML) analysis using

4 metropolis-coupled Monte Carlo Markov Chains (MCMC) was performed setting the

temperature of the chains to 0.2 and sampling once each 100 generations. The analysis

was run until convergence parameters were met in two simultaneous runs, after about

2M generations. A full search for the highest scoring ML trees was also performed in

RAXML (Stamatakis 2006) using the standard search algorithm, a thousand bootstrap

replications, and allowing an independent evolutionary model for each locus.

Eremiomyces magnisporus sp. nov. (Spain) ... 105

Molecular results

The final alignment of the ITS region included 327 of 658 variable sites,

288 of which were also parsimony-informative. In turn, nLSU alignment

included 114 parsimony-informative among 151 variable sites from a total 592

bases. Both ITS and nLSU data were best described by the evolutionary model

GTR+G+I.

Taxonomy

Eremiomyces magnisporus G. Moreno, P. Alvarado, Manjon & Sanz sp. nov.

MycoBank MB561665 FIGS. 3-11

Ascomata subglobosa, 5 cm longa, peridio pallide brunneo passim atrorufo. Colores iuvenum

speciminum permanent post siccationem. Peridium 0.1-0.3 mm crassum, angularis

structura, magnis cellulis 25-45 x 5-20 um composita, praeditum. Gleba griseobrunnea

vel griseostraminea numerosis albidis venis incomposite distributis, formantibus varium

reticulum. Sporae 14-17 um sine ornamentis, globosae, hyalinis vel pallide luteolis vacuolis

praeditae. Sporarum ornamenta ex obtusis bacillis vel conis dense distributis, in mente

revocantibus ornamenta Terfeziae arenariae, sed perspicue breviora (1.5-2 um longa,

I um lata) et magis dense disposita, efformata. Asci cylindrici difficiles inventu aetate

provecta speciminis, videntur una serie dispositi. Paraphyses haud observatae. Odor fortis

revocans caseum domesticum.

Type — Spain, Madrid, Alcala de Henares, Mount San Juan del Viso, 750 m a.s.l.,

10.VII.2010, leg. M.A. Sanz (Holotype AH 38981). [GenBank JN032128 (ITS), JN032129

(28S nLSU)].

ErymMo.ocy — Latin, magni- (big) + -sporus (spores), referring to the larger size relative

to those in E. echinulatus.

Ascoma subglobose 5 cm diam., without an obvious attachment point to the

substrate, peridium smooth, pale brown with reddish to reddish-black areas

where damaged, fresh colours retained after drying. PERrpIuM 0.1-0.3 mm

thick, with an angular structure formed by entangled cylindrical hyphae; cells

25-45 x 5-20 um, becoming more globose toward the interior; cell walls < 1.5

um, hyaline to slightly yellowish; outermost cell layer 10-40 um thick, red-

brownish vacuolar pigments present; external layer of amorphous debris < 25

uum thick. GLEBA beige to dun straw with abundant 0.05-0.4 mm broad whitish

veins irregularly distributed forming a variable reticulated mesh with multiple

attachment points to the peridium, colour appearing darker where bitten by

animals; once dried, small [0.5-1.5 mm diam.] (sub)globose or subglobose

spore-containing pockets protrude. Ascospores globose, 14-17 um (average

15.875, SD 0.545) without ornamentation, hyaline to slightly yellowish,

vacuolate; ornamented by densely arranged 1.5-2 x 1 um blunt-edged rods

and cones reminiscent of Terfezia arenaria (Moris) Trappe, but clearly shorter

and more densely arranged. Asci and pARAPHYSES could not be found due to

the advanced maturity of the sample, but spores remained linearly arranged,

commonly in groups of 4(-5) spores. ODouR strong, similar to cottage cheese.

106 ... Alvarado & al.

Fics 3-11. Eremiomyces magnisporus (Holotype: AH 38981). 3. Ascoma peridium and gleba.

4, Peridium. 5. Detail of angular peridial cells. 6-7. Spores under LM. 8. Linearly arranged spores.

9-10. Spores under SEM. 11. Detail of spore ornamentation under SEM.

Scale bars: 3 = 1 cm, 4= 100 m, 5 = 25 um, 6-7 = 10 um, 8 = 20 um, 9-10 = 5 um, 11 = 2 um.

ECOLOGY & DISTRIBUTION. Eremiomyces magnisporus is a rare species

fruiting in steppe-like areas of the marl-gypsum hills surrounding Alcala de

Henares (central Spain).

ComMENTSs. A single ascoma was located by the truffle-hunting dog of Miguel

Angel Sanz under Macrochloa tenacissima and Pinus halepensis in July 2010.

Eremiomyces magnisporus sp. nov. (Spain) ... 107

FIGURE 2 shows the exact point of collection under M. tenacissima, without any

sign of the typical ‘burr’ associated with other hypogeous fungi. It cannot be

concluded which plant species was host of this presumably mycorrhizal fungus.

Kalahari truffles have been said to fruit under poaceous species: Enneapogon

cenchroides (Licht.) C.E. Hubb., Eragrostis rigidior Pilg., and Stipagrostis

uniplumis (Licht.) De Winter (Trappe et al. 2008). Interestingly, E. echinulatus

was collected in June during the austral winter (Trappe et al. 2010), while

E. magnisporus appeared in northern hemisphere during the summer.

Eremiomyces magnisporus is characterized by its pale brown peridium with

reddish areas that do not blacken after drying, large-sized spores (14-17 um

without ornamentation), and a spore ornamentation formed by very densely

arranged blunt-edged hyaline to pale yellow cones. Eremiomyces echinulatus

is distinguished by a peridium that turns black after drying, dark brown glebal

veins, and smaller spores (10-14 um without ornamentation).

Discussion

Phylogenetic similarities between South African and Mediterranean

fungi have been reported before. The recently proposed inocybeoid genus

Tubariomyces Esteve-Rav. & Matheny is represented in both Spain and Italy

and Zambia (Alvarado et al. 2010). Tubariomyces species apparently associate

with Cistaceae Juss. in the Mediterranean and with Phyllanthaceae Martinov

and Fabaceae Lindl. in Zambia.

Macrochloa tenacissima, the putative host plant of E. magnisporus, is

commonly found in semiarid basic soils of the Mediterranean basin in Turkey,

Italy, Spain, Morocco, Algeria, Tunisia, and Libya. In the western Mediterranean,

it reaches central Spain and several more scattered spots of northeastern Spain,

yet it is not found beyond the high plateaus preceding the Sahara desert to the

south and southwest of Morocco. Up to now, M. tenacissima has been regarded

as an exclusively endomycorrhizal plant, hosting glomalean species such

as Glomus aggregatum N.C. Schenck & G.S. Sm. and Funneliformis mosseae

(T.H. Nicolson & Gerd.) C. Walker & A. Schiissler (Roldan-Fajardo 1994).

Interestingly, the hypogeous fungus Gastrosporium simplex has also been found

in association with poaceous species of the Arrhenathero-Stipetum tenacissimae

association in the hills around Alcala de Henares (Moreno et al. 1991) as well as

with Festuca ovina L. (Montecchi & Sarasini 2000).

Of the poaceous genera indicated by Trappe et al. (2010) as the most

probable hosts of the Kalahari truffles, those most closely related to Macrochloa

are Stipagrostis Nees, Eragrostis Wolf and Enneapogon Desv. ex P. Beauv.

Although widely distributed, Eragrostis is considered typical of subtropical

and xerophilous habitats and is common in Spain. Likewise, Enneapogon has

a split Australian—African distribution, with only E. desvauxii P. Beauv. being

108 ... Alvarado & al.

AF 276681 Mattirolomyces terfezioides

AJ305045 Mattirolomyces terfezioides

AJ272444 Mattirolomyces terfezioides

AY789329 Peziza phyllogena

400 100| AF 435829 Eremiomyces echinulatus

100 109 *°l AF 435825 Eremiomyces echinulatus

ae AH38981 Eremiomyces magnisporus

10 100 | AY818334 Peziza ostracoderma

1 1FJ537076 Peziza ostracoderma

GQ231749 Ulurua nonparaphysata

100 100 f AY830852 Cazia flexiascus

°° | EU834194 Cazia flexiascus

100 1007 HE U819535 Peziza depressa

a DQ200837 Peziza depressa

EF644112 Peziza badia

100 100 | 22888695 Tirmania pinoyi

*9%°°| GQ888697 Tirmania pinoyi

2° | ao a00 AF 276667 Tirmania nivea

°F J197820 Tirmania nivea

too100 | AF 276678 Terfezia leptoderma

wi | AF276679 Terfezia leptoderma

sony AF396862 Terfezia leptoderma

sooltoo | ~ AF 276676 Terfezia trappei

mo" |» AF387657 Terfezia olbiensis

AF276677 Terfezia‘trappe?

95 79 AF092097 Terfezia boudieri

AF301419 Terfezia boudieri

toqft00-] | AF276672 Terfezia boudieri

AF092096 Terfezia boudieri

AF092098 Terfezia boudieri

| naan) AF276674 Terfezia arenaria

wml AF276675 Terfezia arenaria

AF387646 Terfezia claveryi

1001001 AF 387647 Terfezia claveryi

101! AF301421 Terfezia claveryi

HM352543 Terfezia claveryi

Fic. 12. Eremiomyces magnisporus and its closest relatives in the Pezizaceae. Highest scoring ITS tree

(maximum parsimony, PAUP*). Only nodes supported by three or more of the inference methods

are annotated. Values represent (from upper left to lower right): MEGA4 neighbour-joining, PAUP

maximum parsimony, RAxML bootstrap proportions, and MrBayes posterior probabilities.

Eremiomyces magnisporus sp. nov. (Spain) ... 109

GQ231755 Mattirolomyces terfezioides

GQ231753 Mattirolomyces austroafricanus

AF335159 Peziza retrocurvata

AY789328 Peziza phyllogena

AF335117 lodowynnea auriformis

AH38981 Eremiomyces magnisporus

100 100

100100 | 4p | AF 439823 Eremiomyces echinulatus

100999 | AY 232725 Eremiomyces echinulatus

AY500533 Hapsidomyces venezuelensis

3600 AY500554 Plicania trachycarpa

AY500553 Plicaria anthracina

AF335156 Peziza phyllogena

AF335155 Peziza phyllogena

AF335139 Peziza ellipsospora

AY500558 Terfezia claveryi

AF435824 Terfezia claveryi

AF435828 Terfezia boudieri

AY500557 Terfezia boudieri

AF435820 Terfezia boudieri

AF435822 Terfezia boudieri

AF 499448 Terfezia boudieri

DQ191679 Pezizaceae sp.

nag 4 fe — AF335147 Peziza limnaea

“08 AF335132 Peziza badiofusca

AF335175 Ruhlandiella berolinensis

GQ231747 Mycoclelandia bulundari

9997

98/100

7575

AF335178 Tirmania pinoyi

AF335177 Tirmania nivea

ae AF335149 Peziza michelii

eee AF335166 Peziza succosa

AF335137 Peziza domiciliana

AF335154 Peziza nivalis

AF335128 Peziza ampliata

AF335174 Pfistera pyrophila

seae| AF335131 Peziza arvernensis

AF335130 Peziza arvernensis

AF335145 Peziza fimeti

AF335170 Peziza sp.

AF335173 Peziza sp. 5

AF335161 Peziza subcitrina

AF335162 Peziza subcitrina

sete AY500548 Peziza lobulata

se ‘al AF335165 Peziza subviolacea

100 100

Fic. 13. Eremiomyces magnisporus and its closest relatives in the Pezizaceae. Highest scoring

nLSU rDNA tree (maximum parsimony, PAUP*). Only nodes supported by three or more of

the inference methods are annotated. Values represent (from upper left to lower right): MEGA4

neighbour-joining, PAUP maximum parsimony, RAxML bootstrap proportions, and MrBayes

posterior probabilities.

110... Alvarado & al.

widespread. Most African Enneapogon species range from south to northeastern

Africa, often reaching Arabia and India (Peterson et al. 2010). Only E. persicus

Boiss. reaches western Mediterranean to southeastern Spain. In turn, Stipagrostis

is found from southern Africa to Middle East and northwestern India, reaching

the western Mediterranean up to Morocco (Le Houérou 2001). It would be

interesting to search for this and other hypogeous genera in the distribution

areas of the poaceous species mentioned above. More research on Eremiomyces

and its unexpected plant hosts is clearly needed to shed light on its disjunct

geographical distribution

Acknowledgements

This work was financed by the Spanish Ministry of Education and Culture for FPU

grant AP2006-00890 and the “Subprograma AGR del Ministerio de Ciencia y Innovacién

(Plan Nacional I+D+I)” for the Research Project AGL2009-12884-C03-03. We express

our gratitude to Dr. G. Pacioni and Dr. M. Castellano for reviewing the manuscript

and adding a number of useful comments and to Mr. D.W. Mitchell for reviewing the

manuscript. We express gratitude to Mr. A. Priego and Mr. J.A. Pérez of the Electron

Microscopy Service of the University of Alcala de Henares for their invaluable help

with the SEM. We also thank L. Monje and A. Puebla of the Department of Drawing

and Scientific Photography at the Alcala University for their help in the preparation

of the digital images, Dr. J. Rejos, curator of the AH herbarium for his assistance with

the specimens examined in the present study, Dr. J. Alvarez and Dr. C. Bartolomé for

their comments on the phanerogamic aspects of the discussion section, and finally

Dr. G. Consiglio for his help with the Latin diagnosis.

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Swofford DL. 2001. PAUP”: phylogenetic analysis using parsimony (and other methods). Version

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Trappe JM, Claridge AW, Arora D, Smit WA. 2008. Desert truffles of the African Kalahari: ecology,

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Trappe JM, Kovacs GM, Claridge AW. 2010. Comparative taxonomy of desert truffles of the

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van Tuinen D, Zhao B, Gianinazzi-Pearson V. 1998. PCR studies of AM fungi, from studies to

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MYCOTAXON

http://dx.doi.org/10.5248/118.113

Volume 118, pp. 113-121 October-December 2011

Peziza paludicola, the correct binomial for

P. udicola nom. inval.

GABRIELE CACIALLI’, ANGELA LANTIERI” & GIANFRANCO MEDARDI?

'Via Goito 25, I-57127 Livorno, Italy

*Dipartimento di Biologia “Marcello La Greca”, Universita di Catania,

Via Antonino Longo 19, I-95125 Catania, Italy

°Via Giuseppe Mazzini 21, I-25086 Rezzato (Brescia), Italy

*CORRESPONDENCE TO: angelalantieri@gmail.com

ABSTRACT — “Peziza udicola’, P. crassipes Quél., and P. paludicola are shown to be conspecific,

with P. paludicola as the correct name for the species. Peziza crassipes var. kilimanjarensis is

recombined as a variety of P. paludicola.

Key worps — Ascomycota, Pezizales, nomenclatural analysis, taxonomy

Introduction

During preparation of a monograph on Peziza Fr. —to be published by

the Bresadola Mycological Association (AMB, Trento, Italy)— the name

Peziza udicola Svréek aroused our attention. This name appears only in some

identification keys (Hohmeyer et al. 1989, Spooner 2001), generally as a short

observation without reference to year and place of publication and sometimes

also without author. No information about its date and place of publication was

available from the INDEX OF FUNGI, INDEXFUNGORUM (www.indexfungorum.

org), or MycoBANK (www.mycobank.org).

A loan request for the holotype from the mycological collections of the

National Museum of Prague (PRM), where Svréek’s specimens are located, was

met with the response that there were no specimens or data relating to this

name.

It was moreover necessary to verify the possible conspecificity of “P. udicola”

with Aleuria paludicola Boud. (assumed by Schmid-Heckel 1990) and also with

Peziza crassipes Quél. (reported by Svréek 1970).

Svréek (1970) provided a detailed description of P crassipes Quél. (free

translation from German):

114 ... Cacialli, Lantieri & Medardi

... Hymenial surface 250-300 um tall, tending to brown. Hypothecium narrow, not

distinctly delimited from the medulla. Asci 250-300 x 14-22 um, 8-spored, with

apical ring strongly amyloid... Spores (19—)22-—24(-25) x (10-)10.5-12.5(-13.5)

um, mostly 21-23 x 11-12 um, long elliptical to cylindrical-elliptical, round to

sharpened at both extremities, homogeneous content, often pale yellow-brown,

totally smooth...on decaying twigs, herbaceous stems, completely rotten wood,

debris, sometimes on necrotic herbs or directly on bare ground, in humid to

swampy places, river banks and near springs, particularly under large herbaceous

plants (e.g. Petasites, Chamaenerion angustifolium)....

Svrcéek (1970) also reported:

...I cannot exclude the possibility that Aleuria paludicola Boudier (1907) and

Peziza crassipes are conspecific. Boudier’s original diagnosis states that this Aleuria

has saucer-shaped apothecia, spores 23-26 x 12-15 um and exceptionally wide

paraphyses (18-22 um). Up to now it has been collected by the author only once,

on marsh stems in a swamp. Maybe it is an abnormal form of Peziza crassipes...

Moser (1963) mistakes the two species and unintentionally uses the epithet

“palustris” for Boudier’s “paludicola?

In 1970, therefore, Svréek did not make any mention of the earlier homonym,

Peziza crassipes Wallr. (Wallroth 1833), an inoperculate ascomycete. This

earlier homonym makes P. crassipes Quél. a nom. illegit. Eleven years later

in a checklist of Czech discomycetes, Svréek (1981) listed his 1970 records of

P. crassipes Quél. under a newly coined name “P. udicola,” noting that there was

an earlier homonym of Quélet’s binomial and referring to the description and

illustration in his previous publication: “Svr. 1970: 63 (c. fig.) (Peziza crassipes

Quél., non Wallroth)”. The name “P. udicola” was apparently intended as a

potential nom. nov. for the illegitimate Quélet binomial, but Svréek failed to

meet the requirements for valid publication when he made no full and direct

reference to the place of publication of the replaced synonym. As a potential

sp. nov., it was also not validly published because of the absence of any Latin

description or reference to a previously published Latin description (McNeill

et al. 2006: Art. 36.1); it may even be argued that it is a nom. nud. (McNeill

et al. 2006: Art. 32.1(d)) despite the direct reference to the previous German

description (Svréek 1970) and the indirect reference to the original French

description (Quélet 1883).

Specimen loans of the holotype, isotype, or other relevant collections of

P. crassipes and Aleuria paludicola were requested from the fungal collections of

the National Museum of Natural History, Paris (PC) but none were found. We

have therefore only been able to compare published information.

TABLE 1 summarizes what each cited author wrote about the three names.

Brummelen (1969) noted that microscopic dimensions given by Boudier

(1907, 1904-10) contain an error and need to be decreased by 10%. This was

taken into account when analysing the spore sizes of A. paludicola reported by

2”

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“Peziza udicola” is P. paludicola. ... 117

Boudier. The corrected dimensions of the spores in question here are 20.7—23.4

x 10.8-13.5 um. Comparing these data with those obtained from our analyses

on “P. udicola” makes clear that the various characteristics are very similar and

that small differences may be considered physiological variations of different

individuals. We believe they are not enough to justify treating the names as

separate species.

Materials & methods

Macro- and microscopic examinations were carried out on both fresh and dried

specimens (rehydrated in water or KOH 5%), with water as a mounting medium and

Melzer’s reagent to observe the iodine reaction of the asci, using an Optika optical

microscope (BK 1301 model), with 40x or 100x (immersion oil) objectives. Spore

dimensions were obtained by measuring of 50 mature spores. Collections were placed

in the fungal reference collection of LAquila (HMA), the Royal Botanic Gardens, Kew

K(M), and the private herbaria collections of F Padovan (FP) and G. Medardi (GM).

Taxonomy

Peziza paludicola (Boud.) Sacc. & Traverso, Syll. Fung. 20: 315, 1911. FIG. 1

= Aleuria paludicola Boud., Hist. Classific. Discomyc. Europe: 46, 1907

= Peziza crassipes Quél., C.R. Ass. Frang. Av. Sci. (La Rochelle

1882) 11: 405, 1883, nom. illegit., non Wallr. 1833.

“Peziza udicola” Svréek, Ceska Mykologie 35(2): 72, 1981

nom. inval., ICBN [Vienna] Art. 36.1.

TYPIFICATION: Lectotype designated here, FRANCE, “sur les feuilles pourries des Carex,

Juillet, dans les marais. Montmorency’, Aleuria paludicola coloured plate, Boudier, Icon.

Mycol., Série 5(24): no. 469 [Tom. 2: pl. 269], 1909. Epitype designated here, ITALY,

Trentino-Alto Adige, Belluno, shore of the river Piave, 10/05/86, leg. e det. F. Padovan

[K(M) 169757].

APOTHECIA up to 20 mm diam., slightly cup-shaped, sessile to sub-stalked.

Hymenial surface smooth, slightly depressed to more or less umbilicate,

sometimes flattened, brownish-ochraceous. RECEPTACLE SURFACE concolorous

or slightly paler than the hymenium and with brown granules, more dense

toward the margin; margin regular, appearing dentate because of the

granulation, but actually entire. FLEsH fragile and waxy, greyish-ochraceous.

ASCOSPORES 19-22(-24.5) x 12-13 um, elliptical, smooth, hyaline, without

oil drops, uniseriate in the ascus. Asc1 250-280(-300) x 15-18 um, sub-

cylindrical. PARAPHYSES sub-cylindrical and 4—5 um wide at the base (often

with 1-2 basal enlarged cells), dilated and irregularly shaped in the upper part,

up to 10 um, simple or forked at the base, septate. HyYMENIUM up to 300 um

thick. SUBHYMENIUM 40-50 um thick; textura globulosa-angularis, composed

of rounded or slightly angular cells, 10-30 um diam. MEDULLARY EXCIPULUM

118 ... Cacialli, Lantieri & Medardi

— = 10 um

10 ym >fS )

Fic. 1. Peziza paludicola (Epitype, K(M) 169757). A. Uppermost zone of the hymenium

(tips of paraphyses and asci with ascospores). B. Released ascospores.

about 500 um thick; textura globulosa, composed of rounded cells up to 100 um

diam., mixed with obvious intricate hyphae, 15-20 um wide. ECTAL EXCIPULUM

20-30 um thick; textura angularis, composed of angular or sub-angular cells,

about 10-15 um diam.

Hasirat: in small groups on degraded wood of deciduous trees.

ADDITIONAL MATERIAL EXAMINED: ITALY, ABRUzzo, LAQuita, Castellafiume,

09/11/84, leg. e det. not declared, sub P ampliata, (HMA 4129); LOMBARDY, BRESCIA,

Breno, tableland of Gaver, 21/08/04, leg. e det. G. Medardi, (in herb. priv. GM); Ponte

di Legno, P. so Gavia, on loam and humid woody herbaceous remnants, at 2800 m of

altitude, 06/08/06, leg. e det. G. Medardi, (in herb. priv. GM).

2”

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Peziza paludicola var. kilimanjarensis (J. Moravec) Cacialli, Lantieri & Medardi,

comb. nov.

MycoBank MB 561192

= Peziza crassipes var. kilimanjarensis J. Moravec, Ceska Mykologie 32(2): 77. 1978.

The Moravec (1978) variety required transfer from the illegitimate species,

P. crassipes Quél., to its legitimate synonym P. paludicola.

Discussion

Peziza paludicola can be distinguished from similar species by the following

characteristics: apothecia with monotonous colours; the outside showing a

dark granulation more evident at the margin; smooth and relatively large

spores; habitat generally related to decaying deciduous wood. Similar species

are Peziza boudieri (Cooke) Donadini, Peziza hortensis P. Crouan & H. Crouan,

and Peziza sciophila Medardi. These can be differentiated by the anatomy of the

excipular tissues and by the habitat. Peziza ampliata Pers. is distinctive in both

the excipular architecture and the position of the layers and the dimensions

of the cells; the substratum is also at times important to separate these fungi.

TABLE 2 summarizes the main characteristics of these species.

Acknowledgments

The authors thank Prof. D.H. Pfister (Massachusetts, USA) and Prof. W-Y Zhuang

(China) for critically reviewing the manuscript, Dr. B. Buyck (Natural History Museum

of Paris), Dr. A. Kubatova, and Dr. M. Chlebicka (National Museum of Prague) for help

in searching for specimens, Dr. M. Svréek (Czech Scientific Society for Mycology) for

information about P. udicola, Dr. J. Moravec (Czech Republic) for information about

P. crassipes, Ing. C. Lavorato (Calabria, Italy) for help in translating some articles from

German, Dr. G. Lalli (Univ. of LAquila, Italy) and Dr. F. Padovan (Veneto, Italy) for

putting their collections at our disposal, and Prof. D. Minter for reviewing the English.

Literature cited

Boudier E. 1904-10. Icones mycologicae ou iconographie des champignons de France, 4 vol. Paul

Klincksieck, Paris.

Boudier E. 1907. Histoire et classification des discomycétes d’Europe. Paul Klincksieck, Paris.

Brummelen J van. 1969. Clues for the determination of the spore-sizes in Boudier’s illustrated

publications. Persoonia 5(3): 233-236.

Hohmeyer H, Ludwig E, Schmid H. 1989. Seltene Ascomyceten in Bayern (2). Uber einige

operculaten Discomyceten (Pezizales). Hoppea 47: 5-36.

McNeill J, Barrie FE Burdet HM, Demoulin V, Hawksworth DL, Marhold K, Nicolson DH, Prado J,

Silva PC, Skog JE, Turland NJ, Wiersema J. 2006. International Code of Botanical Nomenclature

(Vienna Code). Adopted by the Seventeenth International Botanical Congress, Vienna, Austria,

July 2005. Regnum Vegetabile 146. 568 p.

Medardi G. 2007. Una nuova Peziza dall’Italia: Peziza sciophila. Rivista di Micologia (50) 4:

333-343.

Moravec J. 1978. Fungi of Kilimanjaro - I. Discomycetes, Pezizales. Ceska Mykologie 32(2): 70-78.

“Peziza udicola” is P. paludicola. ... 121

Quélet L. 1883. Quelques espéces critiques ou nouvelles de la Flore Mycologique de France.

Comptes-rendus de l’'Association Francaise pour l’Avancement des Sciences 11: 387-412.

Schmid-Heckel H. 1990. Pilze in den Berchtesgaden Alpen. Nationalpark Berchtesgaden

Forschungebericht 15, 2. Auflage: 3-136.

Spooner B. 2001. The larger cup fungi in Britain - part 3. The genera Peziza and Plicaria. Field

Mycology 2 (2): 51-59. http://dx.doi.org/10.1016/S1468-1641(10)60516-6

Svréek M. 1970. Uber einige Arten der Diskomyzetengattung Peziza [Dill.] L. ex St-Amans. Ceska

Mykologie 24(2): 57-77.

Svréek M. 1981. Katalog operkulatnich diskomycetti (Pezizales) Ceskoslovenska II. (O-W). Ceska

Mykologie 35(2): 64-89.

Wallroth KFW. 1833. Flora cryptogamica Germaniae. Pars posterior, continens algas et fungos.

Norimbergae, sumtibus J.L. Schragii.

MY COTAXON

http://dx.doi.org/10.5248/118.123

Volume 118, pp. 123-129 October-December 2011

Two new species of Corynespora

from northeastern Uttar Pradesh, India

RAGHVENDRA SINGH & KAMAL

Department of Botany, D.D.U. Gorakhpur University, Gorakhpur, U.P, India- 273 009

*CORRESPONDENCE TO: drsinghtaxon@gmail.com

ABSTRACT — Two Corynespora species collected from northeastern Uttar Pradesh, India,

are described and illustrated: C. carrisae sp. nov. from living leaves of Carissa spinarum

(Apocynaceae) and C. peristrophicola sp. nov. from living leaves of Peristrophe bicalyculata

(Acanthaceae).

Key worps — biodiversity, hyphomycetes, foliar diseases, phytopathogenic fungi,

taxonomy

Introduction

Indian researchers have added several novel species to the anamorphic

genus Corynespora during the past few years (Meenu et al. 1997, Meenu &

Kamal 1998, Sharma et al. 2002, Jain et al. 2002, Pal et al. 2007, Singh et al.

2007, Kumar et al. 2008). During our recent survey, we encountered several

plant species exhibiting leaf blights. Upon critical examination and a thorough

survey of the literature, we found that two blights were caused by two new

Corynespora species: C. carrisae on Carissa spinarum L. (Apocynaceae) and

C. peristrophicola on Peristrophe bicalyculata (Retz.) Nees. (Acanthaceae). These

taxa are described and illustrated below.

Materials & methods

Infected leaf samples from different parts of northeastern Uttar Pradesh (U.P.)

were placed in separate polythene bags and taken to the laboratory. Suitable mounts

of surface scrapings and free-hand cut sections were prepared from infected portions

of the leaf samples. The material was mounted in a cotton-blue lactophenol mixture on

microscope slides. The slides were examined, the specimens were measured and camera

lucida drawings were made. Morphotaxonomic determinations were made with the help

of current literature and available resident expertise. Holotypes have been deposited

in HCIO (Herbarium Cryptogamiae Indiae Orientalis), Indian Agricultural Research

124 ... Singh & Kamal

Institute, New Delhi; isotypes were retained in the herbarium of the Department of

Botany, D.D.U. Gorakhpur University, Gorakhpur, for further reference.

Taxonomy

Corynespora carrisae R. Singh & Kamal, sp. nov. FIG. 1

MycoBank MB 519052

Maculae amphigenae, circulares vel subcirculares, 3-5 mm in diam., brunneae vel atrae.

Coloniae amphiphyllae, effusae, brunneae. Mycelium internum, tenuitnicatum, glabrum,

ex hyphis ramosis, olivaceo brunneis vel brunneis. Stromata nulla. Conidiophora singularia,

macronematosa, mononematosa, erecta vel procumbenta, recta vel flexuosa, nonramosa,

cylindrica, glabra, crassitunicata, 4-8-septata et 1-15 successivas proliferationes, 250-675

x 4-10 um, cellula basalis inflatis, brunneis vel atro brunneis. Cellulae conidiogenae in

conidiophoris integratae, terminales, monotreticae, cicatrices non incrassates. Conidia

acrogenaes, solitaria, simplicia, non ramosa, tenui tunicata, glabra, recta vel curvata,

obclavato-cylindrica, 4-17-distoseptata, 75-242 x 6-14 wm, ad apicem obtusa vel

rotundata, olivacea vel luteo brunnea, hilo in crassato, germinato conidium notatum.

Type: On living leaves of Carissa spinarum (Apocynaceae), Kusumhi forest, Gorakhpur

(U.P.), India, 19 September 2007, coll. Raghvendra Singh, HCIO No. 48276 (holotype),

GPU Herb No. KR-10 (isotype).

EryMo_oey: the epithet is derived from the genus name of the host.

Infection spots amphigenous, circular to subcircular, 3-5 mm in diameter,

brown to black. Colonies amphiphyllous, effuse, brown. Mycelium internal,

thin-walled, smooth, branched, olivaceous brown to brown. Stromata absent.

Conidiophores arising singly, macronematous, mononematous, erect to

procumbent, straight to flexuous, unbranched, cylindrical, smooth, thick-

walled, 4-18-septate and 1-15 successive cylindrical proliferations, 250-675

x 4-10 um, basal cell swollen, brown to dark brown. Conidiogenous cells

integrated, terminal, monotretic, scars unthickened. Conidia acrogenous,

solitary, simple, unbranched, thin-walled, smooth, straight to slightly curved

to obclavate-cylindrical, 4-17-distoseptate, 75-242 x 6-14 um, apex obtuse to

rounded, olivaceous to very light brown, hilum thickened, germinating conidia

present.

REMARKS— Only one Corynespora species, C. alstoniae Meenu et al. (Meenu

et al. 1997), has been described on Apocynaceae. Compared with the present

collection, C. alstoniae has shorter and broader conidiophores (121-473.5 x

6.0-13.5 um) and conidia (48.5-154 x 8.5-21.5 um). The hilum in C. carrisae is

thickened while in C. alstoniae it is unthickened.

Unbranched to branched and longer conidiophores (110-850 x 4-11 um)

easily distinguish C. carrisae from C. cassiicola (Berk. & M.A. Curtis) C.T. Wei

(Wei 1950), which has unbranched, shorter conidiophores. Its 4-17-distoseptate

longer thinner conidia also contrast with the 4-20-distoseptate shorter thicker

(40-220 x 9-22 um) conidia in C. cassiicola.

Corynespora carrisae & C. peristrophicola spp. nov. (India) ... 125

Fic. 1. Corynespora carrisae.

a: symptoms; b: conidia, germinating conidia, and conidiophores.

(Scale bars: a = 20 mm, b = 20 um).

126 ... Singh & Kamal

The new species appears to be most similar to C. euphorbiacearum Meenu

et al. (Meenu et al. 1997). Corynespora carrisae has more septa (4-18), more

successive cylindrical proliferations and longer conidiophores compared with

C. euphorbiacearum with 3-7-septate, shorter conidiophores (100-358 x 6-8

um), and thickened noncatenate and narrower conidia separate C. carrisae

from C. euphorbiacearum conidia that are unthickened, solitary to catenate,

and broader (59-235 x 11.5-22.5 um).

One other possible close relative is C. gigaspora (Berk. & Broome) M.B. Ellis

(Ellis 1957), which produces conidia that are longer and broader (100-270 x

19-28 um) with more distosepta (9-52).

Corynespora peristrophicola R. Singh & Kamal, sp. nov. FIG. 2

MycoBank MB 519051

Maculae epigenae, circulares vel subcirculares/irregulares, 1-18 mm in diam., brunneae

vel atrae. Coloniae epiphyllae, effusae, grisae. Mycelium internum, tenui tunicatum,

glabrum, ex hyphis ramosis, olivaceo-brunneis vel brunneis. Stromata nulla. Conidiophora

superficialia, ex hyphis oriunda singulata vel 2-3-fasciculata, macronematosa,

mononematosa, cylindrica, recta vel procumbenta, erecta vel flexuosa, non ramosa, glabra,

crassitunicata, 5-18-septata et 1-3 successivas proliferationes, medio brunnea, 120-325

x 5-10 um, cellula basalis inflatis. Cellulae conidiogenae in conidiophoris incorporatae,

terminales, monotreticae, cicatrices incrassatae. Conidia acrogena, sicca, solitaria,

simplicia, non ramosa, tenui tunicata, glabra, recta vel leniter curvata, obclavata vel

obclavato-cylindricata, 5-12-distoseptata cum 0-1 angulis distoseptis simulatibus, 60-135

um longa et 5-16 um lata, apicem obtusa vel rotundata, olivacea vel luteo-brunnea, hilo

crassato praedita.

Type: On living leaves of Peristrophe bicalyculata (Acanthaceae), Gorakhpur University

campus, Gorakhpur (U.P.), India, 2 December 2007, coll. Raghvendra Singh, HCIO No.

48278 (holotype), GPU Herb No. KR-12 (isotype).

Erymo_oey: the epithet is derived from the genus name of the host.

Infection spots epigenous, circular to subcircular/irregular, 1-18 mm in

diameter, brown to blackish. Colonies epiphyllous, effuse, grayish. Mycelium

internal, thin-walled, smooth, branched, olivaceous to brown. Stromata absent.

Conidiophores arising singly as lateral branches from superficial hyphae,

solitary or in fascicle of 2-3, macronematous, mononematous, cylindrical, erect

to procumbent, straight to flexuous, unbranched, smooth, thick-walled, 5-18-

septate with 1-3 successive cylindrical proliferations, mid brown, 120-325 um

long and 5-10 um wide, basal cell swollen. Conidiogenous cells integrated,

terminal, monotretic, scars unthickened. Conidia acrogenous, dry, solitary,

simple, unbranched, thin-walled, smooth, straight to slightly curved, usually

obclavate to obclavate-cylindrical, 5-12- distoseptate with 0-1 transverse band-

like distosepta, 60-135 um long and 5-16 um wide, apex obtuse to rounded,

olivaceous to very light brown, hilum thickened.

Corynespora carrisae & C. peristrophicola spp. nov. (India) ... 127

Fic. 2. Corynespora peristrophicola.

a: symptoms; b: conidia and conidiophores.

(Scale bars: a = 20 mm, b = 20 um).

128 ... Singh & Kamal

REMARKS— Only C. barleriicola N. Sharma et al. (Sharma et al. 2002) has is

known to occur on members of Acanthaceae. Corynespora peristrophicola has

shorter unbranched 5-18-septate conidiophores while those in C. barleriicola

are branched, 3-7-septate, and longer (253-479 x 7-9 um). Furthermore,

C. barleriicola has longer conidia (41-246 x 10-18.5 um), which are 3-17-

distoseptate with 0-5 distinct transverse band-like distosepta, and its hila are

unthickened.

The conidia of C. peristrophicola appear most similar to those of C. siwalika

(Subram.) M.B. Ellis (Ellis 1961: 88-140 x 15-20 um), C. leptoderridicola M.B.

Ellis (Ellis 1957: 70-120 x 14-17 um), and C. combreti M.B. Ellis (Ellis 1963b:

40-122 x 8-11 um) but lack the rostrate morphology typical of the other

species. The 5-12 conidial distosepta in C. peristrophicola are slightly more

numerous than in C. combreti (4-10-distosepta) and less numerous than in

C. leptoderridicola (6-16-distosepta) and C. siwalika (9-19-distosepta).

The new species also resembles C. alstoniae (Meenu et al. 1997), which,

however, has longer broader conidiophores (121-473.5 x 6-13.5 um) and

broader, catenate unthickened conidia (48.5-154 x 8.5-21.5 um).

Other possible closely related species are separated by number of distosepta:

C. bdellomorpha M.B. Ellis (Ellis 1963a) with 12-19 and C. eranthemi J.M. Yen

& Lim (Yen & Lim 1980) with 5-25.

Acknowledgments

The authors thank Dr Rafael F. Castafieda-Ruiz and Dr Eric H.C. McKenzie for

reviewing the manuscript. We also express our deep thanks to Dr Shaun Pennycook

for nomenclatural review. Thanks are also due to the Curator, HCIO, New Delhi for

accepting holotype specimens and providing accession numbers thereof.

Literature cited

Ellis MB. 1957. Some species of Corynespora. Mycological Papers 65: 1-15.

Ellis MB. 1961. Dematiaceous hyphomycetes: HI. Mycological Papers 82: 1-55.

Ellis MB. 1963a. Dematiaceous hyphomycetes: IV. Mycological Papers 87: 1-42.

Ellis MB. 1963b. Dematiaceous hyphomycetes: V. Mycological Papers 93: 1-33.

Jain SL, Rai AN, Mehta P. 2002. Additions to the genus Corynespora from India. Indian

Phytopathology 55(1): 51-56.

Kumar S, Singh R, Pal VK, Singh DP, Agarwal DK. 2008. Novel additions to Corynespora Giissow

from India. Indian Phytopathology 61(1): 111-117.

Meenu, Kamal. 1998. New species of Corynespora. Mycological Research 102: 344-345.

http://dx.doi.org/10:1017/S0953756297005455

Meenu, Singh A, Singh SK. 1997. Some new forms of genus Corynespora. Indian Phytopathology

50(1): 17-24.

Pal VK, Akhtar M, Agarwal DK, Chaudhary RK, Ahmad N. 2007. Diversity of foliar fungi

in the forest flora of North-Eastern U.P: Five new species of Corynespora Gussow. Indian

Phytopathology 60(3): 330-340.

Corynespora carrisae & C. peristrophicola spp. nov. (India) ... 129

Sharma N, Chaudhary RK, Kamal. 2002. Five undescribed species of Corynespora. Indian

Phytopathology 55(4): 458-463.

Singh R, Kumar S, Pal VK, Upadhyaya PP, Agrawal DK. 2007. New Taxa of foliicolous

hyphomycetes-Cercospora, Corynespora and Phaeotricoconis from North Eastern U.P. India.

Indian Phytopathology 60(4) 506-512.

Wei CT. 1950. Notes on Corynespora. Mycological Papers 34: 1-10.

Yen JM, Lim G. 1980. Etude sur les champignons parasites du Sud-Est asiatique. 37. Les Corynespora

de Malaisie. Cryptogamie Mycologie 1: 83-90.

MY COTAXON

http://dx.doi.org/10.5248/118.131

Volume 118, pp. 131-136 October-December 2011

New lichenicolous fungi records for

Kyrgyzstan, Uzbekistan, and Ukraine

OLGA NADYEINA'& MEHMET GOKHAN HALIcI?*

'M.G. Kholodnyi Institute of Botany, National Academy of Science of Ukraine,

Tereschenkivska str., 2, 01601 Kyiv, Ukraine

?Erciyes Universitesi, Fen Edebiyat Fakiiltesi, Biyoloji Boliimti, 38039 Kayseri, Turkey

CORRESPONDENCE TO *: 'nadyeina@gmail.com & ’*mghalici@erciyes.edu. tr

ABSTRACT — Gemmaspora lecanorae and Rosellinula haplospora are newly reported for

Uzbekistan (Asia), Rosellinula frustulosae for Kyrgyzstan (Asia), and Muellerella ventosicola

and Weddellomyces heterochrous for Ukraine (Europe). The different ascospore sizes reported

for W. heterochrous are also briefly discussed.

Key worps — Ascomycota, cephalothecioid plates, lichens

Introduction

More than 1500 species of lichenicolous fungi had been described by

2003 (Lawrey & Diederich 2003), and 3000-4000 are estimated worldwide

(Hawksworth 2001). Lichenicolous fungi have been one of the least explored

fungi because of the inadequate literature and a confused taxonomy based

largely on nineteenth century concepts (Hawksworth 1983). But with the recent

publication of new keys (Foucard 2001, Nash et al. 2004, Halici 2008, Ihlen

& Wedin 2008) and checklists (e.g. Kocourkova 2000, Diederich & Sérusiaux

2000, Scholz 2000, Faltynowicz 2003, Hawksworth 2003), these organisms have

started to receive more attention. Here, we report new records of lichenicolous

fungi for Ukraine (Europe) and Kyrgyzstan and Uzbekistan (Asia).

Material & methods

Specimens reported are deposited in KW (Kyiv, Ukraine) or KHER (Kherson,

Ukraine). Specimens were examined with a Leica DM 1000 research microscope.

Microphotographs were taken with a Leica DFC 420 digital microscope camera with

c-mount interface and with a 5 megapixel CCD. Sections were prepared by hand and

examined in Merck Lugol’s iodine (I) and water. Ascospores were measured in water.

132 ... Nadyeina & Halici

Ascospores and asci measurements are given by the arithmetic mean flanked by the

mean + standard deviation and parenthetical minimum and maximum values. All

measurements and ratios include the halo in ascospores. The length/breadth ascospore

ratio is indicated as l/b.

Taxonomy

Gemmaspora lecanorae (Werner) D. Hawksw. & Halici

A detailed description is provided by Hawksworth & Halici (2007).

The species is lichenicolous on the thallus of Aspicilia species and evidently

commensalistic, as no damage or discoloration was observed in the host thallus.

Perithecia are black, >300 um wide, semi-immersed to almost superficial. Asci

are 8-spored and ascospores (0—)1-septate, with very thick cell walls, which are

2-layered, and very broadly ellipsoid to globose, (12—)12.5-13-14 x 8.5-9 um

with 1/b ratio (1.35-)1.4-1.5-1.6(-1.65).

Gemmaspora lecanorae was previously known from just two localities: Syria,

on the thallus of Aspicilia cf. farinosa, and Turkey, on the thallus of Circinaria

calcarea (L.) A. Nordin & al. (Hawksworth & Halici 2007). New to Uzbekistan

on A. thjanschanica Oxner.

UZBEKISTAN. ‘TASHKENT REGION, Western Tian-Shan, Velykyi Chymgan

Mountain, 1800 m alt., granite outcrop, on the thallus of Aspicilia thjanschanica with

Rosellinula haplospora. A. Lazarenko, 28.09.1929 (KW 23638, isolectotype of Aspicilia

thjanschanica).

Muellerella ventosicola (Mudd) D. Hawksw.

A detailed description (as M. pygmaea var. ventosicola) is provided by Triebel (1989) and

Triebel & Kainz (2004).

The species is lichenicolous on the thallus of on a wide range of crustose

saxicolous lichens including Rhizocarpon, Ophioparma, and Protoparmelia

(Triebel 1989, Ihlen & Wedin 2008, Halici 2008) and apparently commensalistic,

as no damage or discolouration was observed in the host thallus. Ascomata

perithecioid, black, semi-immersed, 250-300 um. Asci (20-)50-spored, 50-55

x 15 um. Ascospores brown, thick-walled, 1-septate, 6-7.5-9(-10) x 4-4.5-5

(-6) um, |/b ratio (1.3-)1.4-1.6-1.8(-2).

Muellerella ventosicola grows on Rhizocarpon geographicum (L.) DC. and

R. grande (Florke ex Flot.) Arnold. New to Ukraine

UKRAINE. ZAKARPATTIA (TRANSCARPATHIAN) REGION, RAKHIV DISTRICT,

Svydovets, Blyzhnytsia Mountain, 1500 m alt., on the thallus of Rhizocarpon grande,

M. Makarevych, 11.07.1947 (KW 27534). ZAPORIZHZHA REGION, CHERNIGYVSKYI

DISTRICT, Novopoltavka Village, Synia outcrop, on the granite, on the thallus of

R. geographicum, O. Khodosovtsev, 02.10.2007 (KHER 1469).

Lichenicolous fungi new to Kyrgyzstan, Uzbekistan & Ukraine ... 133

Fic. 1. Weddellomyces heterochrous. A, Perithecium; B, Cephalothecioid plates; C, Ascospores and

pseudoparaphyses; D, Ascospores; E, 6-spored and 8-spored asci; F, Ascospores.

134 ... Nadyeina & Halici

Rosellinula haplospora (Th. Fr. & Almq.) R. Sant.

A detailed description is provided by Hafellner (1985).

The species has previously been reported as lichenicolous on the thalli of

Circinaria caesiocinerea (Nyl. ex Malbr.) A. Nordin et al., Aspicilia cinerea

(L.) Koérb., A. intermutans (Nyl.) Arnold, and Bellemerea alpina (Sommerf.)

Clauzade & Cl. Roux (Hafellner 1985; Ihlen & Wedin 2008; Halici 2008). It

is evidently commensalistic, as no damage or discolouration was observed in

the host thallus of our collections, where it grew together with Gemmaspora

lecanorae and Lichenostigma elongatum Nav.-Ros. & Hafellner (on Aspicilia

spp.). Ascomata perithecia 200-230 um, 20-50 spores per ascus, (5-)6-6.5-

7.5(-8) x (4-)4.5-5-5.5(-6) um, with 1/b ratio (1-)1.1-1.3-1.5(-2).

New to Uzbekistan on Aspicilia thjanschanica and Aspicilia species in our

report.

UZBEKISTAN. TASHKENT REGION, Western Tian’-Shan, Velykyi Chymgan Mountain,

1800 m alt., granite outcrop, on the thalli of Aspicilia thjanschanica and Aspicilia sp.

A. Lazarenko, 28.09.1929 (KW 23638, isolectotype of Aspicilia thjanschanica).

Rosellinula frustulosae (Vouaux) R. Sant.

A detailed description is provided by Hafellner (1985).

The species is lichenicolous on the thallus of Lecanora frustulosa and L.

argopholis and evidently commensalistic, as no damage or discolouration was

observed in the host thallus. Perithecia, 220-280 um, c. 100 spores per asci.

Spores (4.5-)5.5-6-7(-7.5) x (3.5-)4-4.5-5(-6) um, with I/b ratio (1-)1.1-

1.3-1.5(-1.8).

Rosellinula frustulosae is specific to Lecanora frustulosa (Dicks.) Ach. and L.

argopholis (Ach.) Ach. (Hafellner 1985). New to Kyrgyzstan on L. argopholis.

Although we have no fresh collections from Ukraine, the holotype was collected

from a L. frustulosa thallus by K. Mereschkowsky near Simpheropol in Crimea

(Hafellner 1985). This is still the only known Ukrainian locality of this species,

but it has not yet been reported in Ukrainian check-lists (Kondratyuk at al.

1998, Kondratyuk at al. 2010).

KYRGYZSTAN. Issyk-Kut’, Pokrovka, Kyrgyz Station, Institute of Geography AS

USSR, in the river valley, on the thallus of Lecanora argopholis, on granite. Sobol;

15.07.1947 (KW 3898 - specimen is kept under name “Rhizocarpon geographicum”).

Weddellomyces heterochrous Nav.-Ros. & Cl. Roux FIGURE 1

A detailed description is provided by Navarro-Rosinés & Roux (1995).

The species is lichenicolous on the areoles of Aspicilia and Circinaria species

and causes a slight bleaching in the infected parts. It is weakly parasitic. The

following description is based on our collection on Aspicilia contorta subsp.

hoffmanniana S. Ekman & Froéberg ex R. Sant.

Lichenicolous fungi new to Kyrgyzstan, Uzbekistan & Ukraine ... 135

Asci cylindrical or largely cylindrical clavate, (110-)111-113-115(-118) x

(18-)19.5-20.5-21.5(-25) um (n = 20), 4-6-8-spored, wall gradually thickened

towards the apex, I-, ocular chamber present. Ascospores 3-septate, brown,

terminal cells concolorous with central cells, surface smooth, halonate, halo ~1

uum, slightly constricted at the septa, but sometimes strongly constricted in the

middle septa, heteropolar, uniseriate or irregularly biseriate in the asci, (21-)23-

25-27(-28) x (8-)9-9.5-10(-11) um (n = 40), I/b = (2.2-)2.4-2.6-2.8(-3.1)

The ascospore sizes in our data and cited in the literature seem very variable.

In the original description, based on a single type on Circinaria calcarea, the

ascospores (primarily 3-septate) measure (25—)26.5-30.4-35.5(-38) x (10-)11-

12-13.5(-14.5) um (Navarro-Rosinés & Roux 1995). In a Turkish collection on

an Aspicilia species, the ascospores measured (32-)36-40 x 11-13 um (Halici

et al. 2007). More collections are needed to reassess the ascospore size and host

variations and to determine whether more than a single taxon exists.

New to Ukraine.

UKRAINE. LuHANsS’K REGION, PEREVAL’S’K DISTRICT, steppe slope between

Mikhailivka and Troits’ke Villages above Isakiivs’ ke water storage, on thallus of

Aspicilia contorta subsp. hoffmanniana on limestone outcrop, O. Nadyeina 09.04.2007

(KW 68135).

Acknowledgements

Sergio Pérez-Ortega (Spain) and Kerry Knudsen (UCR) are thanked for reviewing

this paper. Fungal Research Trust Foundation is acknowledged for supporting the visit

of the first author (ON) to Erciyes University, Turkey.

Literature cited

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MY COTAXON

http://dx.doi.org/10.5248/118.137

Volume 118, pp. 137-146 October-December 2011

New records of Agaricales from Atlantic Forest fragments

of Pernambuco, Northeast Brazil

FELIPE WARTCHOW™, LEONOR C. MAIA?

& M. AUXILIADORA Q. CAVALCANTY’

"Universidade Federal da Paraiba, Departamento de Departamento de Sistematica e Ecologia,

CEP: 58051-970, Jodo Pessoa, PB, Brazil

? Universidade Federal de Pernambuco, Departamento de Micologia/CCB,

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

* CORRESPONDENCE TO: fwartchow@yahoo.com. br

ABSTRACT — Some interesting fungi were collected during recent expeditions to Atlantic

Forest fragments. Entoloma aripoanum is recorded for the first time from Brazil. Crepidotus

flavus, E. tucuchense, and Lepiota erinana are new records from Brazil's northeast region,

and Trogia cantharelloides is new from Pernambuco State. Drawings and descriptions of the

species are provided.

KEY worpDs — Agaricaceae, Crepidotaceae, Entolomataceae, neotropics, taxonomy

Introduction

The Atlantic Forest, which is a priority area for conservation (Myers et al.

2000), has been drastically diminished by human activity since the beginning

of Portuguese colonization (Kimmel et al. 2008, Trindade et al. 2008). Only

11-16% remains of the original forest (Ribeiro et al 2009). Research on fungal

diversity is urgently needed in this threatened biome.

In this continuation of previous reports on agarics from Atlantic Forest

fragments of Pernambuco (Wartchow 2006, Wartchow & Maia 2007, Wartchow

et al. 2007a,b, 2008a,b), we present new records of Crepidotus (Fr.) Staude,

Entoloma (Fr.) P. Kumm., Lepiota (Pers.) Gray, and Trogia Fr.

Material & methods

Collections were undertaken at the Usina Sao José (7°50'20"S 35°00'10” W), an area

covered by variously sized (12-380 ha) Atlantic Forest fragments among sugar cane

fields (Alves-Araujo et al. 2008, Trindade et al. 2008). These preserved fragments shelter

650 species (379 genera, 105 families) of trees, with Fabaceae, Poaceae, Cyperaceae,

138 ... Wartchow, Maia & Cavalcanti

Asteraceae, Euphorbiaceae, Myrtaceae, Rubiaceae, Melastomataceae, Araceae, Malvaceae,

Apocynaceae, Sapindaceae, and Sapotaceae as the most diverse (Alves-Aratjo et al.

2008).

The usual methodology for studying agarics was followed (Singer 1986). Color codes

and names follow Maerz & Paul (1950; ‘M&P’); x = average dimension. Exsiccates are

deposited at URM Herbarium (Thiers 2011).

Taxonomy

Crepidotus flavus Capelari, Mycotaxon 115: 146. 2011. FIG. 1

PiLEus 5-14 mm, uniformly sulfur yellow (M&P 10J1 “Sulphur Y, Citrus”),

convex with incurved margin in young specimens, then more or less straight,

moderately tomentose/pubescent when young, smooth and glabrous in age;

context thin, fleshy, cream to pale yellow, unchanging. LAMELLAE radiating

from an attachment point, narrow (<0.5 mm broad) concolorous with pileus,

then ferruginous in older basidiomata, crowded, with lamellulae. Stipe absent

or very rudimentary.

BASIDIOSPORES 6.5-8.5 x 6-8 um, x = 7.6 x 7.2 um, Q = 1.00-1.09(-1.11),

Qx = 1.05, globose to occasionally subglobose, distinctly punctuate/ echinulate,

slightly thick-walled, yellowish brown to pale brown in KOH. Basip1a 17-25

x 7-8.5 um, clavate, mainly 2-spored. PLEURocystTip1A absent. Lamella edge

sterile, with crowded cheilocystidia. CHEILOCYsTIDIA 20-35 x 8-10 um, mostly

fusoid to utriform/lageniform, very infrequently ellipsoid to somewhat more

or less sinuous, thin walled, hyaline. PILEIPELLIS a cutis with hyphae approx. 5

um wide, radially oriented, walls moderately thick, hyaline, colorless. LAMELLA

TRAMA regular, < 5 um wide. CLAMP CONNECTIONS numerous.

HABITAT: gregarious on rotten wood in a fragment of tropical rain forest.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO, Igarassu, Usina Sao José (“Mata de

Piedade”), 25.viii.2005, F. Wartchow s.n. (URM 80088); Moreno, v.2008, RPPN Garnijé,

G.M. Mueller et al. 27 (URM 80089).

Note: The macroscopic features were noted from both collections; microscopic

characters were taken almost entirely from URM 80088, except for observations of

cheilocystidia in URM 80089.

REMARKS: This C. flavus collection is characterized by small, sulfur yellow

basidiomes with slightly tomentose to smooth pilei, globose to occasionally

subglobose basidiospores, clamped hyphae, exclusively 2-spored_basidia,

and absence of “antler-like” cheilocystidia (Capelari 2011). The last feature

is consistent in the current collections as well as in material from Sao Paulo

(Capelari 2011). All features fit with the species concepts given by Senn-Irlet &

Immerzeel (2003) for this group.

In the protologue of C. flavus, Capelari (2011) tried to segregate this species

from C. sulphurinus Imazeki & Toki, described from Japan, based on the two-

spored basidia described for C. sulphurinus, although apparently no basidia

Agaricales new to Atlantic Forest fragments (Brazil) ... 139

FiGuRE 1. Crepidotus flavus.

A. Habit. B. Basidiospores. C. Cheilocystidia. D. Basidia.

Scale bars: A = 10 mm; B-D = 10 um.

had been found in the original collection of C. flavus. According to Imazeki &

Toki (1954), C. sulphurinus is a disjunct taxon that differs in the less globose

basidiospores and somewhat clavate cheilocystidia. On the other hand, our

materials agree with the protologue of the Sao Paulo material in the yellowish

pileus and utriform/lageniform cheilocystidia (which Capelari interpreted as

ventricose), although larger basidiospores (8.7 x 8.7 um) were reported in the

Sao Paulo material. Hongo (1961), who described C. sulphurinus specimens

from Japan, referred to the cystidia as broadly clavate and ventricose, but

the basidiospores as subglobose to globose (7.5-9 x 6.5-7.5 um). Senn-Irlet

& Immerzel (2003), however, reported utriform cystidia for other Japanese

materials.

Another allied species is C. citrinus Petch from Sri Lanka. A type study

by Pegler (1986) referred to clavate cylindric and often constricted cystidia,

and apparently exclusively 4-spored basidia. Earlier, Singer (1973) reported

C. citrinus as growing in montane subtropical forest in Argentina; he did

mention the type from Sri Lanka but did not describe it. The Argentinean

material and our C. flavus collection have a similar spore size (6.8-8.3 um)

but differ in the clavate to versiform cheilocystidia with frequent projections

(“antler-like” in Senn-Irlet & Immerzeel 2003) and the 2-4-spored. Although

Pegler (1986) did not report “antler-like” cystidia in the type and Pilat (1950)

140 ... Wartchow, Maia & Cavalcanti

had not referred to that feature, Senn-Irlet & Immerzeel (2003), who studied

C. citrinus collections distant from the type locality (La Réunion and Puerto

Rico), observed this feature. Special attention must be given to de Meijer (2008:

321), who reported rare antler-like cystidia in a Parana collection of C. citrinus

(he described them as having a “contorted apical appendage”). We follow the

Senn-Irlet & Immerzeel (2003) species concept of this group and agree with

Capelari (2011): we regard C. flavus as distinct from —but very similar to—

C. citrinus and C. sulphurinus mainly by the cystidial shape. Morphological,

phylogeographical, and molecular studies on a worldwide scale are necessary

to elucidate whether or not all of these yellow species are different.

Crepidotus brunswickianus Speg. is another South American yellow species

closely related to C. flavus. It was first described from Southern Argentinean

forests by Spegazzini (1887) with a “pallide melleus” to “fulvo lutescens” pileus,

concolorous lamellae, and basidiospores measuring 5-6 x 3-4 um. It was

later described from a montane zone in Venezuela as having cadmium yellow

basidiomata, larger (7-8.5 x 6-7 um) subglobose basidiospores, and vesicular

to clavate cheilocystidia (Dennis 1961, 1970). Despite the close similarities

in the descriptions, we believe that C. brunswickianus and C. flavus represent

distinct species, although a revision is needed for the material identified in

Dennis (1961).

Pereira (1990), Pegler (1997), Senn-Irlet & de Meijer (1998), de Meijer (2006,

2008) and Capelari (2007, 2011) reported several species of Crepidotus, and

now we have the opportunity to record C. flavus for the first time in Northeast

Brazil.

Entoloma aripoanum Dennis, Bull. Soc. Mycol. Fr. 69: 196. 1953. FIG. 2

PiLEus < 17 mm, plane-convex, with a central depression, surface white,

glabrous, smooth; margin smooth, entire, slightly involute; context thin, fleshy,

white, unchanging. LAMELLAE adnate to short decurrent, white, membranous,

moderately crowded, with lamellulae. Stipe 36 x 3 mm, central, cylindrical,

white, glabrous, smooth, with rhizomorphs, mycelium present at base. ODOR

strong, pleasant.

BASIDIOSPORES 10-11.5(-12.5) x 7-7.5 um, x= 10.7 x 7.3 um, Q= 1.30-1.50(-

1.66), Qx= 1.45, heterodiametric-elliptical, with 6-8 depressed facets, pink and

moderately thick-walled. Basip1a 32-42 x 10-11 um, clavate, 2- or 4-spored.

PLEUROCYSTIDIA absent. CHEILOCYSTIDIA 47-72 x 9-17 um, cylindrical and

some clavate, hyaline, thin walled. PrILerrrama of exclusively hyphae similar

to ones of lamella trama; vascular hyphae occasional, 5 um wide. PILEIPELLIS a

cutis with erect hyphae 3.5-10 um wide, hyaline. LAMELLA TRAMA regular with

hyphae ranging to 3.5-8.5 um wide, apparently with yellowish brown granular

cytoplasmatic contents. CLAMP CONNECTIONS present.

Agaricales new to Atlantic Forest fragments (Brazil) ... 141

FIGURE 2. Entoloma aripoanum.

A. Basidiome. B. Basidium. C. Basidiospores. D. Cheilocystidia.

Scale bars: A = 10 mm; B-D = 10 um.

Hasirat: Solitary on rotten decayed wood in a tropical forest fragment.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO, IGARASSU, “USsINA SAO JOSE”

(“MATA DE PIEDADE’), 25.VIII.2005, F WARTCHOW S.N. (URM 80085).

REMARKS: This is an interesting species recognized by the pure white basidiome

with slightly depressed pileus centre, adnate to short decurrent lamellae,

lignicolous habit, basidiospore size and shape, and clavate cheilocystidia (Baroni

& Lodge 1998, as Alboleptonia aripoana (Dennis) Pegler). Cheilocystidia in

our collection are slightly shorter than those in the Trinidad type as revised by

Baroni & Lodge (1998), who reported cystidia 46-110 um long, but the other

features agree with the original description. Baroni & Lodge (1998) suggest

that A. aripoana sensu Pegler (1983) represents a distinct species, based on

the number of spore facets (8-10 in Pegler’s Antillean specimen versus 6-8 in

E. aripoanum). Entoloma aripoanum is known from Trinidad (Dennis 1953,

Horak 1977), possibly also from Martinique and Dominica (Pegler 1983; but

see Baroni & Lodge 1998), and now is reported for the first time from Brazil.

Entoloma tucuchense Dennis, Bull. Soc. Mycol. Fr. 69: 196. 1953. FIG.3

PitEus 35 mm, plane slightly convex, surface dark brown (M&P 8A12

“Autumn”) radially striate-rimose showing the white context, disrupting in

small granular squamules; context thin, fleshy, unchanging. LAMELLAE adnate,

142 ... Wartchow, Maia & Cavalcanti

FiGuRE 3. Entoloma tucuchense.

A. Habit. B. Pileipellis showing the inflated cells and tip on one of the erect cylindrical hyphae.

C. Basidiospores. D. Basidium. E. Hyphoid terminal projections from lamella edge.

Scale bars: A = 10 mm; B-E = 10 um.

pale cinnamon pink, membranous, sub distant, with lamellulae. STIPE 66 x 4

mm, central cylindrical, surface pale grey, smooth, white at base because of the

basal mycelium.

Agaricales new to Atlantic Forest fragments (Brazil) ... 143

BASIDIOSPORES 8-9.5(-10) x 6.2-7.5 um, x = 8.7 x 7 um, Q = (1.13-)

1.18-1.30(-1.35), Qx = 1.24, subisodiametric-angular, with 5-6 facets, with thin

pink walls. Basip1a 35-40 x 11-12 um, clavate, 4-spored. PLEUROCYSTIDIA

absent. CHEILOCYSTIDIA absent, although pronounced catenate hyphae 27-52

x 7-13 um, frequently cylindrical, hyaline to light yellowish brown pigmented

sometimes arising from the lamellar trama. PILEITRAMA hyphae filamentous

or inflated, < 11 um diam.; 5-um vascular elements occasionally present.

PILEIPELLIS an epithelium composed of subglobose to ovoid elements 18-37

x 15-26 um intermixed with dark brown erect cylindrical hyphae 52-82 x

7-15 um. LAMELLA TRAMA regular, hyphae filamentous, < 4 um diam. CLamp

CONNECTIONS absent.

Hasirat: solitary on soil near an unidentified angiosperm in a fragment of

tropical rain forest.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO, Igarassu, Usina Sao José (“Mata de

Piedade’), 22.vii.2005, EF Wartchow 18/2005 (URM 80086).

REMARKS: This species and Calliderma fibulatum Karstedt & Capelari,

C. rimosum Karstedt & Capelari, C. caeruleosplendens Largent et al., Entoloma

foldatsii (Dennis) E. Horak, and E. pruinatocutis E. Horak are all characterized

by a granulose (rimose in this case) pileus and distinctly hymeniform pileipellis,

a very infrequent feature in Entoloma sensu lato (Horak 1977, Aime et al. 2010,

Karstedt & Capelari 2010). Except for the lack of cystidioid projections in the

type specimen, other features (i.e. radially sulcate pileus, basidiospore size)

our material matches the type as analyzed by Horak (1977), Pegler [1983, as

Inopilus tucuchensis (Dennis) Pegler], and Karstedt & Capelari (2010).

The recently described Calliderma rimosum, the most macroscopically

similar taxon, differs from E. tucuchense in a pileus that cracks more strongly

with age, a revolute pileal margin, sinuate (not adnate) lamellae, and cylindric,

clavate to ventricose cheilocystidia (Karstedt & Capelari 2010).

Entoloma tucuchense, previously known from Trinidad (Dennis 1953) and

Amazonas State, Brazil (Horak 1982), is newly reported from Pernambuco

State, northeast Brazil.

Recent studies place E. tucuchense and other taxa with hymeniform pileipelli

in Calliderma (Romagn.) Largent (Aime et al. 2010, Karstedt & Capelari 2010).

However, other recent molecular studies support Calliderma as an integral part

of a monophyletic Entoloma (Co-David et al. 2009).

Lepiota erinana Dennis, Kew Bull. 7: 484. 1952. Fic. 4

PILEUS 6mm, convex, surface covered with appressed vinaceous brown (M&P

4K11 “Lacquer R”) squamules, cracking to reveal a pale cream background, but

remaining entire at centre, margin entire, not sulcate, nor striate; context very

thin, submembranous. LAMELLAE free, membranous, white, crowded, with

lamellulae. Stipe 9 x 1 mm, central, cylindrical, surface concolorous with the

144 ... Wartchow, Maia & Cavalcanti

O00 a

aorpeannnscarmeey

C = Ss “i

Ficur™ 4. Lepiota erinana.

A. Basidiome. B. Basidiospores. C. Basidia. D. Cheilocystidia.

Scale bars: A = 5 mm; B-D = 10 um.

pileus, with loose, floccose vinaceous brown squamules, rhizomorphs present.

ANNULUS superior, ephemeral, and indistinct.

BASIDIOSPORES 4.5-5 X 2.5-3 um, x = 4.6 x 2.7 um, Q = (1.53-)1.62-1.78

(-1.88), Qx = 1.24, ellipsoid to elongate, thin walled, smooth, dextrinoid,

hyaline. Bastp1a 15-17 x 5-6 um, clavate, 2- or 4-spored. PLEUROCYSTIDIA

absent. CHEILOCYSTIDIA 18-20 x 6-9 um, inflate-clavate to clavate, thin walled,

hyaline. PILEUS COVERING a trichodermium comprising chains of cylindrical

to subclavate hyphae with slightly thick walled light grayish terminal elements

37-47 x 5-6 um. LAMELLA TRAMA regular. CLAMP CONNECTIONS present.

Habitat: solitary on humus in a fragment of tropical rain forest.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Igarassu, Usina Sao José (“Mata de

Piedade’), 22.vii.2005, EF Wartchow 22/2005 (URM 78709).

REMARKS: As far as we know, this small fragile species with a pileus < 10 mm

is known only from the Neotropics (Dennis 1952, Pegler 1983, de Meijer 2006,

Rosa & Capelari 2009) and the very small size of our basidiome adds to what

is known for Lepiota erinana. Known previously from Trinidad (Dennis 1952),

Martinique and Venezuela (Pegler 1983), and Parana (de Meijer 2006) and

Minas Gerais (Rosa & Capelari 2009) states in Brazil, L. erinana is reported

here for the first time from northeast Brazil.

Trogia cantharelloides (Mont.) Pat., Essai Taxon. Hymén.: 129. 1900.

SOLITARY on leaves and soil arising from a dense mycelium in litter in a

tropical rain forest fragment.

Agaricales new to Atlantic Forest fragments (Brazil) ... 145

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Igarassu, Usina Sao José (“Mata de

Piedade’), 18.viii.2005, F Wartchow (URM 78707).

REMARKS: Singer (1965) previously reported T: cantharelloides was from Paraiba

State, northeast Brazil. This is the first record from Pernambuco.

Acknowledgments

The authors thank Dr. Clark L. Ovrebo and Dr. Marcelo A. Sulzbacher for critically

reviewing the manuscript, and Dr. Vagner G. Cortez for preparing the plates. This

contribution to the “Sustainability of remnants of the Atlantic rainforest in Pernambuco

and its implications for conservation local development’, a Brazilian-German scientific

cooperation within “Science and Technology for the Atlantic Rainforest,’ was funded

by CNPq (590039/2006-7) and BMBF (01 LB 0203 A1), permitted and logistically

supported by Usina Sao José $.A/Grupo Cavalcanti Petribu. CNPq is also acknowledged

for providing grants to L.C. Maia (PP-Proc. 301126/2005-4) and sholarship to

FE, Wartchow (PROTAX/CNPq/MCT Proc. 141073/2006-3). This project was supported

by CNPq (Proc. 170067/02-5). FACEPE (Proc. BFP 0100-2.03/09).

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MY COTAXON

http://dx.doi.org/10.5248/118.147

Volume 118, pp. 147-157 October-December 2011

Tremelloscypha gelatinosa (Sebacinales) from tropical deciduous

Gymnopodium forests in southern Mexico

VicTorR M. BANDALA (*, LETICIA MONTOYA? & RAFAEL VILLEGAS?

'? Red Biodiversidad y Sistemdatica & Red de Ambiente y Sustentabilidad,

Instituto de Ecologia, A.C., RO. Box 63, Xalapa, Veracruz 91000, Mexico

CORRESPONDENCE TO *: *victor.bandala@inecol.edu.mx, * leticia.montoya@inecol.edu.mx

& * rafael.villegas@inecol.edu.mx

ABSTRACT — Populations of Tremelloscypha gelatinosa were found growing on the soil in

Gymnopodium floribundum associations of the tropical deciduous forest in southern Mexico

(State of Chiapas). Documentation of the different developmental coriaceous and spongy

stages revealed a widely variable gross morphology that ranges from stipeless pulvinate forms

to rudimentarily stipitate pseudoinfundibuliform forms that appear polyporoid, steroid, or

gomphoid. The studied collections are described and their taxonomically distinctive macro-

and micromorphological characters are illustrated. The possible mycorrhizal association of

T. gelatinosa with Gymnopodium floribundum and traditional use of the basidiomes as a wild

edible fungus by the local population are also discussed.

Key worps — edible mushrooms, Sebacinaceae, taxonomy, tropical fungi

Introduction

During the last few years we have sampled ectomycorrhizal macromycetes

within the Gymnopodium floribundum Rolfe (Polygonaceae) association, a type

of tropical deciduous vegetation established in the Central Chiapas basin (State

of Chiapas) in southern Mexico [Miranda 1952, as G. antigonoides; Reyes-

Garcia & Souza 1997]. Several species of Amanita, Cantharellus, Russula and

other ectomycorrhizal genera have been found, all possibly associating with

G. floribundum. Lactarius chiapanensis Montoya et al. was previously recorded

from this ecosystem (Montoya et al. 1996) and recently several new fresh

collections were gathered.

An interesting find in the Gymnopodium forest was numerous specimens

of a fungus with coriaceous soft spongy rudimentarily stipitate sporophores.

Mature specimens, which were somewhat trumpet-shaped, resembled a

stout Gomphus species. After examination, we identified these specimens as

148 ... Bandala, Montoya & Villegas

Tremelloscypha gelatinosa, a heterobasidiomycete related to members of order

Sebacinales M. Weif et al. (WeifS et al. 2004a). Molecular-based studies indicate

that T. gelatinosa belongs to Sebacinales group A and is phylogenetically

related to proven ectomycorrhizal species (Weif’ & Oberwinkler 2001, Weif

et al. 2004a, Selosse et al. 2002). There is still no documentation about the

mycorrhizas of T. gelatinosa, but its phylogenetic placement among recognized

ectomycorrhizal sebacinoid species suggests that it could be a potential fungal

symbiont of plant roots (Glen et al. 2002, Selosse et al. 2002, Urban et al. 2003,

Weifé et al. 2004a,b). Considering the abundance of T. gelatinosa and its possible

ectomycorrhizal status in the Gymnopodium forest that we visited, we collected

mycorrhizas from soil samples, including some occurring directly under

T. gelatinosa sporophores. Morphological examination revealed the constant

presence of at least two different morphotypes, but as PCR amplification from

extracted genomic DNA failed, it was not possible to determine whether the

collected mycorrhizas involved T. gelatinosa or other fungi. New collections

of root tips of Gymnopodium floribundum-Tremelloscypha gelatinosa will be

studied in the future.

Tremelloscypha gelatinosa is rare not only in Mexico but also worldwide,

and its morphological variation still deserves to be documented and illustrated.

Therefore we present important information on the extent of morphological

variation exhibited by fresh basidiomes collected at different developmental

stages. Today the species concept of T: gelatinosa is based on the type specimen

(a portion of a basidiome) and some single collections from Jamaica (type

locality), Florida (USA), Yucatan and Quintana Roo (Mexico) (Burt 1915,

Guzman 2004, Roberts 2006, Wells 1961, Wells & Oberwinkler 1982). Here we

describe our new collections in detail, providing photographs and illustrations

of basidiomes and distinctive micromorphological characters and taxonomic

observations.

Tremelloscypha gelatinosa is an economically valuable wild edible fungus

in the Central Chiapas basin, particularly near Tuxtla Gutierrez, Chiapas.

In this region T. gelatinosa is locally known as “nangafiafa” and —with the

so-called “moni” Lactarius chiapanensis (first reported by Miranda 1952 and

formally described by Montoya et al.. 1996)— is commonly collected by the

local population for personal consumption and/or for sale. Gymnopodium

floribundum is a dominant or co-dominant species in the deciduous tropical

forest established at SE of Tuxtla Gutierrez (Miranda 1952, Reyes-Garcia &

Souza 1997); the G. floribundum forest, which (although now fragmented) covers

several hectares, is collectively called “nanganal” (“aguanal” or “aguanacatonal”

in Miranda 1952). More will be published on traditional use of T: gelatinosa in

the future.

Tremelloscypha gelatinosa in Gymnopodium forests (Mexico) ... 149

Materials & methods

Observations were made from fresh basidiomes collected in a forest dominated by

Gymnopodium floribundum, a tropical deciduous component of vegetation in Suchiapa,

municipality of Suchiapa, SE of Tuxtla Gutierrez, Chiapas. Color codes in descriptions

refer to Kornerup & Wanscher (1967; e.g. 10C2) and Munsell (1994; e.g. SYR 4/1). For

microscopic observations, hand sections of dried samples were mounted in 3% KOH

and stained with both 1% Congo red aqueous solution and phloxine. Line drawings were

made with the aid of a drawing tube. Measurements were made in KOH; 35-45 spores

were measured per collection; Xr = range of means of length x width of n collections;

Qr = range of means of basidiospore length/width ratios in n collections. Herbarium

acronyms follow Holmgren et al. (1990).

Identification of collection VB4212 was confirmed by extraction and amplification

of the ITS rDNA region according to Montoya et al. (2010). The obtained sequence was

edited with Sequencher Ver. 4.1 (Gene Codes, Ann Arbor, Michigan) and deposited

in GenBank database (http://www.ncbi.nlm.nih.gov/) under accession number

T. gelatinosa (VB 4212) JQ012947. The ITS rDNA sequence from VB4212 was compared

with T. gelatinosa AF490394 (GenBank) using the BLAST sequence similarity search

tool (Altschul et al. 1997), revealing a 99% maximum similarity.

Taxonomy

Tremelloscypha gelatinosa (Murrill) Oberw. & K. Wells,

Mycologia 74: 325, 1982. PLATES 1-4

= Eichleriella gelatinosa Murrill, Ann. Missouri Bot. Gard. 2: 748, 1915.

BASIDIOMES (25-)40-165 x 30-110 mm, pulvinate or irregularly subglobose

in young stages, becoming more or less trumpet-shaped in profile and then

pseudoinfundibuliform, rudimentarily stipitate, soft and spongy when wet

but coriaceous and even fibrous in dried condition. PILEUS (25—)40-165 mm

wide, subglobose, gradually plano-convex, plane and finally plane-depressed

(not deeply depressed or perforated), then pseudoinfundibuliform, irregularly

circular, frequently becoming flabellate or spathulate, often confluent forming

an irregular mass of spathulate or flabelliform pilei, with somewhat crenate or

more or less lobed margin, this latter thin or broad, slightly elevated, obtuse;

surface dry, zonate, at first velvety, with age appearing fibrillose, villose or

strigose, at times somewhat floccose-scaly (mainly in old specimens); whitish

or grayish when young, at times whitish with pale gray-blue or pinkish areas,

becoming brown-yellow, brownish (5D5-7), brown-reddish or brown-orange

(6D4-6), rarely with greenish shades, with darker concentric zones (6E6-7,

6F5-6), in dried material pale brown to brown, with spongy aspect, dry; margin

whitish becoming brownish or paler, in some stages showing pale vinaceous

tinges. HYMENOPHORE decurrent, smooth, plane to irregularly rugulose or with

short bulges, then at times appearing faintly verruculose, continuous, rarely

cracked, cartilaginous in aspect, tough when dried, whitish or with yellow to

150 ... Bandala, Montoya & Villegas

PLaTE 1. Tremelloscypha gelatinosa. (a: Bandala 4210; b: Bandala 4212). Scale bar = 20 mm.

Tremelloscypha gelatinosa in Gymnopodium forests (Mexico) ... 151

PLaTE 2. Tremelloscypha gelatinosa. (a: Bandala 4361; b. Bandala 4363). Scale bar = 20 mm

152 ... Bandala, Montoya & Villegas

gray tints (SYR 4/1), dark or light grayish-vinaceous (9D3, 9E4, 10C2, 11D2),

gray with pink tinges (SYR 2/5-6) or grayish-brown (5D2-3, 5E3, 6C2, 6D2-3),

occasionally pale orange-brown (5YR 3-4/3-4) or pale orange (7B5), whitish,

pale vinaceous or pale vinaceous-brown towards or on the margin, often with

obscure concentric zones. STIPE as an attenuated end of the hymenophore

attached to the substratum, at times with age becoming poorly defined, around

15-25 x 15-30 mm, more or less cylindrical to attenuate, stout, velutinous,

somewhat hirsute to glabrous at base, same color with pileus or hymenophore,

whitish at base, compact. CONTEXT pale to more or less grayish-white, often

with pale vinaceous or pink tinges, showing concentric growth lines, dry,

fibrous, unchanging on exposure, 6-21 mm thick in pileus. Odor strong, almost

farinaceous (recalling some stipitate hydnoids), taste mild.

BASIDIOSPORES 8-13 x (5.5-)6-9 um, (Xr = 9.1-11.5 x 6.8-7.4 um;

Qr = 1.28-1.58), ellipsoid to broadly ellipsoid, some subglobose, at times

adaxially weakly depressed, apiculus conspicuous, more or less truncate, thin-

walled, smooth, hyaline, guttulate, germination not observed. BAsrp1a 14-18 x

9-12 um, ovate to obovate, at times clavate or subglobose, forming 2-4 basidial

segments, clampless, guttulate; epibasidia cylindric or faintly tapered apically,

up to 45 x 2-4 um; fertile hyphae faintly tortuous, usually branching, forming

basidia by proliferating near subbasidial hyphal segment. DIKARYOPHYSES

simple to shortly branched, 2-4 um diam., guttulate, abundant, not forming

a distinct layer above the basidia; the basal portion of the hymenial elements

consisting of cylindrical, septate and compactly arranged hyphae; the general

aspect of the hymenial stratum is of a gelatinous tissue, with its elements very

compactly arranged and with refractive contents, more or less differentiated

into several growth layers, each consisting of prostrate and ascending elements.

CONTEXT consisting of cylindric, yellowish-brown or pale brown hyphae,

3-5(-9) um wide, simple or bifurcate, shortly or more or less spacing segmented,

thick-walled, loosely interwoven or in some areas with many hyphae arranged

in tight fascicles forming a compact tissue; a narrow, prostrate layer composed

of indistinct, agglutinated hyphae giving rise to the hymenial elements. CLamp

CONNECTIONS absent.

EcoLocy— Scattered or gregarious (occasionally solitary) in soil, commonly

among fallen dead leaves and twigs, the basidiomes often enclosing small,

dead twigs or thin stalks of live plants, in tropical deciduous Gymnopodium

floribundum forest.

MATERIAL STUDIED — MEXICO. Cuiapas: Suchiapa, Mpio. de Suchiapa, 4.X1.2003,

Bandala 3838; 5.VI1I.2005, Montoya 4353; 5.I1X.2006, Bandala 4210, 4212 (GenBank

JQ012947); 14.VI.2008, Bandala 4361, 4363; 15.V1.2008, Bandala 4366, 4370, 4374 (all

at XAL). JAMAICA. Troy and Tyre, Cockpit Country, W.A. Murrill & W. Harris (Fungi

of Jamaica 1087) (Holotype, NY).

Tremelloscypha gelatinosa in Gymnopodium forests (Mexico) ... 153

PiatE 3. Tremelloscypha gelatinosa.

a. Basidiospores. b. Basidia and dikaryophyses. c. Section of basal portion of hymenium.

(Bandala 4210). Scale bar = 10 um

154 ... Bandala, Montoya & Villegas

Prate 4. Tremelloscypha gelatinosa.

a. Context hyphae. b. Basidia and dikaryophyses. c. Basidiospores.

(a: Bandala 4210; b-c: Bandala 4274). Scale bar = 10 um.

Tremelloscypha gelatinosa in Gymnopodium forests (Mexico) ... 155

Discussion

Recognition of Tremelloscypha gelatinosa as a non-resupinate fungus (i.e.

different from members of Eichleriella Bres.; cf. Wells & Raitviir 1980) was

inferred not from the type but from a specimen from Florida gathered by K.

Lampe & A.L. Welden in 1957 (Wells 1961). The holotype (from Jamaica) in fact

consists of only a pressed 35 x 18 mm portion apparently separated from the rest

of the basidiome and superficially bearing a resemblance to an effuse-reflexed

fructification as indicated both by the diagnosis (Burt 1915) and the field label

accompanying the type noting “a specimen of Stereum or Cladoderris? Using

data from the previously cited Florida collection and microscopic examination

of the holotype, Wells & Oberwinkler (1982) recorded T. gelatinosa as growing

on decayed wood and described the basidiomes as 35-75 x 20-55 mm, 1-6

mm thick, flabellate, infundibuliform or pseudoinfundibuliform in shape, and

with a poorly defined stipe.

Our fresh collections covered many different developmental stages, thus

providing more information on the variability of the gross morphology.

The fructifications were irregularly subglobose, pulvinate, lacking a stipe or

rudimentarily stipitate and pseudoinfundibuliform, more or less trumpet-

shaped and sometimes flabellate or spathulate, not with umbilicate, depressed

or deeply depressed pileus, in some stages the sporophores then superficially

resembled polyporoid, stipitate-steroid or gomphoid forms.

Sporophores of both T’ australiensis D.A. Reid, the type species of the

genus (Reid 1979), and our T: gelatinosa collections grew on soil, not wood.

Tremelloscypha australiensis differs morphologically from T. gelatinosa in its

smaller (7-20 x 8-12 mm) distinctly infundibuliform stipitate basidiomes that

are yellow when fresh and yellowish- or ochraceous-brown when dry and a

pileus surface with an almost waxy-cartilaginous appearance (Reid 1979).

Spore characters are particularly important: T: australiensis basidiospores

are subcylindrical, narrowly elliptical to suballantoid (Reid 1982: 10-13.2 x

4.5-5 um; Wells & Oberwinkler 1982: 10.5-13 x 5-5.5 um), while T: gelatinosa

basidiospores are ellipsoid to broadly ellipsoid (Burt 1915: 8-10 x 6 um;

Wells 1961: 7.5-11 x 5.5-8 um; Wells & Oberwinkler 1982: 9-13.5(-14.5) x

5.5-8(-9.5) um; Guzman 2004: (7-)9-13 x 6-8 um; this study: 8-13 x (5.5-)

6-9 um).

Acknowledgments

We are grateful to Dr. D.J. Lodge (USDA Forest Service, Forest Products Laboratory

in Puerto Rico) and Dr. M. Weif} (Institute of Evolution and Ecology, University of

Tubingen, Germany) for critical comments to improve the manuscript. We thank the

Curator from NY for the loan of type material. Thanks to Biols. T.G. Cabrera, T. Acero,

and M.J. Gutierrez (Botanical Garden of Tuxtla Gutierrez Chiapas) for their support

and assistance during visits by V.M. Bandala and L. Montoya to Tuxtla Gutierrez and

156 ... Bandala, Montoya & Villegas

for providing information on local use of T. gelatinosa as a wild edible fungus. We

appreciate the collaboration of Biols. P. del Moral and D. Ramos in the laboratory and in

the field, and MSc E. Garay by assistance during the molecular analysis (all at Instituto

de Ecologia, Xalapa). This work was partially supported by project Monitoreo de aves

en lineas de transmision fase III. Chicouasen, Chiapas, Juile, Veracruz (directed by

R. Villegas).

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MY COTAXON

http://dx.doi.org/10.5248/118.159

Volume 118, pp. 159-176 October-December 2011

New records of polypores from southern Florida

J. VLASAK?”, J. KouT?, J. VLASAK JR. & L. RYVARDEN‘

"Biol. Centre of the Academy of Sciences of the Czech Republic &

*University of South Bohemia, Faculty of Science,

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

°University of West Bohemia, Faculty of Education, Department of Biology,

Klatovska 51, CZ-306 19 Pilsen, Czech Republic

‘Biological Insitute, RO. Box 1045, Blindern, N-0316 OSLO, Norway

*CORRESPONDENCE TO: vlasak@umbr.cas.cz

ABSTRACT — Fifteen new records of polypore species are reported from the southern Florida,

USA, with short comments to their key features, ecology, and distribution. The combination

Fomitiporia apiahyna is proposed.

Key worps — Basidiomycota, Hymenochaetaceae, Polyporaceae, taxonomy, invasive species

Introduction

The southernmost part of Florida south of Tamiami Trail (U.S. 41, which

bisects Miami) lies in the tropical hardwood forest region of the state (Young

& Giese 2003). This relatively small area contributes substantially to poroid

xylophilous fungi diversity in the USA, and about 15% of USA polypore species

published in a compendium by Gilbertson & Ryvarden (1986, 1987) probably are

found only here. Although Ryvarden (2004) and Decock et al. (2007) published

new data on southern Florida Ganodermataceae and Hymenochaetaceae, to our

knowledge no new records have been added for the Polyporaceae since 1987.

Most of the territory belongs to Everglades National Park and is covered

with regularly flooded wetland or with extensive mangrove stands along the

coast. The rather uniform mangrove polypore flora from Brazil treated by

Sotao et al. (2002) and Trierveiler-Pereira et al. (2009) is not the primary focus

of our study.

Much higher polypore diversity is found in tropical hardwood forest areas

with red-barked gumbo-limbo (Bursera simaruba (L.) Sarg. (Burseraceae)) the

typical tree species. This forest-type, however, is confined to small tear-shaped

160 ... Vlasak& al.

islands of “hardwood hammocks” that occur on raised peaty platforms above

surrounding wetlands. As most are inaccessible by foot, the casual visitor is

restricted to only a handful of such localities in the Everglades and several Miami

city parks. Another distinctive Everglades locality comprises the remnant pine

rockland fragments dominated by Pinus elliottii Engelm. (Pinaceae).

Several small state parks were also established in the Florida Keys archipelago.

Their polypore flora is not as rich because many temperate tree species, such as

southern live oak (Quercus virginiana Mill. (Fagaceae)) and sugarberry (Celtis

laevigata Willd. (Ulmaceae)), together with their associated polypores reach

their southern limits on the mainland. Also, Florida Keys hardwood hammocks

are drier because of ocean breezes and low rainfall (Whitney et al. 2004). On

the other hand, there are several tropical trees that occur only in the Keys and

some polypore species seem to be growing only on them.

After exploring all these small localities several times, we have discovered

some tropical polypores not yet recorded from the USA. We describe these

below and also propose one new combination. Critical Florida collections were

compared with our finds of the same or similar species from northern Florida,

US Virgin Islands, Mexico, Belize, and Guatemala. Some species could be

determined only after ITS rRNA region sequencing. Surprisingly, sequencing

confirmed some of the most common polypores as new records.

Materials & methods

During December 2003, April 2009, August 2010, and December 2010, polypore

specimens were photographed in situ, collected, dried, and examined microscopically

mounted in Melzer’s reagent or 10% KOH. All specimens were deposited in private

herbarium of the first author, with duplicates of some specimens maintained in the

Prague Museum Herbarium (PRM).

DNA was isolated from selected polypore samples and the ITS rRNA region was

sequenced to confirm the determination. 0.25 g of the context tissue from dried

specimens was disintegrated 60 s with a steel ball in mixer mill MM301 RETSCH at

room temperature. DNA was isolated using CTAB/NaCl extraction buffer (Murray &

Thompson 1980), followed by repeated extraction with chloroform and isopropanol

precipitation. Crude DNA was dissolved in 100 ml of sterile water and further purified

using Wizard Clean Up kit PROMEGA. Resulting DNA solution (50 ul) was diluted

ten times and 1 ul was used as template for amplification with ITS5 and ITS4 primers

(White et al. 1990) in 25 ml reaction mixture using 55 °C annealing temperature.

Amplified DNA was sequenced in the Genomics laboratory of Biology Centre, Academy

of Sciences of the Czech Republic, Ceské Budéjovice, on ABI 3730xl DNA analyzer,

using BigDye Terminator 3.1 kit.

Results & discussion

Noteworthy collections are listed alphabetically by family, genus, and

species. Descriptions are included for some species, with additional comments

Polypores new to Florida (U.S.A.) ... 161

on distribution, ecology, and diagnostic or critical characters. All species are

new records to Florida.

Hymenochaetaceae Donk

Fomitiporia apiahyna (Speg.) Vlask & Kout, comb. nov. PHOTO 1

MycoBank MB 519982

= Fomes apiahynus Speg., Bol. Acad. Nac. Ci. 11: 438, 1889.

Basidiomes perennial, sessile, triquetrous, ungulate, 3 x 2 x 1 cm; upper surface

glabrous, sulcate, zonate, dark brown to black, margin acute, pore surface

grayish-brown, the pores circular, 7-9 per mm, with thick, entire dissepiments;

tubes 2 mm long each year, context yellowish brown, woody hard, up to 0.2 cm

thick. Hyphal system dimitic, contextual skeletal hyphae brown in KOH, thick-

walled, rarely branched, 2.5-5 um in diam.; generative hyphae hyaline, thin-

walled, 1.5-3 um in diam., with simple septa. Basidia broadly ellipsoid, 7-10 x

4.5-6 um, simple septate at the base, with 4 sterigmata. Basidiospores globose,

hyaline, thick-walled, dextrinoid in Melzer’s reagent, 5-6 x 4.5-5 um.

SPECIMENS EXAMINED — USA. FLORIDA: COLLIER COUNTY, Fakahatchee Strand

Preserve, East Main Tram, 23. IV. 2009, sabal palm, leg. J. Vlasak Jr., det. J. Vlasak

(JV 1008/113), (JV1008/114); MonROE County, Florida Keys, Long Key State Park, 27.

VIII. 2010, shrub, leg. & det. J. Vlasak (JV 1008/46); Sarasota County, Myakka River

State Park, 24. IV. 2009, sabal palm, leg. & det. J. Vlasak (JV 1008/1474).

DISTRIBUTION & ECOLOGY - from northern Argentina to Costa Rica (Ryvarden 2004).

In southern Florida not rare, most often on living sabal palm (Sabal palmetto (Walter)

Lodd. ex Schult. (Arecaceae)) but also on other substrates.

; 5)

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PHOTO 1: Fomitiporia apiahyna JV 1008/113 on living sabal palm.

~ i

“>< i

162 ... Vlasak& al.

ComMENTS - This species is remarkable for its very small thumbnail-sized

applanate (more often ungulate) pilei. Its microscopic features are reminiscent

of Fomitiporia robusta (P. Karst.) Fiasson & Niemela: globose thick-walled

strongly dextrinoid spores that are, however, distinctly smaller in Fo. apiahyna

and an absence of setae. As molecular analysis clusters the species with

Fo. robusta/punctata (Goes-Neto et al. 2002), we transfer this species to

Fomitiporia Murrill as defined by Fiasson & Niemela (1984).

Fomitiporia maxonii Murrill

DESCRIPTION — Ryvarden (2004:188).

SPECIMENS EXAMINED — USA. FLORIDA: DADE County, Miami, Fairchild Botanical

Garden, 24. XII. 2003, leg. J. Vlasak Jr., det. J. Vlasak (JV0312/24.16-J, PRM915956);

US VirGIN IsLANDs: St. John, 3. IX. 2004, leg. J. Vlasak Jr., det. J. Vlasak (JV0409/19-J,

PRM915955).

DISTRIBUTION & ECOLOGY - Costa Rica, Belize, Galapagos (Ryvarden 2004). Probably

quite rare in the USA: we collected it only once in Florida (in Fairchild Botanical Garden)

and also on US Virgin Islands. On dead hardwoods.

ComMENTSs - The species resembles Fo. punctata (P. Karst.) Murrill but has

somewhat larger pores and smaller spores. It can be positively determined only

after DNA sequencing. Sequences of our collections deposited in the GenBank

(GU136210, GU136211) correspond perfectly to other published Fo. maxonii

sequences (Decock et al. 2007).

Inonotus micantissimus (Rick) Rajchenb. PHOTO 2

DESCRIPTION — Ryvarden (2004: 136).

Mae Sy Saye

° . y % ‘ \

Sein, AS Ngee

Epon ee Bh.

“4

PHOTO 2: Inonotus micantissimus JV 1008/80 on living Ocotea coriacea.

Polypores new to Florida (U.S.A.) ... 163

SPECIMENS EXAMINED — USA. FLORIDA: DADE County, Miami, Matheson Hammock

Nature Area, 30. VIII. 2010, Ocotea coriacea, leg. J. Vlasak, det. L. Ryvarden (JV 1008/80,

ITS rRNA JF692194); US VirGIN IsLANDs: St. John, 3. IX. 2004, leg. K. Vlasakova., det.

J. Vlasak (JV0409/3-K, PRM902083). JAMAICA. Ocho Rios, 12. VI. 2008, leg. J. Vlasak

Jr., det. J. Vlasak (JV0806/18-J).

DISTRIBUTION & ECOLOGY - Argentine, Brazil, and Dominican Republic (Ryvarden

2004). We have collected this species before on US Virgin Islands and Jamaica; the ITS

rRNA region sequences of these collections are nearly identical. All collections are from

living hardwoods.

ComMENTs - Inonotus micantissimus may be recognized by its resupinate or

somewhat bulbous, very dark brown basidiomata, abundant setal hyphae as

well as hymenial setae and large, globose, hyaline basidiospores, 10-13 um in

diam.

Fuscoporia callimorpha (Lév.) Groposo, C.L. Leite & Gées-Neto PHOTOS 3-5

DEscRIPTIONS — Ryvarden & Johansen (1980:145); Loguercio-Leite & Wright (1995).

PHOTO 3: Fuscoporia callimorpha JV0904/87, in situ.

SPECIMENS EXAMINED — USA. FLorIDA: BROWARD County, Davie, Tree Tops Park,

26.1V.2009 (JV0904/174, 178); 28.XII.2003 (JV0312/28.5-J); Everglades Nat. Park,

Long Pine Key Campground, 27.VHI.2010 (JV1008/43); 29.VIII.2010 (JV 1008/63);

Everglades Nat. Park, Royal Palm, 19.IV.2009 (JV0904/33); 22.IV.2009 (JV0904/87,

ITS rRNA JF692190&191, JV0904/88); 20.XII.2003 (JV0312/20.1-J, 20.5-J, ITS rRNA

JF692192); 28.XII.2003 (JV0312/28.5-J); DapE County, Miami, Matheson Hammock

Nature Area, 26.VIII.2010, leg. & det. J. Vlasak (JV 1008/10); 19.IV.2009 (JV0904/13-

16); 24.X1I.2003 (JV0312/24.2-J,24.8-J); H1iGHLANDs County, Highlands Hammock

164 ... Vlasak& al.

PHoTO 4: Fuscoporia callimorpha JV0904/155, in collection.

St. Park, 25.IV.2009 (JV0904/155); Saint JOHN Counry, St. Augustine, Washington

Oaks St. Park, 20.XII.2002 (JV0212/7-J); Sarasota County, Myakka River St. Park,

24.1V.2009 (JV0904/143, 144); US VirGIN IsLANDs, St. John, 4.1X.2004, leg. J. Vlasak

Jr., det. J. Vlasdk, (JV0409/14-J, ITS rRNA JF692193, 15-J). BELIZE. Cockscomb Basin,

30.X.2006, leg. J. Kout, det. J. Viasak (JV0610/13P-K). GUATEMALA. Tikal, 4.X1.2006,

leg. J. Kout, det. J. Vlasak (JV0611/1P-K, K3C-K). MEXICO. VERAcRUuz: Los Tuxtlas,

12.X.2006, leg. J. Kout, det. J. Viasak (JV0610/S-K); Cutapas: Palenque, Campground,

19.X.2006, leg. J. Kout, det. J. Viasak (JV0610/6P-K). VENEZUELA. Cueva el Guacharo,

22.11.2004, leg. J. Kout, det. J. Viasak (JV0402/41, 42).

DISTRIBUTION & ECOLOGY - Africa (Ryvarden & Johansen 1980), Central America

(Lowe 1957), Brazil (Loguercio-Leite & Wright 1995). On dead hardwoods.

Comments - This annual, sometimes reviving, corky hard species is by far

the most common pileate Phellinus in southern Florida. Accordingly, it is

also a very variable species forming usually flat applanate pilei but sometimes

also triquetrous or effused-reflexed basidiomes or fully effused (on the log

underside) with detached sharp margin. No cuticle or black line develops

under the pileus tomentum, which is quite persistent, usually structured in

sharp zones (agglutinated in some), exuding a thin waxy layer that is somewhat

shiny-glossy and silvery. This important field trait is, however, not obvious in old

pilei with hardened surfaces. The pore surface is much more constant: fulvous

Polypores new to Florida (U.S.A.) ... 165

PuHoTo 5: Fuscoporia callimorpha JV0312/20.1 and JV0312/20.5, in collection.

with a purplish tint and pores that are very small (7-9 per mm), very regular,

round, and with thick dissepiments. Hymenial setae are short (< 25 um), not

much contrasting, abundant in some specimens but very rare and difficult to

find in others. In the hymenium are thin-walled, ventricose cystidioles (necks

< 10 x 1.5-2 um) that are not mentioned in descriptions but well pictured in

Loguercio-Leite & Wright (1995). Basidiospores (oblong ellipsoid, hyaline,

thin-walled, 3.5-4.2 x 2.2-2.9 um) are always collapsed and difficult to find in

winter and early spring collections.

This species is evidently a taxonomical problem. The ITS rRNA region

sequence is identical with the GenBank sequence AY558649 from culture

CBS 182.34. In 1934 Overholts isolated the culture in the USA as Phellinus

torulosus (Pers.) Bourdot & Galzin. TomSovsky & Jankovsky (2007) showed,

however, that this sequence differs from the European Fuscoporia (Phellinus)

torulosa (Pers.) T. Wagner & M. Fisch. Moreover, Fu. torulosa develops much

thicker basidiomes that are very persistent, even on dead wood for more than

25 years. Fu. torulosa probably does not occur in the USA, for there are no

reliable records (see also Rizzo et al. 2003) and we were not able to find it

despite numerous excursions in Pennsylvania and Virginia where it should

have optimal conditions for growth.

Phellinus roseocinereus (Murrill) D.A. Reid, described from North America

and widespread in Central America (Lowe 1957), corresponds very well to

this common Florida polypore. However, we agree with Ryvarden & Johansen

(1980) and Loguercio-Leite & Wright (1995), who consider P_ roseocinereus

166 ... Vlasak& all.

identical to Fu. callimorpha, described originally from Madagascar. Corner

(1991) was unable to detect any difference between Phellinus senex (Nees &

Mont.) Imazeki from Asia and P callimorphus, except for slightly narrower

spores of the latter. The sequence of Fuscoporia senex (Nees & Mont.) Ghob.-

Nejh. in GenBank AY558647 (isolate CBS 442.76 of B.K. Bakshi from India),

which is really much more similar to our sequences than Fu. torulosa, still

clusters separately (with 99% bootstrap support, not shown). Judging from the

literature and our examinations of two Fu. senex specimens from China, we

think that Fu. senex has only an indistinctly zonate pileus surface, lacks glossy

zones, and produces broader spores (>3.5 um; compared to always <3 um in

Fu. callimorpha). Also, older Fu. senex spores are sometimes slightly thick-

walled and yellowish, never true in Fu. callimorpha.

Phellinus neocallimorphus Gibertoni & Ryvarden, known only from the

type locality in Brazil, differs from Fu. callimorpha only by the absence of

setae. Nevertheless, we also could not find any setae in some of our specimens

(e.g. 0312/20.5) although in other respects, including rDNA ITS region

sequence, the specimens were typical. More detailed study of P. neocallimorphus

is needed, evidently.

Phellinus gilvus (Schwein.) Pat. is another quite similar species found in

southern Florida, although not as often as elsewhere in the USA. The similar

basidiomes are, however, glabrous, without tomentum, azonate, and softer.

Although also hyaline and thin-walled, Ph. gilvus spores are ovoid and 3-3.5

um broad.

We have sequenced nine collections of Fu. callimorpha from southern

Florida, US Virgin Islands, and Mexico with markedly different pileus forms,

and, except for a few mutations, all sequences are the same. We infer that there

is no other similar species in the region.

Phellinus calcitratus (Berk. & M.A. Curtis) Ryvarden PHOTO 6

DESCRIPTION — Ryvarden (2004: 162).

SPECIMENS EXAMINED — USA. FLorIDA: BROWARD County, Davie, Tree Tops Park,

26.1V.2009 (JV0904/167); DapE County, Miami, Matheson Hammock Nature

Area, 30.VIHI.2010, leg. & det. J. Vlasak (JV1008/83, ITS rRNA JF894114, JF894115);

26.VUI.2010 (JV1008/11); 19.IV.2009 (JV0904/12).

DISTRIBUTION & ECOLOGY - South America, West Indies (Ryvarden 2004). Locally

abundant in the Matheson Hammock Nature Area but rare elsewhere. Always on rather

thick, old, decorticated logs, probably oaks.

ComMENtTs ~ This is a characteristic species with a broadly attached, applanate

pileus with a sharp margin and thick crustulose black zone under hard

tomentum. The very thin-walled tubes are unique for Phellinus species in the

region. Spores are thick-walled, globose, yellow to brown. The sequence shows

no homology with GenBank sequences.

Polypores new to Florida (U.S.A.) ... 167

PHOTO 6: Phellinus calcitratus JV1008/83.

Phellinus caribaeo-quercicola Decock & S. Herrera PHOTO 7

DESCRIPTION — Decock et al. (2006).

SPECIMENS EXAMINED — USA. FLorIDA: BROWARD County, Davie, Tree Tops Park,

26.1V.2009, leg. & det. J. Vlasak (JV0904/177, ITS rRNA GU594159); Everglades Nat.

* ~ s ra

PuHoTo 7: Phellinus caribaeo-quercicola JV0904/28-J.

(The photo was taken one year after breaking off the original basidiome.)

168 ... Vlasak& al.

Park, Royal Palm, 19.IV.2009, leg. J. Vlasak Jr., det. J. Viasak (JV0904/28-J, PRM915960,

ITS rRNA GU594158); 20.X1I.2003 (JV0312/20.7-J).

DISTRIBUTION & ECOLOGY - Described relatively recently from western Cuba as growing

exclusively on Quercus cubana A. Rich. (Decock et al. 2006). Nevertheless, it seems not

to be rare around Miami on dead stems and branches of Q. virginiana.

Comments - Phellinus caribaeo-quercicola produces large perennial bulbous

or semipileate basidiomes, usually with several layers of rather long tubes. The

rare to abundant thick-walled setae with hooked tips and subglobose thick-

walled light yellowish spores are characteristic. The ITS rRNA of our collections

corresponds perfectly to the published sequence (Decock et al. 2006).

Phellinus nilgheriensis (Mont.) G. Cunn. PHOTO 8

DESCRIPTION — Ryvarden & Johansen (1980:187).

SPECIMENS EXAMINED - USA. FLoripa: DapE County, Hwy 41, Kirby Storter

Boardwalk, 28.VIII.2010, leg. & det. J. Vlasak (JV1008/49); (JV1008/50); 23.IV.2009

(JV0904/112, PRM915957).

DISTRIBUTION & ECOLOGY - Southeast Asia, Africa, Cuba (Ryvarden & Johansen

1980), French Antilles, and Guiana (David & Rajchenberg 1985), Brazil (Groposo et

al. 2007); always on hardwoods. Not previously published from the USA. However,

around the Kirby Storter boardwalk at a Hwy 41 wayside, the fungus grows on several

living Taxodium distichum (L.) Rich. trees in the slough, approximately at the height of

summer-time water level.

Comments - The applanate pilei, not as hard as in similar Phellinus species, are

typically more or less glabrous with a narrow rounded margin and rather bright

PuotTo 8: Phellinus nilgheriensis JV 1008/50 on living Taxodium distichum.

Polypores new to Florida (U.S.A.) ... 169

brown context. Lack of setae and subglobose thick-walled, yellow to rusty

brown spores (in Melzer’s) may remind of Fomitiporia when viewed in Melzer’s

reagent. It seems to be annual but reviving species— new tubes overgrow dead

the previous year’s tubes with the tube layers separated by a context tissue layer.

The ITS rRNA of our collections corresponds perfectly to published sequence

AY558633.

Polyporaceae Fr. ex Corda

Coriolopsis hostmannii (Berk.) Ryvarden PHOTO 9

DESCRIPTION — Ryvarden (2007).

SPECIMENS EXAMINED - (all on living buttonwood mangrove) USA. FLorIDA:

COLLIER County, Hwy 41, Collier-Seminole State Park, 24.XII.2003, leg. J. Vlasak,

det. L. Ryvarden (JV 1008/56, 57); 23.1V.2009, leg. & det. J. Viasak (JV0904/125);

25.XII. 2003, leg. J. Vlasak Jr., det. J. Vlasak (JV0312/25.7-J); MONROE COUNTY,

Florida Keys, Long Key State Park, 27.VIII.2010, leg. J. Vlasak, det. L. Ryvarden

(JV 1008/42).

DISTRIBUTION & ECOLOGY - Pantropical. This species is probably confined to

living buttonwood (Conocarpus erectus L. (Combretaceae)). Its ecology was

extensively treated in several papers about buttonwood polypore populations

(Bergeman et al. 2009, Parrent et al. 2004) using the name “Datronia caperata

(Berk.) Ryvarden from buttonwood’, even though the authors were aware that

their fungus is not conspecific with typical D. caperata. Our ITS rRNA sequence

Ri:

x»

PHOTO 9: Coriolopsis hostmannii JV 1008/42 on living buttonwood.

170 ... Vlasak& al.

corresponds very well with their published sequences and differ from an authentic

D. caperata sequence.

ComMENTs - This is a very typical and relatively common species. Nonetheless,

it appears in the mycological literature quite rarely under its proper name. It

seems to be often misinterpreted, for example as Datronia caperata, a South

American species with similar spores and pores but with a dark brown context,

tough trametoid consistency, and coarsely hirsute pileus. Coriolopsis hostmannii

looks more like a pileate Inonotus species because of its relatively soft context

and bright brownish colors in all parts of basidiome. The pileus surface is

glabrous to shiny-glossy, the pores are very small and regular, and the pore

surface is characteristically uneven.

Coriolopsis polyzona (Pers.) Ryvarden

DESCRIPTION — Ryvarden & Johansen (1980: 291).

SPECIMENS EXAMINED - USA. FLorIDA: DADE Country, Florida City, irrigation canal

dam, 29.VIII.2010, Ficus sp., leg. J. Vlasak Jr., det. J. Vlasak (JV1008/64-J). BELIZE.

Cockscomb Basin, 30. X. 2006, hardwood, leg. J. Kout, det. J. Vlasak (JV0610/A8-

K). JAMAICA. Ocho Rios, 12.V1.2008, hardwood, leg. J. Vlasak Jr., det. J. Vlasak

(JV0806/12-J). MEXICO. VERAcRUz: Montepio, 14.X.2006, hardwood, leg. J. Kout, det.

J. Vlasak (JV0610/A16-K); Los Tuxtlas, 12.X.2006, hardwood, leg. J. Kout, det. J. Vilasak

(JV0610/A14A-K).

DISTRIBUTION & ECOLOGY - Pantropical, on dead hardwoods (Ryvarden & Johansen

1980:291). We collected this species many times in Mexico, Belize, and Jamaica but only

once in Florida, in a typical man-influenced locality.

COMMENTS — Common in South America and resembling the boreal Trametes

hirsuta (Wulfen) Lloyd but with even more pronounced and more hirsute zones

on pileus surface and larger, less regular pores that are often a bit elongated in

various directions. The broader spores of C. polyzona are narrowly elliptic but

not cylindric.

Navisporus floccosus (Bres.) Ryvarden PHOTO 10

DESCRIPTION — Torres-Torres et al. (2007).

SPECIMEN EXAMINED — USA. FLoripA: DapDE County, Miami, Old Cutler road alley,

hardwood stump, 26.VIII.2010, leg. & det. J. Viasak (JV 1008/30, ITS rRNA JF692195).

DISTRIBUTION & ECOLOGY - Africa, Southeast Asia (Ryvarden & Johansen 1980), Cuba,

Mexico (Torres-Torres et al. 2007), Costa Rica (Mata et al. 2007).

COMMENTS - Remarkable, very large (< 50 cm wide x 10 cm thick) pilei

reminding large Ganoderma P. Karst. distinguish this species. The glabrous

pileus surface has a patchy black cuticle or thin azonate crust. The thick light

brown context has a relatively soft consistency and is strikingly lightweight

when dry. Because of large naviculate spores, this species is traditionally

classified close to Navisporus sulcatus (Lloyd) Ryvarden, also a South Florida

Polypores new to Florida (U.S.A.) ... 171

s f %, ’ \ . 7

{ ‘ied wy : . ; <P

HOTO 10: Navisporus floccosus JV 1008/30.

species, but very different, with small, effused-reflexed pilei and relatively large

pores. Decock (2007) has commented critically on the taxonomical position of

N. floccosus.

Skeletocutis diluta (Rajchenb.) A. David & Rajchenb. PHOTO 11

DEscRIPTION — David & Rajchenberg (1992).

SPECIMENS EXAMINED — USA. FLoripA: DAabDE County, Everglades Nat. Park, Long

Pine Key, Campground, 29.VII.2010, Pinus elliottii log, leg. & det. J. Viasak (JV 1008/61,

ITS rRNA JF692198); Long Pine Key, Pinelands Trail, 20.IV.2009, Pinus elliottii log,

leg. & det. J. Vlasak (JV0904/44); (JV0904/45). BELIZE. Cockscomb basin, 30.X.2006,

hardwood, leg J. Kout, det. J. Vlasak (JV0610/16-K, ITS rRNA JF692197); 29.X.2006,

hardwood, leg J. Kout, det. J. Vlasak (JV0610/G4-K).

DISTRIBUTION & ECOLOGY - Argentina, Africa (David & Rajchenberg 1992), Panama

(Nufiez & Ryvarden 1999). Mostly on hardwoods but the type was collected in Argentina

on Pinus taeda L. (Rajchenberg 1983). In Florida it occurs on logs of slough pine (Pinus

elliottii) in seasonally flooded parts of the Everglades Long Pine Key region. Probably

not rare as we noted several quite disintegrated basidiomes in dry wintertime. During

the next summer the whole area was a half meter under water and we could collect only

one well developed specimen just on the edge of a water basin.

ComMENTS - Dextrinoid to weakly amyloid skeletals that dissolve in KOH

and the tiny spores make this species unmistakable. The species was originally

described as “resupinate or effused-reflexed.” Although our collections from

Belize on hardwoods are always pileate, our Florida specimens (collected on

pine) are strictly resupinate. Also, from the microphotography, the spores

172 ... Vlasak& al.

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PHoTo 11: Skeletocutis diluta JV1008/61 on Pinus elliottii log.

seem to be only 2.5 x 0.4 um, smaller than published (3.1-3.5 x 0.5-0.8 um).

Nevertheless, the ITS rRNA sequences of Belize and Florida samples are nearly

identical. Though originally described as a variety of S. nivea (Rajchenberg

1983), the ITS rRNA sequence shows no significant homology with S. nivea

(Jungh.) Jean Keller but low homology with S. chrysella Niemela (91%) and

S. kuehneri A. David (90%).

Trametes lactinea (Berk.) Sacc.

This species is treated separately in Vlasak & Kout (2011).

Trametes ochroflava Cooke PHOTO 12

= Daedalea microsticta Cooke

DESCRIPTION — Fidalgo & Fidalgo (1967: 848).

SPECIMEN EXAMINED — USA. FLoRIDA: DADE County, Miami, Fairchild Botanical

Garden, Albizia stump, 24. XII. 2003, leg. J. Vlasak Jr., det. J. Vlasak (JV0312/24.17-J,

PRM915968).

DISTRIBUTION & ECOLOGY — South and Central America, Mexico, West Indies. Dead

deciduous trees (Ryvarden & Aime, unpublished).

ComMMENTS - Trametes ochroflava is characterized by large, flat, tough,

whitish pilei with labyrinthine pores and brownish context and trama. Pores

often appear also on pileus surface. The species was long known as Daedalea

microsticta and only recently has the older and more appropriate name been

Polypores new to Florida (U.S.A.) ... 173

PHOTO 12: Trametes ochroflava JV0312/24.17 in collection.

applied (Ryvarden & Aime, unpublished). The type of rot is unknown but the

sequence shows similarity to Trametes Fr., not Daedalea Pers. The sequence of

our specimen is identical with the published sequence FJ403209.

Ceriporiopsis cystidiata C.L. Leite, G.V.C. Gong. & Ryvarden

DESCRIPTION — Loguercio-Leite et al. 2001.

Inconspicuous, white resupinate polypore with 2-3 mm long tubes and very

thin subiculum, margin and touched places somewhat yellowish when dry.

SPECIMEN EXAMINED — USA. FLoRIDA: DADE County, Miami, Matheson Hammock

Nature Area, 26. VIII.2010, hardwood, leg. & det. J. Viasak (JV 1008/26).

DISTRIBUTION & ECOLOGY - up to now known only from the type locality: Brazil, dead

hardwood.

Comments ~The abundant hymenial cystidia and cylindrical but rather thick

straight spores are diagnostic.

Wrightoporia bracei (Murrill) I. Lindblad & Ryvarden PHOTO 13

= Amylosporus bracei (Murrill) A. David & Rajchenb.

DESCRIPTION — Murrill 1921, Rajchenberg 1983.

Strikingly pinkish-violet resupinate polypore, widely effused on the substrate,

with tiny pores (5-7 per mm). Spores amyloid, ovoid, 3.5-4 x 2.5-3 finely

174 ... Vlasak& al.

PHOTO 13: Wrightoporia bracei JV1008/77 in situ.

echinulate, hyphal system dimitic, skeletal hyphae weakly dextrinoid, generative

hyphae with simple septa.

SPECIMEN EXAMINED — USA. FLORIDA: MONROE Country, Florida Keys, Key Largo,

John Pennekamp Coral Reef State Park, 29. VIII.2010, hardwood, leg. & det. J. Vlasak,

(JV 1008/77, ITS rRNA JF692199).

DISTRIBUTION & ECOLOGY - Argentina (Rajchenberg 1983), French Antilles (David &

Rajchenberg 1985), Bahamas, Puerto Rico (Murrill 1921). Tropical hardwood forest.

CoMMENTS— Contrary to original description, this specimen is strongly

rhizomorphic, perhaps due to the growth on an underside of a log and its violet

colors are unchanged after drying. The species was transferred to Amylosporus

Ryv. by David & Rajchenb. (1985), but the rRNA ITS sequence shows only low

homology with Wrightoporia lenta (Overh. & J. Lowe) Pouzar and W. luteola

B.K. Cui & Y.C. Dai (GenBank) and none at all with Amylosporus campbellii

(Berk.) Ryvarden (our sequences JF692200, JF692201).

Acknowledgments

Dr. W. Spirin and Dr. Sergio Pérez Gorjon have kindly acted as presubmission

reviewers and their help is acknowledged. We are grateful also to Dr. Shaun Pennycook

for his help in the correction and improvement of the present paper. This research was

supported by CEZ: AV0Z50510513 fund.

Literature cited

Bergemann SE, Smith MA, Parrent JL, Gilbert GS, Garbelotto M. 2009. Genetic population

structure and distribution of a fungal polypore, Datronia caperata (Polyporaceae) in mangrove

forests of Central America. J. Biogeogr. 36: 266-279.

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MY COTAXON

http://dx.doi.org/10.5248/118.177

Volume 118, pp. 177-180 October-December 2011

Paraphysoderma sedebokerense, gen. et sp. nov.,

an aplanosporic relative of Physoderma (Blastocladiomycota)

TIMOTHY Y. JAMES’ , YORAM HOFFMAN’, ALIZA ZARKA” & SAMMY BOUSSIBA’S

"Department of Ecology and Evolutionary Biology, University of Michigan

Ann Arbor, MI 48109, USA

?Microalgal Biotechnology Laboratory, the Jacob Blaustein Institutes for Desert Research,

Ben Gurian University of the Negev, Sede-Boker Campus 84990, Israel

CORRESPONDENCE TO: ‘tyjames@umich.edu & Ssammy@bgu.ac.il

ABSTRACT — A monocentric blastocladialean fungus was identified as a parasite of

the green alga Haematococcus pluvialis in aquaculture and isolated into pure culture.

Phylogenetic analysis suggests the fungus is a sister taxon to the obligately biotrophic

plant parasitic genus Physoderma. The parasite, which has not been observed to reproduce

by zoospores, instead forms amoeboid aplanospores. We formally describe the new genus

and species Paraphysoderma sedebokerense to accommodate this unusual member of the

Blastocladiomycota.

Key worps — algae, Blastocladiales, cultures, nomenclature

Introduction

A fungal parasite of the green alga Haematococcus pluvialis has recently

been characterized (as “Haematococcus parasite” in Hoffman et al. 2008)

and provisionally referred to as “Paraphysoderma sedebokerensis” (Gutman

et al. 2009; Porter et al. 2011). The parasite has the appearance of a typical

chytridiomycete zoosporic fungus with an epibiotic globular sporangium

subtended by an intracellular rhizoidal system. It was first observed during

aquaculture of H. pluvialis, an important producer of the carotenoid astaxanthin

used in cosmetics and pharmaceuticals (Del Campo et al. 2007). Neither pure

nor dual cultures of the alga and fungus produced a zoosporic stage; instead

sporangia released amoeboid aplanospores that were infective to H. pluvialis

(Hoffman et al. 2008) but not other green algae (Gutman et al. 2009). In a

molecular phylogeny based on the 18S ribosomal RNA gene the novel parasite

was closely related but clearly basal to the plant parasitic zoosporic fungus

Physoderma (Blastocladiomycota) (Hoffman et al. 2008). Its phylogenetic

178 ... James & al.

position, the absence of a zoospore stage, and its occurrence in an algal host

suggest that the parasite should be placed in a separate genus.

A detailed description of the infection cycle and growth characteristics of

the parasite has been published (Hoffman et al. 2008). The goal of this paper is

to provide a formal description of the parasite.

Taxonomy

Paraphysoderma Boussiba, Zarka & T.Y. James, gen. nov.

MycoBank MB 561750

Sporangium monocentricum, eucarpicum, epibioticum. Aplanosporae paucae usque ad

plurimae per sporangium, sine flagellis, congregentes antequam liberati, per incisuram in

sporangio exeuntes, 3-6 um diametro pseudopodiis filosis, in superficie cellulae nutricis

repentes antequam incystati et endogene evolventes. Systema rhizoidale e puncto unico

in sporangio evolvens, intra hospitem bulbosum. Sporangium dormiens epibioiticum,

fuscatius, pariete crassiore quam zoosporangium, germinans ut aplanosporas liberarent.

In alga viridi parasitica.

TYPE SPECIES — Paraphysoderma sedebokerense Boussiba, Zarka & T. James

EryMoLocy — The new genus is closely related to, but distinct morphologically and in

host preference, from the genus Physoderma.

SPORANGIUM monocentric, eucarpic, epibiotic. APLANOSPORES few to dozens

per sporangium, lacking flagella, swarming before being released, exiting

through tear in sporangium, 3-6 um diameter with filose pseudopodia,

crawling on surface of host cell before encysting and developing endogenously.

RHIZOIDAL SYSTEM developing from single point on the sporangium, forming

an apophysis inside host. RESTING SPORANGIUM epibiotic, darker and with

thicker wall than zoosporangium, germinating to release aplanospores. Parasitic

on green algae.

Paraphysoderma sedebokerense Boussiba, Zarka & T.Y. James, sp. nov.

MycoBank MB 561751

Descriptio ad genus. In Haematococcus pluvialis parasitica.

Ho.otype — Hoffman et al. 2008, Mycological Research 112: Fic. 5D.

ErymoLocy — Referring to Sede Boker, Israel, the locality where the species was

originally isolated.

Description as for genus. Sporangium size 20-35 um in dual culture on host

(Hoffman et al. 2008); aplanospores with numerous refractive globules that

coalesce upon encystment; parasitic on non-motile Haematococcus pluvialis

Flot. (Haematococcaceae, Chlorophyceae).

Discussion

Paraphysoderma is differentiated from Physoderma by the absence of a

motile spore, the presence of an epibiotic resting sporangium, culturability,

Paraphysoderma sedebokerense, gen. et sp. nov. ... 179

and host range. However, it has a general appearance of a typical monocentric,

eucarpic chytrid or blastocladialean fungus, and is similar to Physoderma in

having an epibiotic sporangium and a limited rhizoid emanating from a single

axis on the sporangium. Its phylogenetic position (Hoffman et al. 2008) implies

that the genus or species has recently lost a flagellated spore stage. Despite the

absence of a motile spore, the aplanospores of the parasite are nonetheless able

to cause severe epidemics in aquaculture.

Our current observations do not exclude the possibility that additional study

of Paraphysoderma will ultimately reveal a zoospore stage. When sexuality is

observed in members of phylum Blastocladiomycota, an alternation of diploid

and haploid generations is observed (James et al. 2006). Thus it is possible that

a thallus or stage of different ploidy on an alternative host may exist, although

infection studies suggest it is not likely to be another green alga (Gutman et al.

2009). However, the observation that the aplanospores from both resting and

thin-walled sporangia are morphologically similar suggests the absence of a

separate gametophyte stage. If the fungus conforms to the life cycles expected

of other blastoclads, then it would be considered to have a ‘Brachyallomyces’

(short) life cycle, which may or may not include meiosis during resting sporangial

germination (Emerson 1941). The basal placement of Paraphysoderma to

Physoderma is consistent with the derivation of the land plant parasitism from

an aquatic one between a chytrid-like fungus and an alga. The culturability of

Paraphysoderma and not Physoderma is consistent with the evolution of an

increasingly obligate parasitic relationship of Physoderma to its host. Indeed,

Physoderma and Paraphysoderma are unique among Blastocladiomycota in its

parasitism of the plant kingdom, although whether the use of plant hosts is a

derived or ancestral trait is unclear, as Physodermataceae occupy the most basal

branch in the Blastocladiomycota (Porter et al. 2011). The distinct differences in

culturability between the two genera suggests that Paraphysoderma might serve

as a model species for the clade in research requiring large numbers of cultured

cells, such as genomics and biochemistry.

The unique nature of the amoeboid swarming aplanospores suggests that

neither genus nor species has been observed before. Chytridiomycota species

that have been found to reproduce solely by amoeboid aplanospores include

Amoebochytrium rhizidioides Zopf (host: Chaetophora elegans (Chlorophyceae)),

Sporophlyctis rostrata Serbinow (host Draparnaldia spp. (Chlorophyceae)), and

Sporophlyctidium africanum Sparrow (host Protoderma sp. (Ulvophyceae)),

all of which differ in several ways (including rhizoidal development) from

Paraphysoderma. An aplanosporic mode of reproduction is unusual among

aquatic fungi, and the factors that favor the loss of a flagellated spore in an

aquatic environment are unclear. The pattern that many aplanosporic chytrids

parasitize green algae suggests that the need for a motile spore may be minimal

180 ... James & al.

in certain well-mixed environments, while the ability to creep over the host to

find an ideal location for rhizoidal penetration may the trait most influenced

by natural selection.

The number of accepted species of Physoderma is ~50 (Kirk et al. 2008). The

number of species and distribution of Paraphysoderma are currently unknown

but will probably be much greater than currently appreciated given the rapid

expansion of chytrid diversity seen in both culture studies and environmental

DNA surveys (Letcher et al. 2008; Freeman et al. 2009).

Acknowledgments

The authors thank Dr. Joyce E. Longcore (University of Maine), Dr. Wallace Martin

(Randolph-Macon College), and Dr. Merlin White (Boise State University) for their

constructive comments on the manuscript. Thanks also to Angela Piper for her help

with the Latin translation.

Literature cited

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carotenoid production: current state and perspectives. Applied Microbiology and Biotechnology

74: 1163-1174. http://dx.doi.org/10.1007/s00253-007-0844-9

Emerson R, 1941. An experimental study of the life cycles and taxonomy of Allomyces. Lloydia 4:

77-144.

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Schmidt SK. 2009. Evidence that chytrids dominate fungal communities in high-elevation

soils. Proceedings of the National Academy of Sciences of the United States of America 106:

18315-18320. http://dx.doi.org/10.1073/pnas.0907303106

Gutman J, Zarka A, Boussiba S. 2009. The host-range of Paraphysoderma sedebokerensis, a

chytrid that infects Haematococcus pluvialis. European Journal of Phycology 44: 509-514.

http://dx.doi.org/10.1080/09670260903161024

Hoffman Y, Aflalo C, Zarka A, Gutman J, James TY, Boussiba S. 2008. Isolation and characterization of

a novel chytrid species (phylum Blastocladiomycota), parasitic on the green alga Haematococcus.

Mycological Research 112: 70-81. http://dx.doi.org/10.1016/i.mycres.2007.09.002

James TY, Letcher PM, Longcore JE, Mozley-Standridge SE, Porter D, Powell MJ, Griffith GW,

Vilgalys R. 2006. A molecular phylogeny of the flagellated fungi (Chytridiomycota) and

description of a new phylum (Blastocladiomycota). Mycologia 98: 860-871.

http://dx.doi.org/10.3852/mycologia.98.6.860

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

Publishing, The Netherlands.

Letcher PM, Velez CG, Barrantes ME, Powell MJ, Churchill PF, Wakefield WS. 2008. Ultrastructural

and molecular analyses of Rhizophydiales (Chytridiomycota) isolates from North America and

Argentina. Mycological Research 112: 759-782.

http://dx.doi.org/10.1016/j.mycres.2008.01.025

Porter TM, Martin W, James TY, Longcore JE, Gleason FH, Adler PH, Letcher PM, Vilgalys R.

2011. Molecular phylogeny of the Blastocladiomycota (Fungi) based on nuclear ribosomal data.

Fungal Biology 115: 381-392. http://dx.doi.org/10.1016/j.funbio.2011.02.004

MY COTAXON

http://dx.doi.org/10.5248/118.181

Volume 118, pp. 181-186 October-December 2011

Microcyclospora rumicis, a new species on Rumex crispus from Iran

MAHDI ARZANLOU*& MOUNES BAKHSHI

Plant Protection Department, Faculty of Agriculture, University of Tabriz,

29" Bahman Boulevard, Tabriz, PRO. Box 5166614766, Iran.

* CORRESPONDENCE TO: Arzanlou@hotmail.com.

ABSTRACT — A new species of Microcyclospora, associated with leaf spots on Rumex crispus,

is fully described and illustrated.

Key worps — hyphomycetes, Pseudocercospora, weed, cercosporoid fungi

Introduction

During the study of alternative weed hosts for Cercospora beticola Sacc.,

a causal agent of cercospora leaf spot of sugar beet in Northern Iran, a

species of the genus Microcyclospora J. Frank et al. was isolated from Rumex

crispus showing leaf spot symptoms in the Talesh region (Guilan province).

Microcyclospora, with M. pomicola J. Frank et al. designated as type species, was

recently segregated from Pseudocercospora Speg. s. str. based on phylogenetic

and morphological differences (Frank et al. 2010). Currently the three species

known in Microcyclospora are associated with sooty blotch and flyspeck (SBFS)

blemishes on the surfaces of pomaceous fruits, specifically apples (Colby 1920;

Frank et al. 2010). Here we describe a new Microcyclospora species from leaf

spots on Rumex crispus that differs morphologically from the three other

known species of the genus.

Materials & methods

Conditions of isolation, culture, and observation

Leaf samples of Rumex crispus showing leaf spot were collected from Rainforest

Mountains in the Talesh region, Guilan province, in northern Iran near the Caspian

Sea. Single conidial isolates were established directly from symptomatic curly dock

leaves according to Bakhshi et al. (2011). A mass of conidia scraped from the lesion

using a sterile inoculation needle under a stereomicroscope was floated in 10 ml sterile

distilled water and spread on 2% malt extract agar (MEA; Fluka, Germany). After plates

182 ... Arzanlou & Bakhshi

were incubated in a slanted position overnight, germinated conidia were transferred

to new MEA plates, and cultures were incubated in the dark at 25 °C. After 30 days of

incubation, samples from the colony were mounted on glass slides in clear lactic acid

for microscopic examination. Thirty measurements were made for each microscopic

structure, and 95% confidence intervals were derived for the measurements with

extreme values given in parentheses. Line drawings were prepared using a BX41 light

microscope (Olympus, Japan) equipped with a drawing tube; photos were captured

using a Leica camera system. Colony colors on MEA and oatmeal agar (OA; Gams et

al. 2007) [surface and reverse] were determined after one month at 25 °C in the dark.

Nomenclature and descriptions were deposited in MycoBank. The holotype voucher

and an ex-type culture are conserved in CCTU, the culture collection housed at the

Plant Protection Department, Agriculture Faculty, University of Tabriz, Iran.

Taxonomy

Microcyclospora rumicis Arzanlou & Bakhshi, sp. nov. PLATE 1-2

MycoBank MB 563723

Microcyclospora rumicis ab aliis speciebus generis chlamydosporis numerosis, majoribus,

ad 20 um diam distinguenda.

Type: Iran, Guilan, Talesh, on leaves of Rumex crispus L. (Polygonaceae), Sep. 2010, M.

Bakhshi (Holotype, CCTU-H-1; ex-type culture: CCTU 1 = CBS 131546).

Erymo.ocy: Named after the host genus, Rumex.

Culture characteristics — On MEA slow growing, reaching 3 mm diam after 7 d,

and up to 10 mm after 2 weeks at 25 °C, raised, unevenly folded, with moderate,

smoke-gray aerial mycelium; surface irregular, with smooth, lobate margins,

iron-grey; iron-grey in reverse. Colonies on OA reaching up to 2 mm diam

after 7 d, and 6 mm after 2 weeks at 25 °C, flat, submerged, with sparse aerial

mycelium and smooth margin. Microcyclic conidiation commonly observed

on all media in culture.

In vitro on MEA — Mycelium consisting of branched, smooth, septate,

pale brown, 2-4 um wide hyphae developing numerous chlamydospores,

intercalary and terminal, medium brown, 6-20 um diam. Conidiophores

reduced to conidiogenous cells, integrated, lateral on hyphae, mono- to

polyblastic, subdenticulate, 2.5-4 um wide, 8-13 um tall, hyaline, smooth.

Conidia scolecosporous, cylindrical, straight to variously curved, guttulate,

apex obtuse, base truncate, hyaline, (1.5-)2.5(-4) x (15-)37-54(-100) um,

1-10-septate; hila neither thickened nor darkened; microcyclic conidiation

commonly observed; older conidia developing intercalary chlamydospores that

are pale brown, < 9 um diam.

Discussion

The cercosporoids are amongst the major groups of fungi and generally

cause leaf spot diseases on almost all families of flowering plants throughout

Microcyclospora rumicis sp. nov. (Iran) ... 183

PLaTE 1. Microcyclospora rumicis.

Conidia, conidiogenous cells and hyphae from MEA colony. Bar = 10um.

the world. Among these, Pseudocercospora is the second largest cercosporoid

genus, with more than 300 published names (Kirk et al. 2008). Recently it was

shown that Pseudocercospora included taxa that vary considerably in their

conidiomatal morphology, ranging from solitary conidiogenous loci, synnemata,

sporodochia to fascicles. Moreover, conidia in some taxa are transversely

euseptate but with some oblique and longitudinal septa or containing a mixture

of eu- and distoseptation. Conidial hila and scars vary from unnoticeable to

slightly thickened along the rim (Stewart et al. 1999). Even though the conidia

184 ... Arzanlou & Bakhshi

PLaTE 2. Microcyclospora rumicis. a. Chlamydospore in hyphae; b-c. Conidiogenous loci

(arrowed) and conidia with chlamydospore; d. Young conidia; e-f. Hyphae. g. 30-day old

colony on MEA. Scale bars = 10 um.

are commonly solitary, they may in some cases also occur in unbranched

chains (Braun 1995). Frank et al. (2010) established Microcyclospora based on

morphological and molecular data. Morphologically, Microcyclospora can be

distinguished from Pseudocercospora by the arrangement of the conidiophores

that are never fasciculate but reduced to solitary conidiogenous loci on

hyphae and conidia that aggregate in mucoid masses, strongly tending toward

microcyclic conidiation. Only three other species described in the genus

—M. malicola J. Frank et al., M. pomicola, and M. tardicrescens J. Frank et

al.— associated with sooty blotch and flyspeck (SBFS) blemishes on surfaces of

pomaceous fruits (Frank et al. 2010). Here, we introduce a new species based

on morphological characteristics. Microcyclospora rumicis, like M. tardicrescens,

forms intercalary chlamydospores (lacking in M. malicola and M. pomicola)

and grows more slowly in culture. Chlamydospores of M. rumicis are longer

(< 20 um) than those of M. tardicrescens (< 5 um long).

Little is known about sexual stages of the cercosporoid fungi including

Microcyclospora species. Cercosporoids have been traditionally treated as

anamorphs of the ascomycete Mycosphaerella Johanson (e.g., Braun & Mel'nik

1997, Kim & Shin 1998, Crous & Braun 2003). However, most cercosporoids

Microcyclospora rumicis sp. nov. (Iran) ... 185

are considered exclusively asexual, and Mycosphaerella teleomorphs have been

confirmed for only a few species.

Phylogenies derived from multiple sequence analyses place Microcyclospora

with other cercosporoid fungi in the Mycosphaerella clade (Mycosphaerellaceae,

Capnodiales, Dothideomycetidae; Frank et al. 2010). Mycosphaerella, one of the

largest genera in the Ascomycota, comprises several thousand species (Crous et

al. 2001, 2009a,b; Aptroot 2006). Contrary to the assumption that Mycosphaerella

as then defined was monophyletic, Crous and co-workers recently revealed

that the genus is polyphyletic and split the former Mycosphaerellaceae into

several families, of which the Mycosphaerellaceae, Teratosphaeriaceae and

Schizothyriaceae have plant-pathological importance (Crous et al. 2007, 2009a).

Among the vast number of anamorphs in these lineages, up to 30 anamorph

genera have been linked to Mycosphaerella (Crous et al. 2007, 2009a; Arzanlou

et al. 2007). Recent phylogenetic analysis based on multiple sequence data sets,

however, indicate that these interpretations are not quite correct (Crous et al.

2009b) and that the Mycosphaerellaceae encompass instead numerous genera

with morphologically conserved Mycosphaerella-like teleomorphs but quite

distinct anamorphs (Crous et al. 2007, 2009b).

Little is currently known about ecology, host ranges, and sexual stages of the

cercosporoid fungi including Microcyclospora species, and more sampling on

different substrates is needed to address these aspects.

Acknowledgments

The authors are grateful to Prof Walter Gams for the Latin diagnosis and critical

reading of this manuscript and his valuable comments and suggestions, as well as to

Prof. Uwe Braun and Dr. Konstanze Bensch for pre-submission reviews.

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23: 99-118. http://dx.doi.org/10.3767/003158509X479487

Frank J, Crous PW, Groenewald JZ, Oertel B, Hyde KD, Phengsintham P, Schroers HJ. 2010.

Microcyclospora and Microcyclosporella: novel genera accommodating epiphytic fungi causing

sooty blotch on apple. Persoonia 24: 93-105. http://dx.doi.org/10.3767/003158510X510560

Gams W, Verkley GJM, Crous PW. 2007. CBS course of mycology, 5th edition. Centraalbureau voor

Schimmelcultures, Utrecht, Netherlands.

Kim JD, Shin HD. 1998. Taxonomic studies on Cercospora and allied genera in Korea (I). Korean

J. Mycol. 26: 327-341.

Kirk PM, Cannon PF, David JC, Stalpers LA. 2008. Dictionary of the fungi, 10th edn. CAB

International, Wallingford, Oxon.

Stewart EL, Liu Z, Crous PW, Szabo L. 1999. Phylogenetic relationships among some cercosporoid

anamorphs of Mycosphaerella based on rDNA sequence analysis. Mycol. Res. 103: 1491-1499.

http://dx.doi.org/10.1017/S0953756299008680

MY COTAXON

http://dx.doi.org/10.5248/118.187

Volume 118, pp. 187-195 October-December 2011

Mycena guldeniana - a new alpine species from Norway

ARNE ARONSEN? & BRIAN A. PERRY?”

'Torodveien 54, N-3135 Torod, Norway

Biology Department, University of Hawai’ at Hilo, 200 W. Kawili St., Hilo, HI 96720, USA.

CORRESPONDENCE TO: ' araronse@online.no & * baperry@hawaii.edu

AsstTRaAct — Mycena guldeniana (section Polyadelphia) is proposed for an agaric found in

alpine areas of south Norway that is here described, illustrated and compared with other

species of the section. The new species grows on fallen, decaying leaves under Salix spp. or

occasionally on small twigs in the same habitat and is characterized by its small size, greyish

brown pileus, 4-spored basidia, smooth cheilocystidia, prominent, wide terminal cells of

the pileipellis hyphae, and clamp connections. Phylogenetic analyses of nucLSU support the

taxon within Mycena and genetically distinct from the morphologically similar M. terena.

KEY worps — taxonomy, alpine mycology, basidiomycete phylogenetics

Introduction

During a stay at ‘Finse Research Centre’ in south Norway, August 2005, the

first author collected and studied several Mycena species. ‘The research centre

is situated at approximately 60°30'N on the mountain plateau Hardangervidda

1200 m above sea level. The area lies entirely within the alpine zone and is

devoid of trees but in parts is covered with Salix shrubs. The research led to the

proposal of two new Mycena species (Aronsen & Gulden 2007). Two collections

were also made of a small Mycena that initially was misinterpreted as M. terena

Aronsen & Maas Geest., a species found on fallen, decaying leaves of Salix caprea

in the coastal area of South Norway (Aronsen & Maas Geesteranus 1992).

In 2008 the ‘Finse research centre’ was visited again, and the main

investigation was done on the south-facing slope at Nordnut at approximately

1400 m. The slope is formed by basic, phyllitic rocks intermixed with Salix shrubs

and some calcicolous vegetation (Gulden & Jenssen 1982). On small, partially

buried twigs on the ground under the Salix shrubs a tiny Mycena species was

collected. Further investigation showed that not only was it identical with the

taxon found in 2005, it clearly differed from M. terena and did not match any

other known species of Mycena.

188 ... Aronsen & Perry

The same year this taxon was also collected at the base of the mountain

Gaustatoppen at approximately 59°51'N at approximately 1150 m. Gaustatoppen

(1883 m) is the peak of a 7 km long mountain spine consisting of quarzite,

and in the lower slopes there are areas with dense Salix shrubs. In these Salix

shrubs the unknown taxon was found in great numbers together with M. exilis

Aronsen & Gulden. The taxon is herein proposed as a new species.

Materials & methods

The description is based on six collections from two different years and two different

localities including specimens in all stages of age. Preliminary observations and photos

were made in daylight in the field, while more thorough observations and descriptions

were made the same day in lamp light in the laboratory of the research centre. The dried

material was rehydrated in 2% KOH and examined in ammoniated Congo Red and

Melzer’s reagent using a Nikon light microscope with high resolution, 100x oil objective.

For each collection 30 spores were measured. The spore length and width ratios (q) were

calculated and the average quotient value (q, ) is given in the description. Measurements

and drawings were made of basidia, spores, cheilocystidia, pileipellis hyphae, and the

stipe cortical layer. Melzer’s reagent was used to check amyloidity of spore walls and

colour reactions in the lamellar trama. Authors abbreviations follow Index Fungorum

(2011), and the collections are deposited in the university museum of Oslo (O).

To assess the phylogenetic position of M. guldeniana within Mycena, and the

relationship of this taxon to the morphologically similar M. terena, sequences of the

5’ end of the nuclear large ribosomal subunit gene (nucLSU) spanning domains D1

and D2 were analyzed within a broader sampling of taxa representative of Mycena and

the tricholomatoid clade sensu Matheny et al. (2006). Two members of Boletales were

included as outgroup taxa for rooting purposes. Genomic DNA was extracted from

dried basidiomes representing each taxon (TABLE 1) using the E.Z.N.A. Forensic DNA

Kit (Omega Bio-Tek, U.S.A.) according to the manufacturer’s protocols. PCR protocols

followed Perry & Pfister (2007) using primers LROR and LR5 (Moncalvo et al. 2007).

Amplified DNA was cleaned with ExoSAP-IT (USB Molecular Biology Reagents and

Biochemical, U.S.A.) and sent to Elim Biopharmaceuticals (http://www.elimbiopharm.

com/) for cycle sequencing and visualization on an ABI 3700 capillary sequencer

(Applied Biosystems, U.S.A.) using the same primers as above. Sequences were edited

and assembled using Sequencher 4.0 (Gene Codes Corp., U.S.A.). The resulting data

matrix was aligned manually with MacClade 4 (Maddison & Maddison, 2000). Edited

sequences have been deposited in GenBank (TaBLE 1), and the aligned dataset is

available via TreeBase (www.treebase.org; $11538).

Maximum likelihood (ML) analyses were conducted in PAUP* version 4 (Swofford

2003) and employed an iterative approach. Starting trees were built via the Neighbor

Joining method and used to estimate parameters of the model of sequence evolution.

Estimated parameter values were then fixed, and a ML search was conducted. Resulting

topologies were then used to re-estimate and fix parameter values. This process was

repeated for a total of three iterations to insure the analyses had converged on the most

likely topology. Clade support was assessed by non-parametric, maximum likelihood

TABLE 1. List of sequences included in this study and associated GenBank

accession numbers.

Mycena guldeniana sp. nov. (Norway) ... 189

SPECIES COLLECTION GENDENK

ACCESSION #

Asterophora lycoperdoides (Bull.) Ditmar CBS 170.86 AF223190

Boletellus projectellus (Murrill) Singer MB 03-118 NG_027638

Clitocybe dealbata (Sowerby) Gillet HC 95.cp3 AF223175

Collybia tuberosa (Bull.) P. Kumm. TENN53540 NG_027631

Entoloma prunuloides (Fr.) Quél. TJB4765 AY700180

Mycena clavicularis (Fr.) Gillet RV87/6 AF042637

Mycena crocata (Schrad.) P. Kumm. BAYER G 057 AY207241

Mycena galericulata (Scop.) Gray GLM 45970 AY207251

Mycena guldeniana A 36/05 JF944831

Mycena haematopus (Pers.) P. Kumm. HKI ST 22545 AY207252

Mycena insignis A.H.Sm. DAOM208539 AF261413

Mycena leaiana (Berk.) Sacc. DAOM167618 AF261411

Mycena maculata P. Karst GLM 4597 AY207254

Mycena niveipes (Murrill) Murrill GLM 45975 AY207242

Mycena olivaceomarginata (Massee) Massee GLM 45976 AY207255

Mycena plumbea P. Karst. PBM 2718 DQ470813

Mycena polygramma (Bull.) Gray GLM 45977 AY207243

Mycena pura (Pers.) P. Kumm. JM98/136 AF261410

Mycena renati Quél. GLM 45979 AY207256

Mycena rubromarginata (Fr.) P. Kumm. GLM 45981 AY207245

Mycena sanguinolenta (Alb. & Schwein.) P. Kumm. GLM 45982 AY207257

Mycena terena A 41/09 JF944832

Mycena tintinnabulum (Fr.) Quél. GLM 4598 AY207258

Mycena viscidocruenta Cleland DUKE3411 AF261414

Mycena zephirus (Fr.) P. Kumm. GLM 45984 AY207259

Suillus pictus (Peck) A.H. Sm. & Thiers MB03-002 AY684154

Termitomyces sp. ZA164 DQ110875

Tricholoma aestuans (Fr.) Gillet PBM2494 AY700197

bootstrap analyses (Felsenstein 1985) as implemented in GARLI (Zwickl 2006), and

consisted of 1000 replicates with all parameter values estimated by the program. All

analyses were performed under a GTR+I+G model of sequence evolution as determined

using the Akaike Information Criterion in MrModelTest 2.3 (Nylander 2004). Bayesian

190 ... Aronsen & Perry

analyses were performed using Metropolis-coupled Markov Chain Monte Carlo

(MCMCMC) methods as implemented in MrBayes 3.1.2 (Huelsenbeck & Ronquist

2001; Ronquist & Huelsenbeck 2003) using the same model as the ML analyses. Analyses

consisted of two parallel searches, run for 3 million generations and initiated with

random starting trees. Chains were sampled every 300 generations for a total of 10,000

trees per search, sampled from the posterior distribution. Trees sampled prior to the

analysis reaching a split deviation frequency of 0.02 were discarded as the burn-in, while

the remaining trees were used to calculate the posterior probabilities of the individual

clades. Default settings in MrBayes were used to set the incremental heating scheme,

chain number, unconstrained branch lengths and uninformative topology priors.

Taxonomy

Mycena guldeniana Aronsen & B.A. Perry, sp. nov. FIG. 1-7

MycoBank MB 561558

Basidiomata solitaria vel gregaria. Pileus usque ad 2 mm latus, hemisphericus, conicus

vel parabolicus, interdum papillatus, plerumque sulcatus, translucente striatus, pruinosus,

glabrescens, pallide brunneus, pallide griseo-brunneus vel obscure griseus, centro obscurior.

Caro tenuis, pallida, odore nullo. Lamellae 4-8 stipitem attingentes, adnatae vel late

adnatae, pallide brunneae, pallide griseae vel albae. Stipes usque 30 x 0.2 mm, brunneo-

griseus vel griseus deorsum albido-pallescens, basi fibrillis albis instructus.

Basidia 18-33 x 7-11 wm, clavata, (2-)4-spora, fibulata. Sporae 7.5-8.7-10 x

4.8-5.5-6.5 um, amyloideae. Cheilocystidia 21-40 x 6-11 um, cylindracea, subclavata,

sublageniformia, fibulata, laevia. Pleurocystidia nulla. Hyphae pileipellis 5-10(-17)

um latae, fibulatae, diverticulatae, cellulae terminales usque 15 um latae, clavatae

vel subcylindraceae, diverticulatae. Hyphae stipitis corticales 1-7 um latae, fibulatae,

diverticulatae.

Ad Salicis folia decisa, in zona alpina.

Type: NORWAY, Telemark, Tinn, Gaustakneet, UTM(WGS84): MM 8249 3560, altitude

ca. 1150 m., 21 Sept. 2008, leg. A. Aronsen A54/08 (HOLOTYPE O-F300034).

Erymo ocy: The species is named after Prof. Gro Gulden, Oslo, in recognition of her

contributions to the knowledge of arctic and alpine fungi.

Pileus 1-2 mm across, hemispherical, then conical to parabolical, flattening to

convex, becoming more or less plano-convex with or without a small papilla

centrally, occasionally somewhat depressed centrally, pruinose, glabrescent,

shallowly sulcate, more or less translucent-striate, pale brown or pale grey-

brown or grey to dark grey, sometimes pale grey, the centre dark brown to

dark grey. Flesh very thin, whitish. Odour and taste indistinctive. Lamellae

4-8 reaching the stipe, with or without lamellulae, usually well developed and

fairly broad, but occasionally only showing as faint ridges, ascending, the edge

concave to convex, narrowly adnate to broadly adnate, sometimes decurrent

with a very short tooth, pale brown, pale grey to white, the edge pallid. Stipe

up to 30 x 0.2 mm, very thin, hollow, equal, terete, firm, pruinose, glabrescent,

becoming shiny, curved to flexuous, brownish grey to watery grey, becoming

more whitish; sometimes dark grey at the apex in younger specimens; often

Mycena guldeniana sp. nov. (Norway) ... 191

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FIGURES 1-5. Mycena guldeniana (holotype). 1. Hyphae of the pileipellis with terminal cells.

2. Hypha of the cortical layer of the stipe. 3. Basidia. 4. Cheilocystidia. 5. Spores.

Bar = 20 um.

conspicuously insititious but occasionally attached to the substratum by a

whorl of radiating, flexuous, white mycelial fibrils.

Basidia 18-33 x 7-11 um, clavate, (2- and) 4-spored, with plump sterigmata

up to 5 um long. Spores 7.5-8.7-10 x 4.8-5.5-6.5 um, q = 1.3-1.9, q,, ~ 1.6,

broadly ellipsoid to pip-shaped, smooth, amyloid. Cheilocystidia 21-40 x 6-11

um, occuring mixed with the basidia, cylindrical, subclavate, sublageniform,

smooth. Pleurocystidia absent. Lamellar trama dextrinoid, reddish brown in

Melzer’s reagent. Hyphae of the pileipellis 5-10(-17) um wide, densely covered

with cylindrical, straight excrescences 1-3.5 x 0.5 um; terminal cells up to 40

x 15 um, clavate to subcylindrical, covered with short, straight, cylindrical

excrescences. Hypodermium of broad, cylindrical to subglobose, smooth cells

15-40 x 13-28 um. Hyphae of the cortical layer of the stipe 1-7 um wide, densely

covered with cylindrical, sometimes more thorn-like, straight excrescences 1-6

x 0.5-1 um; terminal cells not typically differentiated but a few observed at

the base of the stipe being clavate, 26-35 x 5-10 um, densely covered with

cylindrical excrescences 1-4 x 0.5-1 um. Clamp connections observed at the

septa of some of the basidia, the cheilocystidia, and the hyphae of the pileipellis

and the stipe cortex.

192 ... Aronsen & Perry

Solitary or in small groups on fallen, decaying leaves of Salix sp.; occasionally

on small, fallen twigs on the ground in dense Salix shrubs.

ADDITIONAL SPECIMENS EXAMINED — NORWAY: HorDALAND, Ulvik, Finse, Nordnut

sor, UTM(WGS84): MN 1667 1903, alt. ca. 1400 m, 27 Aug. 2005, leg. A. Aronsen A 34/05

and A 36/05, GENBANK JF944831; 23 Aug. 2008, leg. A. Aronsen A 30/08; TELEMARK,

Tinn, Gaustakneet, UTM(WGS84): MM 8249 3560, alt. ca 1150 m, 14 Sept. 2008, leg. A.

Aronsen A 44/08; 21 Sept. 2008, leg. A. Aronsen A 63/08.

Discussion

Mycena guldeniana belongs to section Polyadelphia Singer ex Maas Geest.

(Maas Geesteranus 1986) based on its small size, adnate lamellae, pileipellis

and stipitipellis hyphae densely covered with short cylindrical excrescences,

absence of pleurocystidia, and occurrence on decaying leaves. The species of

this section usually possess cheilocystidia that are densely covered with short,

cylindrical excrescences, but Aronsen & Maas Geesteranus (1992) described the

new species M. terena with smooth cheilocystidia. Another species in the same

section known to have more or less smooth cheilocystidia is M. querciphila

Esteve-Rav. & M. Villarreal (Esteve-Raventos & Villarreal 1997).

The smooth cheilocystidia of M. guldeniana suggest a close relationship

with both M. terena and M. querciphila, with M. terena seeming the most

similar. Mycena querciphila differs in the pale olive-yellow to olive-brown

colours of pileus, lamellae, and stipe and in having some cheilocystidia

sparsely diverticulate by warts or short cylindrical excrescences. Additionally,

M. querciphila is typically found growing on fallen, decaying leaves of Quercus

ilex subsp. ballota. Mycena terena, recorded on fallen decaying Salix caprea

leaves in two coastal lowland localities in South Norway, has a pale beige or

very pale grey pileus that soon turns white, a black stipe apex (when young),

somewhat narrower spores, and pileipellis hyphae that lack prominent terminal

cells. The differences are shown in TABLE 2 below.

TABLE 2. A comparison between Mycena guldeniana and M. terena.

: MYCENA GULDENIANA

: MYCENA TERENA

Pateus pale brown to grey-brown pale beige or pale grey,

: soon fading to white

Gar Bc ste Se white to dark grey black

: in young specimens : in young specimens

SPORES q,y ~1.6 q,,~ 1.8

HYPHAE OF THE

PILEIPELLIS

HABITAT

with prominent, inflated

terminal cells

on fallen leaves and twigs under

Salix spp. in alpine area

: without prominent

terminal cells

on fallen leaves of Salix caprea

in coastal lowland

Mycena guldeniana sp. nov. (Norway) ... 193

FiGuRE 6. Mycena guldeniana (holotype).

On 11 Aug. 1979, Gulden & Jenssen (1982) reported an unknown species of

Mycena, collected at Finse, Nordnut, on a dead leaf. Although their material was

‘too scanty for a formal description of a new species, the description indicates

that their unknown species was identical with M. guldeniana.

Phylogenetic analyses

The nucLSU dataset comprises 887 aligned positions for 26 ingroup taxa

and contains 131 parsimony informative characters. Maximum likelihood

analyses converged on a common topology, with each iteration producing trees

that did not differ significantly in their likelihood values. The final ML tree

recovered is shown in FiGuRE 7 (-/nL = 3527.88466). Bayesian analyses reached

a split deviation frequency below 0.02 after approximately 880,000 generations,

and the first 2930 trees sampled were discarded as the burn-in. All Mycena

sequences included in the analyses form a well-supported (ML bootstrap = 100,

posterior probability = 1.0) lineage, sister to the remaining taxa sampled from

the tricholomatoid clade. Although relationships within the Mycena lineage

are not resolved with significant support, M. guldeniana and M. terena are

194 ... Aronsen & Perry

99/1.0

100/1.0

0.05

Mycena clavicularis

94/1.0

Mycena tintinnabulum

Mycena plumbea

-/0.80

Mycena rubromarginata

Mycena zephirus

Mycena insignis

Mycena crocata

Mycena leaiana

Mycena maculata

-/0.89

Mycena niveipes

Mycena polygramma

Mycena guldeniana

Mycena terena

Mycena haematopus

76/0.92

Mycena sanguinolenta

Mycena olivaceomarginata

Mycena viscidocruenta

Mycena renati

Mycena pura

Mycena galericulata

Entoloma prunuloides

Termitomyces sp.

Tricholoma aestuans

Collybia tuberosa

Asterophora lycoperdoides

Clitocybe dealbata

Boletellus projectellus

Suillus pictus

FIGURE 7. Maximum likelihood topology of Mycena guldeniana and other Mycena species inferred

from nucLSU sequence data (-InL = 3527.88466). Numbers separated by / represent maximum

likelihood bootstrap proportions and bayesian posterior probabilities greater than 70% and 0.70,

respectively (- designates a value below 70% / 0.70).

resolved as sister taxa in all topologies recovered by the ML analysis. Although

the relationship between these taxa is not well supported by ML bootstrap or

bayesian posterior probabilities, these results do support our recognition of

M. terena and M. guldeniana as distinct phylogenetic species. While these two

taxa are very similar morphologically, the nucLSU data indicate that genetically

they are as distinct from one another as are the majority of other closely related

Mycena species included in our analyses.

Mycena guldeniana sp. nov. (Norway) ... 195

Acknowledgments

The authors would like to thank Dr. J. Miersch (Halle, Germany) and Dr. T. Leess@e

(Copenhagen, Denmark) for critical revision of the manuscript.

Literature cited

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Progress 6: 1-6. http://dx.doi.org/10.1007/s11557-006-0518-5

Aronsen A, Maas Geesteranus RA. 1992. Mycena terena, a new member of section Polyadelphia

from Southern Norway. Persoonia 15(1): 105-107.

Esteve-Raventdés F, Villarreal M. 1997. Mycena quercophila, a new species of Mycena section

Polyadelphia growing on Quercus ilex leaves. Osterr. Z. Pilzk. 6: 67-70.

Felsenstein J. 1985. Confidence limits on phylogenies - an approach using the bootstrap. Evolution

39: 783-791.

Gulden G, Jenssen KM. 1982. Mycena and related genera in alpine habitats of South Norway. Arctic

and Alpine Mycology (Symposium) 1: 164-197.

Index Fungorum. 2011. http://www.speciesfungorum.org/ (viewed online on 12 Mar 2011).

Heulsenbeck JP, Ronquist F. 2001. MRBAYES: Bayesian inference of phylogeny. Bioinformatics 17:

754-755.

Maas Geesteranus RA. 1986. Conspectus of the Mycenas of the Northern Hemisphere - 6. Sections

Polyadelphia and Saetulipedes. Proc. K. Ned. Akad. Wet. (Ser. C) 89(2): 159-182.

Maddison DR, Maddison W. 2000. MacClade 4: Analysis of phylogeny and character evolution.

Version 4.0. Sinauer Associates, U.S.A.

Matheney PB, Curtis JC, Hofstetter V, Aime M.C, Moncalvo J-M, He Z-W, Yang Z-L, Slot JC,

Ammirati JF, Baroni TJ, Bougher NL, Hughes KW, Lodge DJ, Kerrigan RW, Seidl MT, Aanen

DK, DeNitis M, Daniele GM, Desjardin DE, Kropp BR, Norvell LL, Parker A, Vellinga EC,

Vilgalys R, Hibbett DS. 2006. Major clades of Agaricales: a multilocus phylogenetic overview.

Mycologia 98(6): 982-995. http://dx.doi.org/10.3852/mycologia.98.6.982

Moncalvo JM, Lutzoni FM, Rehner SA, Johnson J, Vilgalys R. 2000. Phylogenetic relationships of

agaric fungi based on nuclear large subunit ribosomal DNA sequences. Systematic Biology 49:

278-305.

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology

Centre, Uppsala University (http://www.abc.se/~nylander/)

Perry BA, Pfister DH. 2007. Chaetothiersia vernalis, a new genus and species of Pyronemataceae

(Ascomycota, Pezizales) from California. Fungal Diversity 28: 65-72.

Ronquist F, Huelsenbeck JP. 2003. MRBAYES 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. http://dx.doi.org/10.1093/bioinformatics/btg180

Swofford D. 2003. PAUP*. Phylogenetic Analysis Using Parsimony (*and other methods). Version

4. Sinauer Associates, U.S.A.

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of Texas at Austin.

MY COTAXON

http://dx.doi.org/10.5248/118.197

Volume 118, pp. 197-201 October-December 2011

Neolecta vitellina, first record from Romania,

with notes on habitat and phenology

VASILICA CLAUDIU CHINAN ™* & DAvID HEwITT ”

‘Faculty of Biology, Alexandru Ioan Cuza University, Bd. Carol I, No. 20A, 700505, Iasi, Romania

*Department of Botany, Academy of Natural Sciences

1900 Benjamin Franklin Parkway, Philadelphia, PA 19103 USA

* CORRESPONDENCE TO: vasilechinan@yahoo.com

Asstract — Neolecta vitellina, one of the rarely collected ascomycete species in Europe,

is reported from Romania for the first time. The species was found on the ground, under

Norway spruce (Picea abies) in 2004, from the Nature Reserve “Tinovul Mare” Poiana

Stampei (Eastern Carpathians, Romania). Subsequent field observations have confirmed the

presence of Neolecta vitellina in the same location, in the period 2005-10. A description and

photographs of the specimens are presented.

Key worps — Ascomycota, Neolectaceae, ITS sequence

Introduction

The genus Neolecta Speg. belongs to the ascomycete family Neolectaceae

(Redhead 1977), order Neolectales (Landvik et al. 1993), class Neolectomycetes

(Taphrinomycotina, Ascomycota). This genus includes three accepted species

—N. flavovirescens Speg., N. irregularis (Peck) Korf & J.K. Rogers, N. vitellina—

with clavate, unbranched to lobed yellow ascomata, up to about 7 cm tall

(Landvik et al. 2003). Of these three species, only N. vitellina has been reported

from Europe (Bresadola 1882, Geitler 1958, Hansen & Knudsen 2000: 48-50,

Krieglsteiner 1993: 421, Landvik et al. 2003, Ohenoja 1975, Redhead 1989).

Work published by European researchers about Neolecta vitellina refers mainly

to phylogeny, morphology and ultrastructure (Landvik 1996, 1998; Landvik

et al. 1993, 2001, 2003). The purpose of this paper is to report the presence

of Neolecta vitellina in Romania and to provide morphological and ecological

notes on this fungus.

Materials & methods

During 2005-2010 the known habitat of Neolecta vitellina in the Nature Reserve

“Tinovul Mare” Poiana Stampei (Eastern Carpathians, Romania) was monitored for

198 ... Chinan & Hewitt

TABLE 1. List of Neolecta vitellina specimens, GenBank (ITS) accession numbers,

and sequence identity compared with the Romanian specimen.

Heras Aceon SequenidentY origin ‘Dat Calle

I 114506 FJ171855.1 N/A Romania 2 July 2005; VC Chinan

FH:DAH-7 FJ171850.1 99% Norway, 14 Aug 2002; S Landvik,

near Oslo DA Hewitt, P Inderbitzin,

S Huhtinen, inter al.

FH:DAH-18 FJ171849.1 99% USA, MA 3 Nov 2005; DA Hewitt, B Wolfe

FH:DAH-22 FJ171847.1 99% USA, MA 15 Nov 2005; WJ Neill

FH:DAH-21A FJ171852.1 99% USA, MA 17 Nov 2005; DA Hewitt

FH:DAH-21B FJ171853.1 99% USA, MA 17 Nov 2005; DA Hewitt

FH:DAH-11 FJ171851.1 99% USA, MA 6 Nov 2003; DA Hewitt, G Riner

FH:KH.04.34 FJ171848.1 100% USA, NM 30 Aug 2004; K Hansen, B Perry

OSC:119159 FJ171854.1 99% USA, OR 3 Oct 2001; M Russell

ascomata. Fresh specimens were photographed in situ prior to collection. Ascomata

were sectioned and mounted in Melzer’s reagent for observation of asci and ascospores

under the light microscope.

The ITS DNA sequence was generated from a dried Neolecta vitellina ascoma

(I 114506) that was crushed using a FASTPREP DNA extraction machine (Qbiogene,

Inc.). DNA was extracted from crushed material using a phenol-chloroform extraction

protocol. The ITS (Internal Transcribed Spacer) region was amplified by PCR using

primers ITS4 and ITS5 (http://www.biology.duke.edu/fungi/mycolab/primers.htm). We

used BlastN (Altschul et al. 1997) to query the GenBank database for similar sequences

and calculate percent identity and gaps. The obtained ITS sequence is deposited in

GenBank (accession number FJ171855.1).

Analyzed specimens are deposited in the Herbarium of Alexandru Ioan Cuza

University, Faculty of Biology, Iasi, Romania (I) and Farlow Herbarium, Harvard

University, Cambridge, USA (FH).

Taxonomy

Neolecta vitellina (Bres.) Korf & J.K. Rogers, Phytologia 21: 204 (1971). Fic. 1

= Geoglossum vitellinum Bres., Rev. Mycol. 4: 212 (1882).

ASCOMATA 20-40 mm long, irregularly clavate, lanceolate or spathulate and

consisting of a sterile zone (stipe) at the bottom and a fertile zone (hymenium)

Neolecta vitellina new to Romania... 199

FiGure 1. Neolecta vitellina.

A-ascomata in habitat; B-ascoma attached to a root of Norway spruce (Picea abies).

Scale bar = 1 cm.

on the top; fertile zone 9-18 mm long, 3-8 mm wide, yellow to bright yellowish,

smooth, sometimes longitudinally subplicate, with margin entire or slightly

irregularly lobate; sterile zone 10-22 mm long, 2-4 mm wide at the fertile zone,

pubescent or tomentose, whitish or pale yellow (Fic. 1A).

Asct 55-75 um long, 4-5.5 um wide above, 3-3.5 um wide below, cylindrical

to cylindrical-clavate, 8-spored. Paraphyses absent. Ascospores uniseriate,

unicellular, 5.5-8 x 3-4 um, reniform, ellipsoid or ovoid, hyaline, smooth.

Phialoconidia not found associated with ascospores.

Hasirat on the ground, among mosses, in coniferous forest under Norway

spruce (Picea abies (L.) H. Karst.) at an altitude of 915 m.

SEQUENCE ANALYSIS BlastN searches of GenBank with the ITS sequence

(646 nucleotides) of the Romanian N. vitellina specimen (I 114506) produced

significant alignments with ITS sequences from N. vitellina specimens collected

from Norway and USA (TABLE 1).

SPECIMENS EXAMINED: ROMANIA. EASTERN CARPATHIANS: Nature Reserve “Tinovul

Mare” Poiana Stampei, 47°18'02.69"N, 25°06'45.59"E, 915 m altitude, on ground under

Picea abies, 4 October 2004, leg. Chinan (I 114507); 2 July 2005, leg. Chinan (I 114506,

FH:DAH-31, GenBank ITS— FJ171855.1); 29 June 2006, leg. Chinan (I 114508); 30

June 2007, leg. Chinan (I 137103); 12 July 2008, leg. Chinan (I 137104); 8 July 2010, leg.

Chinan (I 137105).

Discussion

Neolecta vitellina is rarely collected in Europe, where it has a boreal-montane

distribution. The species has previously been reported in Norway, Sweden,

Finland (Hansen & Knudsen 2000), and Italy (Bresadola 1882, Geitler 1958).

In Romania, N. vitellina was found in the northern part of the country at the

periphery of “Tinovul Mare” Poiana Stampei peat bog on the ground among

200 ... Chinan & Hewitt

mosses under Norway spruce (Picea abies). Our report represents the first

record of this species for Romania and the Carpathian Mountains.

Two ascomata were first found on October 2004. In July 2005, more specimens

were in the same location with 53 ascomata inventoried in approximately a 10

m/’ area. Subsequent monitoring during 2006-10 has confirmed its presence in

the same place and shown that late June and early July seem to be the optimal

time for Neolecta vitellina to form ascomata in Romania. For comparison,

Landvik et al. (2003) reported N. vitellina from Northern Europe in August

while Redhead (1977) reports the species from North America in August-

October. Additionally, a Bresadola collection of Geoglossum vitellinum from

Southern Tyrol (Italy) in the Patouillard collection at the Farlow Herbarium is

dated ‘Aug. 1889; later than the fruiting time recorded herein for the Romanian

populations.

The macroscopic and microscopic characters of the specimens collected

from Romania are in accordance with the literature (Redhead 1977, Hansen &

Knudsen 2000, Landvik 2003) and the ITS sequence derived from one ascoma

(I 114506) matches those of other specimens identified as Neolecta vitellina.

We also confirm that Neolecta vitellina ascomata are attached to Norway

spruce roots (Fic. 1B), on which was observed a whitish sleeve consisting

of mycelium. Regarding this aspect, Redhead (1979) mentions that Neolecta

vitellina is possibly a root parasite, based on morphology and the close

association of the hyphae and the roots to which the hyphae are attached.

The presence of this species in the Nature Reserve “Tinovul Mare” Poiana

Stampei, part of the EUROPEAN ECOLOGICAL NATURA 2000 NETWORK,

emphasizes the importance of this site for macrofungi conservation.

Acknowledgements

The ITS sequence was generated in the laboratory of Joey Spatafora (OSU) with

the invaluable aid of Conrad Schoch, funded by the Assembling the Fungal Tree of

Life project, NSF grant EF-0228671, and NSF grant #0090301 Phylogeny of Kingdom

Fungi. We thank Michaela Schmull (FH) for assistance in locating references. The

authors are grateful to Donald H. Pfister (Harvard University, USA), Conrad L. Schoch

(NIH/NLM/NCBI, USA), and Shaun Pennycook (Manaaki Whenua Landcare Research,

New Zealand) for reviewing the manuscript.

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BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids

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Bresadola J. 1882. Discomycetes nonnulli Tridentini novi. Rev. Mycol. 4: 211-212.

Eriksson OE, Winka K. 1997. Supraordinal taxa of Ascomycota. Myconet 1: 1-16.

Geitler L. 1958. Konidienbildung aus Ascosporen bei Geoglossaceen. Osterreichische botanische

Zeitschrift (Pl. Syst. Evol.) 105(1-3): 159-166. http://dx.doi.org/10.1007/BF01289007

Neolecta vitellina new to Romania... 201

Hansen L, Knudsen H. 2000. Nordic Macromycetes, Vol. 1: Ascomycetes. Nordsvamp-

Copenhagen.

Krieglsteiner GJ. 1993. Verbreitungsatlas der Grofspilze Deutschlands (West), Band 2, Schlauchpilze.

Ulmer Verlag, Stuttgart.

Landvik S. 1996. Neolecta, a fruit-body-producing genus of the basal ascomycetes, as shown by SSU

and LSU rDNA sequences. Mycological Research 100(2): 199-202.

http://dx.doi.org/10.1016/S0953-7562(96)80122-5

Landvik S. 1998. Hall 6gonen 6ppna for Neolecta. Jordstjarnan 19(2): 16-19.

Landvik S, Eriksson OE, Gargas A, Gustafsson P. 1993. Relationships of the genus Neolecta

(Neolectales ordo nov., Ascomycotina) inferred from 18S rDNA sequences. Syst. Ascomycetum

11: 107-115.

Landvik S, Eriksson I, Berbee ML. 2001. Neolecta-a fungal dinosaur? Evidence from B-tubulin

amino acid sequences. Mycologia 93(6): 1151-1163. http://dx.doi.org/10.2307/3761675

Landvik S$, Schumacher TK, Eriksson OE, Moss ST. 2003. Morphology and ultrastructure of

Neolecta species. Mycological Research 107(9): 1021-1031.

http://dx.doi.org/10.1017/S0953756203008219

Ohenoja E. 1975. Leotia, Cudonia, Spathularia and Neolecta (Ascomycetes) in Finland. Annales

Botanici Fennici 100: 193-196.

Redhead SA. 1977. The genus Neolecta (Neolectaceae fam. nov., Lecanorales, Ascomycetes) in

Canada. Can. J. Bot. 55: 301-306. http://dx.doi.org/10.1139/b77-041

Redhead SA. 1979. Mycological observations: 1, on Cristulariella; 2, on Valdensinia; 3, on Neolecta.

Mycologia 71: 1248-1253. http://dx.doi.org/10.2307/3759112

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3003-3062. http://dx.doi.org/10.1139/b89-384

MY COTAXON

http://dx.doi.org/10.5248/118.203

Volume 118, pp. 203-211 October-December 2011

Epitypification, morphology, and phylogeny of Tothia fuscella

Harixia Wu’, WALTER M. JAKLITSCH’,

HERMANN VOGLMAYR’ & KEVIN D. HYDE”* **

‘International Fungal Research and Development Centre, Key Laboratory of Resource

Insect Cultivation & Utilization, State Forestry Administration, The Research Institute

of Resource Insects, Chinese Academy of Forestry, Kunming, 650224, PR China

? Department of Systematic and Evolutionary Botany, Faculty Centre of Biodiversity,

University of Vienna, Rennweg 14, A-1030 Wien, Austria

3 School of Science, Mae Fah Luang University, Tasud, Muang, Chiang Rai 57100, Thailand

4 Botany and Microbiology Department, College of Science, King Saud University,

Riyadh, 11442, Saudi Arabia

*CORRESPONDENCE TO: kdhyde3@gmail.com

ABSTRACT — The holotype of Tothia fuscella has been re-examined and is re-described and

illustrated. An identical fresh specimen from Austria is used to designate an epitype with

herbarium material and a living culture. Sequence analyses show T. fuscella to be most closely

related to Venturiaceae and not Microthyriaceae, to which it was previously referred.

Key worps — Dothideomycetes, molecular phylogeny, taxonomy

Introduction

We have been re-describing and illustrating the generic types of

Dothideomycetes (Zhang et al. 2008, 2009, Wu et al. 2010, 2011, Li et al. 2011)

and have tried where possible to obtain fresh specimens for epitypification and

use molecular analyses to provide a natural classification.

Our previous studies of genera in the Microthyriaceae, a poorly known

family within the Dothideomycetes, have resulted in several advances (Wu et

al. 2010, 2011). The Microthyriaceae are characterized by superficial, flattened,

dimidiate ascomata with cells of the upper wall that radiate and open by an

ostiole. Asci are bitunicate and fissitunicate and ascospores are hyaline to brown

(Hawksworth et al. 1995, Kirk et al. 2008).

Tothia is a poorly known monotypic genus forming thyriothecia currently

classified in the Microthyriaceae (Lumbsch & Huhndorf 2010). To reassess the

morphology of T. fuscella, we examined its holotype and re-collected the species

204 ... Wu &al.

from stalks of Teucrium chamaedrys in two different regions of Austria. Cultures

of these fresh collections were prepared to extract DNA and to sequence LSU

and ITS rDNA in order to establish the phylogenetic position of this fungus. In

this paper, we provide a detailed description and illustrations of T: fuscella and

establish its familial placement through nuLSU rDNA sequence analyses.

Materials & methods

Type material of Tothia fuscella was borrowed from URM. Ascomata were rehydrated

in 3% KOH prior to examination and sectioning. Specimens were examined under

a Leica MZ16A stereomicroscope, and fine forceps were used to remove one or two

ascomata, which were mounted in water, Melzer’s, Congo red, or cotton blue reagents.

Observations and photographs were made under a Nikon E800 or Nikon 80i light

microscope. For some hyaline structures differential interference contrast microscopy

was used. Sections were cut by hand with a sharp razor blade or into 8-1m thick slices

using a Leica CM1100 freezing microtome and then were transferred to a drop of water

or cotton blue for examination and photography. Measurement ranges cover a minimum

of 30 ascospores, 25 asci, or 10 ascomata. Microtome sections were made from one or

two ascomata, and ten measurements were made from whole ascoma and peridia.

Fresh specimens were collected in Austria and herbarium material is deposited in

IFRD and WU. Single ascospore isolates were obtained following Choi et al. (1999) and

grown on 2% malt extract agar (MEA). Cultures are deposited at the CBS.

Total DNA was extracted from liquid cultures according to Voglmayr & Jaklitsch

(2011). A ca. 1600 bp fragment containing partial SSU, ITS1, 5.88, ITS2 and partial

LSU rDNA was amplified with primers V9G (de Hoog & Gerrits van den Ende 1998)

and LR5 (Vilgalys & Hester 1990). PCR products were purified by 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 V9G, LR3 (Vilgalys & Hester 1990) and

LRS5, 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 TABLE 1.

One Tothia fuscella sequence was aligned with 12 sequences downloaded from

GenBank (TaBLE 1). Preliminary multiple alignments were generated using BioEdit

(Hall 1999) and ClustalX v. 1.83 (Thompson et al. 1997) and then checked visually and

manually optimized.

The alignment was analyzed using PAUP* 4.0b10 (Swofford 2002) and MrBayes v.

3.0b4 (Ronquist & Huelsenbeck 2003). Schismatomma decolorans (Arthoniomycetes),

which is basal to other ascomycetes in a recent study (Schoch et al. 2009), was selected

as outgroup.

A Maximum Parsimony (MP) tree was inferred with PAUP* using the heuristic search

option with 10000 random taxa addition and tree bisection and reconnection (TBR) as

the branch-swapping algorithm. All characters were unordered and of equal weight;

gaps were treated as missing data. Maxtrees were unlimited, zero-length branches were

collapsed, and all most parsimonious trees were saved. Maximum parsimony bootstrap

Tothia fuscella, epitypified ... 205

TABLE 1. Dothideomycetes and Arthoniomycetes specimens analysed

(Newly deposited sequences shown in bold).

(Vuill.) Fabric.

Saueris | CULTURE | FAMILY | GENBANK ACCESSION

| | (Schoch et al. 2009) | nuLSU rDNA

Apiosporina collinsii | CBS118973 | Venturiaceae | GU301798

(Schwein.) Hohn. |

Apiosporina morbosa | dimosp | Venturiaceae | EF114694

(Schwein.) Arx. | |

Dothidea insculpta | CBS189.58 | Dothideacae | DQ247802

Wallr. |

Dothidea sambuci | DAOM231303 | Dothideacae | AY544681

(Pers.) Fr. |

Microthyrium microscopicum | CBS115976 | Microthyriaceae | GU301846

Desm.

Myriangium duriaei | CBS260.36 | Myriangiaceae | DQ678059

Mont. & Berk.

Myriangium hispanicum | CBS247.33 | Myriangiaceae | GU301854

J.B. Martinez |

Schismatomma decolorans |

i DUKE0047570 i Roccellaceae i AY548815

(Turner & Borrer ex Sm.) |

Clauzade & Vézda | | |

Tothia fuscella | (TE)CBS130266 | - | JE927786

Tothia fuscella | (TF1)CBS130267 | - | JE927787

Trichodelitschia bisporula CBS262.69 | Phaeotrichaceae GU348996

(PB Crouan & H. Crouan) Munk |

Trichodelitschia munkii Kruys201 | Phaeotrichaceae DQ384096

N. Lundq. | |

Venturia inaequalis CBS594.70 | Venturiaceae GU301879

(Cooke) G. Winter |

Venturia populina CBS256.38 Venturiaceae | GU323212

analyses were done with the same settings, with ten rounds of random sequence addition

(Hillis & Bull 1993).

The consistency index (CI; Kluge & Farris 1969), retention index (RI; Farris 1989),

and rescaled consistency index (RC; Farris 1989) were obtained from PAUP*.

For Bayesian analyses, evolution was estimated by using MrModeltest 2.2 (Nylander

2004). Posterior probabilities (Rannala & Yang 1996, Zhaxybayeva & Gogarten 2002)

were determined by Markov Chain Monte Carlo sampling (BMCMC) in MrBayes v.

3.0b4 (Huelsenbeck & Ronquist 2001). Six simultaneous Markov chains were run for

1 million generations; trees were sampled every 100th generation, resulting in 10 001

saved trees.

206 ... Wu & al.

The first 2000 trees were discarded and the remaining 8001 trees were used for

calculating posterior probabilities in the majority rule consensus tree (Cai et al. 2006,

2008, Liu et al. 2010). The phylogenetic tree was visualized with TREE-VIEW (Page

1996).

Results

Because the ssu-1Ts-Lsu rDNA sequences obtained for the two Tothia

fuscella isolates are identical, only one (1535 bp) was included in the analyses.

Of the 735 nucleotides analyzed, 478 were constant and 182 variable characters

were parsimony informative. Maximum parsimony analysis produced a single

Schismatomma decolorans § Outgroup

Tothia fuscelia

Venturiaceae

100 F

700 Phaeotrichaceae

‘ roti

100 :

700 Dothideaceae

: Dothidea insculpta

| Miriangium duriaei

100 Myriangiaceae

10 nucleotide substitutions Myriangiwn hispanicum

Fic. 1. Phylogram showing the single MP tree of 487 steps, inferred from a heuristic parsimony

analysis of the nuLSU rDNA using PAUP*. MP bootstrap support values = 50 % and Bayesian

values = 90 % are shown above and below branches, respectively.

Tothia fuscella, epitypified ... 207

MP tree of 487 steps (Fic. 1), with CI = 0.729, RI = 0.735, RC = 0.536 and

HI = 0.271. The overall topology of trees sampled in the Bayesian analysis

was compatible with the MP tree. MP bootstrap support values and posterior

probabilities are shown in Fic. 1.

In the phylogenetic analyses (Fic. 1), Tothia fuscella was remote from the

Microthyriaceae clade but clustered near the Venturiaceae clade with 97%

bootstrap support and 1.0 posterior probability.

Taxonomy

Tothia Bat., in Toth, Annls hist.-nat. Mus. natn. hung. 52: 105 (1960).

Saprobic on stems. Ascomata, superficial, thyriothecial; in section conical,

relatively small, opening with a minute flat or slightly papillate ostiole. Asci 8-

spored, bitunicate, fissitunicate, cylindrical or obclavate. Ascospores, 1-septate,

light brown.

ANAMORPHs: None reported for the genus (Hyde et al. 2011).

Tothia fuscella (Sacc.) Bat., in Toth, Annls hist.-nat. Mus. natn. hung.

52: 106 (1960). Fic. 2

= Microthyrium fuscellum Sacc., Michelia 2(6): 57 (1880).

Saprobic on stems of Teucrium chamaedrys. Superficial mycelium absent.

Ascomata, solitary or gregarious, superficial, appearing as small black dots on

the host surface, thyriothecial; in section 130-270 um diam, 60-122 um high,

dome-shaped or flat-conical, dark brown, membranaceous, opening by a short

papillate ostiole. Peridium 7-15 um wide above, dark brown, comprising a

single upper layer of cells of textura angularis; ascoma base, comprising hyaline

ellipsoidal cells. Hamathecium of pseudoparaphyses 1.5-3 um wide, longer

than asci. Asci 57-71 x 9-13 um (mean = 64 x 10.5 um, n = 25), 8-spored,

bitunicate, fissitunicate, obclavate, with a knob-like short pedicel about 4-6 x

5-7 um, lacking an ocular chamber. Ascospores 24-28 x 4-6 um (mean = 25.7

x 4.7 um, n = 30), 2-3 seriate, fusiform or oblong-ellipsoid, light brown, one-

septate, slightly constricted at the septum, upper cell slightly wider than the

lower, with four guttules, smooth-walled.

After critical morphological comparisons showed that the fresh collections

were identical with the holotype, we designated one specimen as the epitype

for T. fuscella.

MATERIALS EXAMINED: AUSTRIA, KARNTEN, St. Margareten im Rosental, Aussicht,

grid square 9452/3, on stalks of Teucrium chamaedrys (Lamiaceae), soc. Ophiobolus

erythrosporus, 3 July 2010, W. Jaklitsch (epitype designated here, WU31396; ex-

epitype culture TF1; LSU-ITS sequence JF927787; iso-epitype IFRD8982); 12 Sep

2010, W. Jaklitsch & O. Siikésd (WU 31397). NIEDEROSTERREICH, Krems, Egelsee, on

stems of Teucrium chamaedrys, soc. Ophiobolus sp., 15 Sep 2010, H. Voglmayr (WU

31398; culture TE; ITS-LSU sequence JF927786). HUNGARY, on stems of Teucrium

chamaedrys, further data not given (holotype, URM 8210).

208 ... Wu &al.

Fic. 2. Tothia fuscella (A, D-E, H, N from epitype; B-C, F-G, I-M, O-P, from holotype).

A. Appearance of ascomata on the host surface. B, C, D. Squash mount of ascoma. E-G. Ascomata

in Section. H-J, L. Asci. K. Hamathecium. M-P. Ascospores. Scale bars: A = 500um, B = 40 um,

C-L= 10 um. M-P =5 um.

ComMMENTS—In this paper we redescribe the type of Tothia fuscella and

designate an epitype with dried material and an associated living ex-type culture.

Ascomata of T. fuscella are morphologically indistinguishable from typical

representatives of Microthyriaceae. Its ascospores resemble Microthyrium in

being one-septate, guttulate, and slightly asymmetrical but differ in their dilute

brown color. Phylogenetic analyses indicate that T: fuscella is most closely

related to the Venturiaceae, to which it should be assigned.

Tothia fuscella, epitypified ... 209

Thyriothecial ascomata are unusual for the Venturiaceae, which illustrates

that this ascoma type has evolved independently several times within

Dothideomycetes. Genera in Venturiaceae usually have yellowish, greenish

brown to brown, one-septate ascospores and obclavate asci (Barr 1968, 1989),

which agrees well with Tothia.

Acknowledgments

The Research Institute of Resource Insects, Chinese Academy of Forestry provided

financial support for the PhD study of Haixia Wu. Funds for research were provided

by the Grant for Essential Scientific Research of National Non-profit Institute (no.

CAFYBB2007002). The curators of herbarium URM are thanked for providing

material on loan for this study. The authors also thank Professor Xiaoming Chen and

Hang Chen (The Research Institute of Resource Insects, Chinese Academy of Forestry,

China), Jiankui Liu; Putarak Chomnunti (Mae Fah Luang University, Thailand, MFU)

and Cai Lei (Institute of Microbiology, Chinese Academy of Sciences, Beijing, China)

for their valuable help. Our deep thanks to Drs Eric McKenzie and Wen- Ying Zhuang

for reviewing the manuscript. K.D. Hyde acknowledges a research grant from the

Biodiversity Research and Training Program (BRT R253012) and Training Program

Grant (BRTR251181) and partial support from the Global Research Network for Fungal

Biology and King Saud University.

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MYCOTAXON

http://dx.doi.org/10.5248/118.213

Volume 118, pp. 213-218 October-December 2011

Exserohilum neoregeliae sp. nov.,

a new pathogen of Neoregelia carolinae

TAKUYA SAKODA'& TAKAO TSUKIBOSHI”

'Kobe Plant Protection Station, 1-1 Hatoba-cho, Chuo-ku, Kobe, 650-0042, Japan.

*National Institute of Livestock and Grassland Science, National Agriculture and Food Research

Organization, Senbonmatsu 768, Nasushiobara, Tochigi, Japan.

*CORRESPONDENCE TO: tuki@affrc.go.jp

Asstract —Exserohilum neoregeliae is described from Neoregelia carolinae leaves imported

from the Netherlands at plant quarantine inspection in Japan. The new fungal species is

characterized by long rostrate conidia with an obconic basal end cell with a distinct hilum.

ITS + 5.8SrDNA sequence analyses revealed that E. neoregeliae forms a monophyletic group

with E. minor.

Key worps — hyphomycetes, Bromeliaceae, leaf blight, phylogeny

Introduction

Neoregelia carolinae (Beer) L.B. Sm. (Bromeliaceae) is an ornamental plant

native to South America (Takabayashi 1994). In May 2006, brown spindle-

shaped lesions were found on N. carolinae leaves imported from the Netherlands

during plant quarantine inspection at Narita International Airport in Japan.

A new species of Exserohilum isolated from the lesions is described based on its

distinctive conidial morphology. In addition, molecular phylogenetic analyses

were carried out on the internal transcribed spacer of ribosomal DNA (rDNA-

ITS), including 5.8S rDNA.

Materials & methods

Isolates (IM201-c, -pD, -E, -F) were obtained from four lesions on the leaves of N.

carolinae and used for morphological observation, pathogenicity tests, and molecular

phylogenetic analyses. For morphological observation, the isolates were incubated

on V8 juice agar under alternating 12 h black light blue (BLB)/12 h darkness at 25°C

for about 20 days, and the resulting anamorph was observed under light microscopy.

Pathogenicity tests were conducted by placing V8 juice agar pieces, including hyphae,

on needle-wounded leaves of the original host that was grown in a greenhouse for four

214 ... Sakoda & Tsukiboshi

TABLE 1. Conidial morphology of Exserohilum neoregeliae and similar species.

SPECIES neoregeliae * longisporum ° rostratum ° longirostratum ° minor ©

SHAPE Narrowly Fusiform, Ellipsoid, Narrowing Fusiform

obclavate narrowly narrowly gradually to

or rostrate obclavate obclavate, long apical

or rostrate or rostrate beak

COoLoR Pale brown Yellowish Brown or Olivaceous Olivaceous

to brown brown olivaceous brown brown

No. OF 6-20 (6-)8-13 < 18 — 5-11

DISTOSEPTA (-26) (-18)

LENGTH 80.5-285 54-240 15-200 60-475 55-135

WIDTH 12.5-27 11.5-21.5 7-29 12-26 12.5-20

SEPTA Concolorous Concolorous Darker & End cells often Concolorous

thicker at cut off by dark,

basal septum thick septum

BASE Basal cell Basal cell Subhyaline Broadest at base —_ Base truncate

obconic; occ. often sl. obconic;

constricted at/ bulging lower half occ.

below septum constricted

*= Isolate No. IM201-p; ’ = Sun et al. (1996); © = Sivanesan(1986)

months. The aseptic V8 juice agar pieces were placed in the same way as a control. The

inoculated plants were kept in a plastic case with a cover to maintain high humidity for

2 days. Then they were kept at room temperature under natural light after uncovering.

The ITS+5.8S rDNA were amplified with ITS1/4 primers (White et al. 1990), sequenced

directly, and analyzed to reveal the phylogenetic relationship of the new species with

other Exserohilum species registered in the DNA Data Bank of Japan (DDBJ; see also

Berbee et al. 1999, Chung et al. 2005). The DNA sequences were aligned using ClustalX

v2.0.7 (Larkin et al. 2007). Phylogenies were generated using distance methods. The

distance matrix for the aligned sequences was calculated using Kimura’s two-parameter

method (Kimura 1980) and was analyzed with the neighbor-joining method (Saitou &

Nei 1987) using ClustalX v2.0.7 (Larkin et al. 2007). The sequences of isolates IM201-p

and IM201-k were registered in DDBJ as AB607956 and AB607957, respectively.

Symptoms

The brown spindle-shaped lesions were usually observed in the center of

the leaves (Fic. 1A, B) but occasionally near the leaf margin (Fic. 1C). Some

lesions elongated along the veins or turned grayish white with brown margins,

producing leaf blight. No sporulation was detected from any lesions under

natural conditions.

Taxonomy

Exserohilum neoregeliae Sakoda & Tsukib., sp. nov. Fic. 1E-G

MycoBank MB 561917

Coloniae in agaro 'V8 succus' cinereae vel brunneae, floccosae. Hyphae pallide brunneae,

ramosae, septatae, laevis. Stromata absentia. Conidiophora singularia, erecta, vel flexuosa,

Exserohilum neoregeliae sp. nov. (Japan) ... 215

Fic. 1. Exserohilum neoregeliae. A-C. Natural symptoms on Neoregelia carolinae; D. Symptoms

on N. carolinae inoculated with IM201-2; E-F Conidia of IM201-D on V8 juice agar;

G. Germinating conidium. Scale bars: E-G, 100 um.

216 ... Sakoda & Tsukiboshi

supra geniculata, laevia vel verruculosa, pallide brunnea, ad 340 um longa, 5-10 ym lata.

Conidia pallide brunnea vel brunnea, concoloria, lepto-obclavata vel rostrata, recta vel

leviter curvata, laevia, ad vel infra septum basale constricta, apex obtusus, basis obconica,

hilo protrudenti, 6-20(-26)-distoseptata, 80.5-285 x 12.5-27 um. In foliis Neoregeliae

carolinae.

Type: NIAESH 20605 (Holotype IM201-D), dry cultura isolated from affected dead

leaves, Narita, Chiba, Japan, 24 May 2006, T. Sakoda, in Herbarium of the Institute

of National Agro-environmental Science, Tsukuba, Japan. Isotype: NIAESH 20606

(IM201-k).

Colonies on V8 juice agar gray to brown, floccose. Hyphae pale brown, branched,

septate, and smooth. Stromata not formed. Conidiophore solitary, straight, or

flexuous, sometimes geniculate above, smooth to verruculose, pale brown, up

to 340 um long, 5-10 um wide. Conidia pale brown to brown, concolorous,

narrowly obclavate or rostrate, straight or slightly curved, smooth, sometimes

constricted at basal septum or below it, apex obtuse, basal cell obconic, with a

distinctly protruding hilum, 6-20(-26)-distoseptate, 80.5-285 x 12.5-27 um

(av. 187.2 x 20.6).

KEY CHARACTERS — The conidial morphology of E. neoregeliae is similar to

that of E. longisporum G.Y. Sun (Sun et al. 1997), E. rostratum (Drechsler) K.].

Leonard & Suggs, E. longirostratum (Subram.) Sivan., and E. minor Alcorn

(Sivanesan 1987) but does show differences (TABLE 1). Although concolorous

and narrowly obclavate or rostrate conidia are common to both, the conidial basal

end cell often bulges slightly in E. longisporum but is obconic in E. neoregeliae.

While the conidial basal end cells are often cut off by thick dark septa in both

E. rostratum and E. longirostratum, in E. neoregeliae all septa are concolorous.

The conidia of E. neoregeliae are broader than those of E. longisporum and

shorter than those of E. longirostratum. The conidia of E. minor are not rostrate

but fusiform, although the obconic basal end cell is especially similar to that of

E. neoregeliae.

Pathogenicity

Approximately twenty days after inoculation, symptoms similar to the

natural ones were reproduced, and inoculated fungi were readily reisolated

from the lesions (Fic. 1D), whereas controls remained symptom-free.

Phylogenetic analysis & discussion

According to the neighbor-joining tree derived from ITS+5.8S rDNA

sequences made in this study, the two E. neoregeliae isolates (AB607956 and

AB607957) form a monophyletic group (100% bootstrap value) close to

E. minor (AF071341) (Fic. 2). However, the 91.6% (489/534 bp) sequence

similarity between E. neoregeliae and E. minor indicates that those two species

clearly differ. Moreover, despite similar conidial morphologies, E. neoregeliae is

Exserohilum neoregeliae sp. nov. (Japan) ... 217

E. fusiforme (AF 163063)

100 E. pedicellatum (AF229478)

5S E. monoceras (AF071340)

E. turcicum (AF 163067)

E. longirostratum (AF 163064)

96 E. gedarefense (AF 163068)

Fe E. meginnisii (AF081453)

E. rostratum (AF071342)

100 | E. rostratum (AF 163066)

E. minor (AF071341)

86 IM201-D (AB607956)

100 | 1M201-E (AB607957)

Cochliobolus heterostrophus (AF 158107)

——} 0.02 substitutions/site

Fic. 2. A neighbor-joining tree inferred from the ITS+5.8S rDNA sequences from 10 Exserohilum

taxa. The number in front of represented isolates shows the bootstrap values in 1000 bootstrap

replicates. The rDNA-ITS accession numbers in DDBJ are shown in parentheses. The length of

branches is proportional to the number of base changes, indicated by the scale bar. Cochliobolus

heterostrophus (AF158107) is outgroup.

in a different clade from E. rostratum and E. longirostratum. Other Exserohilum

species with relatively short or non-rostrate conidia, including E. turcicum

(Pass.) K.J. Leonard & Suggs and E. pedicellatum (A.W. Henry) K.J. Leonard

& Suggs (Sivanesan 1987), were considerably distant from E. neoregeliae. Both

218 ... Sakoda & Tsukiboshi

morphological and phylogenetic evidence supports E. neoregeliae as a new

species of Exserohilum.

Acknowledgments

The authors are grateful to Dr. Meng Zhang (Henan Agricultural University) and

Dr. Wen-Hsin Chung (National Chung Hsing University) for serving as pre-submission

reviewers. We also express our deep thanks to Dr. Shaun R. Pennycook (Manaaki

Whenua Landcare Research) for nomenclatural review.

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Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG. 2007. ClustalW and

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101: 776-779. http://dx.doi.org/10.1017/S0953756297003547

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California.

MYCOTAXON

http://dx.doi.org/10.5248/118.219

Volume 118, pp. 219-230 October-December 2011

Species of Rhytismataceae on Camellia spp.

from the Chinese mainland

JIANG-LIN CHEN’ YING-REN LIN”? CHENG-LIN Hou? & SHI-JUAN WANG?

' School of Life Science, Anhui Agricultural University, &

? School of Forestry & Landscape Architecture, Anhui Agricultural University,

West Changjiang Road 130, Hefei, Anhui 230036, China

* College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

ABSTRACT—Six species in five different genera of the Rhytismataceae are reported

from Camellia spp. on the Chinese mainland. Among them Lophodermium sinense on

Camellia sinensis is described as a new species, and Terriera camelliae on C. octopetala is

a new combination, while Bifusella camelliae, Coccomyces sinensis, Hypohelion durum, and

Lophodermium jiangnanense are already known for China. This paper provides descriptions

and a key for all these species as well as illustrations for the new species. All examined

specimens are deposited in the Reference Collection of Forest Fungi of Anhui Agricultural

University, China (AAUF).

Key worps—Rhytismatales, taxonomy, plant pathogens, Theaceae

Introduction

Members of Camellia L. (Theaceae) are evergreen shrubs or trees, with a

total of 280 species distributed in bilateral zones of the Tropic of Cancer in

east Asia, including 238 species in southwestern and south China (Zhang &

Ren 1998). Camellias are economically valuable as ornamentals and sources of

beverages, tea oil, and medicine.

Of the 1072 fungal species known to inhabit Camellia worldwide, only

10 belong in the Rhytismataceae (Farr & Rossman 2011). Cryptomyces theae

Sawada, the first rhytismataceous species recorded on C. sinensis (L.) Kuntze,

was described from Taiwan by Sawada (1919) and later found on the same

host in Japan (Hara 1936). Minter (1982) reported Lophodermium camelliicola

Minter on C. sinensis from India. Kobayashi (2007) recorded L. hysterioides

220 ... Chen & al.

(Pers.) Sacc. (= L. foliicola (Fr.) RE Cannon & Minter) and two Coccomyces

species on C. japonica L. in Japan. Up to now, five species of Rhytismataceae

have been described on Camellia from mainland China (Teng 1933; Hou 2000;

Lin et al. 2001a, 2004a,b). These fungi are mostly plant pathogens leading to

different degrees of economic loss. Lophodermium jiangnanense can cause a

leaf cast of C. oleifera C. Abel (Lin et al. 2004a), while Bifusella camelliae and

Hypohelion durum on C. sinensis can cause serious branch rot (Hou 2000, Lin

et al. 2004b). The present study, based on specimens collected by the authors,

reports one hitherto undescribed species, makes one new combination,

and discusses the four other species known on Camellia from the Chinese

mainland. These records may not be complete, as more rhytismataceous species

on camellias can be expected in this region.

Materials & methods

Macroscopic appearance was described from observations made under the dissecting

microscope at 10-50 x magnification. Reference collection material was rehydrated in

water for 15 min, and 10-15 um thick sections of the fruit bodies were cut using a

freezing microtome. For the observations of outlines of ascomata and conidiomata in

vertical section, sections were mounted in lactic acid or cotton blue with pretreatment

in water. The color of the various structures and of ascospore contents was observed in

water. Gelatinous sheaths surrounding ascospores and paraphyses were examined in

water or 0.1% (w/v) cotton blue in lactic acid. Measurements were made using material

mounted in 5% KOH or Melzer’s reagent and from 30 asci, ascospores, and paraphyses

for each specimen. Line and point integrated illustrations of external shapes and internal

structures of fruit bodies were drawn using a microscopic drawing tube (Panasonic

XSJ-2, Japan).

Taxonomy

Bifusella camelliae C.L. Hou, Mycosystema 19: 7, 2000.

TyPE: CHINA, ANHUI, Yuexi, Wen/ao, alt. ca 1100 m, on Camellia sinensis, 10 May 1992,

C.L. Hou 0210 (AAUF 90085).

ILLUSTRATION: Hou 2000: 8, Fig. 1.

ZONE LINES brown to black-brown, infrequent, thin or slightly broad, sometimes

not closed.

CONIDIOMATA on twigs, scattered. In surface view conidiomata 90-190 x

100-170 um, circular to elliptical, black-brown in the centre and at the edge

of the conidioma, brown elsewhere, flattened or slightly raising the substratum

surface, opening by one apical ostiole. In vertical section subcuticular. CONIDIA

4-6 x ca 1.2 um, cylindrical, hyaline, aseptate.

ASCOMATA scattered in similar positions on the host. In surface view

ascomata 500-1260 x 230-420 um, elliptical, shinning black, edge defined,

slightly raising the substratum surface, opening by a somewhat irregular

Rhytismataceae on Camellia (China) ... 221

longitudinal split nearly extending to the edge of ascoma. Lips absent. In

median vertical section ascomata subcuticular. COVERING STROMA 20-30

um thick near the opening, gradually thinner towards the edge, connecting

to the basal stroma, composed of black-brown textura globulosa-angularis

with cells of 3-5 um diam. BASAL STROMA 5-7 um thick, composed of dark

brown, thick-walled globular and angular cells 3-5 um diam. SUBHYMENIUM

10-15 um thick, consisting of hyaline textura intricata. PARAPHYSES slightly

exceeding the asci, 1.5-2 um wide, filiform, septate, sometimes swollen to 3-5

um at the apex, covered with a thin mucous coating. AscI ripening sequentially,

70-100 x 12.5-14 um, clavate, short-stalked, nearly truncate-conical to obtuse

at the apex, J-, 8-spored. AscosporEs biseriate or multiseriate, 19-25 x 2.5-4

um, bifusiform, isthmus ca 1 um wide, hyaline, aseptate, with a 0.8-2 um thick

gelatinous sheath.

HOsT SPECIES, HABITAT, AND DISTRIBUTION: Camellia sinensis; producing

conidiomata and ascomata in lesions on decaying and dying twigs. Known only

from Anhui Province, China.

SPECIMENS EXAMINED: On Camellia sinensis: CHINA, ANHUI Mt Guniujiang, alt. ca

1600 m, 2 July 2006, Y.R. Lin, S.J. Wang 2104 (AAUF 68212); Mt Tiantangzhai, alt. ca

1200 m, 23 June 2009, J.L. Chen, X.M. Gao 5294 (AAUF 71402).

CoMMENTS—Bifusella tsugae H.S. Cao & C.L. Hou, closely related to B. camelliae,

differs in having ascospores with a wider isthmus, paraphyses not swollen at

the apex, absence of zone lines, and occurrence on needles of Tsuga chinensis

(Franch.) E. Pritz. (Cao et al. 1996, as T! tchekiangensis).

Bifusella camelliae can cause a serious branch rot of the tea plant in regions

where the specimens were collected, the damage of branch tips exceeding 85%

in the tea garden where the disease occurs. It may therefore pose a threat to the

tea-growing industry. According to our observations, this fungus infects new

twigs, gradually causing brown to grey-yellow lesions where conidiomata and

ascomata are successively produced. Between May and June of the second year,

ascomata mature to discharge ascospores, leading to the next infection.

Coccomyces sinensis Y.R. Lin & Z.Z. Li, Mycosystema 20: 3, 2001.

TyPE: CHINA, HUNAN, Changsha, Yuelushan, alt. ca 300 m, on Camellia oleifera, 24 June

1990, Y.R. Lin et al. 0521b (AAUF 66629b).

ILLUSTRATION: Lin et al. 2001a: 4, Fig. 2.

ZONE LINES grey-black, thin, entirely or partly surrounding the bleached spots.

CONIDIOMATA not observed.

Ascomata on both sides of leaves, mostly on the lower side, crowded, in

subcircular bleached spots. In surface view ascomata 600-1150 um diam.,

triangular to pentagonal, black-brown to black, raising the substratum surface

but depressed in the central region, opening by radial splits to expose a light

222 ... Chen & al.

orange-yellow hymenium. Lips absent. In median vertical section ascomata

intraepidermal. COVERING STROMA 25-32 um thick near the opening,

slightly thinner towards the edge, extending to the basal stroma, black-brown,

composed of 3-6 um diam., thick-walled angular cells. Periphysoids absent.

BASAL STROMA 12-18 um thick, composed of dark brown textura angularis

with thick-walled cells 5-8 um diam. ExcipuLum arising from the inner cells

of the covering stroma, 25-30 um wide above, consisting of several rows of

2-3 um diam., multi-septate hyphae. INTERNAL MATRIX STROMA 20-35 um

thick, gelatinized, consisting of loose textura intricata. SUBHYMENIUM 10-15

um thick, composed of textura porrecta-intricata. PARAPHYSES filiform, 2—2.5

um wide below, abruptly enlarged to 4—6.5 um and subfusoid-ventricose above,

with a subcylindrical, ca 1.5 um wide pointed apex, capped with a mass of

yellow-brown, subcylindrical gel 8-15 x 6-8 um, forming a solid refractive

epithecium 18-25 um thick. Asci ripening sequentially, 110-130 x 5.5-6.5 um,

cylindrical, short-stalked, rounded at the apex, with circumapical thickening,

J-, 8-spored. Ascosporss fasciculate, 60-95 x 1-1.2 um, filiform, hyaline,

aseptate, covered by an inconspicuous gelatinous sheath.

HOsT SPECIES, HABITAT, AND DISTRIBUTION: Camellia cf. japonica (Kirschner

et al. 2009), C. chekiangoleosa Hu, C. oleifera; producing ascomata on dead

leaves. Known from southern China and Taiwan (Kirschner et al. 2009).

SPECIMENS EXAMINED: On Camellia chekiangoleosa: CHINA, ZHEJIANG, Hangzhou,

Liuxia, alt. ca 60 m, 6 August 1990, Y.R. Lin 0727 (AAUF 66835).

On C. oleifera: CHINA, ANHUI, Huangshan Arboretum, alt. ca 550 m, 5 Sep. 2009,

J.L. Chen, S.J. Wang 5317 (AAUF 71425); Fuy1an, Fuzhou Arboretum, alt. ca 320 m,

2 July 1990, Y.R. Lin 0644 (AAUF 66752); GUANGDONG, Guangzhou, South China

Botanical Garden, alt. ca 382 m, 22 June 1990, Y.R. Lin 0515 (AAUF 66623); JIANGXI,

Nanchang People Park, alt. ca 25 m, 28 June 1990, Y.R. Lin 0587 (AAUF 66695).

COMMENTS—Coccomyces sinensis is common on camellias in southern

China and was also found recently on fallen leaves of Camellia cf. japonica

in Taiwan (Kirschner et al. 2009). It is very similar in the shape of its

paraphyses (subfusoid-ventricose and pointed apices) to C. mucronatus Korf &

W.Y. Zhuang on Fagaceae, which differs in polygonal to subcircular ascomata,

much narrower asci (4.3-5 um) and ascospores (0.6-0.8 wm), dense

periphysoids, and paraphyses lacking refractive solid gel at the apex (Korf &

Zhuang 1985). Coccomyces urceoloides Spooner, which also somewhat resembles

C. sinensis in paraphyses capped with a mass of subhyaline or yellowish solid

gel and measuring 6-12 x 5-8 um, differs in other aspects (Spooner 1990).

Hypohelion durum Y.R. Lin, C.L. Hou & S.J. Wang, Mycosystema 23: 169, 2004.

TyPE: CHINA, ANHUI, Yuexi, Wen/ao, alt. ca 1100 m, on Camellia sinensis, 27 May 1994,

C.L. Hou 0014 (AAUF 66122).

ILLUSTRATION: Lin et al. 2004b: 170, Fig. 1.

Rhytismataceae on Camellia (China) ... 223

ZONE LINES brown to dark brown, infrequent, narrow to broad, somewhat

diffused.

CONIDIOMATA on twigs, usually crowded. In surface view conidiomata

90-230 um diam., subcircular, black-brown in the centre and at the perimeter

line, brown elsewhere, somewhat raising the substratum surface. In vertical

section subcuticular. UPPER WALL 3-5.5 um thick, dark brown, consisting of

small angular cells. BASAL WALL 5-7 um thick, black-brown. CONIDIOGENOUS

CELLS 7.5-12 x 2—2.5 um, flask-shape. CONIDIA 4.5—-6.5 x 0.8-1 um, cylindrical,

hyaline, aseptate.

Ascomarta in similar positions to conidiomata on the substratum, scattered

to crowded in yellow-brown to grayish-white lesions. In surface view ascomata

450-950 x 260-400 um, elliptical or broad-elliptical, black, with a clear outline,

slightly raising the substratum surface, opening by a longitudinal split. Lips

absent. In median vertical section ascomata subcuticular. COVERING STROMA

20-28 um thick near the opening, slightly thinner towards the edge and

extending to the basal stroma, composed of textura angularis-epidermoidea

with black-brown or brown, thick-walled cells 4-6 um diam. BASAL STROMA

12-18 um thick, composed of 4.5-7 um diam., black-brown, thick-walled

angular and elongate cells. The triangular space between the covering and

basal stroma is filled with grey-brown rather thin-walled, large angular cells.

SUBHYMENIUM 18-25 um thick, consisting of textura porrecta. PARAPHYSES

110-135 x 1.5-2 um, filiform, swollen to 2.5-3.5 um above, surrounded by a

ca 1.5 um thick gelatinous matrix. Asci ripening sequentially, 75-115 x 13-16

um, clavate, somewhat long-stalked, apex rounded to subtruncate, J-, 8-spored.

AscosporEs more or less biseriate, 18-25 x 3-4 um, cylindrical to subclavate,

hyaline, aseptate, with a conspicuous gelatinous sheath 4-6 um thick.

HOST SPECIES, HABITAT, AND DISTRIBUTION: Camellia sinensis; producing

conidiomata and ascomata in lesions on decaying and dying twigs. Common

in southern China.

SPECIMENS EXAMINED: On Camellia sinensis: CHINA, ANHUI, Huangshan Arboretum,

alt. ca 550 m, 20 Oct. 2001, Y.R. Lin et al. 1785 (AAUF 67893); Huangshan Arboretum,

alt. ca 550m, 5 Sep. 2009, J.L. Chen, S.J. Wang 5322 (AAUF 71430); JIANGSU, Nanjing,

alt. ca 90 m, 10 April 1973, DJ. Lu 1308 (AAUF 67416).

CoMMENTS— Hypohelion durum is rather distinctive. It is consistent with

typical features of the genus except for the presence of basal stroma and yet is

far from genera such as Hypoderma De Not. Hypohelion scirpinum (DC.) P.R.

Johnst. is similar but differs by its much larger ascomata (800-3000(-—7000) x

500-800 um) and ascospores with a median septum (40-75 x 4.5-6.5 um), the

absence of dark basal stroma, and its occurrence in marshes on Scirpus spp.

(Johnston 1990). A strong parasite that is common on tea plants in southern

China, H. durum can cause different degrees of branch rot (Lin et al. 2004b).

224 ... Chen & al.

Lophodermium jiangnanense Y.R. Lin & S.J. Wang, Mycosystema, 23: 15, 2004.

Type: CHINA, HUNAN, Changsha, Yuelushan, alt. ca 300 m, on C. oleifera, 24 June 1990,

Y.R. Lin 0521a (AAUF 66629a).

ILLUSTRATION: Lin et al. 2004a: 16, Fig. 2.

ZONE LINES absent.

ConripDIOMATA mostly hypophyllous, usually crowded. In surface view

conidiomata 130-220 um diam., subcircular, black-brown in the centre and at

the perimeter line, brown elsewhere. In vertical section subepidermal. CONIDIA

not observed.

Ascomarta on both sides of leaves, predominantly on the lower side, crowded

in grey-yellow, circular to irregular, bleached spots. In surface view ascomata

600-990 x 280-360 um, elliptical or occasionally 3-lobed, black, slightly shiny,

edge defined, raising the substratum surface, opening by a longitudinal split

which is sometimes branched. Lips absent. In median vertical section ascomata

subepidermal. COVERING STROMA 18-30 um thick, extending to the basal

stroma, composed of grey-brown, thick-walled, angular and elongate cells

3-3.5 um diam., markedly black and brittle near the opening, with several rows

of almost colorless, septate, cylindrical thin-walled cells occurring on the inner

side of the upper margin of the covering stroma. BASAL STROMA 15-28 um

thick, composed of 4—7 um diam., black-brown, thick-walled angular cells. The

triangular space between the covering and basal stroma is filled with a hyaline,

gelatinized reticulate tissue. SUBHYMENIUM 15-20 um thick, consisting of

textura porrecta. PARAPHYSES ca 1.5 um wide, filiform, septate, often gradually

swollen to 2—2.5 um, twisted and intertwined above to form a yellow epithecium

10-15 um thick. Asci ripening sequentially, 95-130 x 6-7.5 um, cylindrical,

rounded at the apex, J-, 8-spored. Ascosporss fasciculate, 60-90 x 1.4-1.6

um, filiform, hyaline, aseptate, with a 0.8-1 um thick gelatinous sheath.

HOsT SPECIES, HABITAT, AND DISTRIBUTION: Camellia oleifera; forming

conidiomata and ascomata on dead part of living leaves or on fallen leaves.

Known from Hunan, Anhui, and Guangdong Provinces, China.

SPECIMENS EXAMINED: On Camellia oleifera: CHINA, ANHUI Guichi, Forest Nursery of

Guichi, alt. ca 600 m, 11 Oct. 1992, Q.S. Liu, Q. Cao 1640 (AAUF 67748); Huangshan,

Renzipu, alt. ca 660 m, 6 Sept. 2009, J.L. Chen, Y.R. Lin 5345 (AAUF 71453);

GUANGDONG, Guangzhou, South China Botanical Garden, alt. ca 382 m, 22 June 1990,

Y.R. Lin 0515b (AAUF 66623b).

CoMMENTS—Lophodermium jiangnanense resembles L. intricatum Spooner,

which differs in its association with zone lines, much larger ascomata

(1000-1200 x ca 500 um) and asci (135-152 x 7-7.5um) with truncate-

conical apices, longer ascospores with a median septum, and much-branched

paraphyses (Spooner 1991). This fungus usually produces fruit bodies on

fallen C. oleifera leaves, but the collection from Anhui Province (AAUF 67748)

shows that it primarily infects living leaves and develops fruit bodies in yellow-

Rhytismataceae on Camellia (China) ... 225

brown to yellowish-white spots near the margin of leaves on which an obvious

brown ridged zone between the healthy and sick areas has formed. It is often

accompanied by Coccomyces sinensis, sometimes growing on the same leaves

(Lin et al. 2004a).

Lophodermium sinense Y.R. Lin, C.L. Hou & Jiang L. Chen, sp. nov. Fics. 1-7

MycoBANnk 561775

Ascomata amphigena, dispersa vel aggregata, (290—)400-730 x (150-)220-290 um,

elliptica, interdum triloba, atra, labiis carentia, ab rima longitudinali vel lobis ternis

aperientia, partim subcuticularia et partim intraepidermalia, cellulis epidermalibus

plus quam 4 successive in stromate basali ordinatis. Paraphyses filiformes, prope apicem

plerumque gradatim tumidae et semel aut iterum ramosae, necnon epithecium formantes.

Asci in successione maturescentes, 80-120 x 6-7 um, cylindrici, brevi-stipitati, J-, 8-spori.

Ascosporae 50-95 x 1.2-1.5 ym, filiformes, hyalinae, aseptatae, vagina gelatinosa 1-1.5

um crassa indutae.

Type: China, Anhui, Mt Guniujiang, alt. ca 1600 m, on leaves of Camellia sinensis, 10

July 2006, Y.R. Lin, S.J. Wang 2103 (Holotype AAUF 68211).

ETyMo_ocy: referring to the country where the specimen was collected.

ZONE LINES absent.

ConipiomaTA on both sides of leaves, scattered to crowded, occasionally

merging into one another. In surface view conidiomata 80-160 um diam.,

circular or subrounded, black-brown in the centre, more or less concolorous

with the substratum surface elsewhere, slightly raising the leaf surface,

discharging spores through a 10-15 um diam. apical ostiole. In vertical section

subcuticular. UPPER WALL only present around the ostiole. BASAL WALL absent.

SUBCONIDIOGENOUS LAYER ca 7 um thick, composed of light thin-walled

angular cells. CONIDIOGENOUS CELLS 6-9 x 2-3 um, cylindrical, slightly

tapering towards the apex, hyaline, proliferating sympodially. Conrp1a 4-6 x 1

uum, cylindrical, hyaline, aseptate.

Ascomarta in similar positions to conidiomata on the substratum, scattered

or crowded in subcircular to irregular, yellow-brown bleached spots 5-15

mm diam. In surface view ascomata (290-)400-730 x (150—)220-290 um,

elliptical, sometimes curved or 3-lobed, with a clear outline, ends rounded or

obtuse, matt black except for a grey-brown area at each end, strongly raising the

substratum surface, opening by a longitudinal split more than 4/5 the length

of the ascoma, which is sometimes branched, to expose a light yellow-brown

hymenium. Lips absent. In median vertical section ascomata subcuticular near

the opening and intraepidermal in the lower part of the covering stroma, more

than four epidermal cells being displaced and lying always successively on the

basal stroma. COVERING STROMA 12-20 um thick, black-brown, connecting

to the basal stroma, composed of textura angularis-globulosa with thick-

walled cells 2-5 um diam., markedly black and brittle near the opening, with

226 ... Chen & al.

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Fics. 1-7. Lophodermium sinense on Camellia sinensis. 1. A leaf bearing fruit bodies. 2. Ascomata

and conidiomata observed under a dissecting microscope. 3. Ascoma in median vertical section.

4, Detail of ascoma in median vertical section. 5. Paraphyses, asci and ascospores. 6. Conidioma in

vertical section. 7. Conidiogenous cells and conidia.

Rhytismataceae on Camellia (China) ... 227

4-5 rows of light thin-walled cells occurring on the inner side of the upper

margin of the covering stroma. BASAL STROMA slightly concave or nearly flat,

10-18 um thick, black-brown, composed of 2-3 layers of thick-walled angular

cells 2.5-5.5 um diam. The triangular space between the covering and basal

stroma filled is with 5-8 um diam., light grey-brown, slightly thick-walled

angular cells. SUBHYMENIUM 10-18 um thick, consisting of textura angularis-

porrecta. PARAPHYSES 1-1.5 um wide, filiform, hyaline, septate, covered in a

thin mucous coating 1.5-2 um thick, often gradually swollen to 2-3 um and

1-2 times branched near the apex, contorted and intertwined to above form

a conspicuous epithecium 12-24 um thick. Ascr ripening sequentially, 80-20

x 6-7 um, cylindrical, short-stalked, apex rounded, thin-walled, without

circumapical thickening, J-, 8-spored. Ascosporgs arranged fasciculately or

somewhat helically, 55-95 x 1.2-1.5 um, filiform, slightly tapering towards the

base, hyaline, aseptate, with a 1-1.5 um thick gelatinous sheath.

HOST SPECIES, HABITAT, AND DISTRIBUTION: Camellia sinensis; ascomata

develop on living leaves but are seen in a mature condition only on fallen leaves.

Known only from the type locality, Anhui, China.

ComMMENTS—Lophodermium sinense is very similar to L. jiangnanense on

the same plant genus, but the latter has larger subepidermal ascomata and

conidiomata, unbranched paraphyses, a covering stroma comprising aliform

and elongate cells, and a well developed basal stroma with hyaline reticulate

tissue in the corner between the covering and basal stroma (Lin et al. 2004a).

Lophodermium intricatum is also similar to the new species but differs in its

much larger ascomata, asci, and ascospores, asci with truncate-conical apices,

ascospores with a median septum and without a gelatinous sheath, and the

presence of zone lines (Spooner 1991). Lophodermium implicatum Y.R. Lin

& Z.S. Xu differs in possessing intraepidermal ascomata associated with dark

brown zone lines, shorter ascospores, and intraepidermal conidiomata with

trichogynes (Lin et al. 2001b).

Lophodermium sinense is a pathogen causing leaf spot. The fungus at first

probably develops within the living leaves but visible symptoms do not show,

after which the anamorphic and teleomorphic fruit bodies sequentially appear

on brown lesions of infected leaves as the vitality of the host declines. Diseased

leaves become reddish-brown and fall between July and August. Mature

ascomata have been observed only on fallen leaves.

Terriera camelliae (Teng) Y.R. Lin & Jiang L. Chen, comb. nov.

MycoBAnk 561774

= Lophodermium camelliae Teng, Sinensia 4: 138, 1933

= Clithris camelliae (Teng) Tehon, Mycologia 31: 675, 1939

= Colpoma camelliae (Teng) Teng, Fungi of China: 759, 1963

228 ... Chen & al.

Type: CHINA, FUJIAN, Fuzhou, on fallen leaves of Camellia sp., Teng 1904 (No.77 in the

Metropolitan Museum, Academia Sinica, Nanking, China).

ILLUSTRATION: Tehon 1939: 682, Fig. 6.

ZONE LINES grey-brown, infrequent, thin, entirely or partly surrounding the

bleached spots.

CONIDIOMATA amphigenous, crowded. In surface view conidiomata 70-130

um diam., subcircular to irregular, flattened, black-brown in the centre and at

the perimeter line of the conidioma, grey-brown elsewhere. In vertical section

intraepidermal to subepidermal. Upper wall only present around the ostiole.

BASAL WALL 8-12 um thick, composed of brown to dark brown tissue with no

obvious structure. CONIDIA 2.5-4 x ca 0.8 um, cylindrical, hyaline, aseptate.

Ascomarta in similar positions to conidiomata on the substratum, scattered

to crowded in grey-yellow, subcircular to irregular bleached spots. In surface

view ascomata 440-870(-1020) x 290-490 um, elliptical or occasionally 3-

lobed, black-brown to black, slightly shiny, edge defined, moderately raising

the substratum surface, opening by a longitudinal split about 4/5-7/8 the length

of the ascoma, which is sometimes branched. Lips absent. In median vertical

section ascomata subepidermal. COVERING STROMA 15-22 um thick near the

opening, slightly thinner towards the edge, extending to the basal stroma,

composed of black-brown, thick-walled angular cells 3-5 um diam. Along the

edge of the ascomatal opening is a 10-15 um thick, flattened extension to the

covering stroma which covers the hymenium, and which comprises markedly

black and brittle carbonized tissue with no obvious cellular structure. BASAL

STROMA composed of 1-2 layers of black-brown, thick-walled, angular to

globular cells. SUBHYMENIUM 12-18 um thick, consisting of textura porrecta.

PARAPHYSES extending 15-30 um beyond the asci, ca 2 um wide, filiform,

sometimes gradually swollen to 2.5-3 um or branched above, septate. Asc

ripening sequentially, 85-120 x 5.5-6.5 um, cylindrical, short-stalked, rounded

at the apex, J-, 8-spored. Ascospores arranged fasciculately or somewhat

helically, 52-80 x 1-1.2 um, filiform, hyaline, aseptate, covered by a ca 0.5 um

thick gelatinous sheath.

HOsT SPECIES, HABITAT, AND DISTRIBUTION: Camellia octopetala Hu,

Camellia sp. (Teng 1933); conidiomata and ascomata were found on fallen

leaves. Known only from Fujian Province, China.

SPECIMENS EXAMINED: On Camellia octopetala: CHINA, Fujian, Fuzhou Arboretum,

alt. ca 320 m, 2 July 1990, Y.R. Lin 0643 (AAUF 66751); 11 June 2009, J.L. Chen, L. Chen

5108 (AAUF 71216).

CoMMENTS—Taxonomic placement of Terrea camelliae has long been

controversial. Teng (1933) originally validly published the fungus as L.

camelliae. Tehon (1939), who examined part of the type (labeled “co-type”

by Teng), felt that it lacked structures characteristic of the Hypodermataceae

Rhytismataceae on Camellia (China) ... 229

(= Rhytismataceae) and transferred it to Clithris (Fr.) Bonord. (= Cenangium

Fr.) in the Helotiaceae Rehm (Kirk et al. 2008). Teng’s later (1963) transfer of

the taxon to Colpoma Wallr. cannot be accepted because Colpoma ascomata

develop on bark (not leaves) and have a well-developed basal stroma.

Eriksson (1970) erected the new genus Terriera B. Erikss. based on the type

species T. cladophila (Lév.) B. Erikss. on Vaccinium myrtillus L. Johnston (2001)

and Ortiz-Garcia et al. (2003) transferred several species to Terriera, including

Clithris arundinacea Penz. & Sacc., C. minor Tehon, Lophodermium fuegianum

Speg., L. javanicum Penz. & Sacc., and L. sacchari Lyon. Terriera differs from

Lophodermium Chevall. by the markedly black and brittle extensions of the

covering stroma and by the thin-walled, prismatic or angular cells in the corner

between the covering and basal stroma. The present species on Camellia sp. and

C. octopetala fits closely the characters of Terriera based on the illustration by

Tehon (1939) and examinations carried out by the authors. Consequently, we

redispose it as T. camelliae.

There are some signs that T: camelliae may be a pathogen associated with

leaf cast of eight-petaled Camellia species.

Key to species of Rhytismataceae on Camellia from the Chinese mainland

LAs PASCOMIATA ONG NIG G4, 44, tale hen naire wea hey Wey ng weet gag nae Bey ngt tiny as bg cake yng gts a 2

LbeAScomate Ghuleaves> Buck. takin wees, eae My ott hohe AN dhs We ih8 Bk Seen Oe Re eens 3

2d ASCOSWORE SIU SH OLE 2% FUN. te ALONG #1 1ane RMS AA Ne A ht Bifusella camelliae

2b. Ascospores cylindrical, elliptical or subclavate .............. Hypohelion durum

3a. Ascomata triangular to pentagonal, opening by radial splits .. Coccomyces sinensis

3b. Ascomata elliptical, opening by a longitudinal split .....................0..0.. 4

4a. Ascomata partly subcuticular and partly intraepidermal § Lophodermium sinense

AD ASCOMALa SHDEPIGCEMIGL: 8.08 wie. 5 UE O2F mE Nt OE tet OL A BO! Ut U8 5

5a. Covering stroma with a markedly black brittle flattened extension covering the

IVT CTUTUTN GS bse as Pe Ss otek = ean 5s leans settee sian le spchea'le We + Terriera camelliae

5b. Covering stroma without such an extension ....... Lophodermium jiangnanense

Acknowledgements

The authors are grateful for the pre-submission comments and suggestions provided

by Dr D.W. Minter and Prof. T.Y. Zhang and for the specimens collected by Dr L. Chen

and Dr X.M. Gao. This study was supported by the National Natural Science Foundation

of China (No. 30870014 and 30770006), the Specialized Research Fund for the Doctoral

Program of Higher Education of China (No. 20070364002), and the Natural Science

Foundation of Anhui Province of China (No. 070411005).

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MYCOTAXON

http://dx.doi.org/10.5248/118.231

Volume 118, pp. 231-235 October-December 2011

A new species of Coccomyces (Rhytismatales, Ascomycota)

from Mt Huangshan, China

Guo-JUN JrA', YING-REN LIN? & CHENG-LIN Hov?

' School of Life Science & ? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

* College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

ABSTRACT—A fungus found on leaves of Osmanthus fragrans from Mt Huangshan in

Anhui Province, China, is described as Coccomyces minimus. The new species is similar

to C. cyclobalanopsis but differs in the extremely small, subepidermal ascomata and in the

presence of conidiomata. The type specimen is deposited in the Forest Fungi Dried Reference

Collection of Anhui Agricultural University, China (AAUF). Both illustration and comments

accompany the description.

KEY woRDs—taxonomy, Rhytismataceae, Oleaceae

Introduction

Coccomyces De Not. is the second-largest genus in the Rhytismataceae

(Rhytismatales, Leotiomycetes, Ascomycota) (Kirk et al. 2008). Members of this

genus are characterized by polygonal to more or less circular ascomata opening

by several radiate or irregular splits, cylindrical to clavate asci, and filiform to

fusiform ascospores, often with gelatinous sheaths (Sherwood 1980; Cannon &

Minter 1986; Johnston 1986, 2000; Spooner 1990; Lin et al. 1994).

Of the 116 Coccomyces species known worldwide (Kirk et al. 2008), 23 have

been reported from China (Korf & Zhuang 1985; Lin 1998; Hou et al. 2006,

2007). They are widely distributed and inhabit leaves, twigs, bark, or wood

of vascular plants, especially Ericaceae, Fagaceae, and Lauraceae (Sherwood

1980). Some species, such as C. guizhouensis Y.R. Lin & B.E. Hu, C. ledi Rehm,

C. leptideus (Fr.) B. Erikss., C. strobi J. Reid & Cain, and C. vilis Syd. et al.,

may cause plant disease of economic significance (Lin et al. 1994, Rehm 1913,

Sherwood 1980, Reid & Cain 1961, Cannon & Minter 1984).

232 ... Jia, Lin & Hou

In the present paper, C. minimus on leaves of Osmanthus fragrans (Thunb.)

Lour. (Oleaceae) from Mt Huangshan in Anhui Province, China, is described

as a new species.

Materials & methods

Pieces of plant material bearing fruit bodies containing asci and ascospores were

selected. Sections of the fruitbodies 10-15 um thick were cut using a freezing microtome

and mounted in water, Melzer’s reagent, 5% KOH, or 0.1% (w/v) cotton blue in lactic

acid for microscopical observation. Gelatinous sheaths surrounding ascospores and

paraphyses were observed in water or cotton blue in lactic acid. Measurements were

made using material mounted in 5% KOH or Melzer’s reagent from more than 20

ascospores, asci and paraphyses per ascoma. Point and line integrated illustrations of

external shapes and internal structures of fruit bodies were prepared using a microscope

drawing device.

Taxonomy

Coccomyces minimus Y.R. Lin, C.L. Hou & GJ. Jia, sp. nov. FIGs. 1-7

MycoBAank 561773

Ascomata amphigena, 300-690 um, trigona usque ad hexagona, per lacinias 3-6

aperientia, subepidermalia. Cellulae labiorum et periphysoidei non visae. Excipulum ex

hyphis septatis aliquantum parallelis constatum, apice 20-25 um crassum. Paraphyses

85-95 x 1.0-1.5 um, filiformes, ad apicem gradatim ad 2-3 ym incrassatae. Asci in

successione maturescentes, 75-88 x 4-5 um, clavati-cylindrici, brevi-stipitati, J-, 8-spori.

Ascosporae 50-72 x 0.8-1.0 um, filiformes, hyalinae, aseptatae, vagina gelatinosa ca 0.8

um crassa indutae.

Type: China, Anhui, Mt Huangshan, Ciguangge, alt. 730 m, on Osmanthus fragrans

leaves, 11 Sept. 2007, Y.R. Lin et al. 2227a (Holotype AAUF 68335a).

ETYMOLOGY: minimus (Latin = very little, smallest), referring to the size of the

ascomata.

ZONE LINES frequent, grey-black or black, thin, entirely or partly surrounding

the bleached spots.

CoNnIDIOMATA developing on both sides of leaves, mostly on the lower

side of the leaf, scattered to crowded. In surface view, conidiomata 100-220

uum diam., rotund or slightly irregular, black-brown in the centre and the

perimeter line of the conidioma, grey-brown elsewhere, somewhat raising the

leaf surface, discharging spores through an apical ostiole. In vertical section,

conidiomata subepidermal, more or less double lens-shaped. UPPER WALL

poorly developed, dark brown, with indefinite structure. BASAL WALL 8-14

um thick, black-brown, strongly black and brittle, consisting of 2-3(—4) layers

of thick-walled angular cells 3-6 um diam. SUBCONIDIOGENOUS LAYER 5-7

um thick, composed of light brown, thin-walled angular cells. TRICHOGYNES

28-36 x 1.5-2 um, subfiliform, septate. CONIDIOGENOUS CELLS 6.5-12 x 2-3

233

Coccomyces minimus sp. nov. (China) ...

7. Coccomyces minimus on Osmanthus fragrans. 1. A leaf bearing fruit bodies. 2. Ascomata

and conidiomata observed under a dissecting microscope. 3. Ascoma in median vertical section.

Fics. 1

4. Detail of ascoma in median vertical section. 5. Paraphyses and asci. 6 Discharged ascospores.

7. Conidioma in vertical section.

234 ... Jia, Lin & Hou

um, flask-shape, holoblastic sympodially proliferating. Conip1a 3-5 x ca 1 um,

cylindrical, hyaline, aseptate.

Ascomara in similar positions on the host, scattered to crowded in yellow-

brown or grey-yellow, irregular bleached spots 5-12 mm diam. In surface view,

ascomata black-brown to black, slightly shiny, triangular to hexagonal, 300-690

um diam., strongly raising the substratum surface but depressed in the central

area, with an obvious performed dehiscence mechanism, opening by 3-6 radial

splits, which nearly extend to the edge of ascoma, to expose the waxy-yellow

hymenium. Lips not seen. In median vertical section, ascomata subepidermal,

150-190 um deep. COVERING STROMA 16-20 um thick near the opening,

slightly thinner towards the edge, connecting to the basal stroma, consisting of

textura angularis-globulosa with dark brown or black-brown, thick-walled cells

3-5 um diam. Periphysoids not seen. BASAL STROMA 12-22 um thick, strongly

black and brittle, composed of textura angularis with 2—3(-4) layers of black-

brown, thick-walled cells 4-6 um diam. INTERNAL MATRIX OF STROMA well

developed, 12-25 um thick, consisting of hyaline, gelatinised textura intricata.

EXCIPULUM arising from the marginal paraphyses, 20-25 um wide above,

becoming thinner towards the base, comprised of rows of hyaline, multi-

septate hyphae. SUBHYMENIUM 10-15 um thick, comprised of hyaline textura

angularis-porrecta. PARAPHYSES 85-95 x 1.0-1.5 um, filiform, not branched,

often gradually swollen to 2-3 um above, covered with aca 1 um thick gelatinous

matrix but not forming an epithecium. ASscI ripening sequentially, 75-88 x 4-5

um, cylindrical-clavate, short-stalked, thin-walled, apex round, J-, 8-spored.

Ascosporss fasciculate, 50-72 x 0.8—1.0 um, hyaline, filiform, aseptate, slightly

tapered towards the base, covered by a ca 0.8 um thick gelatinous sheath.

HOST SPECIES, HABITAT AND DISTRIBUTION: Osmanthus fragrans; producing

conidiomata and ascomata on fallen leaves. Known only from the type locality,

Anhui, China.

COMMENTS—Coccomyces minimus resembles C. cyclobalanopsis Y.R. Lin &

Z.Z. Liin the shape of paraphyses, asci, and ascospores as well as in the color of

the exposed hymenium. However, C. cyclobalanopsis has larger, intraepidermal

ascomata (400-900 um diam.) without conidiomata, a thinner basal stroma

(7-10 um thick), a well-developed subhymenium (20-28 um thick), much

longer paraphyses (110-130 um) and asci (90-115 um), and an upper part of

the excipulum 30-50 um thick arising from inner cells of the covering stroma

(Lin et al. 2000).

Coccomyces multangularis Y.R. Lin & Z.Z. Li is easily distinguished from

C. minimus by its 3-9-sided much larger (500-1100 um diam.) ascomata, the

35-50 um thick upper portion of the excipulum, infrequent periphysoids, and

slightly longer asci (85-105 um) (Lin et al. 2001).

Coccomyces minimus sp. nov. (China) ... 235

The similar C. concolor Sherwood differs in its orbicular ascomata lacking a

performed dehiscence mechanism, the covering stroma comprising 1-2 layers

of loosely interwoven brown hyphae 2-3 um diam. that are not black and

brittle, the thicker (40-50 um) excipulum, the swollen (5-6 um) paraphyses

apices, and the much longer asci (80-120 um) and ascospores (70-100 um)

(Sherwood 1980).

Acknowledgements

Weare grateful to Dr D.W. Minter and Prof. T.Y. Zhang for serving as pre-submission

reviewers, and to Dr L.H. Wang for the identification of the associated plant. The study

was supported by the National Natural Science Foundation of China (No. 30870014

and 30770006), and the Specialized Research Fund for the Doctoral Program of Higher

Education of China (No. 20070364002).

Literature cited

Cannon PF, Minter DW. 1984. Coccomyces vilis. CMI Descr. Path. Fungi & Bact. no. 792.

Cannon PF, Minter DW. 1986. The Rhytismataceae of Indian subcontinent. Mycol. Pap. 155:

1-123.

Hou CL, Piepenbring M. 2007. Two new species of Rhytismataceae on twigs of conifers from

Yunnan Province, China. Mycotaxon 102: 165-170.

Hou CL, Kirschner R, Chen CJ. 2006. A new species and new records of Rhytismatales from Taiwan.

Mycotaxon 95: 71-79.

Johnston PR. 1986. Rhytismataceae in New Zealand 1. Some foliicolous species of Coccomyces de

Notaris and Propolis (Fries) Corda. New Zealand J. Bot. 24: 89-124.

Johnston PR. 2000. Rhytismatales of Australia: the genus Coccomyces. Aust. Syst. Bot. 13: 199-243.

http://dx.doi.org/10.1071/SB98034

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

10" ed. CAB International. Wallingford. 771 p.

Korf RP, Zhuang WY. 1985. Some new species and new records of Discomycetes in China.

Mycotaxon 22: 483-514.

Lin YR. 1998. Studies on Coccomyces de Notaris and Neococcomyces gen. nov. in China. [Ph.D.

thesis; in Chinese]. Northeast Forestry University. Harbin. 98 p.

Lin YR, Liu HY, Tang YP, Hu BE 1994. Two new species of the genus Coccomyces from China [in

Chinese]. Acta Mycol. Sin. 13: 8-12.

Lin YR, Li ZZ, Huang CC, Xiang CT. 2000. Studies on the genus Coccomyces from China II [in

Chinese]. Mycosystema 19: 297-301.

Lin YR, Li ZZ, Xu ZS, Wang JR, Yu SM. 2001. Studies on the genus Coccomyces from China IV [in

Chinese]. Mycosystema 20: 1-7.

Sherwood MA. 1980. Taxonomic studies in the Phacidiales: The genus Coccomyces (Rhytismataceae).

Occ. Pap. Farlow Herb. Crypt. Bot. 15: 1-120.

Spooner BM. 1990. Coccomyces and Propolis (Rhytismatales) from Mt Kinabalu, Borneo. Kew

Bulletin 45: 451-484. http://dx.doi.org/10.2307/4110513

Rehm H. 1913. Ascomycetes novi VI. Ann. Mycol. 11: 150-155.

Reid J, Cain RE. 1961. The genus Therrya. Can J.Bot. 39: 1119-1129. http://dx.doi.org/10.1139/b61-098

MYCOTAXON

http://dx.doi.org/10.5248/118.237

Volume 118, pp. 237-244 October-December 2011

New records of poaceous rusts from Pakistan

A. IsHAQ’, N.S. AFSHAN”* & A.N. KHALID’

"Department of Botany &*Centre for Undergraduate Studies,

University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

*CORRESPONDENCE TO: pakrust@gmail.com

ABSTRACT — Puccinia stipae var. stipae and Uromyces sporobolicola are described and

illustrated as new records for Pakistan. Calamagrostis pseudophragmites for Puccinia sessilis,

Agrostis gigantea for P. recondita, and Dactylis glomerata for P. striiformis are new poaceous

hosts for these rust fungi from Pakistan.

KEY worps — graminicolous, Uredinales, weeds

Introduction

Poaceous weeds are an important competitor of all crop plants worldwide

as well as in Pakistan. Economically important grass species are threatened by

invasive weeds, and several host-specific rust fungi have been used as biological

control agents against certain noxious weeds (Yandoc-Ables et al. 2006). There

are about 158 genera (492 species) of plants from the family Poaceae in Pakistan.

Among the 100 rust species known to infect 130 plant species (Afshan et al.

2010) are 70 species of Puccinia Pers. and 14 species of Uromyces (Link) Unger

(Afshan & Khalid 2009; Afshan et al. 2009, 2010a,b, 2011; Iqbal et al. 2009;

Khalid & Afshan 2009).

In this paper we report Puccinia stipae var. stipae on Stipa sp. and Uromyces

sporobolicola on Sporobolus marginatus as new records and identify new

poaceous hosts for Puccinia sessilis (Calamagrostis pseudophragmite), P. recondita

(Agrostis gigantea), and P. striiformis (Dactylis glomerata) in Pakistan.

Materials & methods

Infected plants with their inflorescences were collected from sampling sites

throughout Pakistan. The rusted specimens were photographed and free hand sections

and scrape mounts of infected portions were observed under the stereomicroscope.

Twenty spores of each spore state found were examined under microscope (Nikon YS

100), and spore dimensions were determined using a Zeiss eyepiece screw micrometer.

238 ... Ishaq, Afshan & Khalid

Sections and spores were photographed by digiporo-Labomed and illustrated using a

Leitz camera lucida (Wetzlar Germany).

Enumeration of taxa

Uromyces sporobolicola J.C. Lindq., Revta Fac. Agron. Vet. Univ. nac. La Plata, Ser.

3, 38: 89 (1963 [“1962”]). Fig. 1

SPERMOGONIA and AECIA not found. URepINniA adaxial, cinnamon-

brown; UREDINIOsPORES light brown to hyaline, echinulate, globose to

subglobose, (19.5-)25-33 x (22-)2-38 um; wall 1.6-2.6 um thick, germ pores

2-4, supraequatorial. TeL1a amphigenous, blackish, covered by the epidermis;

TELIOSPORES 1-celled, brown, ovate, 19-21.5 x 21.5-27 um; apex 2.4-5.4 um

thick, wall 1.4-2 um thick; pedicel short, < 11 um long.

MATERIAL EXAMINED: PAKISTAN, PunyjaB, Sialkot, at 234 m a.s.l., on leaves of

Sporobolus marginatus Hochst. ex A. Rich., stages II + HI, 20 April 2011, AIM # 27

(LAH Herbarium No.1130).

CoMMENTs: Cummins (1971) reported U. sporobolicola, here a new record for

Pakistan, on Sporobolus pyramidatus. Other rusts known to infect Sporobolus

Fic. 1: Uromyces sporobolicola (lucida drawing; scale = 10 um).

A. Urediniospores showing germ pores (arrowed); B. One-celled teliospores.

Poaceous rusts new to Pakistan ... 239

species (S. arabicus, S. coromandelianus) in Pakistan include Puccinia sporoboli-

arabici Afshan et al., P sporoboli-coromandeliani Afshan et al., Uromyces ignobilis

(Syd. & P. Syd.) Arthur and U. tenuicutis McAlpine from Lahore (Ahmad

1956a,b, 1976, Masood et al. 1995) and NWFP (Afshan et al. 2008, 2010a).

Puccinia stipae (Opiz) Arthur, Bull. lowa Agric. Coll. Dept. Bot.: 160 (1884)

var. stipae Fic. 2

Fic. 2: Puccinia stipae var. stipae (lucida drawing; scale = 13 um).

Teliospores showing germ pores (arrows).

SPERMOGONIA, AECIA, & UREDINIA not found. Text1a blackish brown,

adaxial, exposed, compact, 0.14-0.27 x 0.13-0.19 mm. TELIOsPoREs golden

brown, broadly ellipsoid, 23-34 x 45-63 um; wall 1.9-2.6 um thick at the sides,

4.5-8 um thick apically; pedicel long, broken mostly, hyaline, 5.5-9 um wide,

< 112 um long.

MATERIAL EXAMINED: PAKISTAN, GiLGiT & BALTISTAN, Skardu, Shigar valley, at

3000 m a.s.L, on leaves of Stipa sp., stage III, 20 July 2010, AIM # 38 (LAH Herbarium

No.1131).

COMMENTs: (Cummins (1971) previously reported P. stipae var. stipae, here a

new record for Pakistan, on Stipa spartea. Ahmad (1969) reported a different

variety, Puccinia stipae var. stipina (Tranzschel) H.C. Greene & Cummins

(= Puccinia stipina Tranzschel), on Stipa himalaica from Quetta in Pakistan.

240 ... Ishaq, Afshan & Khalid

Fic. 3: Puccinia sessilis var. sessilis (lucida drawings; scale = 14 um).

A. Urediniospores showing echinulate surface ornamentation and germ pores; B. Teliospores.

Puccinia sessilis W.G. Schneid., Jahresber. Schles. Ges. Vaterl. Cult. 49: 19 (1871)

var. sessilis Fic. 3

SPERMOGONIA and AECIA not found. UREDINIA on stipe, brown, naked.

UREDINIOSPORE light brown to hyaline, globose to subglobose, 21-32 x 24-36

um; wall 1.9-2.4 um; germ pores up to 4, scattered or tending to be equatorial.

Poaceous rusts new to Pakistan ... 241

TELIA amphigenous, blackish, covered by epidermis. TELIOsPORE chest nut

brown, 23-32 x 47-68 um; wall 1.5-3 um thick at sides, 6.5-13 um thick

apically, pedicel < 77 um long.

MATERIAL EXAMINED: PAKISTAN, GILGIT & BALTISTAN, Skardu, Shigar valley, at 3000

maz.s.l., on leaves of Calamagrostis pseudophragmites (Haller f.) Koeler, stages II + IIL, 21

July 2010, AIM # 39 (LAH Herbarium No.1132).

ComMENtTs: Khalid & Iqbal (1996) previously reported P. sessilis on leaves

of Phalaris minor from Deosai Plains near Pakora Lake. Calamagrostis

pseudophragmites is a new host for the variety in Pakistan.

Puccinia striiformis Westend., Bull. Acad. Roy. Sci. Belgique 21: 235 (1854)

var. striiformis Fia. 4

SPERMOGONIA and AECIA not found. UREDINIA light brown, adaxial, in linear

series. UREDINIOSPORES globose to subglobose, hyaline, echinulate, 24-31 x

27-36 um; Wall 1.4-2.8 um thick, germ pores 2-many, scattered. PARAPHYSES

Fic. 4: Puccinia striiformis var. striiformis (lucida drawings; scale = 13 um).

A. Paraphyses; B. Urediniospores showing germ pores (arrowed); C. Teliospores.

242 ... Ishaq, Afshan & Khalid

capitate, having cap of 6.5-8.0 um diameter. TELIA blackish, abaxial, sheaths in

linear series, covered by epidermis, with hyaline paraphyses, forming locules.

TELIOSPORES clavate, light brown, 2-celled, 13-19(-22.0) x 34.0-50.0 um;

sometimes upper cell thicker than lower, apex 3.0-7.0(-9.0) um thick, wall

1-2 um thick, pedicel very delicate.

MATERIAL EXAMINED: PAKISTAN, GILGIT & BALTISTAN, Fairy Meadows, at 3036

m a.s.l, on Dactylis glomerata L., stages II + IH, 12 August 2007, AIM # 20 (LAH

Herbarium No.1133).

Comments: Numerous authors have reported P. striiformis var. striiformis

on Triticum aestivum, Hordeum vulgare, and Poa sp. from Quetta, Kalat,

Tandojam, Rawalpindi, Nathia Gali, Shogran (Kaghan valley), and Lahore

(Ahmad 1956a,b, Khan & Kamal 1968, Malik et al. 1968, Malik & Virk 1968,

Kakishima et al. 1993) and on Leersia oryzoides from Fairy Meadows (Afshan

2009). Dactylis glomerata is a new host for the variety in Pakistan.

Puccinia recondita Desm., Bull. Soc. bot. Fr. 4: 798 (1857) Fic. 5

SPERMOGONIA and AEcIA not found. Urepinia abaxial, brown.

UREDINIOSPORES globose to subglobose, hyaline, 15-21 x 17-26 um; walls

1.4-2.6 um thick, echinulate; germ pores 2—many, scattered. TeL1A abaxial,

black, covered by epidermis, 0.1-0.2 x 0.04-0.05 mm. TELIospores light

Fic. 5: Puccinia recondita (lucida drawings; scale = 13 um).

A. Urediniospores showing germ pores; B. Teliospores.

Poaceous rusts new to Pakistan ... 243

brown to hyaline, clavate, 2-celled, 13-19(-21) x 38-52 um; wall 1.6-2.4 um

thick, apex 3-6 um thick; pedicel very delicate.

MATERIAL EXAMINED: PAKISTAN, KHYBER-PAKHTUNKHAWAH (KPK), Sharan, at

2752 m a.s.l., on Agrostis gigantea Roth, stages II + HI, 27 July 2007, AIM # 19 (LAH

Herbarium No.1134).

COMMENTS: Agrostis gigantea is a new host from Pakistan for Puccinia recondita,

which has previously been reported on Agropyron sp., Aquilegia fragrans,

A. pubiflora, Brachypodium sp., Eremopyrum bonaepartis, Lolium perenne,

Triticum aestivum, and Thalictrum minus from Swat, Kaghan valley, Ayubia,

Tandojam, Shogran to Sari, Sharan, Murree, Faisalabad, and Quetta (Ahmad

1956a, Hasnain et al. 1959, Jorstad & Iqbal 1967, Ghaffar & Kafi 1968, Khan &

Kamal 1968, Malik & Virk 1968, Malik et al. 1968, Ahmad & Arshad 1972, Ono

1992, Ono & Kakishima 1992).

Acknowledgements

We sincerely thank Dr. Abdul Rehman Niazi (Department of Botany, University

of the Punjab, Lahore, Pakistan) and Dr. Omar Paino Perdomo (Dominican Society

of Mycology, Santo Domingo, Dominican Republic) for their valuable suggestions to

improve the manuscript and acting as presubmission reviewers. We are also thankful to

Dr. Shaun Pennycook, Nomenclature Editor, for reviewing the manuscript critically.

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Northern Areas of Pakistan. Ph.D. Thesis, Department of Botany, University of the Punjab,

Lahore, Pakistan.

Afshan NS, Khalid AN. 2009. New records of Puccinia and Pucciniastrum from Pakistan. Mycotaxon

108: 137-146. http://dx.doi.org/10.5248/108.137

Afshan NS, Khalid AN, Niazi AR. 2008. New records and distribution of rust fungi from Pakistan.

Mycotaxon 105: 257-267.

Afshan NS, Khalid AN, Iqbal SH, Niazi AR, Sultan A. 2009. Puccinia subepidermalis sp. nov. and

new records of rust fungi from Pakistan, Mycotaxon 110: 173-182.

http://dx.doi.org/10.5248/110.173

Afshan NS, Khalid AN, Niazi AR. 2010a. Three new species of rust fungi from Pakistan. Mycological

Progress 9: 485-490. http://dx.doi.org/10.1007/s11557-010-0655-8.

Afshan NS, Khalid AN, Iqbal SH, Niazi AR, Sultan A. 2010b. Puccinia anaphalidis-virgatae, a new

species and a new variety of rust fungi from Fairy Meadows, Northern Pakistan. Mycotaxon

112: 483-490. http://dx.doi.org/10.5248/112.483

Afshan NS, Khalid AN, Niazi AR, Iqbal SH. 2011. New records of Uredinales from Fairy Meadows,

Pakistan. Mycotaxon 115: 203-213. http://dx.doi.org/10.5248/115.203

Ahmad 8.1956a. Uredinales of Pakistan. Biologia 2: 29-101.

Ahmad S. 1956b. Fungi of Pakistan. Biological Society of Pakistan, Lahore Monograph 1: 1-126.

Ahmad S. 1969. Fungi of Pakistan. Biological Society of Pakistan, Lahore, Monograph 5, Suppl. 1.

110 p.

Ahmad S. 1976. Contribution to the fungi of Pakistan. XVII. Sultania 2: 17-21.

Ahmad §, Arshad M. 1972. Contribution to the fungi of West Pakistan. XV. Biologia 18: 113-119.

244 ... Ishaq, Afshan & Khalid

Cummins GB. 1971. The rust fungi of cereals, grasses and bamboos. Springer Verlag, Berlin-

Heidelberg—New York.

Ghaffar A, Kafi A. 1968. Fungi of Karachi. Pak. J. Sc. 20: 5-10.

Hasnain SZ, Khan A, Zaidi AJ. 1959. Rust and smut of Karachi. Department of Botany, University

of Karachi, Monograph 2, 36 p.

Iqbal SH, Afshan NS, Khalid AN, Niazi AR, Sultan A. 2009. Additions to the rust fungi of Fairy

Meadows, Northern Areas of Pakistan. Mycotaxon 109: 1-7. http://dx.doi.org/10.5248/109.1

Jorstad I, Iqbal SH. 1967. Uredinales from West Pakistan. Nytt. Mag. Bot. 14: 31-38

Kakishima M, Okane I, Ono Y. 1993. Graminicolous rust fungi (Uredinales) from Pakistan.

181-186, in: Nakaike & Malik (eds). Cryptogamic flora of Pakistan, vol.2. Nat. Sci. Mus.,

Tokyo.

Khalid AN, Afshan NS. 2009. Additions to the graminicolous rust fungi of Pakistan. Mycotaxon

108: 175-183. http://dx.doi.org/10.5248/108.175

Khalid AN, Iqbal SH. 1996. Addition to the rust flora of Pakistan. Pak. J. Bot. 28(1): 115-117.

Khan SA, Kamal M. 1968. The fungi of South West Pakistan. Part I. Pak. Jour. Sci & Ind. Res. 11:

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Malik SA, Virk. 1968. Contribution to knowledge of parasitic fungi of Quetta-Kalat Region.

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Malik SA, Javaid MT, Khan MA. 1968. Uredinales of Quetta-Kalat region of Pakistan. Biologia 14:

37-46.

Masood A, Khalid AN, Iqbal SH. 1995. New records of graminicolous rust fungi (Uredinales) from

Pakistan. Sci. Int. 7(3): 415-416.

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

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Yandoc-Ables CB, Rosskopf EN, Charudattan R. 2006. Plant pathogens at work: progress

and possibilities for weed biocontrol. Plant Pathology Department, University of Florida,

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MY COTAXON

http://dx.doi.org/10.5248/118.245

Volume 118, pp. 245-250 October-December 2011

New records of smut fungi. 5

CvVETOMIR M. DENCHEV' , TEODOR T. DENCHEV'

& BRIAN M. SPOONER?

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

2 Gagarin St., 1113 Sofia, Bulgaria

*Mycology Section, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3AB, U.K.

* CORRESPONDENCE TO: cmdenchev@yahoo.co.uk

ABSTRACT — Morphological study of an anthericolous smut fungus on Scilla sardensis

(Chionodoxa sardensis) and S. luciliae (Chionodoxa luciliae) from UK has shown it to

represent Antherospora scillae. This is also known in UK on Scilla verna and S. forbesii,

new host records for this species. The new species, Urocystis bolboschoeni on Bolboschoenus

maritimus, is described and illustrated from UK.

Key worps — taxonomy, Urocystidales, Ustilaginomycetes

Introduction

In the current taxonomic scheme of the Ustilaginomycetes, the genus Ustilago

is restricted to species parasitizing plants of Poaceae (Vanky 1999, 2002).

For species on Liliaceae s. lat. previously referred to Ustilago, Ershad (2000)

erected a new genus, Vankya, with three species: V. heufleri (Fuckel) Ershad

on Erythronium and Tulipa; V. ornithogali (J.C. Schmidt & Kunze) Ershad on

Gagea; and V. vaillantii (Tul. & C. Tul.) Ershad on Albuca, Bellevalia, Eucomis,

Hyacinthus, Muscari, Puschkinia, Scilla (including Chionodoxa), and Urginea.

Vankya was later restricted to the species on leaves and stems of liliaceous

plants: V. heufleri, V. ornithogali, and the recently described V. lloydiae Vanky

on Lloydia triflora (Ledeb.) Baker (Vanky 2009a). For the anthericolous smut

fungus Vankya vaillantii s. lat. on plants of Asparagaceae (syn. Hyacinthaceae;

Liliaceae p.p.), Bauer et al. (2008) described a new genus, Antherospora, with

seven species: A. albucae (Syd. & P. Syd.) R. Bauer et al. on Albuca, A. peglerae

(Bubak et al.) R. Bauer et al. on Ornithogalum, A. scillae on Scilla bifolia L.,

A. tourneuxii (A.A. Fisch. Waldh.) R. Bauer et al. on Bellevalia, A. urgineae

(Maire) R. Bauer et al. on Urginea, A. vaillantii (Tul. & C. Tul.) R. Bauer et al.

246 ... Denchey, Denchev & Spooner

on Muscari, and the cryptic species A. vindobonensis R. Bauer et al. on Scilla

vindobonensis Speta. An eighth species, A. eucomis Vanky on Eucomis, was

added by Vanky (2009b). The narrow host association of these anthericolous

smut fungi (Bauer et al. 2008, Vanky 2009b) warranted closer study of the

morphology of Antherospora species on Chionodoxa (currently treated as

members of Scilla). The result of this study is presented here.

In June 2010, two specimens of Urocystis in leaves of Bolboschoenus maritimus

were collected by Mr N.W. Legon in South Devon, England. No Urocystis has

been previously recorded on Bolboschoenus nor on plants of related genera

(Scirpus, Holoschoenus, Isolepis, Schoenoplectus, Trichophorum). We consider

those specimens as belonging to a new species, described below.

Material & methods

Dried specimens from the mycological collection of the Royal Botanic Gardens, Kew

[K(M)] were examined under light (LM) and scanning electron (SEM) microscopes. For

LM observations, spores were mounted in lactophenol solution on glass slides, gently

heated to boiling point to rehydrate the spores, and then cooled. Spore measurements

are given in the form: min-max [mean + | standard deviation]. In the description, the

total number of spores (n) from all collections (x) measured are given in the form “(n/x).

For SEM, spores were attached to specimen holders by double-sided adhesive tape and

coated with gold with an ion sputter. The surface structure of spores was observed at 10

kV and photographed with a JEOL JSM-5510 scanning electron microscope.

Taxonomy

Antherospora scillae (Cif.) R. Bauer et al., Mycol. Res. 112: 1301, 2008. FIGs 1-4

Sori in the anthers and on the surface of anther filaments. Spore mass

powdery, dark olivaceous brown. INFECTION systemic, all anthers are infected.

Spores variable in shape, globose, subglobose, broadly ellipsoidal, ovoid,

elongated or slightly irregular, sometimes pyriform, 7-12.5 x 6.5-9.5 [9.4 + 1.2

x 7.8 + 0.8] um (n/, = 200), light olivaceous brown; wall even, 0.7—1.0 um thick,

densely verruculose.

SPECIMENS EXAMINED — On Scilla sardensis (Whittall ex Barr & Sugden) Speta: UK,

ENGLAND, Surrey, Kew, Royal Botanic Gardens, 21 April 1947, leg. G.M. Waterhouse

[ex herb. University of Sheffield, no. 764; IMI 15342] (as Ustilago vaillantii on Chionodoxa

sardensis Whittall ex Barr & Sugden, K(M) 116304). — On Scilla luciliae (Boiss.) Speta:

UK, ENGLAND, Surrey, Kew, Royal Botanic Gardens, near Lion Gate, 23 March 1954,

leg. R.W.G. Dennis (as Ustilago vaillantii on Chionodoxa luciliae Boiss., K(M) 69569).

ComMENts — ‘The study of this fungus on these two hosts was provoked by

the high host specialization amongst Antherospora species infecting Scilla

with the presumption that A. scillae is restricted to Scilla bifolia as suggested

by Bauer et al. (2008: 1302). Bauer et al. (2008) described both Antherospora

scillae on Scilla bifolia and A. vindobonensis on Scilla vindobonensis as having

Smut fungi 5 — Urocystis bolboschoeni sp. nov. ... 247

ve)

6 .

"Ss oe.

6S.

Figs 1-4. Spores of Antherospora scillae. 1-2 on Scilla luciliae (K(M) 69569) in LM,

3 on S. sardensis (K(M) 116304) in SEM, 4 on S. luciliae (K(M) 69569) in SEM.

Scale bars: 1-2 = 10 um, 3 = 2 um; 4=5 um.

248 ... Denchey, Denchev & Spooner

Fics 5-8. Spores of Urocystis bolboschoeni on Bolboschoenus maritimus.

1-2 in LM (holotype), 3 in SEM (K(M) 166454), 4 in SEM (holotype).

Scale bars: 5-8 = 10 um.

Smut fungi 5 — Urocystis bolboschoeni sp. nov. ... 249

spore walls with a thickness of ca 0.5 um. The specimens we examined on Scilla

sardensis and S. luciliae have spores with a thicker wall (0.7-1.0 um wide), and

it seemed warranted to make additional observations of the spore wall on S.

bifolia. We found that the spore wall thickness of two specimens on S. bifolia

measured 0.7-0.9 um (Vanky Ustilag. exs. no. 57) and 0.8-1.1 um (SOMF

19206). Additionally, Vanky (2009b) cited a spore wall thickness of 0.8-1.5 um

for Antherospora scillae. Our spore measurements from four specimens of A.

scillae on Scilla bifolia (Vanky Ustilag. exs. no. 57; SOMF 2859, 2888, & 19206)

(7-14.5 x 6.5-10 [9.8 + 1.1 x 8.5 + 0.7] um; n/, = 350) resembled those of the

British specimens we examined.

ADDITIONAL SPECIMENS EXAMINED — On Scilla bifolia: HUNGARY, prope pag.

Makad insulae Csepel sziget, ca. 98 m, 6 April 1965, S. Toth (Vanky Ustilag. exs., no. 57);

BULGARIA, |.d. Papazova Korija prope urb. Elhovo, 21 March 1962, C. Hinkova (SOMF

2859); Ld. Gorna Top¢ija, distr, Jambol, 22 March 1961, C. Hinkova SOMF 2888); Pirin

Mts, Sinanitsa, ca. 2000 m, 24 May 1985, C.M. Denchev (SOMF 19206).

Currently, there is no molecular phylogenetic inference for distinguishing

the anthericolous smut fungus on Scilla sardensis and S. luciliae from that on

S. bifolia. Because no morphological differences were found, we identified

Antherospora scillae as the fungus on the British specimens that we examined.

This anthericolous smut fungus known in UK on Scilla verna Huds. (several

collections) has been recently collected also on S. forbesii (Baker) Speta (=

Chionodoxa forbesii Baker) [K(M) 169982]. Scilla sardensis, S. luciliae, S. verna,

and S. forbesii are new host records for Antherospora scillae.

Urocystis bolboschoeni Denchey, T. Denchev, Spooner & Legon, sp. nov. Fics 5-8

MycoBank MB 561704

Sor! in foliis et vaginis foliorum strias formati. GLOMERULI SPORARUM 16-40 x 14.5-28.5

um, e sporis 1-3 (-5) compositi [numeri sporarum: 1 = 44.6%, 2 = 39.2%, 3 = 11.2%, 4=

4.2%, 5 = 0.8%], strato cellularum sterilium perfecte circumdatis composite. SPORAE 12.5-

18.5 x 10-15 [15.2 1.1 x 12.7 + 1.1] um, rufobrunneae, pariete 0.6-0.9 um crasso.

Type on Bolboschoenus maritimus (L.) Palla: UK, England, South Devon, Budleigh

Salterton, Otter Estuary, 29 June 2010, leg. N.W. Legon, as Urocystis sp. (holotype, K(M)

166455).

EryMo_oey: the name refers to the host genus.

Sori in leaves and leaf sheaths between the veins, as streaks, sometimes

confluent, initially covered by the epidermis, rupturing longitudinally to expose

a powdery, dark reddish brown mass of spore balls. SPORE BALLS subglobose

to ellipsoidal or slightly irregular, composed of 1-3 (-5) central teliospores [1

= 44.6%, 2 = 39.2%, 3 = 11.2%, 4 = 4.2%, 5 = 0.8%; n/, = 950] and a continuous

layer of peripheral sterile cells; 16-25 x 14.5-22.5 um [with 1 teliospore],

21-33.5 x 15.5-24.5 um [with 2 teliospores], 25-40 x 17-28.5 um [with 3

teliospores]. STERILE CELLS broadly elliptical, elliptical, suborbicular or ovate

250 ... Denchev, Denchev & Spooner

in outline, or collapsed, sometimes slightly irregular, 5-11.5 um long, pale or

yellowish, wall 0.6-1.0 um thick, smooth. Sporss globose, subglobose, broadly

ellipsoidal or ovoid, 12.5-18.5 x 10-15 [15.2 + 1.1 x 12.7 + 1.1] um (n/, = 200),

medium reddish brown; wall 0.6-0.9 um thick, finely verruculose.

DISTRIBUTION — On Cyperaceae: Bolboschoenus maritimus, Europe

(England). Known only from the type locality.

ADDITIONAL SPECIMENS EXAMINED — On Bolboschoenus maritimus: UK, England,

SouTH Devon, Budleigh Salterton, Otter Estuary, 22 June 2010, leg. N.W. Legon (as

Urocystis sp., K(M) 166454); 12 May 2011, leg. N.W. Legon (K(M) 170764).

COMMENTS — On Cyperaceae three Urocystis species have been described: (i)

U. fischeri Korn. ex G. Winter on species of Carex, with type on C. acuta L.

from Germany; distributed in Europe, Asia, and North America (sori in leaves

and culms, spore balls 20-40 um long, composed of 1-3 (-4) fertile spores);

(ii) U. littoralis (Lagerh.) Zundel on Carex maritima Gunnerus from Norway

(sori in leaves, spore balls 25-50 um long, composed of (1-) 2-6 (-9) fertile

spores); and (iii) U. chorizandrae Cunningt. et al. on Chorizandra enodis Nees

from Australia (sori in leaves and sterile culms, spore balls 25-50 um long,

composed of 1-5 (-8) fertile spores) (Vanky & Shivas 2003, Vanky 2009c).

It is remarkable that no smut has been previously reported on this common

and widely distributed host species.

Acknowledgements

We gratefully acknowledge Dr Kalman Vanky (Herbarium Ustilaginales Vanky,

Tubingen, Germany) and Dr Roger G. Shivas (Agri-Science Queensland, Australia) for

critically reading the manuscript and serving as pre-submission reviewers.

Literature cited

Bauer R, Lutz M, Begerow D, Piatek M, Vanky K, Bacigalova K, Oberwinkler F. 2008. Anther smut

fungi on monocots. Mycological Research 112: 1297-1306.

http://dx.doi.org/10.1016/j.mycres.2008.06.002

Ershad D. 2000. Vankya, a new genus of smut fungi. Rostaniha 1: 65-72 [in English], 151-161 [in

Farsi], Figs 1-6.

Vanky K. 1999. The new classificatory system for smut fungi, and two new genera. Mycotaxon 70:

35-49.

Vanky K. 2002. Illustrated genera of smut fungi. 2"* edn. APS Press, St. Paul, Minnesota, USA.

238 pp.

Vanky K. 2009a. The genus Vankya (Urocystidaceae) revisited. Mycologia Balcanica 6: 73-78.

Vanky K. 2009b. Taxonomic studies on Ustilaginomycetes - 29. Mycotaxon 110: 289-324.

Vanky K. 2009c. Keys to smut fungi of selected host plant families and genera. Mycologia Balcanica

6: 1-36.

Vanky K, Shivas RG. 2003. Further new smut fungi (Ustilaginomycetes) from Australia. Fungal

Diversity 14: 243-264.

MY COTAXON

http://dx.doi.org/10.5248/118.251

Volume 118, pp. 251-256 October-December 2011

A new species of Marasmius from northern Argentina

BERNARDO LECHNER”* & LEANDRO PAPINUTTI

PROPLAME-PRHIDEB (CONICET), DBBE, Facultad de Ciencias Exactas y Naturales,

Ciudad Universitaria, (C1428EHA) Ciudad de Buenos Aires, Argentina.

*CORRESPONDENCE TO: blechner@bg.fcen.uba.ar

ABSTRACT — Marasmius tyrius is proposed as a new species. It is characterized by collariate

lamellae, insititious stipe, violet pileus, pileipellis with Siccus-type broom cells, and clavate to

fusoid spores. Detailed description and illustrations of macro- and microscopic characters

are provided.

Key worps — Argentinean mycobiota, Marasmiaceae, taxonomy

Introduction

The genus Marasmius (Marasmiaceae, Agaricales) is represented by a large

number of species, especially in the tropics (Singer 1976, 1986). Itis characterized

by a hymeniform pileipellis composed of broom cells of Siccus- or Rotalis-

type, or of exclusively smooth cells. Siccus-type cells are characterized by apical

appendages or more or less erect setulae, while the Rotalis-type usually have

short appendages and that descend from the apex down towards the middle of

the cell (Singer 1986).

Spegazzini (1880a,b, 1883, 1887, 1891, 1898, 1902, 1909, 1925, 1926a,b),

Singer (1950, 1959, 1965, 1969, 1976, 1989), and Singer & Digilio (1953)

described species of Marasmius from Argentina, Chile, and Paraguay. Singer

(1953) later studied several types of species described by Spegazzini and

included his observations in the exhaustive monographs on Marasmius in

South America and Neotropics (Singer 1959, 1965, 1976). After these reports,

mycological literature on the genus in the region has been scant. Recently, there

has been a renewed worldwide interest in Marasmius (Desjardin et al. 2000,

Antonin 2003, 2004a,b, 2007, Wannathes et al. 2004, 2009, Antonin & Buyck

2006, Desjardin & Ovrebo 2006, Puccinelli & Capelari 2006, Tan et al. 2009,

Antonin & Noordeloos 2010). In Argentina, Lechner et al. (2006) reported new

records of Marasmius collected in the northern region, and several species have

252 ... Lechner & Papinutti

been described and illustrated in the Pictorial Atlas of Iguazi National Park

(Wright et al. 2008).

According to our knowledge, one specimen collected during an expedition

in northern Argentina does not match any described Marasmius. Thus, we

propose Marasmius tyrius as a new species.

Materials & methods

The specimen was photographed and its macroscopic features were recorded when

fresh. Tissues were mounted in 5% KOH + 1% aqueous phloxine or Melzer’s reagent and

observed under a Nikon E-600 microscope. For basidiospore measurements, the length

: width quotient (Q) was calculated, with Qm indicating the mean value of Q quotients.

Line drawings were made with the aid of a light tube. Basidiomata were dried, kept

frozen for a week, and deposited in the BAFC herbarium (Department of Biodiversity

and Experimental Biology, Faculty of Exact and Natural Sciences, Universidad de

Buenos Aires).

Taxonomy

Marasmius tyrius B.E. Lechner & Papinutti, sp. nov. Fics. 1-2

MycoBank MB 518617

Pileo 4-5.5 mm lato, 2-2.7 mm alto, hemisphaerico, centro depresso, violaceo, glabro,

sulcato. Lamellis albis, subdistantibus (proxime 17), collariatis. Stipite castaneo, glabro,

31-37 x 0.2-0.5 mm. Basidiosporis 11.1-14.1 x 2.5-3.7 um, clavatis ad fusiformibus,

laevibus, hyalinis, inamyloideis. Basidiis clavatis, 23-28 x 4-6 um, 1-3 sterigmata.

Cheilocystidiis difformibus: (1) elementis typi Marasmii sicci, 17.3-20.5 x 3.7-9.5 um,

setulis 2-4 um longis, tenuitunicatis vel crassitunicatis, castaneis in KOH, et (2) clavatis,

22.5-27.4 um longis, tenuitunicatis, hyalinis. Elementis epicuticularibus plerumque typi

Marasmii sicci, 11.9-22.5 x 11.2-13.1 um, setulis 4-5 um longis.

Type: Argentina, Chaco, Colonia Benitez, 27°26'19.20"S 58°50'51.39"W, on fallen leaves,

19-IX-2007, col. L. Papinutti, G. Rol6n (HOLOTYPE BAFC 51726).

Erymo.oey: The epithet refers to the similarity of the pileus color to Tyrian purple, the

dye famously produced in the ancient Phoenician city of Tyre (Latin: Tyrus).

Piteus 4-5.5 mm in diam x 2-2.7 mm high, hemispheric, depressed at the

centre, glabrous, deeply sulcate, violet (Plate 42, K10K11, Maerz & Paul 1930),

centre light brown with an outer light violet ring. PILEUS CONTEXT white,

very thin (less than 1 mm). LAMELLAE (Fig. 1c) collariate, subdistant (ca. 17),

whitish, entire, lamellulae absent; edge broad, concolorous with the pileus.

STIPE 31-37 x 0.2-0.5 mm, chestnut-brown, creamy to whitish at the apex,

glabrous, insititious, cylindrical. RHIzomorpPus frequently present, thin, black.

STIPE CONTEXT thin, white.

BASIDIOSPORES (9.3—)11.1-14.1(-15.4) x 2.5-3.7 um, Q = 3.24-4.97, Qm = 4.23,

n = 30, clavate to fusoid, hyaline, inamyloid, smooth, thin-walled. BAsrp1a

clavate, 23-28 x 4-6 um, 1-3 spored. Basidioles clavate, 19.2-25.0 um long.

PLEUROCYSTIDIA absent. LAMELLA EDGE Sterile, with crowded cheilocystidia.

Marasmius tyrius sp. nov. (Argentina) ... 253

Fic. 1. Basidiomes of Marasmius tyrius.

Scale bar = 1 cm.

CHEILOCYSTIDIA of two types: (1) Siccus-type broom cells, body 17.3-20.5 x

3.7-9.5 um, setulae 2-4 um long, thin- to thick-walled, castaneous in KOH, and

(2) clavate, 22.5-27.4 um long, thin-walled, hyaline. HYMENOPHORAL TRAMA

regular to subregular; hyphae hyaline, clamped, 2.1-6.0 um diam. PILEIPELLIS

composed of Siccus-type broom cells, body 11.9-22.5 x 11.2-13.1 um, setulae

4-5 um long, thin- to thick-walled, castaneous in KOH, mostly subvesiculose;

and of clavate, some bifurcate, 20.1-34.1 x 5.4-7.0 um, thin- to thick-walled

elements (wall up to 2.3 um thick), castaneous in KOH. HYPHAE OF PILEUS

CONTEXT hyaline, thin-walled, clamped, 2.6-6.0 um diam, weakly dextrinoid.

HYPHAE OF STIPE CONTEXT generative, hyaline, clamped, thin-walled, 2.6-5.2

um diam, and thick-walled (up to 1.5 um), 4.2-6.3 um diam, scant in upper

zones of the apex; non-dextrinoid.

ECOLOGY AND DISTRIBUTION — Gregarious, abundant in wet forest soil and

litter at Colonia Benitez. Growing on fallen leaves of Phoebe porphyria (Griseb.)

Mez (Lauraceae). Known only from type locality.

COMMENTS — Marasmius tyrius belongs to sect. Marasmius, subsect. Sicciformes

Antonin based on the presence of a collarium, the insititious stipe, and

Siccus-type broom cells in the pileipellis. In his monograph on Neotropical

254 ... Lechner & Papinutti

MONO Ni

Fic. 2. Marasmius tyrius, micromorphology.

a: Spores. b: Basidia. c: Cheilocystidia. d: Elements of the pileipellis.

Scale bar = 10 um.

marasmioid fungi, Singer (1976) did not present any violet-colored species

with the characteristics of the subsect. Sicciformes, but several purple colored

ones. We will mention the three most similar species. Marasmius marthae

Singer has a purple-red pileus, black stipe at maturity, somewhat longer spores,

and grows on wood pieces; M. rubromarginatus Dennis has a carmine-purple

to brown-red pileus, more distant lamellae (11-15), and longer spores; and M.

sanguirotalis Singer has a dark purple pileus, distant lamellae (11-13), much

longer spores, and grows on branches and woody matter. Among the few other

purple or purple tinged species not treated by Singer (1976), the most similar

is M. purpureobrunneolus Henn. from Indonesia. It differs by reddish-brown to

purplish-brown pileus and more distant lamellae (9-14).

The only two violaceous species in subsect. Sicciformes were described by

Singer (1989) from Brazil, both with significantly smaller spores: M. iodactylus

Singer (spores 6-7.5 x 2.5-3.3 um) and M. izonetae Singer (spores 5 x 2.2 um).

Marasmius tyrius sp. nov. (Argentina) ... 255

In sect. Marasmius, subsect. Marasmius (= Pararotulae Singer), Singer (1976)

described M. violeorotalis Singer with a violet-lilac pileus. It has the same

habitat as M. tyrius (on leaves of dicotyledonous trees) but differs in smaller

spores (6.2-9.5 x 2.5-3 um), Rotalis-type broom cells in the pileipellis, and

more distant lamellae (ca. 12). Other Marasmius species with violaceous or

violaceous tinged pileus belong to other sections of the genus.

Acknowledgments

We wish to express our gratitude to Dr. Zdenko Tkaléec and Dra. Marina Capelari

for reviewing the manuscript and offering useful comments. We thank Guillermo Rolén

for the technical assistance with the figures. This work was supported by CONICET

(Consejo Nacional de Investigaciones Cientificas y Técnicas) Argentina and Universidad

de Buenos Aires.

Literature cited

Antonin V. 2003. New species of Marasmius (Basidiomycetes, Tricholomataceae) from tropical

Africa - I. Sect. Epiphylli, Fusicystides, Globulares, Hygrometrici and Neosessiles. Mycotaxon

85: 109-130.

Antonin V. 2004a. New species of marasmioid genera (Basidiomycetes, Tricholomataceae) from

tropical Africa - III. Marasmius sect. Sicci. Mycotaxon 89: 399-422.

Antonin V. 2004b. New species of marasmioid genera (Basidiomycetes, Tricholomataceae) from

tropical Africa - IV. Four new taxa of the genus Marasmius and one new combination.

Mycotaxon 89: 423-431.

Antonin V. 2007. Monograph of Marasmius, Gloiocephala, Palaeocephala and Setulipes in tropical

Africa. Fungus flora of tropical Africa 1. Meise, National Botanic Garden (Belgium). 177 p.

Antonin V, Buyck B. 2006. Marasmius (Basidiomycota, Marasmiaceae) in Madagascar and the

Mascarenes. Fungal Diversity 23:17-50.

Antonin V, Noordeloos ME. 2010. A monograph of marasmioid and collybioid fungi in Europe.

Eching, IHW Verlag. 480 p.

Desjardin DE, Ovrebo CL. 2006. New species and new records of Marasmius from Panama. Fungal

Diversity 21: 19-39.

Desjardin DE, Retnowati A, Horak E. 2000. Agaricales of Indonesia. 2. A preliminary monograph

of Marasmius from Java and Bali. Sydowia 52: 92-194.

Lechner BE, Wright JE, Popoff O. 2006. New taxa and new records for Argentina of fungi from

Iguazu National Park, Misiones. Fungal Diversity 21: 131-139.

Maerz A, Paul M. 1930. Dictionary of color. New York. 207 p.

Puccinelli C, Capelari M. 2006. Two new species of Marasmius (Basidiomycota, Marasmiaceae)

from Brazil. Mycotaxon 95: 295-300.

Singer R. 1950. Die hoheren Pilze Argentiniens. Bulletin Suisse de Mycologie 28: 181-196.

Singer R. 1953. Type studies on agarics III. Lilloa 25: 463-514.

Singer R.1959. Studies toward a monograph of South American species of Marasmius. Sydowia

12: 54-148.

Singer R. 1965. Monographic studies on South American Basidiomycetes, especially those of the

east slope of the Andes and Brazil 2. The genus Marasmius in South America. Sydowia 18:

106-358.

Singer R. 1969. Mycoflora australis. Nova Hedwigia 29: 1-405.

256 ... Lechner & Papinutti

Singer R. 1976. Marasmieae (Basidiomycetes - Tricholomataceae). Flora Neotropica 17: 1-347.

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

Books. 981 p.

Singer R. 1989. New taxa and new combinations of Agaricales (Diagnoses fungorum novorum

Agaricalium IV). Fieldiana Botany 21: 1-133.

Singer R, Digilio APL. 1953. Prodromo de la flora agaricina Argentina. Lilloa 25: 5-461.

Spegazzini CL. 1880a. Fungi Argentini. Pugillus 1. Anales de la Sociedad Cientifica Argentina 9:

158-192.

Spegazzini CL. 1880b. Fungi Argentini. Pugillus 3. Anales de la Sociedad Cientifica Argentina 10:

122-142.

Spegazzini CL. 1883. Fungi Guaranitici. Pugillus I. Anales de la Sociedad Cientifica Argentina 16:

272-284.

Spegazzini CL. 1887. Fungi Fuegiani. Boletin de la Academia Nacional de Ciencias de Cordoba

11: 135-311.

Spegazzini CL. 1891. Fungi Guaranitici nonnulli novi vel critici. Revista Argentina de Historia

Natural 1: 101-111.

Spegazzini CL. 1898. Fungi Argentinici novi vel critici. Anales del Museo de Historia Natural de

Buenos Aires 6: 81-288.

Spegazzini CL. 1902. Mycetes Argentinenses. Series 2. Anales del Museo de Historia Natural de

Buenos Aires 8: 49-89.

Spegazzini CL. 1909. Mycetes Argentinenses. Series 4. Anales del Museo Nacional de Historia

Natural de Buenos Aires 19: 257-458.

Spegazzini CL. 1925. Séptima contribucién a la micologia Chilena. Revista Chilena de Historia

Natural 29: 58-64.

Spegazzini CL. 1926a. Contribucién al conocimiento de la flora micolégica de las Sierras de

Cordoba. Boletin de la Academia Nacional de Ciencias de Cordoba 29: 113-190.

Spegazzini CL. 1926b. Observaciones y adiciones a la micologia argentina. Boletin de la Academia

Nacional de Ciencias de Cordoba 28: 267-406.

Tan Y-S, Desjardin DE, Perry BA, Vikineswary S, Noorlidah A. 2009. Marasmius sensu stricto in

Peninsular Malaysia. Fungal Diversity 37: 9-100.

Wannathes N, Desjardin DE, Retnowati A, Tan YS, Lumyong S. 2004. A redescription of Marasmius

pellucidus, a species widespread in South Asia. Fungal Diversity 17: 203-218.

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.

Wright JE, Lechner BE, Popoff O. 2008. Atlas pictérico de los hongos del Parque Nacional Iguazu.

Editorial LOLA, Buenos Aires. 227 p.

MY COTAXON

http://dx.doi.org/10.5248/118.257

Volume 118, pp. 257-264 October-December 2011

Arrasia rostrata (Basidiomycota),

a new corticioid genus and species from Italy

ANNAROSA BERNICCHIA*', SERGIO P. GORJON? & KAREN K. NAKASONE?

"Dipartimento di Scienze e Tecnologie Agroambientali,

Universita degli Studi di Bologna, Via Fanin 42, 40127 Bologna, Italy

*Centro de Investigacion y Extension Forestal Andino Patagonico,

Area de Proteccién. CC 14, 9200 Esquel, Chubut, Argentina

°Center for Forest Mycology Research, U.S. Forest Service,

One Gifford Pinchot Drive, Madison, WI 53726-2398 USA

*CORRESPONDENCE TO: annarosa. bernicchia@unibo. it

ABSTRACT — An unusual corticioid species with distinctive large basidiospores that develop

a distal refractive rostrum when fully mature is described as new. It grows on living bark

of Juniperus phoenicea on the Italian island of Sardinia. Because it is morphologically

distinct from any known genus of corticioid fungi, the new genus Arrasia is proposed to

accommodate it.

Key worps — dendrotheloid fungi, Italy

Introduction

Many new species of polypores and corticioids have recently been described

from Sardinia: Aleurodiscus ilexicola Bernicchia & Ryvarden, Antrodiella

ichnusana Bernicchia et al. Antrodia sandaliae Bernicchia & Ryvarden,

Echinodontium ryvardenii Bernicchia & Piga, Neolentiporus squamosellus

(Bernicchia & Ryvarden) Bernicchia & Ryvarden, Phellinus juniperinus

Bernicchia & S. Curreli, and Vararia maremmana Bernicchia. Sardinia may

be a refugium from the last glacial period, as demonstrated by the presence

of Piloporia sajanensis (Parmasto) Niemela, previously known as a boreal

species, or the occurrence of Echinodontium ryvardenii, while other species of

Echinodontium Ellis & Everh. are known from North America and Asia.

Conservation International (CI 2007) lists the Mediterranean Basin as

a “Biodiversity Hotspot.’ Several basidiomycete species associated with

Mediterranean juniper forests are Echinodontium ryvardenii, Hyphoderma

etruriae Bernicchia, Lenzitopsis oxycedri Malencon & Bertault, Peniophora

258 ... Bernicchia, Gorjon & Nakasone

junipericola J. Erikss., Phellinus juniperinus, Trametes junipericola Manjon et al.,

and Vararia maremmana. Recently, a striking corticioid species was discovered

from Sardinia growing on living bark of Juniperus phoenicea L. (Cupressaceae).

This new species is here fully described and illustrated; because it has no close

relatives in any described corticioid genus, a new genus is proposed.

Materials & 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

cyanophily of basidiospore walls. Crystalline deposits were dissolved in a 50% HCl

solution to individualize microscopical elements. Line drawings were made with a

camera lucida attachment. Specimens are deposited in HUBO, CFMR, and SALA.

Taxonomy

Arrasia Bernicchia, Gorjon & Nakasone, gen. nov.

MycoBank MB 561760

Basidiomata effusa, adnata, tenuissima, levia, tenuiter farinosa, margine distincto.

Systema hypharum monomiticum; hyphae generativae fibulatae, tenuitunicatae et

ramosae. Dendrohyphidia filamentosa, ramosa, fibulata. Basidia suburniformia deinde

subclavata, flexuosa, fibulata, tetraspora. Basidiosporae hyalinae, leviter crassitunicatae,

leves, cyanophylae, inamyloideae, indextrinoideae, subfusoideae vel biapiculatae, parte

distali se extendente ad rostrum crassitunicatum et refractivum.

TYPE SPECIES: Arrasia rostrata Bernicchia, Gorjon & Nakasone

ErymMo_oey: the genus is dedicated to Luigi Arras, mycologist and friend, who is always

present during the mycological excursions across Sardinia.

BASIDIOMATA effuse, adnate, thin, white, smooth, finely farinaceous, with a

distinct margin.

HyPHAL SYSTEM monomitic, hyphae clamped. DENDROHYPHIDIA

filamentous, branched, clamped. Basip1a suburniform at first, then flexuous,

clavate to obclavate, sometimes with a basal lobe, basally clamped, with 4

sterigmata. BAstp1ospores broadly subfusiform to biapiculate, distal end

elongating into a thick-walled rostrum, walls hyaline, slightly thickened,

smooth, cyanophilous, inamyloid, nondextrinoid.

REMARKS — ‘The distinctive feature of the new genus is the rostrate

basidiospores.

Arrasia rostrata Bernicchia, Gorjon & Nakasone, sp. nov. PLATES 1-4

MycoBank MB 561761

Basidiomata resupinata, effusa, adnata, tenuissima, 30 mm longa et 5 mm lata, laevia

sed farinosa vel subgausapata in maturitate, subalba vel albocinerea, margine distincto.

Systema hypharum monomiticum; hyphae generativae tenuitunicatae, fibulatae, 1.8-2.3

um latae. Dendrohyphidia filamentosa, ramosa, fibulata ad basim. Cystidia desunt.

Basidia primo suburniformia deinde subclavata, flexuosa, fibulata ad basim, tenuitunicata,

Arrasia rostrata gen. & sp. nov. (Italy) ... 259

PiaTE 1. Echinodontium ryvardenii (center) surrounded by circular to linear patches of

Arrasia rostrata (Bernicchia 8087, holotype). Scale bar = 5 cm.

50-90 x 10-16 ym, tetraspora. Basidiosporae hyalinae, laeves, leviter crassitunicatae,

cyanophylae, inamyloideae, indextrinoideae, late subfusoideae vel biapiculatae, parte

distali se extendente ad rostrum crassitunicatum et refractivum, 27-40 x 10-15 um. Ad

corticem arborum coniferarum viventis.

TyPE: Italy, Sardinia, Nuoro province, Lanaittu valley, 180 m a.s.1., on bark in trunk and

old branches of living Juniperus phoenicea, 30.11I.2010, leg. A. Bernicchia, coll. 8087.

Holotype in HUBO. Isotype in SALA et CFMR.

EryMoLocy: the name rostrata refers to the long refractive rostrum on the

basidiospores.

BASIDIOMATA resupinate, effuse, small, circular to linear patches, becoming

confluent, up to 30 x 5 mm, thin, up to 180 um thick, soft, white, smooth,

initially finely farinaceous to subfelty, finally thickly farinaceous; margin

abrupt, distinct.

HYPHAL SYSTEM monomitic with clamped generative hyphae. Subiculum

thin, a dense tissue of hyphae and crystals; subicular hyphae 1.8-2.3 um in

diam., clamped, moderately branched, walls hyaline, thin, encrusted with

hyaline crystals. Subhymenium not observed. Hymenium a palisade of

dendrohyphidia, basidia, and indistinct, collapsed hymenial elements obscured

by crystals. DENDROHYPHIDIA filamentous, irregular, with short branches near

260 ... Bernicchia, Gorjon & Nakasone

apex, 40-62 x 1.5-2 um, clamped at base, walls hyaline, thin. Cystrp1a absent.

Basip1A obclavate to suburniform at first, then flexuous, narrowly clavate

to obclavate, occasionally with a lateral lobe near base, 50-90 x 10-16 um,

clamped at base, containing resinous globules, often lower part collapsed below

a secondary septum, walls hyaline, thin, smooth, (2-)4-sterigmate, sterigmata

stout, up to 29 x 5 um. Basrprospores broadly subfusiform to bi-apiculate,

initially distal end obtuse, later developing an extended, thick-walled, refractive

rostrum or beak, 27-40 x 10-15 um, rostrum 8-14 x 1.2-2 um long, with a

distinct, refractive, thick-walled apiculus, containing resinous material, walls

hyaline, with distinct walls to slightly thick-walled, smooth, cyanophilous, not

reacting in Melzer’s reagent.

HABITAT AND DISTRIBUTION — Known from Sardinia growing on bark

of living Juniperus phoenicea, frequently in association with Echinodontium

ryvardenii.

ADDITIONAL SPECIMENS EXAMINED — ITALY. SARDINIA, NUORO PROVINCE, Lanaittu

valley, 180 m a.s.l., on trunk and old branches of living Juniperus phoenicea, 14.X1.2009

leg. A. Bernicchia, coll. 8086, 8097; 03.11.2010, leg. L. Arras coll. 8557; 30.11.2010: leg. A.

Bernicchia, coll. 7998, 8070, 8071, 8072, 8073, 8074, 8075, 8076, 8077, 8080, 8085.

ComMENTsS — ‘The most remarkable feature of Arrasia rostrata is its large,

beaked basidiospores. In developing basidiospores still attached to the

basidium, a distal knob forms that will develop into the rostrum. When fully

mature, the rostrum is a straight, thick-walled, refractive structure. By the time

the basidiospore is mature and the rostrum is fully developed, the basidium

is empty and collapsed. The indistinct remnants of post-mature basidia can

be observed if the crystalline matter is dissolved. The apiculus of mature

basidiospores is refractive and thick-walled with a notched appearance.

Arrasia rostrata is probably related to the corticioid genus Dendrothele

Hohn. & Litsch., which shares the same ecology (inhabiting bark of living trees),

crustose basidiomata, suburniform basidia, and hymenial structure with many

dendrohyphidia and abundant crystalline deposits (possibly an adaptation to

dry and exposed habitats). Dendrothele is a polyphyletic genus with species

distributed among several lineages in the hymenochaetoid, russuloid, corticioid,

and agaricoid clades (Goranova 2003, Goranova et al. 2003, Bodensteiner et al.

2004). The type species, Dendrothele papillosa Héhn. & Litsch. [= D. griseocana

(Bres.) Bourdot & Galzin], is included in the Niaceae Julich within the

Agaricales Underw. and closely related to the cyphelloid genera Lachnella Fr.

and Cyphellopsis Donk. Recently, Nakasone & Burdsall (2011) and Gorjén et

al. (2011) observed navicular basidiospores in Dendrothele species from New

Zealand and Argentina, a feature previously also known in some cyphelloid

genera. Convergences in morphological traits and habit seem to have occurred

repeatedly, characterizing the artificial dendrotheloid group. However, no other

Arrasia rostrata gen. & sp. nov. (Italy) ... 261

PLATE 2. Arrasia rostrata. Hymenial elements (Bernicchia 8087, holotype).

a) immature basidiospores, b) basidia, c) generative hyphae, d) dendrohyphidia

basidiospores with a refractive rostrum are known in Dendrothele or any other

corticioid species.

262 ... Bernicchia, Gorjon & Nakasone

PLaTE 3. Arrasia rostrata. Basidia and immature basidiospores.

(Bernicchia 8087, holotype)

It is also interesting to note that some basidiospores in Vararia P. Karst.

(such as in Vararia investiens (Schwein.) P. Karst.) develop an empty amyloid

part separated by a septum in the proximal region (close to the sterigma

attachment). The biological function, if any, of this structure is unknown.

Arrasia rostrata gen. & sp. nov. (Italy) ... 263

NS 0 10 20pm

PLATE 4. Arrasia rostrata. Mature basidiospores with a well developed rostrum and immature

(right and center right) basidiospores. (1: Bernicchia 8087, holotype; 2: Bernicchia 8074; 3:

Bernicchia 8071)

264 ... Bernicchia, Gorjon & Nakasone

Morphological similarities may also be drawn with some genera belonging to

Vuilleminiaceae Maire, Punctulariaceae Donk, and Corticiaceae Herter. Among

the Vuilleminiaceae, Vuilleminia Maire, Australovuilleminia Ghobad-Nejhad &

Hallenb., and Cytidia Quél. all share the saprophytic habit growing on attached

recently dead angiosperm wood (not known from coniferous substrata),

basidiomes that are usually gelatinous and decorticating, and allantoid to

ellipsoid basidiospores. Members of the Punctulariaceae, Punctularia Pat.,

Punctulariopsis Ghobad-Nejhad, and Dendrocorticium M.J. Larsen & Gilb.,

grow on fallen angiosperm wood and all species have ellipsoid basidiospores.

Genera in the Corticiaceae, as delimited by Ghobad-Nejhad et al. (2010), show

a more ecological and morphological complexity, but none display the features

that characterize Arrasia rostrata. Preliminary molecular studies indicate that

Arrasia rostrata fits no existing homobasidiomycete genus or order thus far

recognized (data not shown); further analyses are still required.

Acknowledgments

Nils Hallenberg and Alina G. Greslebin acted as presubmission reviewers and their

comments are acknowledged. We thank Luigi Arras and Marco Facchini for their help,

Giovanni Consiglio for revision of Latin diagnosis, and Cristina Spinelli for the colour

photo. The Council of Lanusei (Sardinia, Italy) supported AB and SPG on some collecting

trips. Karl-Henrik Larsson and Ellen Larsson sequenced Arrasia rostrata and conducted

a preliminary molecular analysis, and we are very grateful for their comments about the

molecular relationship of the new species.

Literature cited

Bodensteiner P, Binder M, Agerer R, Moncalvo JM, Hibbett DS. 2004. Phylogenetic diversity of

cyphelloid forms in the Homobasidiomycetes. Molecular Phylogenetics and Evolution 33(2):

501-515. http://dx.doi.org/10.1016/j.ympev.2004.06.007

CI [Conservation International] 2007. Biodiversity hotspots. (accessed online 09.Jun.2011).

http://www. biodiversityhotspots.org/xp/hotspots/mediterranean/Pages/default.aspx

Ghobad-Nejhad M, Nilsson RH, Hallenberg N. 2010. Phylogeny and taxonomy of the genus Vuille

minia (Basidiomycota) based on molecular and morphological evidence, with new insights into

the Corticiales. Taxon 59: 1519-1534. http://dx.doi.org/10.1007/s11557-010-0674-5

Gorjén SP, Greslebin AG, Rajchenberg M. 2011. Dendrothele latenavicularis sp. nov. (Lachnellaceae,

Basidiomycota) from the Patagonian Andes. Mycotaxon 117: 101-108

http://dx.doi.org/10.5248/117.101

Goranova G. 2003. Phylogenetic analyses of rDNA sequences indicate the corticioid genus

Dendrothele is highly polyphyletic. Master of Arts, Clark University, Worcester

Goranova G, Binder M, Hibbett DS. 2003. Molecular phylogenetics indicate that the corticioid

genus Dendrothele is highly polyphyletic. Inoculum 54: 22.

Nakasone KK, Burdsall HH Jr. 2011. The genus Dendrothele (Agaricales, Basidiomycota) in New

Zealand. New Zealand Journal of Botany 49(1): 107-131.

http://dx.doi.org/10.1080/0028825X.2010.512636

MY COTAXON

http://dx.doi.org/10.5248/118.265

Volume 118, pp. 265-272 October-December 2011

Cortinarius mikedavisii sp. nov. from northern California

DIMITAR BOJANTCHEV

MushroomHobby.com, 345 Shipwatch Lane, Hercules, CA 94547, USA

CORRESPONDENCE TO: dimitar@pontix.com

ABSTRACT — A new Cortinarius (subg. Phlegmacium) species is described from northern

California. Cortinarius mikedavisii is a brightly colored bulbopodium in sect. Laeticolores.

Phylogenetically it belongs in the /cupreorufus clade.

Key Worps — Cortinariaceae, fungal taxonomy, nrITS data

Introduction

Cortinarius sect. Laeticolores M.M. Moser ex Moénne-Locc. & Reumaux

was erected to accommodate some of the brightly colored bulbopodiums of

subg. Phlegmacium. The section type was originally designated as “Cortinarius

orichalceus” (a misapplied name, not conspecific with Agaricus orichalceus

Batsch); its valid name is Cortinarius cupreorufus Brandrud (Brandrud et al.

1994).

Recent molecular studies (Fraslev et al. 2007, Garnica et al. 2009), congruent

with my own phylogenetic analyses, show that the broad circumscription of

sect. Laeticolores by Moser (1960) and Bidaud et al. (2004) is polyphyletic.

However, a group of species around C. cupreorufus forms a well-supported

clade including C. rufo-olivaceus (Pers.) Fr. and C. prasinus (Schaeff.) Fr. The

/cupreorufus clade, which represents the core of sect. Laeticolores, is well

represented in western North America with Cortinarius mikedavisii, described

here as new, and other undescribed species (Fic 1).

Materials & methods

Methods for morphological studies and DNA extraction, PCR conditions and

primers, PCR product clean up, and sequencing were employed as outlined in Bojantchev

& Davis (2011). Color codes follow Munsell™ soil color charts (Anonymous 2000).

Collections are stored in the private herbarium of the first author or at the University of

California herbarium in Berkeley (UC) where noted.

266 ... Bojantchev

C. prasinus (AY174835) Germany

C. prasinus (AY174843) Germany

C. prasinus (DQ663390) Denmark

C. rufoolivaceus (AY174845) Germany

C. rufoolivaceus (EU088219) Germany

C. rufoolivaceus (DQ663410) Denmark

100) Cortinarius mikedavisii DBB40100 (JF795384) California

Cortinarius mikedavisii DBB12455 (JF795389) California

Cortinarius sp. RMD101203 (JF8&28732) California

57| C. aff. cupreorufus (FJ717580) British Columbia, Canada

C. aff. cupreorufus (FJ039645) British Columbia, Canada

C. cupreorufus (JF828733) Bulgaria

C. cupreorufus (AY174831) Austria

C. cupreorufus (EU056992) Germany

C. piceae (EU056956) Europe

C. sodagnitus (AY174829) Germany

apesa snynsioaidn3/

Fic 1. Phylogenetic tree inferred by maximum parsimony analysis of 16 Cortinarius subg.

Phlegmacium nrITS sequences. The tree shows the position of C. mikedavisii relative to its

closest neighbors in the /cupreorufus clade. The percentages of clustered replicate trees based

on the bootstrap test (1000 replicates) are shown above the branches while the branch lengths

representing estimated nucleotide substitutions are shown below. GenBank accession numbers

are enclosed in brackets.

PHYLOGENETIC ANALYSIS—During these studies all Cortinarius nrDNA sequences from

the public databases GenBank (http://www.ncbi.nlm.nih.gov) and UNITE (http://unite.

ut.ee/) were downloaded and reviewed. Preliminary phylogenetic analysis (not shown)

of 1012 Phlegmacium nrITS sequences from the northern hemisphere, including

312 sequences from the author's own collections, clarified the closest relatives of

C. mikedavisii and resolved its position within the /cupreorufus clade of the calochroid

super-clade as defined by Froslev et al. (2007) and Garnica et al. (2009).

A set of sixteen Phlegmacium sequences representing eight taxa was selected for

a detailed phylogenetic analysis. Twelve sequences were sourced from GenBank and

four sequences are from the author's collections. Fourteen sequences were from the

/cupreorufus clade, represented by three well known European taxa combined with

C. mikedavisii and two western North American taxa close to C. cupreorufus. Cortinarius

sodagnitus Rob. Henry and C. piceae Froslev et al. {formerly known as C. calochrous

var. coniferarum (M.M. Moser) Nezdojm.} were selected as the outgroup because they

represent the core of the calochroid cortinarii but are distant from the /cupreorufus

clade.

Cortinarius mikedavisii sp. nov. (California) ... 267

Sequence alignments were generated with MAFFT v6.821b (Katoh et al. 2002) with

the G-INS-i global alignment iterative refinement strategy. Minimal gap opening and

extension penalties were set for better resolution of the more variables sectors within the

nrITS. The alignments were visually inspected and corrected where needed.

The evolutionary history was inferred using the Maximum Parsimony method as

implemented by MEGAS (Tamura et al. 2007). The MP trees were generated by the

Close-Neighbor-Interchange algorithm with search level 0 in which the initial trees

were obtained with the random addition of sequences (10 replicates). The percentage

of replicate trees in which the associated taxa clustered together was calculated from

a bootstrap test with 1000 replicates. The search resulted in thirty most parsimonious

trees (length = 97), which differed mainly in the topology of the terminal nodes. One

tree is given in Fie. 1.

Taxonomy

Fic 2. Cortinarius mikedavisii (UC 1860820, holotype).

Cortinarius mikedavisii Bojantchev, sp. nov. FIGs 2-6

MycoBank MB 561221

Pileo 60-220 mm lato, hemispherico, dein planoconvexo, planoconcavo, glutinoso,

roseobrunneo, cum KOH purpureobrunneo, margine involuto. Lamellis emarginatis,

caesiis dein flavidis, purpureobrunneis, in statu senili. Stipite 50-140 mm longo, cylindrico,

bulbo marginato, 30-50 mm lato. Velo universale roseobrunneo. Velo partiale copioso,

flavido. Carne caesio dein albido. Sapore miti. Sporis 10-13 x 6-7 um, amygdaliformibus,

verrucosis, basidiis 28-42 x 8-9 um, tetrasporigeris, fibulis praesentibus.

Type: USA. California: Mendocino County, Caspar, Caspar Cemetery, under Picea

sitchensis, 27 Nov 2010, Bojantchev DBB40100 (Holotype UC 1860820; Genbank nrITS

JF795384).

268 ... Bojantchev

Erymo.oey: In honor of Prof. Mike Davis whose encouragement and support in the

area of molecular genetics made these Cortinarius studies possible.

STATURE pileocarpous bulbopodium. PILEus 60-220 mm diam., hemispherical

to convex when young, plano-convex to plano-concave at age; margin involute

then straight; colors intense, mostly reddish to orange brown on the disk,

varying from carmine brown to rose brown (10R 3/6-4/8), paler on the margin,

copper brown to sulphur yellow (10R 6/8-7.5YR 7/8); surface very glutinous

when wet, glabrous to dull glossy when dry. LAMELLAE crowded, 10-22 mm

broad, blue (GLEY2 6/10G-8/10G) at first, turning olive (SY 6/4-8/4) then

yellowish clay (SY 8/6-8/8) with purplish tinges, then purplish brown as the

spores mature; edge even; attachment sinuate; lamellulae abundant. STIPE

50-140 mm long, 15-30 mm wide, cylindrical to subclavate above the bulb,

pale yellow, with rose brown streaks from universal veil. BuLB 30-80 mm diam

at the widest point, well-developed, abruptly emarginate, tapering below, rose

to purplish brown around margin; the subterrestrial part with a pale to sulphur

yellow cottony mycelial felt and rhizomorphs. UNIVERSAL VEIL rose to purplish

brown, leaving copious remnants on the pileus and bulb margin. CorTINA

yellow, leaving an annular zone of dense fibrils on the stipe. CONTEXT white to

bluish at first, paling to white at maturity with yellow tinges in the bulb. ODoR

earthy. Taste mild, earthy. MACROCHEMICAL REACTIONS 5% KOH strong and

complex, dark purple red on the pileus; on the context pale olive at first, soon

lilac on pileus context, strongly purplish on bulb context and paler on stipe

context. SPoRE Deposit deep rusty brown.

BASIDIOSPORES (9.5—)10.5-13.2(-15.5) x (5.7—)6.2-7.2(-7.8) um (mean 11.5

x 6.7 um), Q = 1.61-1.78, Q. = 1.72 (N = 178, 7 basidiomata, four collections),

amygdaliform, some with apical papilla, strongly verrucose. BAsip1a 28-42

x 8-9 um, 4-spored, cylindro-clavate, clamped. GILL EDGE sparsely fertile.

Fic 3. Cortinarius mikedavisii. a) Basidiospores (UC 1860820, holotype) b) Cuticle hyphae with an

olive to purplish intracellular pigment in 5% KOH (UC 1860820, holotype)

Cortinarius mikedavisii sp. nov. (California) ... 269

Fic 4. Cortinarius mikedavisii. 5% KOH reaction a) UC 1860820, holotype b) DBB12455

CysTIDIA not observed. PILEIPELLIS an ixocutis, simplex, no hypodermium

detected, composed of parallel to interwoven hyphae in a dense gelatinous

matrix 230-300 um thick, made up of 4-12 um wide, irregular hyphae, with

greenish to purplish intracellular pigment when mounted in 5% KOH. No

distinct reactions to Melzer’s reagent were observed. CLAMP CONNECTIONS

common in all parts.

HABITAT AND DISTRIBUTION — Cortinarius mikedavisii appears to be an

uncommon species. Currently it is known from only three locations, at two of

which it fruits regularly. The habitat is mixed woods with Sitka spruce (Picea

sitchensis (Bong.) Carriére) being the one common symbiont in all locations.

The author has never seen it south of Mendocino. Its distribution appears to

be limited to northern California and southern Oregon, where it was collected

once. There are no matching records from Washington and British Columbia

despite the intensive collecting and molecular cataloguing that has taken place

in these regions.

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: MENDOCINO COUNTY,

Caspar, Caspar Cemetery, under Picea sitchensis, 23 Dec 2008, Bojantchev DBB12455,

(UC 1860821, Genbank nrITS JF795389); Jackson State Demonstration Forest, under

Picea sitchensis, 22 Nov 2009, Bojantchev DBB28043; OREGON: CurRY CounrTy, Samuel

Boardman State Park, under Picea sitchensis, 11 Nov 2009, Bojantchev DBB27700.

Discussion

Cortinarius mikedavisii is a beautiful species that so stands out with its large

size and bright colors that it should present no difficulty for field identification.

The distinctive alkaline color reaction indicates the presence of rufoolivacin,

an anthraquinone compound, which Steglich & Oertel (1985) detected in other

members of the /cupreorufus clade.

Phylogenetically, C. mikedavisii (Fic 1) stands almost equidistant in the

clade between the two main branches of C. cupreorufus and C. rufo-olivaceus.

Cortinarius rufo-olivaceus, a beech associate from Europe, is not known from

270 ... Bojantchev

Fic 5. Cortinarius mikedavisii. a) Mature basidiomata can become very large (DBB12455). The

scale is a US quarter dollar coin (~25mm) b) Basidiomata in a button stage (DBB28043)

/ ye REE

4 if / Z

| vy ;

y ? ae E™ , ‘4

es,

ZAI

- * . ers

4 >

Fic 6. Cortinarius mikedavisii — a study in the development of a pileocarpous bulbopodium. Note

the changing color of the gills. a) DBB28043 b) DBB28043 c) DBB27700

Ds ne" “ . x q < r

‘ 4 4 “50 . ae v ‘ Lae

ee wa es, A ov ‘ =

— GN ra ‘ Ob Sad) ‘ ¥ ; \

US ee es , aI

Fic 7. Cortinarius mikedavisii. a) DBB12455. The scale is a US quarter dollar coin (~25mm) b)

Yellow mycelial strands (DBB27700)

western North America. Cortinarius cupreorufus (Fic 8a) and its western

American ally differ in the straw yellow lamellae and generally more pale olive

pileus, colors noticeable in all stages of development. The alkaline reactions in

C. mikedavisii are more intense than seen in C. cupreorufus, and the context

turns lilac to purplish red immediately, bypassing the pale olive stage. There is

Cortinarius mikedavisii sp. nov. (California) ... 271

Fic 8. Related species: a) Cortinarius cupreorufus DBB19840 from Europe; b) Cortinarius

sp. RMD101203, an undescribed species from the xeric oak stands of central and southern

California.

an undescribed species in the same clade, represented by coll. “RMD101203”

in Fic. 1, which is only found in xeric oak stands of the central valley and

southern California and differs significantly in colors (Fic. 8b). All members

of the /cupreorufus clade share large (10-13 um) amygdaloid spores with very

rough ornamentation.

Complete iconography of C. mikedavisii and a comparative image study is

available on http://www.mushroomhobby.com.

Acknowledgements

The author is grateful to Dr. Else C. Vellinga and Dr. Boris Assyov for their

presubmission reviews and comments. Dr. Shaun Pennycook was very helpful in

answering nomenclatural questions during the preparation of this manuscript. Boris

Assyov reviewed and corrected the Latin diagnosis.

Literature cited

Anonymous. 2000. Munsell” soil color charts, revised edition. Munsell Color, New Windsor, NY.

Bidaud A, Carteret X, Eyssartier G, Moénne-Loccoz P, Reumaux P. 2004. Atlas des Cortinaires

Vol. 14. Editions Fédération mycologique Dauphiné-Savoie, Lyon.

Bojantchev D, Davis RM. 2011 Cortinarius xanthodryophilus sp. nov. — a common Phlegmacium

under oaks in California. Mycotaxon 116: 317-328. http://dx.doi.org/10.5248/116.317

Brandrud TE, Lindstrém H, Marklund H, Melot J, Muskos S. 1994. Cortinarius Flora Photographica

Vol. 3. Cortinarius HB, Matfords, Sweden.

Froslev TG, Jeppesen TS, Lzessge T, Kjoller R. 2007. Molecular phylogenetics and delimitation of

species in Cortinarius section Calochroi (Basidiomycota, Agaricales) in Europe. Mol. Phylogenet.

Evol. 44: 217-227. http://dx.doi.org/10.1016/j.ympev.2006.11.013

Garnica S, Weif’ M, Oertel B, Ammirati J, Oberwinkler F. 2009. Phylogenetic relationships in

Cortinarius, section Calochroi, inferred from nuclear DNA sequences. BMC Evol. Biol. 9(1):

[1]-[17]. http://dx.doi.org/10.1186/1471-2148-9-1

Katoh K, Misawa K, Kuma KI, Miyata T. 2002. MAFFT: a novel method for rapid multiple

sequence alignment based on fast Fourier transform. Nucleic Acids Res. 30: 3059-3066.

http://dx.doi.org/10.1093/nar/gkf436

272 ... Bojantchev

Moser MM. 1960. Die Gattung Phlegmacium (Schleimk6pfe). Die Pilze Mitteleuropas, Vol. 4.

J. Klinkhart, Bad Heilbrunn.

Steglich W, Oertel B. 1985. Untersuchungen zur Konstitution und Verbreitung der Farbstoffe von

Cortinarius, Untergattung Phlegmacium (Agaricales). Sydowia 37: 284-295.

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis

(MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596-1599.

http://dx.doi.org/10.1093/molbev/msm092

MY COTAXON

http://dx.doi.org/10.5248/118.273

Volume 118, pp. 273-282 October-December 2011

Observations on gasteroid Agaricomycetes

from the Brazilian Amazon rainforest

LARISSA TRIERVEILER-PEREIRA™, ALLYNE CHRISTINA GOMES-SILVA?

& IURI GOULART BASEIA?

"Departamento de Micologia, Universidade Federal de Pernambuco,

Av. Nelson Chaves s/n, Recife -PE, 50670-420, Brazil

?Departamento de Botanica, Ecologia e Zoologia, Universidade Federal do Rio Grande do Norte,

Campus Universitario, Natal - RN, 59072-970, Brazil

*CORRESPONDENCE TO: /t_pereira@yahoo.com.br

ABSTRACT — Field trips carried out in an indigenous protected area in the states of Rondénia

and Mato Grosso (Brazil) in 2009 revealed new records: Geastrum albonigrum, new from

South America; and Geastrum lageniforme, Mutinus caninus, and Tulostoma exasperatum,

new from the Brazilian Amazon rainforest. Descriptions, photographs, and line drawings of

the specimens are presented.

Key worps — Basidiomycota, fungal taxonomy, gasteromycetes, neotropical mycota

Introduction

The Amazon rainforest is recognized as one of the areas with higher

biodiversity of the globe. Interesting data on macrofungal taxonomy from

the Brazilian Amazon rainforest have been published recently by Desjardin &

Braga-Neto (2007), Martins-Junior et al. (2008), Gibertoni (2008), Gibertoni et

al. (2008), Gomes-Silva et al. (2008, 2009, 2010a,b, 2011), Sotao et al. (2008),

Gomes-Silva & Gibertoni (2009), Trierveiler-Pereira et al. (2009a), and Baltazar

et al. (2010). However, gasteromycete diversity is poorly known in this Brazilian

ecosystem.

Although more than two hundred species of gasteroid fungi have been

acknowledged from Brazil (Trierveiler-Pereira & Baseia 2009), fewer than

fifteen species are known to occur in the Amazon rainforest, an unexplored

highly diverse area.

Species of gasteroid fungi previously recorded from the Brazilian Amazon

forest are listed in Trierveiler-Pereira et al. (2009a), including the description of

the new species, Cyathus amazonicus Trierveiler-Pereira & Baseia.

274 ... Trierveiler-Pereira, Gomes-Silva & Baseia

“66° =AMAZONAS “60°

PORTO VELHO

RONDONIA

Fic. 1. Location of the “Terra Indigena Sete de Setembro’ (hatched area)

in the states of Rondénia and Mato Grosso (Brazil).

Materials & methods

Fungi were collected along pre-existing trails in a Brazilian indigenous protected area

called “Terra Indigena Sete de Setembro’ (TISS). This territory shelters ten indigenous

tribes distributed in the states of Rondénia (RO) and Mato Grosso (MT) (Fie. 1). In this

area there are about 1200 indigenous individuals from the ethnic group of Paiter-Surui.

The TISS extends over approximately 249 thousand hectares and its territory is mostly

covered by the Amazonian dense ombrophilous forest (RADAMBRASIL 1978, Ferronato

& Nunes 2010).

Field trips were carried out from May 28" to June 06" 2009 in the tribal areas

‘Sertanista Apoena Meirelles’ (Rondolandia, MT; 10°10'15"S 59°28'14"W) and

‘Lapetanha’ (Cacoal, RO; 11°26'19"S 61°26'50"W).

The collected basidiomata were placed in paper bags or plastic containers prior to

analysis. Macroscopic descriptions are based on observations of fresh and dried material

(Miller & Miller 1988). Colors are coded according to Kornerup & Wanscher (1978).

Microscopic observations of tissues from dried specimens mounted in 5% KOH were

made under a light microscope.

Gasteroids from the Amazon (Brazil) ... 275

Voucher specimens were deposited in the Herbarium HFSL and duplicates were sent

to Herbarium URM. Additional specimens deposited in MA-Fungi, ICN, and MBM

were also examined. Gasteromycete specimens cited in Capelari & Maziero (1988) were

borrowed from the Herbarium SP to confirm identification. Herbarium acronyms are

according to Thiers (2011).

Taxonomy

Geastrum albonigrum Calonge & M. Mata, Bol. Soc. Micol. Madrid

28: 332. 2004. FIG 2, 4A

SPECIMENS EXAMINED: BRAZIL. Mato Grosso, RONDOLANDIA, Terra Indigena Sete

de Setembro, Aldeia Apoena Meirelles, 30.V.2009, leg. Gomes-Silva et al. 209 (URM

82098).

IMMATURE BASIDIOMATA epigeous, globose to ovoid, 2.1 cm high x 1.5 cm

broad, grayish brown (4C5) to yellowish brown (5F5), hirsute, with a single

robust rhizomorph attached at the base, rhizomorph ramified or not, 0.6-1.8

cm length, with debris strongly attached. EXPANDED BASIDIOMATA 0.7-1.9 cm

high x 2-3.4 cm broad. ExopEeRipIuM non-hygroscopic, split into 6-7 rays,

saccate or with rays becoming involute, more rarely arched; external layer

hirsute, grayish brown (4C5) to yellowish brown (5F5), peeling off at maturity;

fleshy layer persistent, brown (6E5) to grayish brown (6E3), with a purplish

shade. ENDOPERIDIUM globose to depressed globose, 0.7-1.6 cm high x

1.2-1.6 cm broad, brown (6E6), sessile, without apophysis; peristome fibrillose,

not delimited, slightly darker than endoperidium. GLEBa pulverulent at

maturity, dark brown (6F4).

BASIDIOSPORES globose, 4-4.5 um diam. (including ornamentation),

yellowish brown to dark brown in KOH, ornamented with short columns.

CAPILLITIAL HYPHAE straight, thick-walled, with narrow lumen, pale brown to

yellowish brown in KOH, 3-6 um diam., not branched.

ECOLOGY & DISTRIBUTION: Gregarious on rotten wood, without subiculum.

The Brazilian material represents the first record from South America.

Previously known from Costa Rica, Mexico, and Panama. It is possible that

the species occurs in neotropical rainforests and warmer regions of temperate

America.

ADDITIONAL SPECIMENS EXAMINED: COSTA RICA. GUANACASTE, TEMPISQUE,

Parque Nacional Palo Verde, 27.1X.2003, leg. I. Lopez 4869 (MA-Fungi 59260, isotype);

20.X.2004, leg. I. Lopez 6277 (MA-Fungi 65423); MEXICO. Curapas, Tapachula,

03.X.1993, leg. G. Guzman 30743 (MA-Fungi 59258); PANAMA. IsLa DE Corsas,

Cerro de la X, 14.XII.1996, leg. Pando & Nunez (MA-Fungi 36140-2).

REMARKS: Geastrum albonigrum is characterized by an epigeous hirsute

form when young, a saccate or involute exoperidium that peels off in age, a

sessile endoperidium, and a non-delimited fibrillose peristome. When the

dark hirsute external exoperidial layer peels off, it reveals a white fibrous layer

276 ... Trierveiler-Pereira, Gomes-Silva & Baseia

where the rhizomorph remains attached. The fleshy layer is purplish and the

endoperidium is dark.

Brazilian material differs from the holotype in spore measurements (3-5 um

in the original description) and the presence of a narrow lumen in the capillitial

threads (absent in the material described by Calonge & Mata 2004).

Geastrum albonigrum belongs to the group of epigeous xylophilous Geastrum

species, such as G. schweinitzii (Berk. & M.A. Curtis) Zeller, G. hirsutum Baseia

& Calonge, and G. javanicum Lév., all of which differ by forming a white

subiculum over the substrata (Ponce de Leén 1968, Baseia et al. 2003, Baseia &

Calonge 2006), a character not seen in G. albonigrum.

Geastrum hirsutum also has a hirsute exoperidium that peels off at maturity,

but lacks a robust rhizomorph, has a delimited peristome, and a light-colored

endoperidium and fleshy layer. Similar to G. hirsutum, G. schweinitzii produces

smaller basidiomata lacking a hirsute external layer. Geastrum javanicum has

no hirsute external layer and it rarely peels off completely (Trierveiler-Pereira

et al. 2011).

Geastrum lageniforme Vittad., Monogr. Lycoperd.: 16. 1842. FIG 3A, 4B

SPECIMENS EXAMINED: BRAZIL. Mato Grosso, RONDOLANDIA, Terra Indigena Sete

de Setembro, Aldeia Apoena Meirelles, 30.V.2009, leg. Gomes-Silva et al. 206, 222 (URM

82099, 82100).

IMMATURE BASIDIOMATA not observed. EXPANDED BASIDIOME 0.7-1.0 cm

high x 2.3-3.0 cm diam. Exoperipium non-hygroscopic, split into 6-8 rays,

saccate, rays long, slender, external layer glabrous, with longitudinal ridges,

grayish yellow (1B5) to blond (4C4); fleshy layer persistent, brownish gray

(6C2). ENDOPERIDIUM globose, 0.7-0.9 cm high x 1.0-1.2 cm broad, grayish

beige (4C2) to brownish gray (5D2), sessile, without apophysis; peristome

fibrillose, grayish brown (5E3), delimited by a whitish line. GLEBA pulverulent

at maturity, brownish gray (5E2).

BASIDIOSPORES globose, 3.5-5 um diam. including the ornamentation,

yellowish brown in KOH, ornamentation columnar. CAPILLITIAL HYPHAE

straight to more or less sinuous, slightly thick-walled, with narrow lumen,

yellowish in KOH, 2.5-8.0 um diam., not branched.

ECOLOGY & DISTRIBUTION: Solitary on forest soil. Widespread (Calonge et

al. 2004). In Brazil, the species has been reported from the states of Rio Grande

do Sul (Rick 1961, Cortez et al. 2008), Rio de Janeiro (Hennings 1904), Bahia

(Trierveiler-Pereira et al. 2009b), and Pernambuco (Trierveiler-Pereira et al.

2011). This is the first record from the Brazilian Amazon rainforest.

ADDITIONAL SPECIMEN EXAMINED: SPAIN. BURGOS, QUINTANA DEL PIDIO, 22.X1.1991,

leg. L.A. Parra (MA-Fungi 30752).

REMARKS: Geastrum lageniforme resembles G. saccatum Fr. in a saccate

exoperidium, sessile endoperidium, and fibrillose delimited peristome (Trierveiler-

Gasteroids from the Amazon (Brazil) ... 277

Fic. 2. Geastrum albonigrum.

A. Mature and immature basidiomata in situ.

B. Basidiomata in different stages of development.

Pereira et al. 2011). According to Sunhede (1989), the two species can be

distinguished by the presence of clamped hyphae in the external mycelial layer,

which occur only in G. lageniforme.

278 ... Trierveiler-Pereira, Gomes-Silva & Baseia

Ponce de Leon (1968) considered G. lageniforme a synonym of G. indicum

(Klotzsch) Rauschert (= G. triplex Jungh.), but there are characteristics that can

separate these two species, since G. triplex is usually larger, with involute rays,

and forms a fleshy collar around the endoperidium.

Mutinus caninus (Huds.) Fr., Summa Veg. Scand. 2: 434. 1849. FIG 3C, 4C

SPECIMEN EXAMINED: BRAZIL. RONDONIA, CACOAL, Terra Indigena Sete de Setembro,

Aldeia Lapetanha, 05.VI.2009, leg. Gomes-Silva et al. 126 (URM 82101).

BASIDIOME 5.5 cm high, with a whitish rhizomorph attached at the base,

rhizomorph ramified, up to 2.2 cm long. VoLva saccate, 1.6 cm high x

1 cm broad, yellowish white (1A2). PseuDosTIPE cylindrical, tapering at the

apex, 4.1 cm high x 0.7 cm broad, spongy, light orange (5A4) at the base and

becoming reddish towards the apex, fertile region reddish orange (7B8). GLEBA

mucilaginous, olive brown (4F5), covering the fertile region.

Basiprosporgs cylindrical, 4.5-5.5 x 1.5 um, hyaline to greenish in KOH,

smooth, biguttulate.

ECOLOGY & DISTRIBUTION: Solitary on forest soil. Cosmopolitan, but rare in

the tropics (Baseia et al. 2006). In Brazil the species was reported only from Rio

Grande do Norte (Baseia et al. 2006). This is the first record from the Brazilian

Amazon rainforest.

ADDITIONAL SPECIMENS EXAMINED: Mutinus caninus — SPAIN. GUIPUZCOA, SAN

SEBASTIAN, 16.X.1976, leg. Sociedad de Ciencias Naturales Arazandi (MA-Fungi 22361).

Mutinus elegans - SPAIN. LuGo, CHANTADA, 03.X1.1999, leg. M.T. Jacobo s/n (MA-

Fungi 42064); BRAZIL. Rio GRANDE DO SUL, VIAMAO, Parque Estadual de Itapua,

22.V.2004, leg. V.G. Cortez 016/04 (ICN 139004).

REMARKS: Mutinus caninus is similar to M. elegans (Mont.) E. Fisch. but forms

a well delimited gleba on the apical zone; the glebal zone is not defined in

M. elegans (Calonge 1996). The pseudostipe in the examined material is

yellowish, but according to Calonge (1996) it may also be whitish or pinkish.

Zeller (1944) described a white variant of this species, M. caninus var. albus

Zeller.

Tulostoma exasperatum Mont., Ann. Sci. Nat., Bot., Sér. 2, 8: 362. 1837. FIG 3B, 4D

SPECIMENS EXAMINED: BRAZIL. RONDONIA, CACOAL, Terra Indigena Sete de Setembro,

Aldeia Lapetanha, 06.VI.2009, leg. Gomes-Silva et al. 100, 145 (URM 82102, 82103).

BASIDIOMATA 1.1-7.0 cm high. SporE sac globose to depressed-globose,

0.6-0.8 cm high x 1.8-2.2 cm broad. Exoperidium spiny, light brown (5E7),

peeling off at maturity. Endoperidium reticulate, papery, yellowish white (2A2)

to pale yellow (4A3); peristome conical, slightly lighter than endoperidium,

fibrillose, delimited. GLEBA dull yellow (3B3). Stipe 0.9-6.1 cm high x 0.2-0.25

cm diam., light brown (5E7), with longitudinally arranged scales.

Gasteroids from the Amazon (Brazil) ... 279

Fic. 3. Gasteroid species from the Brazilian Amazon rainforest.

A. Geastrum lageniforme. B. Tulostoma exasperatum. C. Mutinus caninus.

BasIpiosPorEs globose to subglobose, 6-7.5 um diam., yellowish in KOH,

with a columnar-reticulate ornamentation. CAPILLITIAL HYPHAE straight to

tortuous, thick-walled, swollen at the septa, branched, light yellow in KOH,

4-7 um diam.

ECOLOGY & DISTRIBUTION: Gregarious on rotten wood. Widely distributed

in tropical and subtropical regions (Wright 1987). In Brazil the species is known

from Rio Grande do Sul (Rick 1961, Cortez et al. 2009), Parana (Meijer 2006),

Paraiba (Silva et al. 2007), Sao Paulo and Pernambuco (Baseia & Milanez 2002).

This is the first record from the Brazilian Amazon rainforest.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. RIO GRANDE DO SUL, VIAMAO, Parque

Estadual de Itapua, 25.VI1.2005, leg. R.M. Silveira 456 (ICN 154635); PARANA, TIBAGI,

Quartela, 06.1X.1992, leg. A.A.R. de Meijer 2341 (MBM).

REMARKS: Among the 14 Tulostoma species recorded from Brazil (Baseia &

Milanez 2002, Meijer 2006, Silva et al. 2007, Cortez et al. 2009), T: exasperatum

is the only one with a lignicolous habit. Tulostoma species usually occur on

sandy soil, pastures, humus, forest soil and even flooded areas (Wright 1987).

280 ... Trierveiler-Pereira, Gomes-Silva & Baseia

Fic. 4. Line drawings of microscopic elements.

A. Geastrum albonigrum: capillitial threads and spores.

B. G. lageniforme: capillitial threads and spores. C. Mutinus caninus: spores.

D. Tulostoma exasperatum: capillitial threads and spores (scale bar = 10 um).

Specimens kept in SP

We reviewed and confirmed the identities of the following gasteromycetes

previously reported from the Brazilian Amazon rainforest by Capelari &

Maziero (1988): SP 211544, SP 211545, SP 211547, and SP 211548 as Cyathus

montagnei Tul. & C. Tul.; SP 214636 as C. limbatus Tul. & C. Tul.; and SP 211223

and 211526 as Morganella fuliginea (Berk. & M.A. Curtis) Kreisel & Dring. The

immature basidiomata in SP 211549 could not be identified morphologically.

We did not review SP 211527 (labelled as Morganella sp.) or SP 211301 (labelled

as Lycoperdon sp.).

Acknowledgments

We thank the people of Paiter; Associacao Metareila do Povo Indigena Surut’;; ‘Equipe

de Conservacao da Amazénia - ACT Brasil’; ‘Associagao de Defesa Etnoambiental

Kanindé and ‘Fundacao Nacional do Indio (FUNAI)’ The United States Agency for

International Development (USAID) is acknowledged for financial support. We

also would like to thank the curators of SP, ICN and MBM for specimen loans and

Gasteroids from the Amazon (Brazil) ... 281

Dr. Francisco D. Calonge (CSIC, RJB de Madrid) for the access to the MA-Fungi

exsiccata. LTP acknowledges PROPESQ (‘Pré-Reitoria para Assuntos de Pesquisa e Pés-

Graduacao, UFPE) and PPGBF (‘Programa de Pés-Graduacao em Biologia de Fungos,

UFPE) for financial support. We are grateful to Admir J. Giachini (Universidade Federal

de Santa Catarina, Brazil) and André A.R. de Meijer (Parana, Brazil) for critically reading

the manuscript.

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MY COTAXON

http://dx.doi.org/10.5248/118.283

Volume 118, pp. 283-288 October-December 2011

Septobasidium saurauiae sp. nov. (Septobasidiaceae)

and S. pseudopedicellatum new to China

SUZHEN CHEN?” & LIN Guo’*

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

Chinese Academy of Sciences, Beijing 100101, China

?Ocean University of China, Qingdao 266003, China

CORRESPONDENCE TO *: Chensz2009@gmail.com & *guol@im.ac.cn

ABSTRACT—A new species, Septobasidium saurauiae on Saurauia tristyla associated with

Chionaspis sp., and a new Chinese record, Septobasidium pseudopedicellatum on Schefflera

octophylla and Psychotria serpens, are described. They were collected from Hainan Province.

Key worps—Pucciniomycetes, Septobasidiales, taxonomy

Hainan Island is located in the South China Sea. It is a tropical area with a

rich fungal diversity. To date, six new species and one new Chinese record of

Septobasidium have been discovered from this Province (Chen & Guo 2011b,

Lu & Guo 2009a, 2010b,c). An additional new species and a new Chinese record

of Septobasidium are reported as follows:

Septobasidium saurauiae S.Z. Chen & L. Guo, sp. nov. Fics. 1-7

MycoBank MB 561871

Basidiomata resupinata, discontinue crescentia, 6-7 cm longa, 3-4 cm lata, cinnamomeo-

brunnea vel griseo-brunnea, margine determinata, superficie laevia, saepe protuberationibus

rotundis praedita, in sectione 900-1330 um crassa. Subiculum brunneum, 30-50 um

crassum, ab subiculo stratum hypharum vel curtam columnam formans. Columnae

brunneae, 20-50 ym altae, 20-70 um latae. Strata hyphararum 780-970 um alta. Ab

strato hymenii hyphae saepe repullulantes tum stratum hypharum et hymenium secundum

formantes. Stratum hypharum secundum 200-240 um altum. Hymenium hyalinum,

40-50 um crassum. Basidia cylindrica, recta vel leviter curvata, 4-cellularia, 42-50 x 6-8

um, hyalina. Sine probasidio. Basidiosporae 10-25 x 4-6 um, hyalinae. Haustoria hyalina,

ex hyphis irregulariter spiralibus constantia.

Type: China, Hainan Province, Limu Mountain, alt. 529 m, on Saurauia tristyla DC.

(Actinidiaceae), associated with Chionaspis sp. (Diaspididae), 23.X1.2010, Y.R Zhu &

FE. He 520 (holotype, HMAS 263145 ).

284 ... Chen & Guo

1 A

- /

10 pm

Fic. 1. Basidia and basidiospores of Septobasidium saurauiae (HMAS 263145, holotype).

Basidiomata on branches, resupinate, growing discontinuously, 6-7 cm long,

3-4 cm wide, cinnamon-brown or grey-brown; margin determinate, surface

smooth, frequently with round protuberance. In section 900-1330 um thick.

Subiculum brown, 30-50 um thick. Forming hyphal layer or short pillars.

Pillars brown, 20-50 um high, 20-70 um wide. Hyphal layer 780-970 um

high. From hymenial layer the fungal hyphae often renew growth to form the

second hyphal layer and hymenium. Second hyphal layer 200-240 um high.

Hymenium hyaline, 40-50 um thick. Basidia arising directly from the hyphae

without a probasidial cell, at first obovoid, later cylindrical, straight or slightly

curved, 4-celled, 42-50 x 6-8 um, hyaline. Basidiospores 10-25 x 4-6 um,

hyaline. Haustoria hyaline, consisting of irregularly coiled hyphae.

REMARKS: Morphologically, Septobasidium saurauiaeis similar to S. meizhouense

C.X. Lu et al., which differs in numerous cracks in the basidioma surface and

a thinner section (400-650 um high) and hyphal layer (220-440 um high)

(Lu et al. 2010).

Fics. 2-7 (right). Septobasidium saurauiae (HMAS 263145, holotype). 2. Basidiomata on branch.

3-4. Sections of basidiomata. 5-6. Basidia (arrows). 7. Haustoria.

Septobasidium saurauiae sp. nov. (China) ... 285

286 ... Chen & Guo

Fic. 8. Basidia and probasidia of Septobasidium pseudopedicellatum (HMAS 242745).

Septobasidium pseudopedicellatum Burt, Ann. Mo. bot. Gdn 3: 327, 1916.

Fics. 8-14

Basidiomata on trunks, branches and leaves, resupinate, 0.3-19 cm long,

0.2-9 cm wide, chestnut brown, brown or smoke grey; margin white,

determinate, surface smooth. The pillars near the margin are visible with naked

eyes. In section 850-1700 um thick. Subiculum white or brown, 30-50 um

thick. Pillars brown, 500-700 um high, 50-170 um wide. The pillars branch out

to form hyphal layer, 400-650 um high. From hymenial layer the fungal hyphae

often renew growth to form the second hyphal layer and hymenium, up to

3-strata. Hymenium brown, 40-60 um thick, with closely packed paraphyses

that are parallel, upright and curved at top. Probasidia hyaline or brownish,

obovoid or ellipsoidal, 16-20 x 10-16 um. Basidia cylindrical, straight or

slightly curved, 4-celled, 52-62 x 5-7.5 um, hyaline, with persisting probasidial

cell. Sterigmata conical, 2.5-8 x 2.5-3.5 um. Basidiospores not seen. Haustoria

consisting of irregularly coiled hyphae.

SPECIMENS EXAMINED: CHINA, HaINAN PROVINCE, Bawangling Natural Reserve,

Nanchahe, alt. 600 m, on Schefflera octophylla (Lour.) Harms (Araliaceae), 14.1V.2011,

L.Guo 11611, HMAS 242745; on Psychotria serpens L. (Rubiaceae), 14.IV.2011, L.Guo

11612, HMAS 251216.

Fics. 9-14 (right). Septobasidium pseudopedicellatum (HMAS 242745). 9. Basidiomata on trunk.

10-11. Sections of basidiomata. 12. Probasidia (arrows). 13. Basidium (arrow). 14. Haustoria.

Septobasidium saurauiae sp. nov. (China) ... 287

288 ... Chen & Guo

Including the two species reported in this paper, 35 Septobasidium species have

now been reported in China (Sawada 1933, Couch 1938, Teng 1963, Tai 1979,

Kirschner & Chen 2007, Lu & Guo 2009a,b,c, 2010a,b,c, 2011, Lu et al. 2010,

Chen & Guo 2011a,b).

Acknowledgements

The authors would like to express their deep thanks to Drs. Eric H.C. McKenzie

(Auckland, New Zealand) and Shuanghui He (Beijing Forestry University) for serving

as pre-submission reviewers, to Dr. Shaun Pennycook (Auckland, New Zealand) for

nomenclatural review, to Prof. Jian-Yun Zhuang (Institute of Microbiology, Chinese

Academy of Sciences) for Latin corrections, to Prof. Zhenyu Li and Mr. Ziyu Cao

(Institute of Botany, Chinese Academy of Sciences) and Mr. Qing Chen (Bawangling

Natural Reserve, Hainan Province) for identifying the host plants, to Prof. Sanan Wu

(Beijing Forestry University) for identifying the scale insect, and to Mrs. Xiangfei Zhu

for inking in line drawings. This study was supported by the Foundation of Ministry of

Science and Technology of the People’s Republic of China (No. 2006FY110500-5).

Literature cited

Chen SZ, Guo L. 2011la. Septobasidium sichuanense sp. nov. (Septobasidiaceae) from China.

Mycotaxon 115: 481-484. http://dx.doi.org/10.5248/115.481

Chen SZ, Guo L. 2011b. Septobasidium atalantiae sp. nov. (Septobasidiaceae) and S. henningsii new

to China. Mycotaxon 117: 291-296. http://dx.doi.org/10.5248/117.291

Couch JN. 1938. The genus Septobasidium. Univ. of North Carolina Press, Chapel Hill. 480 p.

Kirschner R, Chen CJ. 2007. New reports of two hypophyllous Septobasidium species from Taiwan.

Fung. Sci. 22(1,2): 39-46.

Lu CX, Guo L. 2009a. Septobasidium maesae sp. nov. (Septobasidiaceae) from China. Mycotaxon

109: 103-106. http://dx.doi.org/10.5248/109.103

Lu CX, Guo L. 2009b. Two new species of Septobasidium (Septobasidiaceae) from China. Mycotaxon

109: 477-482. http://dx.doi.org/10.5248/109.477

Lu CX, Guo L. 2009c. Septobasidium annulatum sp. nov. (Septobasidiaceae) and Septobasidium

kameii new to China. Mycotaxon 110: 239-245. http://dx.doi.org/10.5248/110.239

Lu CX, Guo L. 2010a. Three new species of Septobasidium (Septobasidiaceae) from Gaoligong

Mountains in China. Mycotaxon 112: 143-151. http://dx.doi.org/10.5248/112.143

Lu CX, Guo L. 2010b. Two new species of Septobasidium (Septobasidiaceae) and S. pallidum new to

China. Mycotaxon 113: 87-93. http://dx.doi.org/10.5248/113.87

Lu CX, Guo L. 2010c. Two new species of Septobasidium (Septobasidiaceae) from Hainan province

in China. Mycotaxon 114: 217-223. http://dx.doi.org/10.5248/114.217

Lu CX, Guo L. 2011. Two new species of Septobasidium (Septobasidiaceae) from Gaoligong

Mountains in China. Mycotaxon 116: 395-400. http://dx.doi.org/10.5248/116.395

Lu CX, Guo L, Wei JG, Li JB. 2010. Two new species of Septobasidium (Septobasidiaceae) from

southern China. Mycotaxon 111: 269-274. http://dx.doi.org/10.5248/111.269

Sawada K. 1933. Descriptive catalogue of the Formosan fungi. Part VI. Rep. Dept. Agric. Govt. Res.

Inst. Formosa 61: 1-99.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing. 1527 p.

Teng SC. 1963. Fungi of China. Science Press, Beijing. 808 p.

MY COTAXON

http://dx.doi.org/10.5248/118.289

Volume 118, pp. 289-292 October-December 2011

Hallenbergia (Agaricomycetes), a new corticioid genus

G.S. DHINGRA* & PRIYANKA

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

* CORRESPONDENCE TO: dhingragurpaul@gmail.com

ABSTRACT - A new corticioid genus, Hallenbergia, and its single species, H. singularis, are

described from Thimphu, Bhutan.

Key worps - Eastern Himalaya, Nawephu, angiosperm host

While conducting a fungal foray in Nawephu of Thimphu, Bhutan, Dhingra

made a collection on decaying angiospermous twigs. On the basis of

macroscopic and microscopic characters it was compared with similar genera

within Corticiaceae s.]. (Rattan 1977, Eriksson & Ryvarden 1976, Hjortstam et

al. 1987) but could not be assigned to any already known, hence the description

of a new genus. Morphological traits show similarities with Hypochnicium and

Intextomyces.

Hallenbergia Dhingra & Priyanka, gen. nov.

MycoBank MB 560467

Basidiocarpum resupinatum, adnatum, effusum, ceraceum; hymenium laevigatum,

farinaceum, subvitrum, continuum, rimosum in sicco; systema hyphale monomiticum;

hyphae tenuitunicatae, septatae, fibulatae; hyphae basilarae irregulariter ramosae, dense

intertextae; hyphae in subhymenio parvae-loculosae, compaginatae et apparenter cellulosae;

cystidia absum; basidia subclavatae ad suburniformes, 4-sterigmatae; basidiosporae

ellipsoidae ad ovoidae vel globosae, laeves, crassitunicatae, cyanophilae, inamyloidae.

Type SpEcIEs: Hallenbergia singularis Dhingra & Priyanka

Erymo.ocy: The name of the genus is in the honour of Prof. Nils Hallenberg.

Basidiocarp resupinate, adnate, effused, thin, ceraceous; hymenial surface

smooth, farinose under the lens, continuous, some cracks developing on

drying; margins not well differentiated. Hyphal system monomitic; generative

hyphae thin-walled, septate, clamped; basal hyphae irregularly branched and

interwoven into a dense texture; subhymenial hyphae short-celled, compactly

packed and appear like a cellular tissue. Cystidia absent. Basidia subclavate to

290 ... Dhingra & Priyanka

Com ayentan

AO / Se

aot st) SSeS

CORT .\ \ \ SN NEEEEY

ae eX

sae

anit

Pret .

| CSUMPURG TA

FO Bat \ Ni)

SAAN

“ere

(}

mT LY \ eS

Ses, wn ee \ 3

Gi A (f ( YA\ \

(

a) :

\ (i

\ \\

CSN :

HOY

-4. Hallenbergia singularis: microscopic structures:

1. basidiospores; 2. basidia; 3. generative hyphae; 4. vertical section through basidiocarp.

sterigmate. Basidiospores ellipsoid to ovoid or subglobose,

Fics 1

, with thickened walls, cyanophilous, inamyloid.

REMARKS—Hallenbergia resembles the genus Hypochnicium in having broadly

ellipsoid to subglobose basidiospores with somewhat thick and cyanophilous

walls. There are also affinities with Intextomyces in having a densely interwoven

texture. The new genus differs from both genera by the peculiar hyphal texture,

suburniform, 4-

smooth

Hallenbergia singularis gen. & sp. nov. (Bhutan) ... 291

Fic. 5. Hallenbergia singularis: basidiocarp showing hymenial surface.

with a densely and irregularly branched lower part and a pseudoparenchymatous

upper part. Moreover, it differs from Intextomyces in the way basidia are

formed. In I. contiguus (P. Karst.) J. Erikss. & Ryvarden, basidia are formed

at the apex of penetrating, sinuous hyphae, while in H. singularis the basidia

are directly produced from the surface of the pseudoparenchymatic tissue.

A sample has been studied by Prof. John Eriksson and Prof. Nils Hallenberg,

who both supported the concept of a new genus.

Hallenbergia singularis Dhingra & Priyanka, sp. nov. Fics 1-5

MycoBaAnk MB 560468

Basidiocarpum resupinatum, adnatum, effusum, ceraceum, ad 250 wm crassum;

hymenium laevigatum, farinaceum, subvitrum, continuum, rimosum in sicco; systema

hyphale monomiticum; hyphae ad 4.5 um latae, tenuitunicatae, fibulatae; hyphae

basilariae irregulariter ramosae, intricatae in textura compacta; hyphae in subhymenio

parvae-loculosae, compaginatae et appariter cellulosae; cystidia absum; basidia 15-33 x

7-10.5 um, subclavatae ad suburniformes, 4-sterigmatae; basidiosporae 6-9 x 4.5-7.5 um,

latae ellipsoidae ad ovoidae vel globosae, laeve, crassitunicatae, cyanophilae, inamyloidae,

un-i vel multiguttatae.

Type: Bhutan, Thimphu, Nawephu, on decaying angiospermous twigs, 31 July 1981,

Dhingra 19548 (PAN, holotype).

EryMoLocy: The epithet refers to the uncommon and strange combination of

microscopic features.

Basidiocarp resupinate, adnate, effused, thin, up to 250 um thick in section,

ceraceous; hymenial surface smooth to farinose under the lens, yellowish gray

to orange gray, continuous when fresh, some cracks developing on drying;

292 ... Dhingra & Priyanka

margins not well-differentiated. Hyphal system monomitic; generative hyphae

up to 4.5 um wide, thin-walled, septate, clamped; basal hyphae irregularly

branched and interwoven into a dense texture, covered in the upper half by

some crystalline material, followed by a narrow zone of compactly packed

horizontal hyphae; subhymenial hyphae short-celled, compactly packed and

appear like a cellular tissue. Cystidia absent. Basidia 15-33 x 7-10.5 um, at first

ellipsoid, then broadly subclavate to suburniform, rarely sinuous, basal clamp

not observed, 4-sterigmate, with oily contents; sterigmata up to 7 um long; anew

basidium generally takes the place of a decomposed basidium. Basidiospores

6-9 x 4.5-7.5 um, broadly ellipsoid to ovate or subglobose, smooth, somewhat

thick-walled, cyanophilous, inamyloid, with one large guttule or many small

oil drops.

REMARKS—As mentioned above Hallenbergia singularis is superficially similar

to Intextomyces contiguus. ‘The latter species is easily distinguished by smaller

spores and basidia.

Acknowledgements

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; Head, Department of Botany, Punjabi University,

Patiala, is thanked for providing research facilities.

Literature cited

Eriksson J, Ryvarden L. 1976. The Corticiaceae of North Europe - IV. Fungiflora, Oslo. pp.

549-886.

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

http://dx.doi.org/10.5248/118.293

Volume 118, pp. 293-302 October-December 2011

Cylindrochytridium johnstonii is a member of the Cladochytriales

REBECCA A. STEIGER, D. RABERN SIMMONS” & JOYCE E. LONGCORE

School of Biology & Ecology, University of Maine, 5722 Deering Hall, Orono, ME 04469 USA

* CORRESPONDENCE TO: david.r.simmons@umit.maine.edu

ABSTRACT — The taxonomy of the Chytridiomycota has been in flux between a classical

system based on thallus morphology and a new system based on zoosporic ultrastructure and

analyses of genetic sequences. Chytridiales sensu Sparrow has been divided into 7 orders plus

undescribed lineages. We found and brought into pure culture Cylindrochytridium johnstonii,

the type species of the genus, which heretofore has not been characterized by molecular

methods. We confirmed that this species is a member of the Cladochytriales, but it does not

lie within a recognized family.

KEY wWORDs — nucLSU rDNA, nucSSU rDNA

Introduction

Molecular studies have led to new orders being segregated from the large

chytridiomycete order Chytridiales sensu Sparrow (Barr 1980; Letcher et al.

2006, 2008; Longcore & Simmons 2012; Mozley-Standridge et al. 2009; Simmons

et al. 2009), yet many genera and species were not represented in these studies

because they were unavailable in culture. Such species, which were classified

in the Chytridiales in the classical literature (e.g., Sparrow 1960, Karling 1977),

remain incertae sedis in the Chytridiomycetes. Therefore, when chytrids that

have not been studied with molecular tools are isolated into pure culture it

is appropriate to determine their taxonomic position in the new system and

provide photographs of developmental morphology. Reporting genetic data for

described species is intended to promote universal taxon concepts. The phylum

Chytridiomycota has previously relied on thallus morphologies and zoosporic

ultrastructure, both of which, at least to some extent, have led to paraphyletic

groupings (e.g. Chytridiales sensu Sparrow 1960, Chytridiales sensu Barr 1980)

compared to molecular phylogenies (James et al. 2000, 2006).

The genus Cylindrochytridium was described by Karling (1941) for C. johnstonii,

a species with unique developmental morphology that occurred on boiled grass

used as bait in samples of aquatic debris. Subsequently, C. johnstonii has been

294 ... Steiger, Simmons & Longcore

reported from sites around the world, including New Zealand (Karling 1968),

South America (Marano et al. 2008a,b; Rocha & Pires-Zottarelli 2002), Europe

(Czeczuga et al. 2007), and India (Karling 1963), but no authors have reported

isolating this fungus into pure culture. Its phylogenetic position therefore has

remained unconfirmed. We found this species growing on the edge of onionskin

bait placed with algae and aquatic debris collected from Smith’s Fen (Schwintzer

1978) during a field trip that was part of the 2008 centennial celebration of the

University of Michigan Biological Station in northern Michigan, USA. Herein

we report that C. johnstonii is a member of the Cladochytriales and support this

conclusion with molecular data. Also, we include photos of the development of

this chytrid in pure culture for comparison with the type description.

Materials & methods

Collection, isolation & morphology

JE Longcore collected algae and plant debris on 22 Sep 2008 from Smith's Fen (pH

= 5.7; Schwintzer 1978), University of Michigan Biological Station, Cheboygan County,

Michigan, USA. The sample was transported at ambient temperature in a Whirlpak®

plastic bag to our laboratory in Maine. We placed the aquatic sample in a finger bowl

at room temperature and baited it with pieces of boiled, white onionskin. After several

weeks we noted thalli with catenulated rhizoids on the bait. Most of the thalli grew at

the edges of the onionskin and produced long, cylindrical zoosporangia. We rinsed

the onionskin with a stream of distilled water and placed it in a depression slide with

distilled water. After ~1 hour, when many active zoospores could be seen, we added

the water to plates of TC agar (0.4 g tryptone, 4 g cellobiose, 10 g agar, 1 L distilled

water) containing 300 mg/L streptomycin and 200 mg/L penicillin G. We monitored

the isolation plates at 100x magnification by inverting them on the stage of a compound

microscope. After nearly two weeks, we selected Cylindrochytridium johnstonii,

recognized by its long, cylindrical zoosporangia, from among the other chytrids on

the isolation plate and removed thalli to clean plates of TC agar. We transferred the

isolate, designated JEL596, to mPmTG slants (Longcore 1992) in screw-topped 125 x

20 mm culture tubes and maintained the stock tubes at room temperature. Stocks were

transferred at 3-month intervals. To document morphology and days to maturity, we

inoculated cultures on mPmTG agar in 9 cm Petri plates stored at 23 °C. To document

morphology on cellulosic substrates, we inoculated a flask of sterilized lake water

containing pieces of white onionskin with thalli near maturity or with motile zoospores

present and incubated the flasks at room temperature. We aseptically removed portions

of agar with thalli from plates and pieces of onionskin from flasks to view the chytrid

by light microscopy. We photographed developmental stages with phase contrast and

Hoffman Modulation Contrast (HMC) optics on a Nikon E400 microscope (Nikon

Instruments, Melville, New York) equipped with a Spot RT digital camera (Diagnostic

Instruments, Sterling Heights, Michigan). Composite images of thalli through multiple

focus planes were assembled from overlapping micrographs of individuals in Adobe

Photoshop 6.0 (Adobe Systems Inc., San Jose, California).

Cylindrochytridium johnstonii (Cladochytriales) ... 295

TABLE 1. Isolates of Cladochytriales used for phylogenetic analyses

of Cylindrochytridium johnstonii

Ae aes ne SOmierk : HABITAT i GENBANK ACCESSION NO.

ISOLATE : NO : ee esata

i ; i NucSSU : NucLSU

INGROUP:

UCC GEAT os oi ats a san OEE, : Soil/detritus ' AF164291S1 | EU828501

expandens : : Carolina, USA : : :

SRE ae ! Ontario, CAN | Sandy soil/detritus { AY349047

age DESHI rs Shera atm, Waseca ona eal ta iets, eI See Ein, Ae SRN Lh

ree a | JELI45 £ Maine,USA | Aquatic/Eriocaulon | EU828475 | EU828503

cae PAGO «2 vargas Maine USA Aquatic/detritus EU828476 EU828504

yes igi «wer gaa | Maine USA Aquatic/detritus EU828478 EU828506

Cladochytrium sp JEL153 Maine, USA Aquatic/maple leaf EU828458 EU828485

Qiindrechy iar cs oi sae: 9 4 Michi enn bate PAquaticldetitus > {P796051

a se ee Ee 3b a 2 Re a Sols it 8 Oe Sd en es

Diplophlyctis sp. : JEL331 : Maine, USA Aquatic/Characeae EU828477. _: EU828505

Endochytrium : JEL402 : Michigan, USA : Aquatic/Cladophora : EU828484 | EU828513

ssl ce LOAN Gt me Aa Nye NR ea items Sah Oe We Mee OE NA ce A ED

ite ae JEL027 Maine, USA Aquatic/Eriocaulon EU828471 EU828498

pe ge JELO70 Maine, USA Aquatic/Eriocaulon EU828472 EU828499

ane JEL324 Maine, USA Aquatic/Elodea EU828473 EU828500

Nephrochytrium : Maine, USA; Aquatic/Sparganium : EU828468 | EU828495

aurantium i ; i

i EU828494

Maine, USA Aquatic/Characeae AF164295S1 } EU828511

: Maine,USA —_ Aquatic/Nitella : BU828467

eee ge JEL046 : Maine, USA Aquatic/detritus EU828463 EU828490

se are JELI27_: Maine, USA Aquatic/Characeae | EU828466 | EU828493

Septochytrium sp. JEL177 Wales, UK : Aquatic/detritus EU828474 EU828502

"Siri Diasec Fae Read caer ult es ane ; : 1 Raat uh ee ma)

Unidentified sp. | JELO72 ! Maine, USA: Aquatic/Eriocaulon : EU828470 : EU828497

OUTGROUP

Karlingiomyces sp. JEL093 Maine, USA Aquatic AF164278 AY349085

Falychy ring : JELI09 : Maine, USA: Aquatic ! AY601711 | AY349084

aggregatum : : i i i

296 ... Steiger, Simmons & Longcore

Sequencing & analyses

DNA of JEL596 was extracted with Whatman® FTA card technology (Whatman

Ltd., Maidstone, Kent, UK; Borman et al. 2006, Simmons 2011). Segments of nucSSU

and nucLSU rDNA were amplified, sequenced, and aligned as in Simmons (2011) with

taxa from Cladochytriales (TABLE 1) after a BLAST search in GenBank grouped JEL596

within that order. A region of nucSSU rDNA was amplified with the NS1/NS4 primer

pair (White et al. 1990), as was a region of nucLSU rDNA with the LROR/LRS primer pair

(Rehner & Samuels 1994, Vilgalys & Hester 1990). The Akaike Information Criterion in

jModeltest 0.1.1 (Guindon & Gascuel 2003, Posada 2008) chose the TIM3+G model of

evolution for the combined dataset. We entered these parameters into MrBayes 3.1.2

(Ronquist & Huelsenbeck 2003), which ran for 1M generations, selecting every 100"

tree. A 50% majority rule phylogram was constructed and MP bootstrap and BPP values

were computed in PAUP* 4.0b10 (Swofford 2002) as in Simmons (2011).

Taxonomy

Cylindrochytridium johnstonii Karling, Bull. Torrey Bot. Club 68: 383 (1941)

FIGURE 1

TYPE: Karling (1941: Figs. 1-16).

SPECIMEN EXAMINED. USA. MICHIGAN. On onionskin substrate added to water culture

of algae and aquatic plant detritus, 22 Sep 2008. JEL596 (nucSSU rDNA sequence

GenBank JF796051; nucLSU rDNA sequence GenBank JF796052).

We emend the description of Cylindrochytridium johnstonii by adding

morphology on agar and axenic onionskin and molecular sequence information

for JEL596.

THALLUS CHARACTERS — In pure culture, isolate JEL596 has morphological

characters similar to Karling’s (1941, 1977) descriptions of C. johnstonii. On

agar, germlings developed endogenously, forming catenulate swellings at a

single axis or multiple rhizoidal axes (Fic. 1A, B). The catenulations and the

spherical base of what would become the zoosporangium developed relatively

FiGurRE 1 (right). Cylindrochytridium johnstonii on mPmTG agar and liquid medium grown at

23 °C and on onion skin grown at room temperature. Micrographs taken with phase contrast

optics unless otherwise noted. A. Germling at 3 days with catenulated rhizoids. B. Developing

thallus at 8 days with two catenulated rhizoidal axes (arrowheads) and diffusely branching

rhizoids. C. Brightfield micrograph of amber, ovoid zoospore cyst on onion skin with ridges

along surface of cell wall and a single, large lipid globule. D. Brightfield composite micrograph

of exogenously developing germling in onion skin, with zoospore cyst (arrow) and catenulated

rhizoids (arrowheads). E. Composite micrograph of broad tube extending from spherical base of

zoosporangium. F. Septum (white arrow) with tapered nipple delimiting zoosporangium base and

apically migrating cytoplasm in thallus. G. Hoffman Modulation Contrast (HMC) micrograph of

zoospores released from apical pore. H. HMC micrograph of spherical zoospores after release from

zoosporangium by dehiscence of operculum (white arrowhead). I. Composite HMC micrograph

of developing thallus with catenulated rhizoids, vacuolated portion of zoosporangium towards

spherical base, and apically migrating cytoplasm. Scale bars = 10 um; bar in A for G-H; bar in

C for D; bar in E for EI.

Cylindrochytridium johnstonii (Cladochytriales) ... 297

298 ... Steiger, Simmons & Longcore

slowly as the rhizoidal system expanded through the agar (Fic. 1B). After nearly

two weeks, one side of the spherical zoosporangium elongated to form a tube

nearly as wide as the spherical base (Fic. 1E). Most of the cytoplasm migrated

toward the apical portion of the broad tube, leaving a highly vacuolated portion

at the zoosporangium base (Fic. 1K, I). At maturity, the basal portion of the

zoosporangium was empty and may have been walled off by a septum (Fie. 1F).

Zoospores exited the zoosporangium through a pore made by the complete

detachment of an apical operculum (Fic. 1H). Zoospores possessed a single

lipid globule (Fic. 1G, H), and flagella were ~35 um long.

Our attempts to reintroduce C. johnstonii to onionskin led to the production

of germlings, but after 1 month, no further development occurred. On onionskin

isolate JEL596 developed exogenously. The amber, ovoid zoospore cyst on the

surface of the onionskin had a cell wall with ridges and generally possessed a

single, large lipid globule (Fic. 1C). Germlings possessed one or two rhizoidal

axes with catenulations (Fic. 1D).

PHYLOGENETICS — A majority rule Bayesian phylogram (Fic. 2) was

constructed from a data matrix with 337 parsimony-informative characters ofa

total matrix of 1410 characters. The phylogeny indicates that Cylindrochytridium

johnstonii is within the Cladochytriales. The species is in a clade that is sister to

a clade containing the type species of Allochytridium Salkin (Salkin 1970) and

Septochytrium Berdan (Berdan 1939), which are the only two genera recognized

in Septochytriaceae (Mozley-Standridge et al. 2009).

Discussion

Our phylogeny (Fic. 2) places Cylindrochytridium johnstonii as sister to an

isolate with Catenochytridium morphology (JEL145). Together these two taxa

are sister to a group of isolates in the Septochytriaceae (Mozley-Standridge

et al. 2009) and are also part of a larger clade (Fic. 2), whose members form

large catenulations in their rhizoids. One species in this clade, Allochytridium

expandens Salkin, also has an elongated zoosporangium, but not to the extent

seen in C. johnstonii. Additionally distinguishing these two taxa, A. expandens

develops exogenously both in natural substrates and on agar medium (Barr

1986) whereas C. johnstonii develops endogenously on agar medium.

Shanor (1944) commented upon the resting spore of C. johnstonii, but he did

not illustrate this feature. He described the resting spore on filter paper as being

light-brown or amber, nearly spherical, with a smooth, thick cell wall and one

large yellow oil globule. This description is very similar to that of the zoospore

cyst that we saw on onionskin, but fine ridges appeared on the surface of most

zoospore cysts we observed. Though the amber zoospore cysts on onionskin

may look like resting spores, their connection to larger portions of the thallus

(Fic. 1D) lead us to conclude they are zoospore cysts. Thus, we believe Shanor

may have mistaken the zoospore cyst for a resting spore.

Cylindrochytridium johnstonii (Cladochytriales) ... 299

o7oof Bart 253 Allochytridium expandens

88/100] = JEL191 Septochytrium variabile

61/98|"™" JEL177 Septochytrium sp.

-/90

JEL145 Catenochytridium sp. 1

JEL596 Cylindrochytridium johnstonii

77/100

CRC = JEL024 Catenochytridium sp. 2

JEL331 Diplophlyctis sp.

ABR JEL044 Catenochytridium sp. 3

98/100

99/100 Barr463 Allochytridium luteum

77/100} YEL324 Endochytrium sp. 1

100 J | | JELO27 Endochytrium sp. 2

— 100] ' JELO70 Endochytrium sp. 3

JELO036 Nephrochytrium aurantium

sahing sootoot YEL127 Nowakowskiella elegans

JELO46 Nowakowskiella elegans

-/100

100/100 JEL327 Nephrochytrium sp. 1

E JELO72 Unidentified sp.

\ JEL402 Endochytrium ramosum

400/100 JEL125 Nephrochytrium sp. 2

C> JEL153 Cladochytrium sp.

165 JEL109 Polychytrium aggregatum

i. JEL093 Karlingiomyces sp.

== 50 changes

FIGURE 2. Majority rule consensus Bayesian phylogram of combined nucSSU and nucLSU

rDNA for 20 taxa of Cladochytriales and 2 isolates in the Polychytrium clade (James et al. 2006).

Labeled nodes correspond to the catenulated rhizoids clade (CRC) and families within the

Cladochytriales (arrowhead): Septochytriaceae (S), Nowakowskiellaceae (N), Endochytriaceae

(E), and Cladochytriaceae (C) (Mozley-Standridge et al. 2009). Branch support values are listed

as maximum parsimony bootstrap values / Bayesian posterior probabilities. Tree length = 1234,

CI = 0.6183, RI = 0.6235.

300 ... Steiger, Simmons & Longcore

Karling (1941) illustrated elongation of C. johnstonii thalli as beginning

during growth of the germling, with some zoosporangia having basal swellings.

The developing zoosporangia we examined on agar were spherical (Fic. 1B) and

grew an apical elongation only when nearly mature (Fie. 1E, FE, I). Additionally,

young thalli of our isolate on onionskin were also spherical (Fic. 1D).

Considering that Karling (1941) observed C. johnstonii in gross culture

and we studied our isolate in pure culture, our observations agree well with

his drawings, which constitute the type of C. johnstonii. Cylindrochytridium

endobioticum Willoughby (Willoughby 1964) is the only other species in the

genus, but Karling (1977) considered it to be a dubious member because of its

considerable variation from C. johnstonii. Our genetic information provides

the basis for adding additional species to the genus as well as evaluating the

suitability of the genus for C. endobioticum, when it is found and brought into

culture.

Acknowledgments

We thank University of Michigan Biological Station visiting phycology professor

RL Lowe of the Bowling Green State University Department of Biological Sciences for

leading the centennial celebration phycology field trip during which this collection was

made. We thank D Cox and P Singer of the DNA Sequencing Facility at the University

of Maine for their services, DV Neace of the Yale University School of Medicine for

document procurement, and J Shepard and H Reyes for their fastidious diligence. We

thank SE Mozley-Standridge and MJ Powell for reviewing this manuscript. This study

was financially supported by the NSF DEB grant PEET 0529694.

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James TY, Letcher PM, Longcore JE, Mozley-Standridge SE, Porter D, Powell MJ, Griffith GW,

Vilgalys R. 2006. A molecular phylogeny of the flagellated fungi (Chytridiomycota) and

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898-915. http://dx.doi.org/10.1016/j.mycres.2006.06.011

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MY COTAXON

http://dx.doi.org/10.5248/118.303

Volume 118, pp. 303-309 October-December 2011

A contribution to the study of smut fungi of Israel

KyrRYLO G. SAVCHENKO?”*, VASYL P. HELUTA?,

SOLOMON P. WASSER?” & EVIATAR NEVO’!

‘Institute of Evolution & Department of Evolutionary and Environmental Biology,

Faculty of Natural Sciences, University of Haifa, Mt. Carmel, Haifa 31905, Israel

2M.G. Kholodny Institute of Botany of the National Academy of Sciences of Ukraine,

2 Tereshchenkivska St., Kyiv 01601, Ukraine

* CORRESPONDENCE TO: savchenko.kyryll@gmail.com

ABSTRACT — Four species of smut fungi, Antherospora vaillantii, Microbotryum holostei,

Urocystis magica, and U. muscaridis, are reported for the first time in Israel. Urocystis magica

was found in the Judean desert on a new host plant, Allium rothii, and M. holostei is new for

Asia.

Key worps — Urocystales, biodiversity, Microbotryales, Middle East

Introduction

The smut fungi that occur in Israel have been studied by mycologists and

phytopathologists since the start of the 1900s (Magnus 1900; Reichert 1921,

1930, 1931; Savulescu & Rayss 1935; Rayss & Zwirn 1944; Rayss 1952; Palti et

al. 1966). Nevertheless, knowledge about species composition and distribution

of these fungi was incomplete until a critical revision of the published accounts

of smut fungi in Israel that showed 59 species were recorded (Savchenko et al.

2010).

During February-March 2011, we collected plants infected by smut fungi,

mainly in the territory of Mount Carmel, a coastal mountain range in northern

Israel that stretches southeast from the Mediterranean Sea. This region is

botanically very rich in species with a typical East-Mediterranean flora with

luxuriant herbaceous components. Some of them are host plants for smut fungi.

In the present study we report three smut species found in this region and one

species from the Judean Desert.

Materials & methods

Sorus and spore characteristics were studied using both fresh and dried herbarium

specimens. Spores were dispersed in a droplet of lactophenol on a microscope slide,

304 ... Savchenko & al.

covered with a cover glass, gently heated to boiling point to rehydrate the spores,

cooled, and then examined by a Carl Zeiss Axiostar light microscope (LM) at 1000x

magnification. For scanning electron microscopy (SEM), spores were attached to

specimen holders by double-sided adhesive tape and coated with gold. The surface

structure of spores was observed at 15 kV and photographed with a scanning electron

microscope JEOL JSM-6700F. Specimens used in this study are stored in the Herbarium

of Haifa University (HAI).

Results & discussion

Descriptions and critical notes of four new Israeli species of smut fungi are given

below.

Antherospora vaillantii (Tul. & C. Tul.) R. Bauer et al., Mycol. Res. 112: 1304 (2008)

FIGS ET=2

Sort in the anthers and on the surface of inner floral organs, producing dark

olive-brown powdery mass of spores, enclosed by the floral petals. Infection

systemic, all flowers of an inflorescence are infected. SporEs variable in shape

and size, globose, subglobose, ovoid, slightly elongated, 7-8.5 x 8-10(-11) um,

olive-brown. SPORE WALL even, ca. 0.5-0.7 tum thick, finely, densely verruculose.

In SEM spores densely irregularly verruculose. SPORE PROFILE appears wavy.

DISTRIBUTION: Europe, Asia, Africa, North America, Oceania.

SPECIMEN EXAMINED: ISRAEL. HAIFA DISTRICT, Carmel National Park, 32°75'79"N,

35°01'50"E, on Muscari comosum (L.) Mill., 2.11.2011, leg. K.G. Savchenko (HAI 2857).

Norte. The genus Antherospora R. Bauer et al. consists of eight species of

smut fungi that infect the anthers and inner floral organs of host plants in the

Hyacinthaceae (Liliaceae s.1.) (Vanky 2009). The genus differs from all other

taxa of Urocystales by the organogenic specialization, lack of sterile cells, and

development of single spores (Bauer et al. 2008). Most of its species are highly

specialized parasite restricted to certain host plant genera. Only one other

Antherospora species, A. urgineae (Maire) R. Bauer et al. on Urginea maritima

(L.) Baker, is known to occur in Israel (Savulescu & Rayss 1935, Savchenko

et al. 2010). Antherospora vaillantii is easily distinguishable from A. urgineae,

which has smaller spores (9.5-15(-17.5) x 7-12 um in diam.) and hosts in

different genera.

Microbotryum holostei (de Bary) Vanky, Mycotaxon 67: 44 (1998) FIGs. 3-4

Sor! in ovules, fill the capsules with a powdery reddish brown spore mass.

Infection systemic. Spores reddish-brown, globose, sometimes ovoid, 9-14

x 11-14 um. SporE WALL reticulate, 4-6 meshes per spore diam., meshes

1.2-2.4 um in diam.; the bottom of the meshes with conspicuous round low

protuberances.

DISTRIBUTION: Europe, Asia.

Smut fungi (Israel) ... 305

15.0kV 6.500 Tye WD 78

Fics. 1-6. 1-2: spores of Antherospora vaillantii in flowers of Muscari comosum. 3-4: spores of

Microbotryum holostei in flowers of Holosteum umbellatum. 5-6: sori and spores of Urocystis

muscaridis on leaves of Muscari comosum. 1, 3, 6 = LM; 2, 4 = SEM. Scale bars: 1, 3, 6 = 10 um;

2,4=1um;5=3 mm.

306 ... Savchenko & al.

SPECIMEN EXAMINED: ISRAEL. HAIFA DISTRICT, Carmel National Park, 34°74'97"N,

35°03'01"E, on Holosteum umbellatum L. (Caryophyllaceae), 3.11.2011, leg. K.G.

Savchenko (HAI 2858).

Norte. Only three species of Microbotryum Lév. were reported from Israel

before our investigations: M. cordae (Liro) G. Deml & Prillinger on Polygonum

acuminatum Kunth (Polygonaceae), M. jehudanum (Zundel) Vanky on Silene

apetala Willd. (Caryophyllaceae), and M. scorzonerae (Alb. & Schwein.)

G. Dem! & Prillinger on Scorzonera papposa DC. (Asteraceae) (Rayss 1952, Rayss

& Zwirn 1944, Savchenko et al. 2010). All three are typical ovaricolous species.

Israel has a great diversity of potential hosts for smuts from Microbotryum,

especially those parasitizing Caryophyllaceae (43 Silene species grow in the

country; Feinbrun-Dothan & Danin 1998). Microbotryum holostei has probably

been overlooked because of the inconspicuous symptoms of infection and pre-

vernal development. To our knowledge, M. holostei has never been reported

from Asia.

Urocystis magica Pass., in Thiimen, Mycoth. Univ. 3: no. 223 (1875) Fics. 7-9

Sor! as pustules in leaves and bulbs, initially covered by an epidermis that

later ruptures to expose the black mass of spore balls. SPORE BALLS globose to

ovoid, 18-25 x 20-40 um in diam., composed of 1 (90%) to 2 (10%) central

spores and continuous layer of peripheral sterile cells. Spores globose,

subglobose, ovoid, 10-15 x 12-16 um in diam., dark reddish-brown. SPORE

WALL ca. | um thick. STERILE CELLS globose, ovoid, irregular, 4-13 um in diam.,

pale yellowish-brown.

DISTRIBUTION: Europe, Asia, North America, South America, Africa,

Oceania.

SPECIMEN EXAMINED: ISRAEL. SOUTH DISTRICT, Judean Desert, near Arad, Wadi Kidot,

31°15'73"N, 35°13'59"E, alt. 569 m., on Allium rothii Zucc. (Alliaceae), 15.11.2011, leg.

K.G. Savchenko (HAI 2860).

Note. During a collection trip to the Judean Desert in March 2011, one Allium

rothii plant was found infected by a smut fungus. Allium rothii belongs to the

subgenus Melanocrommyum and is distributed in Israel in the Judean Desert,

North and South Negev, and the Dead Sea area on rocky slopes between shrubs

(Kamenetsky 1994). Microscopic examination revealed that the smut was

U. magica s.l., not previously recorded in Israel (Savchenko et al. 2010). Allium

rothii represents a new host plant for this fungus.

Allium L. is a major genus of the monocot family Alliaceae naturally

distributed throughout the northern hemisphere that is represented by

ca. 750 species worldwide (Stearn 1992). Almost 40 species are known in

Israel (Feinbrun-Dothan & Danin 1998). According to recent taxonomical

changes based on nrDNA ITS sequence analyses, Allium is divided into 15

well-delimited monophyletic subgenera (Friesen et al. 2006). Species from

Smut fungi (Israel) ... 307

Figs. 7-9. Urocystis magica on Allium rothii. 7: infected plant; 8: sori: 9: spore balls (in LM).

Scale bars: 7 = 1 cm; 8 = 3 mm; 9 = 10 um.

six subgenera —Allium, Amerallium, Cepa, Melanocrommyum, Polyprason,

Rhizirideum— have been found as hosts for Urocystis Rabenh. ex Fuckel. At

least five different Urocystis taxa have been described from onion plants during

the last two centuries (see Vanky 1994): U. magica, U. cepulae Frost, U. colchici

f. allii-subhirsuti Beltrani, U. allii Schellenb., and Tuburcinia oblonga Massenot

(= U. oblonga (Massenot) H. Zogg). As these taxa are not easily distinguished

from one another, they have been treated asa single taxon, U. magica s.1. However,

these taxa were described from different subgenera of Allium: U. magica s.str.

from subg. Melanocrommyum, U. cepulae from subg. Cepa, U. colchici f. allii-

subhirsuti from subg. Amerallium, and U. allii and T. oblonga from subg. Allium.

Specialization on certain host plant subgenera or even species is not unusual

among smut fungi (cf. Entyloma species on Eryngium L.; Vanky 2009). Thus, it

is not inconceivable that these Urocystis taxa may be recognized as “good” or at

least “cryptic” species after intensive molecular-phylogenetic research.

308 ... Savchenko & al.

Urocystis muscaridis (Niessl) Moesz, Kaérpat-mend. Uszég.: 199 (1950) FIGs. 5-6

Sor! in leaves as ellipsoidal pustules, variable in size, visible on the outer

surface along the viens, filled by a black powdery mass of spore balls. SPORE

BALLS globose, subglobose, ovoid to irregular, 18-40 x 20-45 um in diam.,

composed of 1-7(-8) spores surrounded by almost a continuous layer of sterile

cells. SporEs globose, ovoid to irregular, 9-17(-18) x 10-20(-21) um in diam.,

dark reddish-brown. SPORE SURFACE smooth. STERILE CELLS variable in shape

and size, globose, ovoid to irregular, 4-9 x 5-14 um, yellowish-brown.

DISTRIBUTION: EUROPE, Asia.

SPECIMEN EXAMINED: ISRAEL. HAIFA DISTRICT, Carmel National Park, 32°75'25"N,

35°02'50"E, on Muscari comosum (L.) Mill. 18.11.2011, leg. K.G. Savchenko (HAI

2862).

Note. Urocystis muscaridis infects different species of Muscari Mill. in Europe

and Asia (Vanky 1994). Muscari comosum, a principal host for this smut, is

quite common in Mediterranean forests and semi-steppe shrub lands of Israel.

In March 2011 we examined a large M. comosum population in Carmel National

Park and found one plant with leaves bearing several swollen sori produced by

U. muscaridis. It is interesting that only a single plant in the whole population

was found to be affected. In the Middle East U. muscaridis is also known from

Iran (Ershad & Deghani 2001).

Acknowledgments

The authors thank Dr. Dominik Begerow and Dr. Roger G. Shivas for peer-reviewing

the manuscript, Dr. Shaun R. Pennycook for some useful comments, Dr. Eli Harlev for

organizing a collection trip to the Judean Desert and Mr. Ivan Hurnenko for help with

the SEM microscopy.

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Ershad D, Deghgani A. 2006. Urocystis muscaridis, a smut fungus new to Iran. Rostaniha 7(1):

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(Alliaceae) based on nuclear ribosomal DNA ITS sequences. Aliso 22: 372-395.

Kamenetsky R. 1994. Life cycle, flower initiation, and propagation on the desert geophyte Allium

rothii. International Journal of Plant Sciences 155: 597-605. http://dx.doi.org/10.1086/297198

Magnus P. 1900. Bornmiiller, Iter syriacum 1897. Fungi. Weiterer Beitrag zur Kenntnis der Pilzen

des Orientes. Verhandlungen der Kaiserlich-K6niglichen Zoologisch-Botanischen Gesellschaft

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Jerusalem Series 5: 229-236.

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Jerusalem Series 3: 114-116.

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MY COTAXON

http://dx.doi.org/10.5248/118.311

Volume 118, pp. 311-323 October-December 2011

Species of Rhytismataceae on Lithocarpus spp.

from Mt Huangshan, China

QIAN ZHENG‘, YING-REN LIN”, SHENG-MING YU? & LI CHEN?

" School of Life Science & ? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

* Management Committee of Huangshan Scenic Area of Anhui, Huangshan 242709, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

ABSTRACT—Seven members of the Rhytismataceae are reported from leaves of tanoaks from

Mt Huangshan, China. Among them Coccomyces mucronatoides and Terriera coacervata

are new species, Terriera illiciicola is a new combination, and the other species are already

known for China. This paper provides descriptions, discussions for these species as well

as a dichotomous key for the eight rhytismataceous species known to occur on tanoaks

worldwide.

Key worps—Rhytismatales, morphology, taxonomy, Fagaceae

Introduction

Worldwide, at least 55 genera (+ 30 synonyms) and 728 species of

Rhytismataceae have been reported (Kirket al. 2008). These fungi are parasites on

leaves, stems, twigs, or fruits of gymnosperms, angiosperms and pteridophytes

or saprobes on plant residues. More than 240 different fungi have been

discovered on Lithocarpus spp. (Fagaceae), but records of the Rhytismataceae

are few (Farr & Rossman 2011). Coccomyces dentatus (J.C. Schmidt & Kunze)

Sacc. on Lithocarpus densiflorus (Hook. & Arn.) Rehd. is known from the USA

(Sherwood 1980), and five Rhytismataceae species —C. delta, C. huangshanensis,

C. mucronatus, Lophodermium agathidis, L. illiciicola— have been reported on

Lithocarpus spp. from China (Lin et al. 2000a,b, 2001, 2005; Wang et al. 2006).

Mt Huangshan in southern Anhui Province is suitable for many

rhytismataceous species because of the warm climate, abundant rainfall, and

high plant diversity. Seven species representing three different genera on

Lithocarpus leaves are described from Mt Huangshan in this paper, including

two new species, one new combination, and four species already known in

China.

312 ... Zheng & al.

Materials & methods

Leaves with bleached spots were collected from plants on Mt Huangshan, and those

with open fruit bodies characteristic of Rhytismataceae were selected. Observations of

the external shape, size, color, opening mechanisms of ascomata and conidiomata, and

zone line characteristics were made using a stereoscope with 10-50x magnification.

Microscopic preparations were made in water, Melzer’s reagent, 5% KOH, or 0.1%

(w/v) cotton blue in lactic acid. For observing the outlines of ascomata and conidiomata,

vertical sections were mounted in lactic acid or cotton blue with water pretreatment.

Gelatinous sheaths surrounding ascospores and paraphyses were observed in water or

cotton blue in lactic acid. The color of structures and ascospore contents were observed

in water. Measurements and drawings of asci, ascospores, and paraphyses were made

from material mounted in 5% KOH or Melzer’s reagent from 30 ascospores, asci and

paraphyses for each specimen. Point and line integrated illustrations of external shapes

and internal structures of the fruit bodies were drawn using a Panasoianic XSJ-2

microscopic painting device.

Taxonomy

Coccomyces delta (Kunze) Sacc., Boletim da Sociedade Broteriana 11: 13, 1893

= Phacidium delta Kunze, Linnaea 5: 551, 1830

Type: MADEIRA, on leaves of Lauraceae. | Without further data. ]

ZONE LINES frequent, black, thin, surrounding the bleached spots.

CONIDIOMATA not observed.

Ascomata on both sides of leaves, scattered in irregular, yellow-brown

bleached spots. In surface view, ascomata (700—)1000-—1600(—1800) um diam.,

triangular or quadrangular, black, shiny, slightly raising the substratum surface,

opening by 3-4 radial splits. Lips present. In median vertical section, ascomata

intraepidermal. COVERING STROMA up to 25-28 um thick near the opening,

black to dark-brown, becoming thinner or firstly thickening then becoming

thinner towards the edge, composed of textura globulosa or angularis with cells

4—7 um diam. Lip cells fringed, 1.5-2.2 um diam., immersed in a gel. BASAL

STROMA Slightly hollow or flat, 4.0-6.5 um thick, dark-brown, composed of

textura globulosa or angularis with cells, connecting to the covering stroma.

INTERNAL MATRIX STROMA only between the covering stroma and basal stroma.

SUBHYMENIUM 15-20 um thick, consisting of textura intricata and porrecta.

PARAPHYSES 100-130 x 1.5-2.0 um, filiform, aseptate, gradually swollen to

2.2-3.0 um above, covered in a thin mucous coating. Asci ripening sequentially,

85-120 x 7.5-9.5(-11) um, cylindric-clavate or subclavate, somewhat long-

stalked, apex thin-walled, J-, 8-spored. Ascosporss fasciculate, 52-85 x

1.5-1.8(—2.0) um, filiform, hyaline, aseptate, with a ca 1 um thick gelatinous

sheath.

ILLUSTRATION: Sherwood (1980: Fig. 18).

Rhytismataceae on Lithocarpus (China) ... 313

ECOLOGY & DISTRIBUTION: Ascomata found on fallen or not yet fallen

diseased leaves. Widely distributed in Europe (Sherwood 1980). In China, it is

known from southern parts (Lin et al. 2000a).

SPECIMENS EXAMINED: On Lithocarpus glaber (Thunb.) Nakai: CHINA, ANHUI, MT

HUANGSHAN, alt. ca 550 m, 11 August 1995, S.M. Yu, Y.R. Lin 1596a (AAUF 67794a);

Ciguangge, alt. ca 800 m, 4 October 2009, Q. Zheng, S.J. Wang 5436 (AAUF 71544).

ComMENTS—The species was originally described from Madeira, and mainly

occurs on plants of the Lauraceae and evergreen Quercus (Sherwood 1980).

Teng (1934) first reported C. delta from China. According to our observation,

it infects living leaves at early stage, and may be followed by other Coccomyces

species as the ascomata mature. Compared with the polygonal species in the

genus, C. delta ascomata are the largest. The asci and ascospores in the Mt

Huangshan specimens are shorter than described by Sherwood (1980).

Coccomyces huangshanensis Y.R. Lin & Z.Z. Li, Mycosystema 19: 449, 2000

Type: CHINA, ANuulI, Mt. HUANGSHAN, Songgu‘n, alt. ca 600 m, on Cyclobalanopsis

glauca (Thunb.) Oerst., 9 August 1995, Y.R. Lin & S.M. Yu 1632 (AAUF 67740).

ZONE LINES frequent, black-brown, thin, surrounding the bleached areas.

ConIDIOMATA with a distribution on the leaf similar to that of the

ascomata. In surface view, conidiomata 180-330 x 150-260 um, subcircular

or elliptical, brown in the centre and at the perimeter line of the conidioma,

concolorous with the substratum surface or light grey-brown over the rest

of the appearance, slightly raising the surface of the leaf. In vertical section,

conidiomata intraepidermal, double lens-shaped. UPPER WALL very thin, brown

to dark-brown, with an indefinite structure. BASAL WALL 4—7 um thick, brown,

composed of thick-walled, angular cells. CONIDIOGENOUS CELLS 5.5-9.0 x

1.5—2.0 um, flask-shape, holoblastic sympodially proliferating. Conip1A 3.5-5.0

x ca 0.8 um, bacilliform, hyaline, aseptate.

AscoMata on both sides of leaves, more on the upper side of the leaf,

scattered in irregular bleached spots. In surface view, ascomata 700-1100 um

diam., triangular or quadrangular, black-brown, slightly raising the substratum

surface, opening by 3-4 radial splits. Lips present at least in the early stages. In

median vertical section, ascomata intraepidermal. COVERING STROMA 15-18

um thick near the opening, becoming thinner towards the edge, extending to

the basal stroma, consisting of brown to dark-brown textura angularis with

cells 3.5-6.5 um diam. The opening often covered by the heavily carbonized

tissue. Lip cells tend to disappear as the ascomata mature. Periphysoids present.

BASAL STROMA 8-12 um thick, dark-brown, consisting of textura angularis or

globulosa with cells 5.5-8 um diam. INTERNAL MATRIX STROMA only between

covering stroma and basal stroma. ExcrpuLuM 22-30 um thick, arising

from the inner layer of the covering stroma. SUBHYMENIUM 10-18 um thick,

314 ... Zheng & al.

consisting of textura porrecta. PARAPHYSES 110-135 x 1.5-2.0 um, filiform,

septate, slightly swollen at the apex. Ascr ripening sequentially, 85-115 x

6.0-7.0 um, cylindrical, short-stalked, apex obtuse, thin-walled, J—, 8-spored.

Ascosporss fasciculate, 60-90 x 1.2-1.5 um, filiform, hyaline, aseptate, with a

thin gelatinous sheath, sometimes with a gelatinous cap at the top.

ILLUSTRATION: Lin et al. (2000b: Fig. 2).

ECOLOGY & DISTRIBUTION: Ascomata found on decaying leaves. Known

only from China (Lin et al. 2000b).

SPECIMENS EXAMINED: On Lithocarpus henryi (Seemen) Rehder & E.H. Wilson: CHINA,

ANHUI, Mt Huancsuan, Tangkou, alt. ca 500 m, 29 September 1993, Y.R. Lin, L. Chen

151la (AAUF 67619a); 2 October 2009, Q. Zheng, X.M. Gao 5389 (AAUF 71497).

COMMENTS—Coccomyces limitatus (Berk. & M.A. Curtis) Sacc. is very similar

to this species but differs in the well developed internal matrix of the stroma, 50

um thick excipulum, and dissimilar asci and ascospores. Its asci are 4.0-5.6 um

wide, and the ascospores are only 0.8-1.0 um wide (Sherwood 1980; Johnston

1986).

Coccomyces mucronatus Korf & W.Y. Zhuang, Mycotaxon 22: 487, 1985

Type: CHINA, SICHUAN, MT QINGCHENG, alt. ca 900 m, on Castanopsis sp., 6 July 1983,

W.Y. Zhuang HMAS 450558 (CUP-CH 2484).

ZONE LINES frequent, black-brown to black, thin, surrounding or separating

the bleached spots.

CONIDIOMATA not observed.

Ascomarta on both sides of leaves, scattered in irregular, grey-yellow bleached

spots. In surface view, ascomata 650-1200 um diam., polygonal to subcircular,

black-brown to black, slightly raising the substratum surface, opening by radial

splits. Lips absent. In median vertical section, ascomata intraepidermal to

subepidermal. COVERING STROMA 15-22 um thick, consisting of grey-brown

to dark-brown textura angularis and globulosa with cells 3.5-7 um diam.,

extending to the basal stroma. Periphysoids, 8-15 x 2.0-3.0 um, cylindrical,

hyaline, 0-3 septate. BASAL STROMA 8-12 um thick, composed of dark-brown

textura angularis and globulose. INTERNAL MATRIX STROMA poorly developed,

10-20 um thick in the center, hyaline, consisting of gelatinous textura intricata.

ExcIPULUM 40-70 um thick in the upper, abruptly thinner towards the base,

arising from the inner layer of the covering stroma. SUBHYMENIUM 15-20

um thick, composed of colourless textura porrecta or angularis. PARAPHYSES

110-140 x 2.0-2.5 um, filiform, abruptly enlarged to 4.5-5.5 um and subfusoid-

ventricose above, with a 2.5-6.0 x 1.0-2.0 um, subcylindrical mucro at the

apex, often agglutinated. Asci ripening sequentially, 104-119 x 4.2-5.0 um,

cylindrical, short-stalked, apex round to obtuse, thin-walled, J-, 8-spored.

Ascosporss fasciculate, 60-86 x 0.6-0.8 um, filiform, hyaline, aseptate, with

an indefinite gelatinous sheath.

Rhytismataceae on Lithocarpus (China) ... 315

ILLUSTRATION: Korf & Zhuang (1985: Fig. 2).

ECOLOGY & DISTRIBUTION: Ascomata found on decaying leaves. Widely

distributed in Indonesia (Korf & Zhuang 1985). In China, it is known from

Sichuan and Anhui provinces (Lin et al. 2001).

SPECIMENS EXAMINED: On Lithocarpus glaber: CHINA, ANHUI, Mt HUANGSHAN,

Tangkou, alt. ca 550 m, 11 August 1995, Y.R. Lin, L. Chen 1596b (AAUF 67704b). On L.

henryi: CHINA, ANHUI, Mt Huancsuan, Tangkou, alt. ca 550m, 29 September 1993,

Y.R. Lin et al. 1511b (AAUF 67619b); Taoyuanting, alt. ca 650 m, 10 June 1994, Y.R. Lin,

S.M. Yu 1569a (AAUF 67677a); Wenquan, alt. ca 700 m, 3 October 2009, Q. Zheng, X.M.

Gao 5411 (AAUF 71519).

ComMENTS— Korf & Zhuang (1985) first described C. mucronatus based

on specimens on fallen leaves of Castanopsis sp. from Sichuan in China and

Java in Indonesia. The Mt Huangshan specimens usually produce fruit bodies

on decaying fallen leaves or leaf fragments, suggesting that the species is not

implicated in any plant disease. The other features of this species agree with

those of the type specimen described by Korf & Zhuang (1985), except for the

inlaid depth of the ascomata and the growth pattern of the excipulum.

Coccomyces mucronatoides Y.R. Lin, Q. Zheng & S.M. Yu, sp. nov. Fics. 1-6

MycoBank MB561883

Ascomata (480-—)600-1100 um, trigona usque ad hexagona, flavo-brunnea, subepidermalia.

Periphysoidei rarissimi. Excipulum 10-15um crassum, ex hyphis hyalinis septatis parallelis

constatum. Paraphyses filiformes, apice abrupte tumidae. Asci 85-120 x 4.5-5.0um,

cylindrico-clavati. Ascosporae 70-105 x 0.8—1.0 um, filiformes, hyalinae, vagina gelatinosa

tenui indutae.

Type: China, Anhui, Mt Huangshan, Tangkou, alt. ca 550 m, on Lithocarpus cleistocarpus

(Seemen) Rehder & E.H. Wilson, 11 September 2007, S.J. Wang, Y.R. Lin 2244b

(Holotype, AAUF 68352b).

EryMo_oey: referring to the similarity of the paraphysis apex to Coccomyces mucronatus

in shape.

ZONE LINES somewhat frequent, black-brown or yellow-brown, thin,

surrounding the bleached spots.

CONIDIOMATA not observed.

AscoMatTa developing on both sides of leaves, more on the upper side of

the leaf, scattered in irregular, yellowish-white or yellowish beached spots. In

surface view, ascomata (480—)600-1100 um diam., triangular to hexagonal,

yellow-brown, not shiny, with a clearly defined edge, somewhat raising the

substratum surface and flattened or slightly concave in the central region, with

an obvious preformed dehiscence mechanism, opening by 3-6 radial splits,

which extend nearly to the edge of ascoma, to expose the sallow hymenium.

Lips absent. In median vertical section, ascomata subepidermal. COVERING

STROMA 25-32 um thick near the opening and 8-15 um thick in other parts,

composed of an upper part of textura globulosa with black-brown cells 5-6 um

316 ... Zheng & al.

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Fics. 1-6. Coccomyces mucronatoides on Lithocarpus cleistocarpus (from holotype). 1. A leaf bearing

ascomata. 2. Ascomata as seen under a dissecting microscope. 3. Ascoma in median vertical section.

4. Detail of ascoma in median vertical section. 5. Paraphyses and asci. 6. Discharged ascospores.

Rhytismataceae on Lithocarpus (China) ... 317

diam., an lower part of textura epidermoidea with dark-brown cells 3.0-4.5 um

diam., and a few yellowish angular cells almost connecting to the basal stroma.

Periphysoids extremely sparse, (5—)8-15 x 2.0-2.5 um, cylindrical, hyaline,

(1-)3-5 septate. BASAL STROMA moderately developed, 10-15(-18) um thick,

composed of black-brown textura angularis or globulosa with thick-walled

cells 5-7 um diam. INTERNAL MATRIX STROMA well developed, 20-30(-45)

um thick, consisting of hyaline, gelatinised textura intricata. ExcIPULUM 10-15

um wide, composed of rows of hyaline, septate, thin-walled, parallel hyphae.

SUBHYMENIUM 15-22 um thick, composed of hyaline textura intricata and

angularis. PARAPHYSES 100-140 x 1.8-2.0 um, filiform, aseptate, abruptly

enlarged to 4.0-6.0 um and elliptical or subfusoid near the apex, cemented in

a thin gel, forming an epithecium 15-20 um thick. Asci ripening sequentially,

85-120 x 4.5-5.0 um, cylindrical, stalked, apex round to obtuse, thin-walled,

J—-, 8-spored. Ascosporgs arranged in a fascicle, 70-105 x 0.8—1.0 um, filiform,

hyaline, aseptate, with a thin gelatinous sheath.

ECOLOGY & DISTRIBUTION: Ascomata found on dead leaves in litter. Known

only from the type locality.

ComMENTS—According to the type descriptions, C. mucronatus differs from

the new species in frequent periphysoids, poorly developed internal matrix that

occurs only between covering stroma and basal stroma, and paraphyses with

an abruptly swollen subfusoid-ventricose upper part with a mucro at the tip

(Korf & Zhuang 1985). Coccomyces mucronatus on Lithocarpus spp. observed

by the authors differs from C. mucronatoides in an excipulum arising from

the inner layers of covering stroma, with 1-2 dark-brown, swollen cells at the

apex. However, the ascomata of the new species from Mt Huangshan have few

periphysoids, a well-developed internal matrix, and non-mucronate paraphyses

with a suddenly enlarged elliptical or subfusoid body near the apex.

Lophodermium agathidis Minter & Hettige, New Zealand Journal of Botany 21: 39,

1983

Type: NEW ZEALAND, AUCKLAND, WAITAKERE RANGES, Oratia, Kelly's Bush, on

Agathis australis (D. Don) Lindl. ex Loudon, 10 November 1979, G. Hettige 259066

(IMI).

ZONE LINES usually frequent, surrounding the bleached spots.

ConipiomarTa with a distribution on the leaf similar to that of the ascomata,

scattered to crowded. In surface view, conidiomata 140-300 um diam.,

somewhat round, black-brown in the centre and at the perimeter line of the

conidioma, concolorous with the substratum surface elsewhere, or grey-brown

to black-brown as a whole. In vertical section, conidiomata subepidermal.

UPPER WALL 4-7 um thick, dark-brown, consisting of thick-walled angular

cells 2-4 diam. BASAL WALL brown to dark-brown, composed of 1-2 layers of

318 ... Zheng & al.

thin-walled angular cells 3-5 um diam. SUBCONIDIOGENOUS LAYER 7-10 um

thick, consisting of thick-walled angular cells. CONIDIOGENOUS CELLS 10-16 x

2.0-3.0 um, flask-shape, proliferating sympodially. Conip1a 3.0-4.0 x ca 1.0

um, elliptical, hyaline, aseptate.

AscomaTa on both sides of leaves scattered in irregular, yellow-brown

bleached spots. In surface view, ascomata 800-1900(—2300) x 390-650 um,

elliptical to elongate-elliptical, black, slightly raising the substratum surface,

opening by a longitudinal split. Lips light reddish-brown. In median vertical

section, ascomata subepidermal. COVERING STROMA consisting of thick-walled

aliform-angular cells 3-7 um diam., 55-70 um thick and heavily carbonized near

the opening, other parts dark-brown, becoming suddenly thinner towards the

edge. Lip cells arranged in a subparallel manner or slightly radially, cylindrical.

BASAL STROMA 6-10 um thick, dark-brown, consisting of thick-walled aliform-

angular cells 4-7 um diam. SUBHYMENIUM ca 12 um thick, hyaline, composed of

angular cells. PARAPHYSES 110-160 x 1.5-2.0 um, filiform, irregularly swollen

to 2.0-3.0 um or occasionally branched above, covered with a mucous coating.

AscI maturing sequentially, 100-145 x 6-7.5 um, cylindrical, short-stalked,

apex round, thin-walled, J-, 8-spored. Ascospores arranged in fascicles

sometimes helically coiled, 70-95 x 1.2-1.6 um, filiform, hyaline, 0-1-septate,

enveloped in a ca 0.5 um thick gelatinous sheath.

ILLUSTRATION: Minter & Hettige (1983: Figs. 1-9).

ECOLOGY & DISTRIBUTION: Ascomata found on dead leaves in litter. Widely

distributed in China (Lin et al. 2005), Malaysia (Spooner 1991) and New

Zealand (Johnston 2001).

SPECIMENS EXAMINED: On Lithocarpus glaber: CHINA, ANHUI, Mr HuANGsHAN,

Tangkou, alt. ca 550 m, 5 April 1994, Y.R. Lin et al. 1655 (AAUF 67763). On L. henryi:

CHINA, Anuul, Mt Huancsuan, Tangkou, alt. ca 550 m, 2 October 2009, Q. Zheng,

X.M. Gao 5385 (AAUF 71493).

ComMMENTS—Lophodermium agathidis occurs on a wide range of gymnosperms

and angiosperms (Minter & Hettige 1983; Johnston 1989a, 2001). Minter &

Hettige (1983) inferred that it is probably a leaf endobiont inhabiting apparently

healthy leaves that fruits only after death of the leaf from some other cause. After

examination of specimens from China, Lin et al. (2005) found that on leaves of

different plants, L. agathidis differs in thickness of the covering stroma, location

of the lip cells, and size of asci and ascospores.

Terriera coacervata Y.R. Lin & Q. Zheng, sp. nov. Fics. 7-13

MycoBank MB561884

Ascomata coalescentia. Singula ascomata (650—)1000-1800(-—2200) x 350-560 yum,

plerumque elliptica, subepidermalia. Extentio valde carbonacea ad apicem stromatis

tegentis affixa. Excipulum maxime debiliter evolutum. Paraphyses filiformes. Asci 90-130

Rhytismataceae on Lithocarpus (China) ... 319

Fics. 7-13. Terriera coacervata on Lithocarpus cleistocarpus (from holotype). 7. A leaf bearing

ascomata. 8. Ascomata as seen under a dissecting microscope. 9. Three coalescent ascomata in

median vertical section. 10. Ascoma in median vertical section. 11. Detail of ascoma in median

vertical section. 12. Paraphyses and asci. 13. Discharged ascospores.

320 ... Zheng & al.

x 6.0-7.0 um, cylindrici. Ascosporae 60-110 x 1.5-1.8 yum, filiformes, hyalinae, vagina

gelatinosa indutae.

Type: China, Anhui, Mt Huangshan, Tangkou, alt. ca 550 m, on leaves of Lithocarpus

cleistocarpus, 11 September 2007, S.J. Wang, Y.R. Lin 2244a (Holotype, AAUF 68352a).

EryMo_oey: referring to the coalescent form of the ascomata.

ZONE LINES infrequent, thin, grey-brown to brown, sometimes not defined,

surrounding or partly surrounding the bleached spots.

CONIDIOMATA absent.

AscomatTa developing on lower side of leaves, 2-9 ascomata coalescent in

irregular, yellowish bleached spots 15-25 um diam. In surface view, ascomata

interlaced or accumulatively coalescent, forming multilocular fruit bodies.

Single ascomata (650—)1000-—1800(—2200) x 350-560 um, elliptical, sometimes

branching into lobed or polygonal shapes, black, slightly shiny, markedly raising

the substratum surface, opening by a longitudinal split more than 4/5 the length

of the ascoma or by not less than 3 lobes. Lips absent. In median vertical section,

ascomata subepidermal with epidermal cells becoming filled with fungal tissue

as ascomata develop. COVERING STROMA 20-30 um thick, composed of black-

brown textura angularis-globulosa with thick-walled cells. Along the edge of

the ascoma opening, there is a short extension, about 12 um thick, adjacent to

the covering stroma, which is comprised of strongly carbonized tissue with no

obvious cellular structure. BASAL STROMA strongly concave, 10-18 um thick,

composed of 3-4 layers of black-brown, angular and globose cells 2.8-4.0 um

diam., extending to the covering stroma. Colorless to light grey-brown textura

prismatica 10-22 um thick exists between the covering stroma and basal

stroma. ExCIPULUM very poorly developed, arising from the inner layer of the

basal stroma, consisting of hyaline textura porrecta. SUBHYMENIUM 8-12 um

thick, composed of hyaline textura angularis. PARAPHYSES 120-160 x 1.4-1.6

um, filiform, usually gradually swollen to ca 2.0 um at the apex, not branched,

covered with a ca 0.8 um thick mucous coating. Asci maturing sequentially,

90-130 x 6.0-7.0 um, cylindrical, short-stalked, apex round, J-, 8-spored.

ASCOSPORES arranged in a fascicle or helically coiled, 60-110 x 1.5-1.8 um,

filiform, hyaline, aseptate, enveloped in a 1.0-1.5 um thick gelatinous sheath.

ECOLOGY & DISTRIBUTION: Ascomata found on dead or fallen leaves and

known only from the type locality.

ComMMENTS— This new species has characteristics typical of Terriera. In each

ascoma, there is a flattened extension comprising strongly carbonized tissue

adjacent to the covering stroma along the edge of the ascomatal opening, a very

poorly developed hyaline textura porrecta excipulum, and a colorless to light

grey-brown textura prismatica between the covering stroma and basal stroma.

Terriera coacervata is unusual and unique in the genus because its ascomata

usually interlace or accumulatively coalesce to form multilocular fruit bodies.

Rhytismataceae on Lithocarpus (China) ... 321

Lophodermium euryae Y.R. Lin & Y.F. He differs from T. coacervata in a different

coalescent form, the oblong, elliptical or broadly elliptical smaller ascomata

(290-650(-870) x 250-350 um), smaller ascospores (45-70 x 0.8-1.2 um),

paraphyses with suddenly swollen to subglobose apices branched to form an

epithecium, and the abundant conidiomata (Lin et al. 2005).

We discovered a hyperparasite — the anamorphic fungus, Cladosporium

lophodermii Georgescu & Tutunaru — in some T! coacervata collections that

infects ascomata to produce dark-brown hyphae and conidia in the interior.

Hyperparasitism is frequent in some Lophodermium taxa (Minter 1981; Lin

1989).

Terriera illiciicola (S.J. Wang, Y.F. He & Y.R. Lin) Q. Zheng & Y.R. Lin, comb. nov.

MycoBank MB561885

= Lophodermium illiciicola S.J. Wang, Y.F. He & Y.R. Lin, Mycosystema 25: 1, 2006.

Type: CHINA, ANHUI, MT HuancsHan, Tangkou,, alt. ca 550 m, on Illicium verum

Hook. f., 25 September 2004, Y.R. Lin et al. 1912 (AAUF 67683).

ZONE LINES frequent, black, thin, surrounding or partly surrounding the

bleached spots.

CONIDIOMATA absent.

AscomMatTa developing on the upper side of leaves, crowded in yellowish to

yellow-brown bleached spots. In surface view, ascomata 300-380 x 280-330

um, subcircular to broad-elliptical, black, shiny, with a clearly defined edge,

moderately raising the surface of the substratum, opening by a longitudinal

split. Lips absent. In median vertical section, ascomata subepidermal. COVERING

STROMA 25-30 um thick, consisting of black-brown textura angularis with

cells 4.5-6.5 um diam. Along the edge of the ascomatal opening, there is

an extension, about 15 um thick, adjacent to the covering stroma, which is

comprised of strongly carbonized tissue with no obvious cellular structure and

tends to disappear as the ascomata mature. BASAL STROMA concave and cup-

shaped, 12-18 um thick, consisting of black-brown textura angularis with 2(-3)

layers of cells 5.0—7.5 um diam., extending to the covering stroma. Colorless or

light brown textura prismatica 15-20 um thick exists between the covering and

basal stromata. SUBHYMENIUM 12-15 um thick, consisting of textura angularis-

porrecta. PARAPHYSES 115-150 x 1.0-1.5 um, filiform, the top not swollen or

branched, covered with an inconspicuous mucous coating. Asci maturing

sequentially, 90-135 x 4.0-5.0 um, narrow-cylindrical or cylindric-clavate,

long-stalked, apex round, J—, 8-spored. Ascosporss fasciculate, 65-95 x ca

1.0 um, filiform, hyaline, aseptate, enveloped in an inconspicuous gelatinous

sheath.

ILLUSTRATION: Wang et al. (2006: Fig. 1).

ECOLOGY & DISTRIBUTION: Ascomata found on fallen leaves and known

only in China (Wang et al. 2006).

322 ... Zheng & al.

SPECIMENS EXAMINED: On Lithocarpus cleistocarpus: CHINA, ANHUI, MT HUANGSHAN,

Shibawan, alt. ca 1000 m, 22 September 2004, S.J. Wang, Y.F. He, Y.R. Lin 1806 (AAUF

67914).

ComMMENTS— This species was originally described as Lophodermium illiciicola

(Wang et al. 2006) and is transferred here because its diagnostic characteristics

agree with those of the genus Terriera. The species resembles T. minor (Tehon)

PR. Johnst. (Ortiz-Garcia et al. 2003), which can be differentiated by oblong

or elliptical larger ascomata (320-800(-1400) x 220-340 um), a flat basal

stroma, and paraphyses that branch 2-3 times at the top, sometimes forming

an epithecium (Johnston 1988, 1989b; Lin et al. 2005).

Key to species of the Rhytismataceae on Lithocarpus spp. worldwide

la. Ascomata polygonal, opening by radial splits .................. 0... eee eee eee 2

1b. Ascomata more or less elliptical or round, opening by a longitudinal split ....... 6

2a. Lip cells:present at-least‘at the:early stage: . foi. wie oe ales Wines lng le etn le 8 ae ea 3

204 Lapreciis absentee oo. Wee ie 0.05 Rk Fe a caer cen oe om, LeBel eae ete ad. 4

3a. Ascomata 700-1100 um diam., with periphysoids ............ C. huangshanensis

3b. Ascomata (700—)1000-1600(-1800) um diam., without periphysoids .... C. delta

4a. Paraphyses gradually swollen at apex ........... 0. cee eee eee eee C. dentatus

4b. Paraphyses‘suddenly swollen’ at‘apex cn... vues ents wast eles Uae oe ee oes 5

5a. Paraphyses swollen to subfusoid-ventricose above, with a subcylindrical

EaL Veh ahr 1G | =>.) A ore Roe, AO Ae AE Sen eee C. mucronatus

5b. Paraphyses swollen to elliptical or subfusoid above, without a mucro at apex

aad ral AEA Re OER A ERATE CARAT SA te AOD LES SD C. mucronatoides

6a. Top of covering stroma with a flattened, strongly carbonized extension ......... 7

6b. Top of covering stroma without an extension ...................06. L. agathidis

7a. Ascomata interlaced or accumulatively coalescent; paraphyses gradually

SWOlleMFADO Ves. p50 Fs oe Rice le Rag ois ape Nae has Eig a Pa T. coacervata

7b. Ascomata crowded; paraphyses not swollen above .................. T. illiciicola

Acknowledgements

The authors are grateful for the pre-submission comments and suggestions provided

by Dr D.W. Minter and Dr C.L. Hou and to Dr S.J. Wang and Dr X.M. Gao for the field

investigations. This study was supported by the National Natural Science Foundation

of China (No. 30870014), the Specialized Research Fund for the Doctoral Program

of Higher Education of China (No. 20070364002), the Natural Science Foundation of

Anhui Province of China (No. 070411005).

Literature cited

Farr DF, Rossman AY. 2011. Fungal Databases, Systematic Mycology and Microbiology Laboratory.

ARS, USDA [http://nt.ars grin.gov/fungaldatabases/fungushost/FungusHost.cfm (viewed

online on 2 April 2011)].

Rhytismataceae on Lithocarpus (China) ... 323

Johnston PR. 1986. Rhytismataceae in New Zealand 1. Some foliicolous species of Coccomyces de

Notaris and Propolis (Fries) Corda. N. Z. J. Bot. 24: 89-124.

Johnston PR. 1988. An undescribed pattern of ascocarp development in some non-coniferous

Lophodermium species. Mycotaxon 31: 383-394.

Johnston PR. 1989a. Rhytismataceae in New Zealand 2. The genus Lophodermium on indigenous

plants. N. Z. J. Bot. 27: 243-274.

Johnston PR. 1989b. Lophodermium (Rhytismataceae) on Clusia. Sydowia 41: 170-179.

Johnston PR. 2001. Monograph of the monocotyledon-inhabiting species of Lophodermium.

Mycol. Pap. 176: 1-239.

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

10" ed. CAB International, Wallingford. 771 p.

Korf RP, Zhuang WY. 1985. Some new species and new records of Discomycetes in China.

Mycotaxon 22: 483-514.

Lin YR. 1989. Preliminary study of a hyperparasite, Cladosporium lophodermii [in Chinese]. Forest

Pest and Disease 8: 33.

Lin YR, Li ZZ, Huang CL, Xiang CT. 2000a. Studies on the genus Coccomyces from China II [in

Chinese]. Mycosystema 19: 297-301.

Lin YR, Li ZZ, Xie YS, Liang SW. 2000b. Studies on the genus Coccomyces from China IL [in

Chinese]. Mycosystema 19: 449-453.

Lin YR, Li ZZ, Xu ZS, Wang JR, Yu SM. 2001. Studies on the genus Coccomyces from China IV [in

Chinese]. Mycosystema 20: 1-7.

Lin YR, Yu SM, He YF, Wang SJ. 2005. One new species and two new Chinese records of

Lophodermium Chev. [in Chinese]. Mycosystema 24: 1-5.

Minter DW. 1981. Lophodermium on pines. Mycol. Pap. 147: 1-54.

Minter DW, Hettige G. 1983. Lophodermium agathidis and Meloderma richeae, two members of the

Rhytismataceae from Australasia. N. Z. J. Bot. 21: 39-48.

Ortiz-Garcia S, Gernandt DS, Stone JK, Johnston PR, Chapela IH, Salas-Lizana R, Alvarez-

Buylla ER. 2003. Phylogenetics of Lophodermium from pine. Mycologia 95: 846-859.

http://dx.doi.org/10.2307/3762013

Sherwood MA. 1980. Taxonomic studies in the Phacidiales: The genus Coccomyces (Rhytismataceae).

Occas. Pap. Farlow Herb. Cryptogram Bot. Harv. Univ. 15:1-120.

Spooner BM. 1991. Lophodermium and Hypoderma (Rhytismataceae) from Mt. Kinabalu, Sabah.

Kew Bulletin 46: 73-100. http://dx.doi.org/10.2307/4110745

Teng SC. 1934. Notes on Discomycetes from China. Sinensia 5: 431-465.

Wang SJ, He YF, Ye GM, Lin YR. 2006. A new species and a new Chinese record of Rhytismataceae

[in Chinese]. Mycosystema 25: 1-5.

MYCOTAXON

http://dx.doi.org/10.5248/118.325

Volume 118, pp. 325-329 October-December 2011

Aschersonia conica sp. nov. (Clavicipitaceae)

from Hainan Province, China

JUN-ZHI Qiu, Yu-BIN Su, CHONG-SHUANG WENG & XIONG GUAN"

Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education,

Fujian Agriculture and Forestry University, Fuzhou, 350002 Fujian, P. R. China

* CORRESPONDENCE TO: guanxfafu@126.com

ABSTRACT — Aschersonia conica, a new anamorphic species of the family Clavicipitaceae,

is described and illustrated based on specimens collected in Hainan Province, China.

The entomogenous fungus is characterized by conical, whitish to pale yellow stromata

that are surrounded by a hypothallus and a wide base, wide ostiolar openings, cylindrical

conidiogenous cells in a compact palisade, filiform sterile elements, and fusiform conidia.

Key worps — morphology, taxonomy, new taxon

Introduction

Aschersonia Mont., a large genus in the family Clavicipitaceae, has been well

studied, especially in America and Europe (Hywel-Jones & Evans 1993; Liu et

al. 2005). Recently, new studies on Aschersonia and Hypocrella were carried out

using both morphological and molecular techniques (Liu et al. 2006; Spatafora

et al. 2007; Sung et al. 2007). Chaverri et al. (2008), who have provided the

most extensive revision of Aschersonia since Petch (1921), accepted 32 species.

They showed that Aschersonia is a form genus that shares similar anamorph

morphologies with Moelleriella and Hypocrella but that both teleomorphic

genera can be distinguished by their ascospore disarticulation and conidial

shape and size. Mongkolsamrit et al. (2009) later published three additional

species based on morphology combined with ITS and B-tubulin sequence

analysis.

In China, Tzean et al. (1997) and Qiu et al. (2009, 2010) studied Aschersonia

and its teleomorphic forms, Hypocrella Sacc., Moelleriella Bres., and Samuelsia

P. Chaverri & K.T. Hodge. During a recent survey on insect pathogenic

fungi in tropical forests in southern China, a previously unknown species

of entomopathogenic Aschersonia was collected in the Jianfengling National

Nature Reserve and is proposed here as a new species.

326 ... Qiu & al.

Materials & methods

Fungal measurements and microscopical observations followed Qiu et al. (2009,

2010). Spore range data exclude 5% of measurements from each end of the range, which

are given in parentheses. Text abbreviations are as follows: CB = Cotton Blue, 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 among all specimens studied,

n = number of spores measured from a given number of specimens. Conidiomata

were carefully dissected with a razor blade and mounted in water or lactic acid mixed

with cotton blue on a slide. Special color terms follow Kornerup & Wanscher (1967).

Specimen vouchers are deposited at the Mycological Herbarium, Fujian Agricultural

and Forestry University (MHFAFU).

Taxonomy

Aschersonia conica Jun Z. Qiu, Y.B. Su & C.S. Weng, sp. nov. Fies. 1

MycoBank MB 561561

Stromata pyriformia vel conico-pulvinata, valde convexa, ex hyphis densis composita,

deorsum effusa, hypothallum membranaceum ad 1.8 mm diam. 0.5 mm altum dilute

luteum formantia, in statu recenti albida, superficie aliquot orificiis ut punctis magnis

visibilibus praedita. Pycnidia plerumque singula, in medio stromatis immersa, 252-306

um alta, 103-141 um diam., elongate ampulliformia. Phialides cylindricae, ad 15 um

longae. Paraphyses praesentes, filiformes, flexuosae, ad 116 um longae, 0.7 um latae.

Conidia fusoidea, utrinque acutata, 8.2-10.5 x 0.9-1.3 um.

Type — China, Hainan Prov., Dongfang County, Jianfengling National Nature Reserve,

alt. 1450 m, on Aleyrodidae, 29.X.2008, J.Z. Qiu, Y.B. Su & C.S. Weng 311 (Holotype,

MHFAFU 20811).

EryMo.Locy — Refers to the conical stroma of this species.

TELEOMORPH: Unknown.

STROMATA—Pyriform, conical-pulvinate, markedly convex, consisting of

dense hyphae, base spreading, forming a pale yellow membranous hypothallus

up to 1.8 mm diam., 0.5 mm high, whitish-coloured when fresh, several ostiolar

openings as large dots visible on the surface.

Pycnip1A—Usually single, embedded in the centre of the stroma, CB-,

252-306 um high, 103-141 um diam, elongate flask-shaped. Conidiogenous

cells phialidic, cylindrical, up to 15 um long.

PARAPHYSES— Present, linear, filiform, flexuous, up to 116 um long, 0.7 um

wide.

Conip1A—Fusoid, sometimes narrowly fusiform, with acute ends, 8.2-10.5

x 0.9-1.3 um, L = 9.5 um, W = 1.1um, Q = 8.6-8.7 (n = 60/2).

ADDITIONAL SPECIMEN EXAMINED — CHINA. HAINAN PROv.: LEDONG COUNTY,

Jianfengling National Nature Reserve, alt. 1350 m, on Aleyrodidae, 30.X.2008, J.Z. Qiu,

Y.B. Su & C.S. Weng 358 (MHFAFU 20828).

CoMMENTS—Aschersonia conica is characterized by the whitish to pale yellow

markedly convex conical stromata, long conidia, wide ostiolar openings, and

Aschersonia conica sp. nov. (China) ... 327

}

FiGuRE 1. Aschersonia conica. A: Stroma; B: Pycnidium; C: Horizontal-section of a flask-shaped

pycnidium; D: Conidiophores and conidiogenous cells; E: Paraphyses; F: Conidia. Scale bars: A = 1

mm; B-C = 100 um; D-E = 30 um; F = 10 um.

328 ... Qiu & al.

presence of paraphyses and hypothallus. Two previously described species, A.

turbinata Berk. and A. juruensis Henn. (Petch 1921, Chaverri et al. 2008), also

have similarly shaped stromata. However, A. turbinata differs in having ovoid

conidia that are longer and wider (10.5-11.2 x 4.5-5.0 um), producing copious

slime, and lacking paraphyses. Aschersonia juruensis differs in pulvinate

stromata with sloping sides (convex), more conidiomata per stroma (>20),

longer wider conidia (14.5-15.5 x 3.7-4.0 um) that are ventricose with acute

ends, and the absence of paraphyses.

Acknowledgements

We express our gratitude to Prof. Guo-Zhong Lt (College of Environment and

Resources, Dalian Nationalities University) and Dr. Ryan Kepler (Department of

Entomology, Cornell University) for serving as pre-submission reviewers. We would

especially like to thank Prof. Wen-Ying Zhuang and Prof. Jian-Yun Zhuang (Institute

of Microbiology, Chinese Academy of Sciences) for providing the Latin diagnoses, and

Dr. Shaun Pennycook for nomenclatural review. This research was supported by the

National Natural Science Foundation of China (30500005, 31070026), the Educational

Programs for Science and Technology Development (JA09085), Fujian Provincial

Science Foundation for Distinguished Young Scholars (2010J06007), and the Key

Program for Constructing the Economic Zone on the Western Side of Taiwan Straits,

Fujian Province (0b08b005).

Literature cited

Chaverri P, Liu M, Hodge KT. 2008. A monograph of the entomopathogenic genera Hypocrella,

Moelleriella and Samuelsia gen. nov. (Ascomycota, Hypocreales, Clavicipitaceae), and their

anamorphs in the Neotropics. Stud. Mycol. 60: 1-66. http://dx.doi.org/10.3114/sim.2008.60.01

Hywel-Jones NL, Evans HC. 1993. Taxonomy and ecology of Hypocrella discoidea and its anamorph,

Aschersonia samoensis. Mycol. Res. 97(7): 871-876.

Kornerup A, Wanscher JH. 1967. Methuen handbook of colour. Methuen, London.

Liu M, Rombach MC, Humber RA, Hodge KT. 2005. What’s in a name? Aschersonia insperata:

a new pleoanamorphic fungus with characteristics of Aschersonia and Hirsutella. Mycologia

97: 246-253.

Liu M, Chaverri P, Hodge KT. 2006. A taxonomic revision of the insect biocontrol fungus

Aschersonia aleyrodis, its allies with white stromata and their Hypocrella sexual states. Mycol.

Res. 110: 537-554. http://dx.doi.org/10.1016/j.mycres.2006.01.013

Mongkolsamrit S, Luangsa-Ard JJ, Spatafora JW, Sung GH, Hywel-Jones NL. 2009. A combined

ITS rDNA and beta-tubulin phylogeny of Thai species of Hypocrella with non-fragmenting

ascospores. Mycol. Res. 113(6-7): 684-699. http://dx.doi.org/10.1016/j.mycres.2009.02.004

Petch T. 1921. Studies in entomogenous fungi. II. The genera of Hypocrella and Aschersonia. Ann.

Roy. Bot. Gard. Peradeniya 7: 167-278.

Qiu JZ, Ma HE, Wang YY, Guan X. 2009. Two Aschersonia species from Fujian new to China.

Mycosystema 28(1): 60-63.

Qiu JZ, Sun CY, Guan X. 2010. A new pathogen of scale insects, Aschersonia fusispora sp. nov.

(Clavicipitaceae) from Guangxi Province, China. Mycotaxon 113: 81-85.

http://dx.doi.org/10.5248/113.81

Aschersonia conica sp. nov. (China) ... 329

Spatafora JW, Sung GH, Sung JM, Hywel-Jones NL, White Jr JE 2007. Phylogenetic evidence for

an animal pathogen origin of ergot and other grass endophytes. Mol. Ecol. 16: 1701-1711.

http://dx.doi.org/10.1111/j.1365-294X.2007.03225.x

Sung GH, Hywel-Jones NL, Sung JM, Luangsa-ard JJ, Shrestha B, Spatafora JW. 2007.

Phylogenetic classification of Cordyceps and the clavicipitaceous fungi. Stud. Mycol. 57: 5-59.

http://dx.doi.org/10.3114/sim.2007.57.01

Tzean SS, Hsieh LS, Wu WJ. 1997. Atlas of entomopathogenic fungi from Taiwan. Council of

Agriculture, Executive Yuan, Taiwan, R.O.C. 214 p.

MYCOTAXON

http://dx.doi.org/10.5248/118.331

Volume 118, pp. 331-336 October-December 2011

Clarkeinda trachodes (Agaricales, Basidiomycetes),

first record from Bangladesh

Mb. IQBAL HOSEN’ & Z.W. GE?

'Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany,

Chinese Academy of Sciences, Kunming 650204, Yunnan, P. R. China

?Graduate University of Chinese Academy of Sciences, Beijing 100039, P. R. China

CORRESPONDENCE TO: ’” iqbalpatho@gmail.com & ' zwge@mail.kib.ac.cn

ABSTRACT — An interesting agaric, Clarkeinda trachodes, is reported for the first time from

Bangladesh. A full description, discussion, and illustrations are provided. The enigmatic agaric

genus, currently known only from south and southeast Asia, is characterized by the presence

of a fawn colored pellicle on the central pileus surface, a stipe with a superior annulus and

basal volva, and thick-walled pigmented spores with a slightly depressed truncated apex. Our

examination of a voucher specimen from Italy indicates that a recent report of Clarkeinda in

Europe was based on a misidentified Agaricus collection.

KEY worps — Agaricaceae, distribution, morphology, taxonomy

Introduction

Taxonomic reports on macrofungi of Bangladesh are rare in the literature.

As a part of an ongoing effort to document the macrofungi of South Asia,

we report the presence of Clarkeinda trachodes from Bangladesh for the first

time. We provide a detailed morphological description, photographs, and line

drawings of this enigmatic agaric.

Materials & methods

During November 2009 and June 2010 specimens were collected several times in

two different locations in Bangladesh: Bhawal National Park, Gazipur (24°45’N 90°50’E;

altitude 20 m), and Akhanagar, Thakurgaon, in northern Bangladesh, (26°03N’ 88°47’E;

altitude 67 m). Specimens were deposited in the SAU Herbarium of Agarics Flora

(SHAF; Plant Pathology Department, Faculty of Agriculture, Sher-e-Bangla Agricultural

University, Dhaka, Bangladesh) and in the Kunming Institute of Botany Herbarium,

Academia Sinica (KUN, with HKAS numbers), Kunming, Yunnan, China.

332 ... Hosen & Ge

Fresh specimens were described and photographed in the field. Small tissue pieces

were dried in silica gel and the remaining basidiomes were dried using an oven. Color

codes follow Kornerup & Wanscher (1978).

Microscopical observations were made in mounts of H,O, glycerin, 5% aqueous

KOH, Congo red solution, and Melzer’s reagent, and at least 20 basidiospores were

measured for each of the six basidiomes collected. Q = average length/width ratio

derived from each basidiospore measured. Line drawings of microstructures were made

from the rehydrated specimens.

To prepare the basidiospores for scanning electron microscopy (SEM), silica dried

basidiome fragments were mounted on aluminum stubs with double-sided tape, coated

with gold palladium, and photographed with a Hiracui S—4800 SEM.

Taxonomy

Clarkeinda trachodes (Berk.) Singer, Lilloa 22: 413, 1951. Fics 1-11

BASIDIOMATA medium to large, fleshy. PILEUs 80 to 110 mm in diameter,

subglobose or hemispherical when young, and becoming convex to applanate

at maturity; pellicle brown (6E8) to coffee (6E5-6) or chocolate brown (6E5,

6F5), thin to thick when young and somewhat brown to grayish brown (7E3-

5) at maturity; the remaining surface covered with grayish brown (7E3-5)

to vinaceous brown (6D8, 8E8) squamules, together with numerous, small,

revolute and loosely floccose, brown squamules; context up to 9 mm thick

in the center of the pileus, white, instantly turning reddish with exposure.

LAMELLAE 47 x 10 mm, free and remote from stipe, white to dirty white when

young, olive brown when mature, becoming red brown after bruising, crowded

with lamellulae, margin entire, concolorous. STIPE 80-121 x (28—)31—47(-61)

mm, central, subcylindrical, solid but fistulose in aged specimens; surface dirty

white to white at the apex, light brown to brown toward the base, glabrous

above the annulus, lower half densely covered with minute, brown, furfuraceous

squamules. ANNULUS present on the upper part of the stipe but not the top, up

to 18 mm, thick and membranous and remaining up to the maturity, sometimes

detached from the stipe when dry, adaxial part glabrous with fine longitudinal

striation but abaxial part rough with squamules. VoLva present, grayish, dirty

white to white, membranous, usually closely appressed to stipe and eventually

inconspicuous. SPORE DEPOSIT not obtained.

BaAsIDIA 18-26 x 6-9 um, mostly clavate to subclavate, thin-walled,

tetrasporic (occasionally 2- or 3-spored), bearing four short sterigmata, 1.1—2.3

x 0.8—1.2 um, hyaline, smooth, lacking incrustations, clamp connection absent.

BASIDIOLES narrowly clavate to clavate. HYMENOPHORAL TRAMA interwoven,

hyphae cylindrical to slightly inflated, up to 14 um wide, thin-walled, hyaline, and

without clamp connections. BAsiDIosPoREs (FIGs 3, 6—7) (5.0—)5.52—5.96(—6.8)

x (3.4—)3.81—-3.98(—4.3) um, mean Q = (1.38—)1.42—1.54(—1.7), ovoid, sometimes

broadly ellipsoid to ellipsoid, glabrous, thick-walled (up to 0.6 um), apiculus

Clarkeinda trachodes, new to Bangladesh ... 333

ad ete

Fics 1-5: Clarkeinda trachodes. 1. Basidiome: (a) immature, (b) mature, (c) longitudinal section of

mature fruiting body. 2. Basidia with basidioles. 3. Basidiospores. 4. Cheilocystidia. 5. Pileipellis.

334 ... Hosen & Ge

5.00um

Fics 6—13: Clarkeinda trachodes. 6—7. Basidiospores showing distinctly truncated and depressed

apical germ pore under SEM. 8-13. Developmental stages of a basidiome fruiting in its natural

habitat. (Photo: M.I. Hosen).

Clarkeinda trachodes, new to Bangladesh ... 335

eccentric, apex or germinating pore prominent and truncate with slightly

depressed, olive brown to dark, umber brown in deposit and maintaining the

same color after desiccation for 1.5 years at room temperature, dextrinoid

in Melzer’s solution, not metachromatic in Cresyl blue. CHEILOCYSTIDIA

24-32 x 10-16 um, abundant, scattered to more or less crowded, narrowly

clavate, clavate to broadly clavate, obpyriform, hyaline, thin-walled, smooth,

lacking incrustations, sometimes long pedicel and narrow up to 3.4 um.

PLEUROCYSTIDIA absent. PILEIPELLIs consisting of short branching chains of

4-7 cells, slightly interwoven, terminal cells 12—22.5 x 8-14 um, dull brown

vacuolar pigment inside the cells in glycerin, water and 5% KOH solutions,

thin-walled, clavate, cylindrical, obpyriform to fusiform or spindle-shaped in

rare cases, occasionally branching with lateral cells that are mostly clavate, basal

cells nearly subglobose to clavate or cylindrical.

HABIT, HABITAT, DISTRIBUTION— The fleshy basidiomes of C. trachodes fruit

as isolated individuals or in groups of two in disturbed habitats and at forest

edges. Our collections were found along the roadside near a Shorea robusta

(Dipterocarpaceae) forest in Bhawal National Park, Gazipur, and on bare soil

near a bamboo fence around a village residence in Akhanagar, Thakurgaon,

Bangladesh. Known also from China, India, Indonesia, Malaysia, and Sri

Lanka.

SPECIMENS EXAMINED — BANGLADESH. Duaka DIvIsIon: Gazipur, Bhawal

National Park, 20 m a.s.l., 30 September 2009, M. I. Hosen 617, 619 (HKAS 62856,

62857); RANGPUR Division: Thakurgaon, Akhanagar, 67 m a.s.l., 16 June 2010, M. I.

Hosen 85-88 (HKAS 62852-62855).

ADDITIONAL MATERIAL EXAMINED — Agaricus sp.: ITALY, South Italy, on sandy

ground next to Castanea sativa, 1100 m a.s.l., 13 October 1994, Carmine Lavorato

(HKAS 49489, as Clarkeinda trachodes).

ComMENTs — Clarkeinda trachodes is distinguished by a large basidiome size,

prominent chocolate or coffee brown to deep brown pellicle on the pileus disc

surface, presence of an annulus, olive brown to umber brown spore deposit,

slightly thick-walled spores with a truncated apex, and a context that changes

from white to reddish brown when cut. Since Berkeley (1847) first described the

species from Sri Lanka, it has been reported from south and southeast Asia by

Petch & Bisby (1950, as Chitoniella), Leelavathy et al. (1981), and Pegler (1985,

1986). Yang (1991) has also reported it from the tropical region of Yunnan,

China.

Examination of the specimen from Italy reported as C. trachodes by Lavorato

& Contu (2002) shows the European record is based on a misidentified Agaricus

specimen. The Italian specimen bears larger, ellipsoid basidiospores (7—8.5 x

4.5—5.5 um) that lack a germ pore and truncated apex, and pileal squamules that

represent a cutis of filamentous cylindrical hyphae 4—8 (—15) um in diameter.

336 ... Hosen & Ge

Vellinga et al. (2011) and our own sequence analyses (Ge unpub.) nest

Clarkeinda phylogenetically in the Agaricus s.l. clade close to Agaricus and

Heinemannomyces (genera within tribe Agariceae).

Acknowledgments

The authors wish to express sincere thanks to Dr. Zhu L. Yang (Kunming Institute

of Botany, Chinese Academy of Sciences, Kunming, China) for providing access

to important literature, improvements to the article, and valuable suggestions. The

authors are very grateful to Dr. Carmine Lavorato (Italy) for providing a specimen of

Clarkeinda trachodes for comparison. The first author is grateful to Prof. Nasim Akhtar,

Dr. Md. Rafiqul Islam, Dr. M. Salahuddin M. Chowdhury, Dr. EM. Aminuzzaman,

and Dr. Nazneen Sultana (Department of Plant Pathology, Sher-e-Bangla Agricultural

University, Dhaka, Bangladesh) and Dr. Ashraf Uddin Ahmed (SSO, Plant Pathology

Division, Bangladesh Agricultural Research Institute, Gazipur) for allowing him to keep

the samples in their lab. This work is partially supported by the National Natural Science

Foundation of China (No. 30800004), the Natural Science Foundation of Yunnan

Province (No. 2008CD164), and the Knowledge Innovation Program of the Chinese

Academy of Sciences (No. 2010KIBA01). Thanks are also extended to Dr. Matthew E.

Smith (Duke University, USA) and Haruki Takahashi (Japan) for serving as the pre-

submission reviewers and their constructive, thoughtful suggestions.

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MYCOTAXON

http://dx.doi.org/10.5248/118.337

Volume 118, pp. 337-347 October-December 2011

Glomus cubense sp. nov.,

an arbuscular mycorrhizal fungus from Cuba

Y. RODRIGUEZ’, Y. DALPE?’, S. SEGUIN?, K. FERNANDEZ’,

F. FERNANDEZ” & R.A. RIVERA’

'Departmento de Biofertilizantes y Nutricion de las Plantas, Instituto Nacional de

Ciencias Agricolas (INCA), Carretera a Tapaste km 3 %, PO Box 1, CP 32700,

San José de las Lajas, Mayabeque, Cuba

Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada,

960 Carling Ave, Ottawa K1A 0C6, Canada

°SYMBORG S.L. Campus de EspinardoNo 7. Edificio CEEIM, CP 30100, Murcia, Espana

* CORRESPONDENCE TO: yolande.dalpe@agr.gc.ca

ABSTRACT —Glomus cubense (Glomeraceae, Glomeromycetes) was isolated from a lagoon

vegetation area on a clay soil deposition environment in the vicinity of San José de las Lajas,

Cuba. The species description is based on spore morphological parameters from in vivo pot

cultures and molecular analyses. The new species is characterized by its small, generally

irregular in shape, 20-48 x (24—)54-72 um hyaline to faintly yellow spores that have a 2-

layered spore wall and arise in clusters. Phylogenetic analyses of the rDNA ITS region and

H+ATPase place the species into the Glomeraceae without close relatives among named

Glomus species. Glomus cubense forms mycorrhizal associations with sorghum and leek

plants under greenhouse pot-culture growing conditions and with a diversity of crop plants

under field conditions. The name cubense refers to Cuba, the country where the species was

found.

Keyworps — Glomeromycota, molecular phylogeny

Introduction

Since 1973, numerous investigations on the diversity of arbuscular

mycorrhizal fungi (AMF) have been performed in Cuba, and several species

have been described and surveyed from this country (Herrera & Ferrer 1980;

Ferrer & Herrera 1981, Herrera-Peraza et al. 2003). With respect to research

and development on the selection of AMF strains with potential for use as

bio-inoculants for agriculture and horticulture productions, AMF species

populations were surveyed and propagated in pure cultures, characterized

morphologically, and simultaneously tested for performance at stimulating

338 ... Rodriguez & al.

plant yields under both controlled and field conditions (Rivera et al. 2003, 2007;

Herrera-Peraza et al. 2011). Among the AMF strains surveyed and tested, there

was an undescribed species, here named Glomus cubense, which formed small,

hyaline, 2-layered glomoid spores in clusters around roots. Morphological

and molecular comparisons with other light-coloured glomoid spore species

(Blaszkowski et al. 2010) clearly distinguished G. cubense from previously

described species. Once propagated in pot cultures, it showed high root

colonization potential and growth benefits over a variety of crop plants tested.

In the present paper, the fungus is proposed as a new arbuscular mycorrhizal

species, G. cubense, in the Glomeromycota.

Materials & methods

AMF strain propagation and evaluation were conducted in the central greenhouse

of the National Institute of Agricultural Sciences (INCA) in San José de las Lajas, Cuba.

Species morphological and molecular characterizations were done at the Eastern Cereal

and Oilseed Research Centre of Agriculture and Agri-Food Canada, Ottawa, Canada.

AMF sampling and propagation

Soil samples were collected at about 50 cm depth from a lagoon area at “San

Rafael” Pond in “Las Papas” area (San José de las Lajas, Cuba), with a Haplic Cambisol

(chromic) soil (IUSS Working Group WRB, 2006), a natural soil deposition environment

occasionally flooded and mainly covered with Cynodon nlemfuensis and Mimosa pigra.

Mixed soil samples were used to establish pot trap-cultures using Sorghum bicolor as host

plant. After a four months growth period, spores of AMF were isolated by morphotypes

and used as inoculant for the establishment of pure cultures in sterilized (121°C for

3 h during three days) clay substrate from a “Gley vértico calcico” soil (Instituto de

Suelos 1999) from “Guayabal” farm in San José de las Lajas, Cuba. Plants were fertilized

and watered at regular intervals (Hewitt 1966). Pure strain cultures were subsequently

maintained as reference culture on sorghum plants under semi-controlled greenhouse

conditions.

AMF isolation and characterization

Spores were extracted from the substrate by wet-sieving (300-38 um mesh sieves)

and centrifugation in sucrose gradient (Dalpé & Hamel 2007). Spores were then

manually isolated from the supernatant under the dissecting microscope (Olympus

SZ-PT) by micropipetting. Plant roots were washed in water, bleached in 2.5% KOH

and stained with fuchsin acid (0.01% in lacto-glycerol) (Kormanik & McGraw 1982).

Once destained in lacto-glycerol, root sections were mounted on microscope slides in

polyvinyl] alcohol-lactic acid-glycerol (PVLG) (Omar et al. 1979), a mixture of PVLG and

Melzer’s reagent (1:1, v/v), or a mixture of PVLG and Cotton Blue (0.15% in lactic acid)

(1:1, v/v). Spore morphology and spore wall architecture were based on examination

of about a 100 spores. Color observations referred to the color chart code of the

International Culture Collection of Arbuscular and Vesicular-Arbuscular Mycorrhizal

Fungi (INVAM; http://invam.caf.wvu.edu/). Microscopic observations were performed

with a Nikon Eclipse 800 compound microscope equipped with Nomarski differential

Glomus cubense sp. nov. (Cuba) ... 339

interference contrast optics and photographs were taken with a Nikon CoolPix 4500

digital camera. Nomenclature and AMF classification follow SchiiSler & Walker (2010).

Reference specimens were deposited at the Cuban National Herbarium, IES-CITMA

(HAC) Cuba, and at the National Mycological Herbarium (DAOM) Canada.

Molecular analyses

Spore clusters were collected under a low magnification binocular, rinsed with ddH,O

and air-dried. DNA extraction was performed using the PrepMan Ultra (PMU) reagent

(Applied Biosystems). A ratio of 10 spores/ul of PMU reagent were crushed using sterile

plastic micropestle. Initial PCR reaction for H* ATPase was performed following Sokolski

et al. (2010) with primer GmossATPup1058 (5'-cGG TAC TTT GTT CTG ATA AGA C-3’)

and primer mix GintraATPlo2663, GmossATP102663, GcoroATPlo2663. Three nested

PCR were performed using primers pairs:1) GintraATPup1104, GmossATPup1104

with GintraATP102663, GmossATP102663, GcoroATP102663 (Sokolski et al. (2010), 2)

GlomusATPup1150 (5'-wTc HGC YGA ACC TGG TGC-3’) with GlomusATP102557 (5'-cTK

GHT TTW GAW GGC CAK AAT-3’) at 52°C or 3) with GintraATP1l02663, GmossATP102663,

GcoroATP1lo2663 at 58°C. The ITS rDNA primers pair used were: GLO2A-forward

(5'-CGT AAC AAG GTT TCC GTA GG-3’), GLO2R-reverse (5'-GCG GGT ACT CCT ACC TGA

TT-3’)(Sokolski et al. 2010). PCR products were cloned using the TOPO TA Cloning

Kits (Invitrogen, USA). Cloned products were purified using the GenElute™ Plasmid

Miniprep Kit (Sigma, USA). Cloning and DNA extraction were performed following

the instructions of the manufacturer for rDNA ITS. Between two and eight rDNA ITS

molecular clones (av. four) were selected. H+ ATPase and ITS were sequenced using

the BigDye Terminator version 3.1 Cycle Sequencing Kit (Applied Biosystems, Foster

City, Calif.) and run on an ABI 3100-Avant automated sequencer (Applied Biosystems/

Hitachi). Sequence editing was done using SeqMan (DNASTAR, www.dnastar.com/),

resulting in 1305 bp and 537, 546 and 549 bp alignments, respectively. The sequences

obtained were compared with those in the GenBank databases using the BLAST program.

Sequences retrieved from GenBank and those in this study were aligned by Muscle

3.8.31 and a neighbour-joining tree inferred using MEGA 5.0 (Tamura et al. 2011) with

1000 replicates. The best-fit model (General Time Reversible plus Gamma) was selected

using the web-based MODELTEST 3.7 (Posada 2006). Sequences were deposited in

GenBank under accession numbers JF510464 for H+ ATPase and JF692724, JF692725,

JF692726 for ITS rDNA.

Taxonomy

Glomus cubense Y. Rodr. & Dalpé, sp. nov. PLATES 1-2

MycoBank MB561650

Sporocarpia ignota. Sporae singulares vel aggregatae. Sporae hyalinae vel luteolae,

ovoideae, vel irregulares, 20-48 x (24-)54-82 um, raro globosae, 24-54 um diam.

Sporae tunica stratis duobus; stratum exterius hyalinum, subflexuosum, ad 0.8-1.0 um

crassum, stratum interius hyalinum vel luteolum, 0.8-1.7 um crassum, rigidum. Hyphae

sustinentes hyalinae, rectae, subcylindricae vel subinfundibuliformes, 4-10 um crassae ad

basim sporae. Porus apertus, raro septo curvo clausus cum strato interiore. Mycorrhizas

vesicular-arbusculares formantes.

340 ... Rodriguez & al.

Type: Cuba: “Las Papas” area, San José de las Lajas, from a Fersialitico Pardo rojizo

mullido carbonatado soil in “San Rafael” Pond. (Holotype, permanent slides mounted

in PVLG deposited at HAC Cuban National Herbarium, Instituto de Ecologia y

Sistematica, CITMA, La Habana, Cuba; isotype, DAOM 241198). Genbank JF510464,

JF692724, JF 692725, JF692726.

Erymo.oey: Latin, cubense, in reference to Cuba, the country where the species was

found.

SPOROCARP unknown, spores mostly found in loose aggregates of 3 to 12 spores

arising blastically at the top of sporogenous hyphae branched from a parent

hypha continuous with an extraradical mycorrhizal hypha, rarely single in the

soil. Spores (FIGS 1-8) hyaline to very light yellow (0/0/40/0); ovoid, ellipsoid,

pyriform to irregular in shape 20-48 x (24-)54-82 um, rarely globose, 24-54

um in diameter with one subtending hypha. SPORE waLL composed of two

layers. Spore wall layer 1 (Swl 1) hyaline, permanent, flexible to semi-flexible,

0.8-1.0 um thick, with a smooth surface, easily separating from layer 2 in

crushed spores. Swl 2 hyaline to very pale yellow, rigid, 0.8-1.7 um thick. Spore

wall layers do not react to Melzer’s reagent, spore wall layer 1 stained light

blue in Cotton Blue. SUBTENDING HYPHA concolorous to spores; cylindrical

to slightly flared, straight or recurved; 4-10 um wide at spore base. WALL OF

SUBTENDING HYPHAE hyaline to very light yellow; 1.6-2.0 um thick at the spore

base; composed of two layers continuous with spore wall layers 1 and 2. PoRE

usually open, occasionally closed by a curved septum located 6-8 um from the

spore base or sometimes occluded by thickening of spore wall layer 2.

INTRARADICAL STRUCTURES — G. cubense forms vesicular-arbuscular

mycorrhizae with intracellular ellipsoid hyphal coils (FIG. 11), globose, 23-38

um, to ovoid to irregular vesicles, 18-25 x 20-60 um (FIGS 10, 12) and a few

arbuscules.

MYCORRHIZAL ASSOCIATION — In the field, found associated with typical

lagoon vegetation plants such as Cynodon nlemfuensis and Mimosa pigra. In

field trials, developed symbiosis with a diversity of crop plants: avocado trees,

banana, cassava, corn, cucumber, forages (Brachiaria decumbens and Panicum

maximum), guava, malanga, mango, pepper, plantain, potato, rice, soybean,

bean, sweet potato, tobacco, tomato, wheat, yam (Rivera et al. 2007). In the

greenhouse, pot cultured for several years with sorghum (Sorghum bicolor and

leek (Allium porrum) plants.

PHYLOGENETIC POSITION — The ITS rDNA and H+ ATPase analyses (Fics

13-14) placed Glomus cubense in the genus Glomus sensu SchiiBler & Walker

(2010) in the family Glomeraceae with no close relationship with known AMF

species of this family. Moreover, no environmental sequences were found to be

closely related to G. cubense.

Glomus cubense sp. nov. (Cuba) ... 341

‘i

tT)

PAS

y v

‘

Fics. 1-8. Spores of Glomus cubense (DIC) from pot-cultures. 1. Juvenile spore with hyphal

attachment. 2. Mature spore with hyphal attachment. 3. Mature spore slightly crushed showing

spore wall layers 1 and 2 (swll, swl2). 4-5. Mature spores with spore wall layer 1 stained with

Cotton Blue compared to spore wall layer 2. 6-8. Irregular shaped spores with subtending

hyphae. Bars: 1-3 =10 um; 4-8 = 20 um.

342 ... Rodriguez & al.

Fics. 9-12. Glomus cubense fungal structures inside colonized roots (DIC). 9. Entry point along

a root. 10. Vesicles. 11. Hyphal coil inside cortical root cell. 12. Vesicles inside cortical root cell. Bars:

9-12 = 20 um.

SPECIMENS EXAMINED: CUBA. Spores and roots isolated from Sorghum bicolor pot

cultures, grown in greenhouse on sterilized clay substrate (Gleyic Calcic Vertisol soil

(IUSS Working Group WRB, 2006) with the following chemical characteristics: Na*

0.81, K* 0.42, Ca** 48.1, Mg’* 6.6 (cmol/kg"); P 30.8 (ppm); organic matter 0.84 (%); pH

Glomus cubense sp. nov. (Cuba) ... 343

8.3. CANADA. Spores and roots isolated from Allium porrum pot cultures, grown on

cultivated Brown Chernozem soil (Saskatchewan, Canada) with a loamy sand texture

and the following chemical characteristics after sterilization: NH,-N 19.7 and NO,-N

14.1 (mg. kg"), P 21.3 and K* 324.5 (mg. kg"), pH 6.5 and EC 0.48 mS.

OTHER SPECIMENS EXAMINED: Type specimens of G. bistratum Btaszk. et al.,

G. perpusillum Blaszk. & Kovacs, provided by J. Blaszkowski, G. canadense (Thaxt.)

Trappe & Gerd. (FH 5045), G. fragile (Berk. & Broome) Trappe & Gerd. (K Berkeley

1182) and of G. viscosum T.H. Nicolson (PI Holotype)

DISTRIBUTION AND HABITAT. Glomus cubense is only known from a single

location: “Las Papas” area, San José de las Lajas, Cuba (22°57'41"N 82°09'04"W).

Spores were originally isolated from Haplic Cambisol (chromic) soil collected

at ca. 50 cm depth. The soil corresponds to a “Fersialitico Pardo rojizo mullido

carbonatado’ soil (Instituto de Suelos, 1999). The species was propagated in pot-

culture using the same type of soil originating from the same area in Cuba.

Notes

The main distinctive morphological properties of G. cubense are its small

hyaline to very light yellow, usually irregularly-shaped spores (Fics 1-2, 6-8)

and its 2-layered spore wall (Fics 3-5) composed of a permanent, flexible to

semi-flexible outer layer staining distinctly in Cotton Blue (Fics 4-5) and a rigid

inner layer, non-reactive to Melzer’s reagent. Under the dissecting microscope,

G. cubense may resemble most glomoid small-sized and pale-colored spores

as treated in the Blaszkowski et al. (2010) identification key. However, when

observed under the compound microscope, only six named species produce

glomoid pale coloured, inamyloid and 2-layered spores that are smaller than 80

um. These are G. bistratum (Btaszkowski et al. 2009b), G. cerebriforme McGee

(McGee 1986), G. indicum Blaszk. et al. (Blaszkowski et al. 2010), G. minutum

Blaszk. et al. (Blaszkowski et al. 2000), G. pallidum LR. Hall (Hall 1977; Oehl et

al. 2003), and G. perpusillum Blaszk. & Kovacs (Blaszkowski et al. 2009a).

The G. cerebriforme and G. perpusillum spore wall layer 1 is laminate (vs.

flexible in G. cubense) and much thicker than spore wall layer 2 (vs. opposite in

G. cubense). Additionally, spore wall layer 2 of G. perpusillum stains in Melzer’s

reagent (vs. no staining in G. cubense). In the spore wall of the other species

listed above, layer 1 is a permanent structure, semi-flexible to rigid, thinner

than layer 2 as in G. cubense but layer 2 is laminate. The G. minutum spores

are only hyaline and of globose to subglobose shape (vs. hyaline to very pale

yellow, and mostly of irregular shape in G. cubense). While spore wall layer 1

easily separates from spore wall layer 2 in freshly crushed G. cubense spores,

that of G. minutum generally remains adherent to the laminate spore wall 2. In

G. minutum, spore wall layer 1 sometimes swells and separates from spore wall

layer 2, but only when spores are mounted in lactic acid-based mountants for

at least some hours.

344 ... Rodriguez & al.

84 AM992829 uncultured Glomus (agricultural soil, Netherlands)

fas [eee rere EF393611 uncultured Glomus (Botrychium virginianum Hungary)

; JF692726 Glomus cubense

mu JF692724 Glomus cubense

_____________ JF@92725 Glomus cubense

88 [oT AF185662 Rhizophagus intraradices

59 —_—————— FM992400 Rhizophagus proliferus

61 | | FJ009614 Rhizophagus irregulare

52 — GQ205070 Rhizophagus diaphanus

a6 ——— GQ205074 Rhizophagus custos

ual GQ205083 Rhizophagus clarus

| 83 | FM253379 Glomus achrum

| 7 GU059547 Glomus indicum

1” | | GQ205036 Glomus cerebriforme

FM253382 Glomus bistratum

EF989114 Funneliformis mosseae

0,02

Fic.13. BIONJ tree based on an alignment of the ITS sequences obtained from Glomus cubense

and related representative sequences from GenBank. Branch support values (only values exceeding

50% are given) on branches refer to BIONJ bootstrap (using GTR + G distances) and maximum

likelihood. Scale indicates nucleotide substitution per site.

99 — GQ205021 Rhizophagus irregulare

60 L GQ205023 Rhizophagus diaphanus

[ GQ205031 Rhizophagus custos

| 98 GQ205032 Glomus sp.

89 | ———~ GQ303448 Rhizophagus intraradices

| Ot GQ205027 Rhizophagus proliferus

_____ 6Q205026 Rhizophagus clarus

| = GQ205025 Glomus cerebriforme

Toners SEB 0464 Glomus cubense

"vr po —- GQ205033 Funneliformis mosseae

84 | - GQ205028 Glomus multiforum

93 - GQ205029 Funneliformis caledaium

—————-—-- GQ205030 Claroideoglomus claroideur

0.05

Fic.14. BIONJ (Neighbour-joining) tree based on an alignment of H+ ATPase sequences of Glomus

cubense and of related representative sequences from GenBank. Branch support values (only values

exceeding 60% are given) on branches refer to BIONJ bootstrap (using GTR + G distances) and

maximum likelihood. Scale indicates nucleotide substitution per site.

Glomus cubense sp. nov. (Cuba) ... 345

Glomus microaggregatum Koske et al. also produces small, 2-layered spores

frequently irregular in shape, but the spores are much darker-colored (yellow

to brownish yellow), their spore wall is thicker (up to 4 um vs. up to 2.7 in

G. cubense) and comprises a rigid outer layer and a flexible inner layer (vs.

flexible to semi-flexible outer layer and rigid inner layer in G. cubense).

The spore wall of G. fragile and G. canadense also consists of a hyaline outer

layer and a pale yellow inner layer. However, observation of their respective type

specimens (K Berkeley 1182, FH 5045) revealed that the inner spore wall layer

in both species is thicker than that of G. cubense and laminate (Gerdemann &

Trappe (1974).

Young or small-sized clustered spores of Glomus viscosum may resemble

those of G. cubense, but they usually are globose, have a 3-layered spore wall, of

which layer 3 is distinctively laminate.

Compared with G. cubense, glomoid spores of Paraglomus spp are hyaline to

pale cream, they form only singly (vs. in cluster in G. cubense) and their spore

wall layers clearly differ phenotypically from those of G. cubense (Blaszkowski

et al. 2010).

Ambispora spp. differentiate hyaline but much larger spores (Walker et al.

2007).

The phylogenetic analyses of sequences of the H+ATPase gene clearly

support morphological analyses and separate G. cubense from the few sequences

actually available and confirmed the efficiency of earlier published primers for

sequencing Glomus species (Sokolski et al. 2010). The analyses of ITS region

confirm that the species G. cubense is a member of the genus Glomus sensu

SchiiBler & Walker (2010) in the family Glomeraceae without having any

close described relatives. When comparing ITS sequence data, the best Blast

alignment with Genbank database sequences indicated a maximum of 83 and

85% similarity with an as yet uncultured Glomus sp. thus supporting the status

of G. cubense as a distinct species.

Acknowledgments

This work was supported by the International Scholarships Program of Canada

Government (Emerging Leaders in the Americas Program) and partially by

“International Foundation for Science” (grant no. C-4463) given to Y. Rodriguez. The

authors want to thank Drs. J. Cayouette, J. Tambong, and S. Redhead (Agriculture and

Agri-Food Canada, Ottawa Canada), Dr. J. Blaszkowski (West Pomeranian University

of Technology, Szczecin Poland), and Dr S. Sokolski (Université Laval Québec Canada)

for their scientific support and Drs Blaszkowski and Redhead for presubmission review.

Special thanks to colleagues of the Mycorrhiza team from the Instituto Nacional de

Ciencias Agricolas, San José de las Lajas, Cuba.

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MY COTAXON

http://dx.doi.org/10.5248/118.349

Volume 118, pp. 349-353 October-December 2011

Two new species of Exserticlava and Spiropes

on decaying wood from Guangdong, China

SHOU-CAI REN?”, JIAN MA’ & XIU-GUO ZHANG"

‘Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*Zaozhuang Vocational College, Zaozhuang, 277800, China

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Two new anamorphic fungi, Exserticlava manglietiae on decaying branches

of Manglietia chingii and Spiropes terminaliae on decaying branches of Terminalia mantaly,

are described and illustrated. The specimens were collected from subtropical rainforests in

Guangdong Province, China. Morphological differences between these two fungi and similar

species are summarized.

KEY worps — anamorph, hyphomycete, microfungi, systematics

Hughes (1978) introduced Exserticlava to accommodate Cordana vasiformis

Matsush. and C. triseptata Matsush., two species originally described on wood

and bamboo from Japan (Matsushima 1975). Exserticlava is characterized

by thick-walled, distoseptate conidia from polyblastic, funnel-shaped

conidiogenous cells borne on mononematous, simple conidiophores (Tsui et al.

2001a). Exserticlava is similar to Cacumisporium Preuss and Chloridium Link

in having sympodulo-phialides (Cole & Samson 1979) and holoblastic conidia

formed from the apex of polyblastic conidiogenous cells. Cacumisporium

differs from Exserticlava in euseptate conidia and cylindrical conidiogenous

cells (Tsui et al. 2001b). Conidia in Chloridium are hyaline and aseptate (Gams

& Holubova-Jechova 1976).

Ciferri (1955) established Spiropes with S. guareicola (F. Stevens) Cif. as type

species. The genus is characterized by macronematous unbranched sometimes

geniculate conidiophores, polyblastic, terminal and intercalary, sympodial

cicatrized conidiogenous cells, numerous and conspicuous scars, and solitary

acropleurogenous most commonly obclavate conidia with 1-9 transverse septa

or distosepta.

350 ... Ren, Ma & Zhang

During continuing research on saprobic fungi in subtropical forests of

Guangdong Province, China, we collected two hyphomycetes that possess

the morphological characteristics of Exserticlava and Spiropes. They differ

morphologically from any described species and are, therefore, proposed as

new to science. The type specimens are deposited in HSAUP (Herbarium of

the Department of Plant Pathology, Shandong Agricultural University) and

HMAS (Mycological Herbarium, Institute of Microbiology, Chinese Academy

of Sciences).

Exserticlava manglietiae S.C. Ren & X.G. Zhang, sp. nov. Fig. 1

MycoBank MB563099

CONIDIOPHORA macronematosa, nonramosa, erecta, laevia, atro-brunnea, apicem versus

pallidiora, 6-11-septata, 360-490 x 5-8.5 um. CELLULAE CONIDIOGENAE polyblasticae,

integratae, terminales, proliferationae sympodialiter. Conip1A acrogena, holoblastica,

solitaria, laevia, ellipsoidea, 1-distoseptata, pallide brunnea, 14-16 x 9-11 um, ad basim

truncata.

Type: CHINA. GUNGDONG PROVINCE: subtropical forest of Nanling, on dead branches

of Manglietia chingii Dandy (Magnoliaceae), 10 Dec. 2010, Sh.C. Ren (Holotype, HSAUP

H8363; isotype HMAS 146154).

ETryMoLoey: in reference to the host genus, Manglietia.

CoLtonies effuse, brown. Mycelium partly immersed, partly superficial,

composed of septate, smooth, brown, branched hyphae. CoNnIDIOPHORES

macronematous, mononematous, unbranched, erect, straight or slightly

flexuous, smooth, thick-walled, dark brown, paler towards the apex,

6-11-septate, 360-490 x 5-8.5 um, terminally swollen, 7.5-11 um wide.

CONIDIOGENOUS CELLS polyblastic, integrated, terminal, clavate, proliferating

sympodially and producing a cluster of conidia. Conidial secession schizolytic.

ConlipliA acrogenous, holoblastic, solitary, dry, smooth, ellipsoid, thick-walled,

1-distoseptate, very pale brown, 14-16 x 9-11 um, base truncate.

Notes: Exserticlava species are separated primarily based on conidial shape

and number of septa (Holubova-Jechova 1990). A key to species and a synopsis

of their characters were provided by Tsui et al. (2001a). Of the six previously

described Exserticlava species, E. manglietiae resembles E. globosa V. Rao & de

Hoog (Rao & de Hoog 1986) in possessing concolorous, thick-walled conidia

with a strictly median septum, but E. globosa produces spherical larger (17 x

21 um diam) verrucose conidia. Exserticlava uniseptata Bhat & B. Sutton and

E. yunnanensis L. Cai & K.D. Hyde also produce 1-septate conidia, but E.

uniseptata can be distinguished by its broadly obovoid, larger conidia (15-20 x

11-14 um) with a smaller basal cell and E. yunnanensis differs in hemispherical,

pale brown to subhyaline, larger conidia (16-22 x 10-13 um) with a smaller

apical cell.

Exserticlava manglietiae & Spiropes terminaliae spp. nov. (China) ... 351

Fic. 1. Exserticlava manglietiae.

A-B. Conidiophores, conidiogenous cells and conidia. C. Conidia.

Spiropes terminaliae S.C. Ren & X.G. Zhang, sp. nov. Fic. 2

MycoBank MB563100

ConipIoPHoRA singula vel fasciculate, superus fertilis parte geniculatis efformata, usque

340 um longa, basi 4.5-6.5 um crassa, apice 7-9 um crassa, CELLULAE CONIDIOGENAE

polyblasticae, integratae, terminales et intercalares, proliferationes sympodialite. CONIDIA

acropleurogena, solitaria, late fusiformia vel oblonga, 5-7-distoseptata, laevia, 27-32 x

8.5-10.5 um.

352 ... Ren, Ma & Zhang

Fic. 2. Spiropes terminaliae.

A-C. Conidiophores, conidiogenous cells and conidia. D. Conidia.

Type: CHINA. GUNGDONG PROVINCE: subtropical forest of Nanling, on dead branches

of Terminalia mantaly H. Perrier (Combretaceae), 15 Dec. 2010, Sh.C. Ren (Holotype,

HSAUP H8218; isotype, HMAS 146155).

ETyMoOLoey: in reference to the host genus, Terminalia.

COLONIES on natural substratum effuse, dark blackish brown to black, hairy.

Mycelium partly immersed and partly superficial, composed of branched, pale

brown, smooth, septate, 2-4 um thick hyphae. ConIDIOPHORES arising singly

Exserticlava manglietiae & Spiropes terminaliae spp. nov. (China) ... 353

or in groups terminally and laterally on the hyphae, erect, sterile lower part

straight or flexuous, upper fertile part with zigzag proliferation, sometimes with

geniculate fertile regions separated by sterile areas, brown to dark brown, paler

toward the apex, with numerous dark conidial scars, 6-14-septate, up to 340 um

long, 4.5-6.5 um wide at the base, 7-9 um wide at the apex. CONIDIOGENOUS

CELLS polyblastic, integrated, terminal and intercalary, sympodial, cylindrical,

cicatrized, scars conspicuous. Conrp1A holoblastic, acropleurogenous, solitary,

dry, broadly fusiform to oblong, brown to dark brown, smooth, 5-7-distoseptate,

27-32 um long, 8.5-10.5 um wide, 2.5-3.5 um wide at the truncate base.

Notes: Of the known Spiropes species, S. terminaliae is closely related to

S. guareicola in possessing macronematous unbranched conidiophores where

the upper fertile part has geniculate proliferations and distoseptate conidia.

However, S. terminaliae differs from S. guareicola in conidial shape, size, and

the number of septa. The conidia of S. guareicola are broadly fusiform, much

larger (25-52 x 10-13 um), and with fewer septa (3-5).

Acknowledgments

The authors express gratitude to Dr Eric H.C. McKenzie and Dr R.F. Castafieda-Ruiz

for serving as pre-submission reviewers and for their valuable comments and suggestions.

This project was supported by the National Natural Science Foundation of China (Nos.

31093440, 30499340, 30770015) and the Ministry of Science and Technology of the

People’s Republic of China (Nos. 2006FY120100, 2006FY110500-5).

Literature cited

Ciferri R. 1955. Observations on meliolicolous Hyphales from Santo Domingo. Sydowia 9:

296-330.

Cole GT, Samson RA. 1979. Patterns of development in conidial fungi. Pitman, London. 190 pp.

Gams W, Holubova-Jechova V. 1976. Chloridium and some other dematiaceous hyphomycetes

growing on decaying wood. Stud. Mycol. 13: 1-99.

Holubova-Jechova V. 1990. Problems in the taxonomy of the dematiaceous hyphomycetes. Stud.

Mycol. 32: 41-48.

Hughes SJ. 1978. New Zealand fungi 25. Miscellaneous species. New Zeal. J. Bot. 16: 311-370.

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Published by the author,

Kobe, Japan. 209 p.

Rao V, de Hoog GS. 1986. New or critical hyphomycetes from India. Stud. Mycol. 28: 1-84.

Tsui CKM, Goh TK, Hyde KD. 2001a. A revision of the genus Exserticlava, with a new species.

Fungal Diversity 7: 135-143.

Tsui CKM, Goh TK, Hyde KD, Hodgkiss IJ. 2001b. New species or records of Cacumisporium,

Helicosporium, Monotosporella and Bahusutrabeeja on submerged wood in Hong Kong streams.

Mycologia 93: 389-397. http://dx.doi.org/10.2307/3761660

MY COTAXON

http://dx.doi.org/10.5248/118.355

Volume 118, pp. 355-359 October-December 2011

Discovery of Geastrum xerophilum from the Neotropics

BIANCA DENISE BARBOSA DA SILVA’, JULIETH DE OLIVEIRA SOUSA?

& IURI GOULART BASEIA’

’ Universidade Federal de Pernambuco, Programa de Pés-Graduagdo em Biologia de Fungos,

Depto. Micologia, Centro de Ciéncias Bioldgicas,

Av. Nelson Chaves s/n, 50670-420, Recife, PE, Brazil

? Universidade Federal do Rio Grande do Norte, Depto. Botanica, Ecologia e Zoologia,

Campus Universitario, CEP: 59072-970, Natal, RN, Brazil

CORRESPONDENCE TO *: biancadeni@yahoo.com.br

ABSTRACT — Geastrum xerophilum, a xerophytic species found in the Brazilian semi-arid

region, is reported for the first time from the Neotropics. Descriptions, taxonomic remarks,

and SEM-photos are provided.

Key worps — Brazil, gasteroid fungi, Geastraceae, taxonomy

Introduction

The biological diversity of the Neotropical region is still poorly known,

when compared with the Palearctic and Nearctic regions. Nevertheless, it is

considered a reference area in terms of rich species diversity (Olson et al. 2001,

Sodhi & Ehrlich 2010), encompassing megadiverse countries such as Brazil

(Brooks 2006).

Most Geastrum species commonly grow in shady, humid habitats (Sunhede

1989), but some taxa, such as G. xerophilum, adapt to desert environments

(Bates 2004). The diversity of these fungi in semiarid Brazilian biomes has

always been underestimated and is practically unknown (Drechsler-Santos et

al. 2008). Only since the turn of the century have research agencies in Brazil

begun to sponsor the study of fungal diversity in the semiarid region of the

country. The implementation of the Semiarid Biological Research Program

(PPBio-Semi-Arido) has led to an increase in the study of these environments.

Material & methods

The studied material was collected in the city of Caicd, Rio Grande do Norte, Brazil.

Macroscopic observations were made from fresh and dried materials. Color references

356 ... Silva, Sousa & Baseia

follow Kornerup & Wanscher (1978). For light microscopy, free-hand sections were

mounted in 5% (w/v) KOH. Basidiospore surface features were observed by SEM

from a small portion of the gleba, which was dusted onto double-sided adhesive tape

on a specimen holder, coated with platinum (Monthoux 1982), and examined with a

Philips - XL30 microscope. Twenty randomly selected basidiospores were measured

using an ocular micrometer and scanning electron microscopy (SEM), and all spore

measurements included surface ornamentation. Statistic measures of spore length and

width are given as x + SD (arithmetic mean + standard deviation) and “Qm” (the ratio

of mean spore length to width). The specimens are deposited in the Herbarium of the

Federal University of Rio Grande do Norte (UFRN), Brazil.

Results

Geastrum xerophilum Long ex Desjardin, Pacific Science 65: 493 (2011).

Fics 1-3

Expanded basidiome saccate, 20 mm wide x 13 mm tall. Exoperidium splitting

into 7 rays, non-hygroscopic. Rays involute rolling up under the endoperidium.

Mycelial layer brown (6E4), felted, thin, encrusted with sand, persistent.

Fibrous layer grayish brown (6D3), papery, thin. Pseudoparenchymatous layer

dark brown (6F4), thick, rigid, persistent. Endoperidial body 16 mm wide x

8 mm tall (including peristome), stalked, depressed globose, grayish orange

(5B3) to brownish orange (5C3), furfuraceous becoming glabrous with age.

Apophysis discrete. Stalk <2 mm tall, concolorous with the endoperidium.

Peristome plicate becoming lacerated with age, not delimited, concolorous with

endoperidium, applanate. Gleba brown (6F5).

Basidiospores globose to subglobose 4.3-6.3 um x 4.3-6.3 um [x = 5.6 + 0.7

x 5.6 + 0.7, Qm = 1.0, n = 20], with pedicel rudimentary, densely verrucose,

yellowish brown in 5% KOH. Eucapillitium 3.8—6.3 um diam., glabrous to lightly

encrusted, lumen absent, walls >1 um thick. Mycelial layer composed of thin

sinuous-walled hyphae, lumen absent, 2.5—5.0 um diam., yellow to hyaline in

5% KOH. Fibrous layer composed of thin straight walled hyphae, lumen absent,

1.6-3.8 um diam., yellow to hyaline in 5% KOH. Pseudoparenchymatous layer

composed of sublobose, citriform to elongated hyphae, 16.5-31.7 um diam.

x 12.7-33.0 um in length, walls >1 um thick, hyaline to pale yellowish in 5%

KOH.

Substrate —Solitary in sandy soil.

SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO NOorTE: Caicdé, 06°14'302"S,

37°02'958"W, 163 ma.s.l., 29.V.2010, col. B.D.B. Silva (UFRN 1508).

TAXONOMIC REMARKS — Geastrum xerophilum, a species typical of xerophytic

environments, was collected in Brazil’s Caatinga domain. ‘The collection site is

characterized by xerophilous vegetation and semiarid climate with low irregular

rainfall and high temperature and evapotranspiration (Leal et al. 2005). The

Geastrum xerophilum from the Neotropics (Brazil) ... 357

10 mm

FIGURE 1. Basidiomes of Geastrum xerophilum.

species is morphologically similar to G. campestre Morgan, which differs in its

hygroscopic rays, delimited peristome, and larger spores (6.5-8.0 um diam.)

(Sunhede 1989). Geastrum kotlabae VJ. Stanék is another proximate species,

but it exhibits strongly hygroscopic rays bent around the endoperidium and a

detached mycelial layer and sessile endoperidium (Sunhede 1989, Bates 2004).

Previously the distribution of G. xerophilum was thought to be restricted to

North America, with records from Arizona (Bates 2004), New Mexico (Ponce

de Leon 1968) and Mexico (Esqueda et al. 1995, 2009, Moreno et al. 2010), and

to Hawaii (Smith & Ponce de Leon 1982, Gilbertson et al. 2001, Hemmes &

Desjardin 2011). Therefore G. xerophilum is recorded here for the first time for

the Neotropical region.

Recently, G. xerophilum was recognized as an invalid name and validated by

Hemmes & Desjardin (2011) because Long’s (1942) original description lacked

a Latin diagnosis (McNeill et al. 2006: Art. 36.1).

Acknowledgments

The authors gratefully acknowledge the CTPETRO-INFRA and FINEP/LIEM for

their collaboration with scanning electron microscopy. The authors express their deep

thanks to Dennis E. Desjardin (State University San Francisco, USA) and Taiga Kasuya

(University of Tsukuba, Japan) for reviewing the manuscript. This work was financed

by the Coordenacao de Aperfeigoamento de Pessoal de Nivel Superior (CAPES) and

Conselho Nacional de Desenvolvimento Cientifico e Tecnolégico (CNPq).

Literature cited

Bates ST. 2004. Arizona members of the Geastraceae and Lycoperdaceae (Basidiomycota, Fungi).

Master Thesis, Arizona State University, U.S.A.

Brooks TM, Mittermeier RA, Fonseca GAB, Gerlach J, Hoffmann M, Lamoreux JF, Mittermeier

CG, Pilgrin JD, Rodrigues ASL. 2006. Global biodiversity conservation priorities. Science 313:

58-61. http://dx.doi.org/10.1126/science.1127609.

358 ... Silva, Sousa & Baseia

Magn

12000x

11000x

FIGURES 2-3. Geastrum xerophilum: basidiospores under SEM.

Geastrum xerophilum from the Neotropics (Brazil) ... 359

Drechsler-Santos ER, Wartchow F, Baseia IG, Gibertoni TB, Cavalcanti MAQ. 2008. Revision of the

Herbarium URM I. Agaricomycetes from the semi-arid region of Brazil. Mycotaxon 104: 9-18.

Esqueda M, Pérez-Silva E, Herrera T. 1995. New records of gasteromycetes for Mexico. Documents

Mycologiques 25(98-100): 151-160.

Esqueda M, Sanchez A, Rivera M, Coronado ML, Lizarraga M, Valenzuela R. 2009. Primeros

registros de hongos gasteroides en la Reserva Forestal Nacional y Refugio de Fauna Silvestre

Ajos-Bavispe, Sonora, México. Revista Mexicana de Micologia 30: 19-29.

Gilbertson RL, Desjardin DE, Rogers JD, Hemmes DE. 2001. Fungi from the Mamane-Naio

vegetation zone of Hawai'i. Fungal Diversity 6: 35-69.

Hemmes DE, Desjardin DE. 2011. Earthstars (Geastrum, Myriostoma) of the Hawaiian Islands

including two new species, Geastrum litchiforme and Geastrum reticulatum. Pacific Science 65:

477-496.

Kornerup A, Wansher JE. 1978. Methuen handbook of colour, 3rd edn., London: Methuen.

Leal IR, Silva JMC, Tabarelli M, Lacher Jr TE. 2005. Mudando o curso da conservacao da

biodiversidade na Caatinga do Nordeste do Brasil. Megadiversidade 1(1): 139-146.

Long WH. 1942. Studies in the Gasteromycetes IV. A new species of Geaster. Mycologia 34: 13-16.

McNeill J, Barrie FR, 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.

Monthoux O. 1982. Micromorphologie des spores et capillitiums des Gastéromycetes dés stations

xériques de la région de Geneve, étudée au microscope életronique 4 balayage (SEM). Candollea

3726399.

Moreno G, Lizarraga M, Esqueda M, Coronado M L. 2010. Contribution to the study of gasteroid

and secotioid fungi of Chihuahua, Mexico. Mycotaxon 112: 291-315.

Olson DM, Dinerstein E, Wikramanayake ED, Burgess ND, Powell GVN, Underwood EC,

D’Amico JA, Strand HE, Morrison JC, Loucks CJ, Allnutt TE, Lamoreux JE, Ricketts TH, Itoua I,

Wettengel WW, Kura Y, Hedao P, Kassem K. 2001. Terrestrial ecoregions of the world: A new

map of life on Earth. BioScience 51: 933-938.

http://dx.doi.org/10.1641/0006-3568(2001)051[0933: TEOT WA]2.0.CO;2

Ponce de Leon P. 1968. A revision of the family Geastraceae. Fieldiana Botany 31: 302-349.

Smith CW, Ponce de Leén P. 1982. Hawaiian geastroid fungi. Mycologia 74: 712-717.

doi:10.2307/3792856

Sodhi NS, Ehrlich PR. 2010. Conservation biology for all. Oxford University Press, Oxford, UK.

344 p.

Sunhede S. 1989. Geastraceae (Basidiomycotina). Morphology, ecology, and systematics with special

emphasis on the North European Species. Synopsis Fungorum 1. 534 p.

MY COTAXON

http://dx.doi.org/10.5248/118.361

Volume 118, pp. 361-381 October-December 2011

A preliminary ITS phylogeny of Melanoleuca (Agaricales),

with special reference to European taxa

ALFREDO VIZZINI’*, ROBERTO PARA”, ROBERTO FONTENLA3,

STEFANO GHIGNONE’, ENRICO ERCOLE’

‘Dipartimento di Biologia Vegetale - Universita degli Studi di Torino

Viale Mattioli 25, I-10125, Torino, Italy

? Via Martiri di Via Fani 22, I-61024, Mombaroccio (PU), Italy

> Via Monte Marino 26, I-60125 Ancona, Italy

* Istituto per la Protezione delle Piante, CNR Sezione di Torino,

Viale Mattioli 25, I-10125, Torino, Italy

CORRESPONDENCE TO ”: alfredo. vizzini@unito.it

ABSTRACT — Ninety-one Melanoleuca collections were chosen to test the agreement of

current and traditional infrageneric tripartite classifications of Melanoleuca (subgenera

Acystis without cystidia, Urticocystis with urticiform cystidia, and Melanoleuca with

macrocystidia) with molecular phylogenetic data (ITS sequences analysis) and to evaluate

the systematic significance of relevant morphological characters. Melanoleuca is found to be

monophyletic, and only two emended subgenera, Urticocystis and Melanoleuca, are supported.

Subg. Urticocystis comprises all taxa with urticoid cystidia plus the macrocystidiate M. cognata

complex. The artificial subg. Acystis is shown to be polyphyletic and no longer tenable. The

entire genus comprises at least 10 clades with 13 subclades. Melanoleuca sublanipes sp. nov.

and new combinations M. exscissa f. iris, M. exscissa f. sarcophylla, M. exscissa f. diverticulata,

and M. paedida f. electropoda are introduced.

Key worps — Basidiomycota, Agaricomycetes, Kinia, pluteoid clade, taxonomy

Introduction

Kirk et al. (2008) state that the basidiomycete genus Melanoleuca Pat., typified

by M. vulgaris (Pat.) Pat. [= M. melaleuca (Pers.) Murrill] and traditionally

placed in subtribe Leucopaxillineae Singer (Tricholomataceae R. Heim ex

Pouzar, Agaricales Underw.) (Singer 1986), comprises approximately 50 saprobic

species worldwide. However, Index Fungorum (http://www.indexfungorum.

org/, accessed 28 June 2011), lists 332 validly published Melanoleuca names

representing 126 European and 206 extra-European taxa (pers. obs.). Of the 164

names that actually represent the genus as presently circumscribed (cf. Pfister

362 ... Vizzini & al.

1984), many are now known as synonyms with others yet to be listed as such.

New taxa are, however, continuously being described, even from well-studied

areas such as Europe (e.g., Bresinsky 2006, this paper). Melanoleuca species

are cosmopolitan and characterised by the following characters: collybioid to

tricholomatoid basidiomata; convex to slightly depressed (often with a shallow

umbo) pilei; emarginated to adnate to shortly decurrent lamellae; absence of

veils; white to pale-yellowish spore print; cutis to trichoderm pileipellis; hyaline

spores with amyloid ornamentations; cheilocystidia (mostly present) of two

types either urticoid, septate, thin-walled or fusiform to lageniform, mostly

aseptate, slightly thick-walled, sometimes with encrusting crystals at apex;

pleurocystidia similar to cheilocystidia; no clamp connections (Pegler & Young

1973, Gillman & Miller 1977, Kithner 1978, Singer 1986, Boekhout 1988, 1999;

Bon 1991, Vesterholt 2008, Watling & Turnbull 2008). Leucopaxillus Boursier,

a morphologically allied genus, differs from Melanoleuca mainly in abundant

clamp connections and (usually) lacking well-developed hymenial cystidia

(Singer 1986, Bon 1991). But, according to recent molecular analyses (Moncalvo

et al. 2002, Matheny et al. 2006), Melanoleuca and Leucopaxillus are not closely

related: Melanoleuca species cluster within the Pluteoid clade (Pluteaceae

Kotl. & Pouzar), whereas Leucopaxillus belongs to the Tricholomatoid clade,

close to Tricholoma (Fr.) Staude. Sequence analyses by Moncalvo et al. (2000,

2002) place Melanoleuca and Pluteus Fr. as sister to Amanitaceae R. Heim ex

Pouzar, while others place the minute uniloculate gasteromycete, Limnoperdon

incarnatum G.A. Escobar, sister to Melanoleuca and Pluteus (Bodensteiner

et al. 2004; Binder et al. 2006; Vizzini et al. 2010) or to Melanoleuca, Pluteus

and Volvariella Speg. (Matheny et al. 2006). Justo et al. (2011) recently place

Melanoleuca as sister to a monophyletic group formed by Pluteus species and

Volvopluteus Vizzini et al. Finally, Vizzini et al. (2010) reduced the puzzling

genus Kinia Consiglio et al. to a subgenus of Melanoleuca with non-amyloid

spores.

Melanoleuca is one of the less appealing fungal genera, whose members

are mostly tedious and drab in appearance and dull in pileus colours. It is a

character-poor genus with many species macroscopically very similar and

differing only in very subtle features (e.g., basidioma colour, odour, stipe

ornamentation) and a morphology strongly influenced by environmental

factors (Bon 1991, Boekhout 1999). Thus far, infrageneric classifications and

species circumscriptions have relied on morphological characters. Identification

especially depends on microscopic observations of a rather limited set of

characters, such as presence/absence of cheilocystidia, cystidial shape, spore size

and ornamentation, and pileipellis structure. Interpretation of some characters

may rely on personal experience (e.g. shape of cystidia); in some cases the

value ranges overlap (e.g. spore size), in others pileipellis structure and spore

ITS sequence analysis of Melanoleuca ... 363

size show great intra-basidiome variability. From a traditional morphological

perspective, this often makes species identification difficult or even daunting.

The paucity of Melanoleuca characters has been detrimental to establishing a

natural taxonomic framework for rationalizing infrageneric relationships.

The lack of a modern Melanoleuca monograph and the existence of

many short, taxonomically and geographically limited publications further

complicate the situation. In addition, there are many controversies concerning

the interpretations of old descriptions and names (Boekhout 1988).

Several different infrageneric classifications of Melanoleuca have been

published. Singer (1935, 1943, 1986), Métrod (1942, 1948), Kithner (1978), and

Moser (1983) proposed schemes based mainly on macromorphological features

(see Boekhout 1988 for an historical review). Singer (1986), for example,

divided Melanoleuca into four sections —Alboflavidae Singer, Humiles Singer,

Oreinae Singer, Melanoleuca— circumscribed only by pileus colour and stipe

ornamentation.

Bon (1978), the first to present an infrageneric classification based equally on

macro- and micro-morphological characters, divided Melanoleuca into seven

sections. Boekhout (1988), focusing mainly on microscopic features, stressed

the importance of absence/presence and shape of cystidia (first noted by Metrod

1948) and divided the genus into three subgenera (Tas. 1): subg. Melanoleuca—

no cystidia; subg. Urticocystis Boekhout—with urticiform cheilocystidia and no

(or very rare) pleurocystidia; subg. Macrocystis Boekhout—with long fusiform

to lageniform cheilocystidia (macrocystidia) and similar pleurocystidia.

Boekhout recognized two urticiform types: the brevipes-type (with narrow

cylindrical upper part, Fic. la) and the exscissa-type (with rather wide upper

part attenuating towards the apex, Fic. 1b).

Bon (1991) proposed a slightly different infrageneric classification (TaB. 1)

by introducing the spore Q value (the ratio of length to width of the spores in

side view) and stressing the importance of fusiform versus lageniform cystidia

(Fic. lc-d) as important key characters for delimiting subsections.

Finally, Boekhout (1999) reintroduced a simplified classification practically

identical to the 1988 scheme except that sect. Grammopodiae was no longer

subdivided into subsects. Grammopodiae and Exscissae (TaB. 1).

There are only two major monographic treatments of Melanoleuca: Bon

(1991) covers over 80 taxa, most reported only from Europe, and Boekhout

(1999), covers only Dutch taxa and unites species into large complexes thereby

recognizing only 14 species and several forms and varieties.

Bon (1991), who covered a larger number of taxa, was used both for selecting

which species to sample and for testing the effectiveness of Bon’s classification

method. Due to the limitation of morphological characters used, several of his

taxonomical units remain controversial.

364 ... Vizzini & al.

TABLE 1. The tripartite infrageneric classifications of Melanoleuca

by Bon and Boekhout.

BOEKHOUT’S CLASSIFICATION (1988, 1999)

SuBG. MELANOLEUCA - without cystidia.

SusG. Urticocystis - cheilocystidia urticiform and pleurocystidia absent or very rare.

Sect. Humiles - stipe squamulose or verrucose throughout; stipe

usually much longer than diameter of the pileus.

Sect. Grammopodiae — stipe smooth or somewhat fibrillose; stipe much

shorter than or equally large as diameter of pileus.

*Subsect. Grammopodiae - urticiform cystidia of the brevipes-type.

*Subsect. Exscissae — urticiform cystidia of the exscissa-type.

SusG. Macrocystis - cheilocystidia fusiform to lageniform and pleurocystidia similar.

Sect. Cognatae - with a bright coloured pileus.

Sect. Alboflavidae - with a white to cream-whitish pileus.

Sect. Strictipedes - with a grey-brown pileus.

BON’ S CLASSIFICATION (1991)

SuBG. ACYSTIS — species without cystidia.

Sect. Acystis —- spores with a Q<1.4(-1.5).

Sect. Decembres - spores with a Q>(1.5-) 1.6.

SusG. Urticocystis — cheilocystidia urticiform to strictly lageniform, septate,

up to 50(-60) um long.

Sect. Humiles — stipe squamulose to dark-dotted.

Sect. Grammopodiae - stipe smooth or striate, sometimes pruinose or flocky.

Subsect. Rasilinae - spores with a Q<1.4(-1.5), with isolated warts,

cystidia typically urticiform.

Subsect. Grammopodiae - spores with a Q>(1.5-)1.6, cystidia typically urticiform.

Subsect. Exscissae — spores with a Q>(1.5-)1.6, urticiform cystidia of the exscissa-type.

SuBG. MELANOLEUCA — macrocystidia fusiform to lageniform, non-septate,

up to (40-)50-90(-110) um long.

Sect. Alboflavidae - basidiomes white to whitish.

Sect. Cognatae — basidiomes with bright colours, pileus and lamellae concolorous.

Sect. Oreinae — basidiomes small (<4(-6) cm), collybioid, often dark coloured.

Sect. Melanoleuca — basidiomes small to medium-sized (>(5-)

6 cm), with variable colourations.

Subsect. Strictipedinae - cystidia mainly lageniform.

Subsect. Vulgarinae - cystidia mainly fusiform.

* = subsections present in the 1988 classification scheme only.

The present study is based on a large ITS sequence dataset and is the first

to examine Melanoleuca extensively. Our aims were to 1) check whether

Melanoleuca is monophyletic as traditionally circumscribed; 2) test Bon’s

(1991) morphologically based taxonomy against molecular phylogenetic data;

3) evaluate whether traditional or other morphological features (e.g., pileus

colour, spore and cheilocystidia shape) reflect phylogenetic relationships.

Materials & methods

Taxon sampling

Samples from 91 Melanoleuca collections (41 taxa, 18 unidentified Melanoleuca sp.;

TaB. 2) were tested. Specimens were collected fresh, dried, and depositedin ANC (Erbario

ITS sequence analysis of Melanoleuca ... 365

FIGURE 1. Types of cystidia in Melanoleuca.

a-b. Urticiform cystidia (a = brevipes-type; b = exscissa-type).

c-d. Macrocystidia (c = fusiform; d = lageniform).

Bar = 20 um.

366 ... Vizzini & al.

TABLE 2. Melanoleuca collections examined (species with multiple collections

numbered consecutively; see also Fic. 2).

COLLECTIONS

* Melanoleuca albifolia 1

* M. albifolia 2

* M. angelesiana 1

* M. angelesiana 2

* M. arcuata

* M. atripes

* M. bataillei

M. brevipes 1

M. brevipes 2

M. cinereifolia 1

M. cinereifolia 2

M. cinereifolia 3

M. cognata 1

* M. cognata 2

* M. decembris 1

* M. decembris 2

* M. decembris 3

M. decembris 4

* M. diverticulata

* M. electropoda

M. evenosa

* M. exscissa 1

* M. exscissa 2

* M. exscissa 3

* M. exscissa 4

* M. exscissa 5

* M. exscissa 6

* M. friesii

* M. graminicola

* M. grammopodia 1

* M. grammopodia 2

* M. grammopodia 3

* M. grammopodia

f. macrocarpa 1

* M. grammopodia

f. macrocarpa 2

M. grammopodia

f. macrocarpa 3

* M. heterocystidiosa 1

* M. heterocystidiosa 2

* M. iris

* M. “lanipes”

* M. melaleuca

* M. microcephala

* M. nivea 1

* M. nivea 2

M. nivea 3

CoOL. ID./ORIGIN

ANC MO0182/Spain

ANC M0184/Italy

ANC M0203/Italy

ANC M0204/Italy

ANC MO0167/Italy

ANC M0180/Italy

ANC M0185/Italy

MCVE 04574/Italy

MCVE 04505/Italy

MCVE 01471/Italy

MCVE 11243/Italy

MCVE 20748/Italy

MCVE 13939/Italy

ANC M0170/Italy

ANC M0197/Italy

ANC M0199/Italy

ANC M0200/Italy

MCVE 01573/Italy

ANC M0206/Italy

ANC MO0187/Italy

MCVE 14576/Italy

ANC M0207/Italy

ANC M0208/Italy

ANC M0210/Italy

ANC M0210B/Italy

ANC M0212/Italy

ANC M0213/Italy

ANC M0186/Italy

ANC M0201/Italy

ANC M0217/Italy

ANC M0218/Italy

ANC MO0219/Slovenia

ANC M0215/Italy

ANC M0216/Italy

MCVE 04410/Italy

ANC MO0174/Italy

ANC MO175/Italy

ANC M0211/Italy

ANC MO0166/Italy

ANC MO0176/Italy

ANC MO0196/Italy

ANC MO0177/Italy

ANC MO0183/Italy

MCVE 09578/Italy

* = collections newly sequenced in this study.

SUBGENERA / SECTIONS

(BON 1991)

Melanoleuca / Oreinae

Acystis / Acystis

Melanoleuca / Cognatae

Melanoleuca / Oreinae

Melanoleuca / Melanoleuca

Urticocystis /Grammopodiae

Melanoleuca / Melanoleuca

Melanoleuca / Cognatae

Acystis / Decembres

Urticocystis /Grammopodiae

Melanoleuca / Oreinae

Melanoleuca / Alboflavidae

Urticocystis /Grammopodiae

Melanoleuca / Melanoleuca

Acystis / Acystis

Urticocystis /Grammopodiae

Melanoleuca / Melanoleuca

Urticocystis /Grammopodiae

Melanoleuca / Melanoleuca

Urticocystis /Grammopodiae

Melanoleuca / Alboflavidae

ITS

Acc. No.

JN616418

JN616419

JN616420

JN616421

JN616422

JN616423

JN616424

JF908352

JF908351

JN052138

JF908356

JN052137

JF908360

JN616425

JN616426

JN616427

JN616428

JF908346

JN616429

JN616430

JN052142

JN616431

JN616432

JN616433

JN616434

JN616435

JN616436

JN616437

JN616438

JN616439

JN616440

JN616441

JN616442

JN616443

JF908350

JN616444

JN616445

JN616446

JN616447

JN616448

JN616449

JN616450

JN616451

JN392452

TABLE 2, concluded

COLLECTIONS

M. oreina

* M. paedida 1

* M. paedida 2

M. “paratristis”

* M. privernensis

* M. pseudoluscina 1

* M. pseudoluscina 2

* M. pseudoluscina 3

* M. pseudoluscina 4

* M. pseudoluscina 5

* M. pseudopaedida

* M. rasilis

* M. robertiana

* M. robusta

* M. strictipes 1

* M. strictipes 2

* M. strictipes 3

* M. stridula

* M. striimarginata

M. subalpina

* M. sublanipes 1

* M. sublanipes 2

* M. sublanipes 3

* M. subpulverulenta 1

* M. subpulverulenta 2

* M. substrictipes 1

M. substrictipes 2

* M. substrictipes

var. sarcophylla

M. verrucipes

* Melanoleuca sp. 1

* M. sp. 2

* M. sp. 3

M. sp. 4

M. sp. 5

M. sp. 6

M. sp. 7

M. sp. 8

M. sp. 9

M. sp. 10

M. sp. 11

M. sp. 12

M. sp. 13

M. sp. 14

M. sp. 15

* M. sp. 16

M. sp. 17

M. sp. 18

CoOL. ID./ORIGIN

MCVE 07839/Italy

ANC MO0189/Italy

ANC MO0190/Italy

MCVE 12645/Italy

GC 08310 holotype/Italy

ANC MO0191/Italy

ANC MO0192/Italy

ANC MO0193/Italy

ANC MO0194/Italy

ANC MO0195/Italy

ANC MO0198/Italy

ANC M0220/Italy

ANC M0205/Italy

ANC MO0179/Italy

ANC MO171/Italy

ANC MO0172/Italy

ANC MO0173/Italy

ANC M0007/Italy

ANC M0202/Italy

MCVE 04112/Italy

ANC M0221/Italy

ANC M0222

holotype/Italy

ANC M0223/France

ANC M0004/Italy

ANC MO178/Italy

ANC M0214/Italy

MCVE 13934/Italy

ANC M0209/Italy

MCVE 09962Switzerland

ANC MO18I1/Italy

ANC MO0188/Italy

ANC M0224/Italy

MCVE 12248/Italy

MCVE 13410/Italy

MCVE 09821/Italy

MCVE 01687/Italy

MCVE 01683/Italy

MCVE 19223/Italy

MCVE 08389/Italy

MCVE 08432/Italy

MCVE 14221/Italy

MCVE 08384/Italy

MCVE 03316/Italy

MCVE 19627/Italy

MCVE 24095/Italy

MCVE 01681/Italy

MCVE 14273/Italy

SUBGENERA / SECTIONS

(BON 1991)

Melanoleuca / Oreinae

Urticocystis /Grammopodiae

Acystis / Acystis

Subgen. Kinia (Vizzini

et al. 2010)

Urticocystis /Grammopodiae

Acystis / Acystis

Urticocystis /Grammopodiae

Acystis / Acystis

Melanoleuca / Melanoleuca

Melanoleuca / Alboflavidae

Acystis / Acystis

Melanoleuca / Alboflavidae

Urticocystis /Grammopodiae

Melanoleuca / Melanoleuca

Urticocystis /Grammopodiae

Urticocystis / Humiles

Acystis / Acystis

Melanoleuca / Melanoleuca

Melanoleuca / Cognatae

Melanoleuca / Melanoleuca

Urticocystis /Grammopodiae

Acystis / Decembres

ITS sequence analysis of Melanoleuca ... 367

ITS

Acc. No.

JN392450

JN616452

JN616453

908357

JN616454

JN616455

JN616456

JN616457

JN616458

JN616459

JN616460

JN616461

JN616462

JN616463

JN616464

JN616465

JN616466

JN616467

JN616468

JN052139

JN616469

JN616470

JN616471

JN616472

JN616473

JN616474

908359

JN616475

908354

JN616476

JN616477

JN616478

JN052141

908358

JN392453

JN392449

JN392448

JN392446

908353

JN052140

JN392454

JN392451

908349

908344

JN616479

JN392447

908362

368 ... Vizzini & al.

dell’Universita di Ancona, Italy) or acquired from MCVE (the Museo di Storia Naturale

di Venezia Herbarium, Italy) (Tas. 2). MCVE collections were chosen, even where

often misdetermined, because DNA had already been bar-coded by Dr. M. Garbelotto

(University of California, Berkeley). All Melanoleuca collections were identified or

redetermined using i) an unpublished key based on the protologues or referenced to

original collections (Fontenla & Para, ined.) and ii) existing monographs (Bon 1991,

Boekhout 1999, Fontenla et al. 2003). Watling & Turnbull (1983), Horak (2005), and

Vesterholt (2008) were also consulted. When not identifiable, collections are cited in

TaB. 2 and Fic. 2 as Melanoleuca sp. Melanoleuca species were selected to represent

subgenera and sections in Bon (1991; Tas. 1). Each section was represented by at least

two species except for sect. Humiles (represented only by M. verrucipes). Melanoleuca

privernensis (Consiglio et al.) Consiglio et al. [= Kinia privernensis] was also included

in the dataset in accordance with Vizzini et al. (2010). In the species descriptions Q =

quotient of length and width of the spores in side view and Qm = average quotient. The

spore Qm value and cystidia shape are reported for each collection in Fic. 2. Herbarium

abbreviations follow Thiers (2011). Author citations follow the Index Fungorum-

Authors of Fungal Names (http://www.indexfungorum.org/authorsoffungalnames.

htm) and the names of new taxa are deposited in MycoBank (http://www.mycobank.

org/DefaultPage.aspx).

DNA extraction, PCR amplification, DNA sequencing

Genomic DNA was isolated, extracted from 62 dried herbarium specimens (Tas. 2)

using the DNeasy Plant Mini Kit (QIAGEN, Milan, Italy). Universal primers ITS1F/

ITS4 were used for ITS region amplification (White et al. 1990, Gardes & Bruns 1993).

Amplification reactions were performed in a PE9700 thermal cycler (Perkin-Elmer,

Applied Biosystems) in 25 mL reaction mixtures with these final concentrations or total

amounts: 5 ng DNA, 13 PCR buffer (20 mM Tris/HCl pH 8.4, 50 mM KCl), 1 mM each

primer, 2.5mM MgCl2, 0.25mM each dNTP, 0.5 unit Taq polymerase (Promega). The

PCR program was 3 min at 95 C for one 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 one cycle. PCR products were resolved on a 1.0%agarose

gel and visualized by staining with ethidium bromide. PCR products were purified with

the AMPure XP kit (Beckman) and sequenced by DINAMYCODE srl (Turin, Italy).

Sequences were assembled and edited with the phred/phrap/consed software suite and

submitted to GenBank (Tas. 2).

Sequence alignment and phylogenetic analysis

Sequences included in the phylogenetic analyses were generated in this study

(TaB. 2) or retrieved from GenBank. GenBank sequences were selected based on other

phylogenetic studies on Agaricales (Moncalvo et al. 2002; Matheny et al. 2006; Justo et al.

2011). Limnoperdon incarnatum (DQ097363) was used as outgroup taxon. Other taxa

belonging to the Pluteoid clade were included in the analysis for testing the monophyly

of Melanoleuca.

Sequences obtained in this study were checked and assembled using Geneious v5.3

(Drummond et al. 2010). Alignment of the ITS dataset was generated using MAFFT

v6.814b (Katoh et al. 2002) with default conditions for gap openings and gap extension

penalties. The alignment was then imported into MEGA 5.0 (Tamura et al. 2011) for

ITS sequence analysis of Melanoleuca ... 369

manual adjustment. Best-fit models were estimated by both the Akaike Information

Criterion (AIC) and the Bayesian Information Criterion (BIC) using jModelTest 0.1.1

(Posada 2008) to provide a substitution model for each single alignment. TPM2uf+I+G

model was chosen. Phylogenetic hypotheses were constructed under Bayesian Inference

(BI) and Maximum Likelihood (ML) criteria.

BI of phylogeny using Monte Carlo Markov Chains (MCMC) was carried out with

MrBayes 3.1.2 (Huelsenbeck & Ronquist 2001). Four incrementally heated simultaneous

MCMC were run over 10.000.000 generations, under model assumption. Trees were

sampled every 1.000 generations resulting in an overall sampling of 10.001 trees. The

“burn-in” value was evaluated using Tracer 1.5 (Rambaut & Drummond 2007). The

first 15% of trees was discarded as “burn-in”. For the remaining trees, a majority rule

consensus tree showing all compatible partitions was computed to obtain estimates for

Bayesian Posterior Probabilities (BPP). Branch lengths were estimated as mean values

over the sampled trees. Only BPP values over 50% are reported in the resulting trees.

This Bayesian analysis was repeated three times, always using random starting trees and

random starting values for model parameters to test the independence of the results

from the revisiting of the prior topologies during chain growth (Huelsenbeck et al.

2002).

ML estimation was performed through RAXML v.7.0.4 (Stamatakis 2006) with 1,000

bootstrap replicates (Felsenstein 1985) using the GTRGAMMAT algorithm to perform

a tree inference and search for a good topology. Support values from bootstrapping runs

(MLB) were mapped on the globally best tree using the “-f a” option of RAxML and

“-x 12345” as a random seed to invoke the novel rapid bootstrapping algorithm. Support

values for major clades that are supported by either BI and ML analyses are visualized

in the resulting tree.

Analysis of the pairwise % identity values (hereafter shortened as P%IV) for the

Melanoleuca sequences were calculated using MEGA 5.0 (Tamura et al. 2011).

Results

Our phylogenetic results are presented in Fic. 2. The general ITS data matrix

comprises a total of 105 sequences (including 14 from GenBank). This 898 bp

dataset contains 621 (69.2%) variable sites, of which 525 (58.4%) are parsimony-

informative. The Bayesian and ML tree topologies are congruent. Our analyses

show that Melanoleuca is clearly monophyletic (1.0 BPP and 89% MLB). Two

major clades, A and B, were distinguished within Melanoleuca. Clade A is

supported by only BI tree with 0.79 BPP, while clade B is well supported with

1.0 BPP and 99% MLB. Clade A consists of 5 clades (A1-A5, with 10 subclades)

and clade B of 5 clades (B1-B5, with 3 subclades).

Discussion

Melanoleuca and infrageneric classification

The Bayesian (1.0 BPP) and ML (89% MLB) analyses strongly support

Melanoleuca as monophyletic and distributed throughout the ingroup within

370 ... Vizzini & al.

2 major clades (A and B) with 10 smaller clades (A1—A5 and B1-B5) (Fie. 2).

The two sequences of M. subsejuncta (Peck) Murrill from GenBank (FJ596898;

FJ596898) cluster outside Melanoleuca, sister to Amanitaceae; Pfister (1984),

who examined its type, referred the collection to Tricholoma. Results of

our phylogenetic analyses and the current morphology-based infrageneric

classification of Melanoleuca are not congruent. In particular, our data (Fic.

2) are incompatible with the tripartite infrageneric Melanoleuca classification

of Bon (1991) and Boekhout (1988, 1999) (Tas. 1). Based on molecular data,

species traditionally ascribed to subgenus Acystis Bon (= subgenus Melanoleuca

sensu Boekhout 1988, 1999) do not form a monophyletic assemblage and are

distributed over the Melanoleuca clade (clade A and B, Fig. 2).

The fact that acystidiate taxa are not phylogenetically related implies that

cystidial acquisition and loss have taken place independently during the

evolution of Melanoleuca and also that this character is homoplastic and

unsuitable for a natural classification of these fungi. Subgenus Acystis should

therefore be considered an artificial superfluous taxon that is no longer tenable.

In addition, in some cases (e.g. subclades A4.1 & A4.2; clade B2, Fic. 2),

cystidiate and non-cystidiate taxa are conspecific (syntaxic): the acystidiate

M. sp. 18, M. sp. 2, and M. sp. 1 represent only forms of the cystidiate taxa

M. pseudoluscina Bon, M. paedida (Fr.) Kihner & Maire, and M. atripes

Boekhout/M. albifolia Boekhout, respectively. It is conceivable that the shift

from cystidiate to non-cystidiate basidiomata and vice versa within a single

species is controlled by a limited number of genes that are switched off by so far

unknown environmental factors.

Most macrocystidiate taxa in subg. Melanoleuca (= Macrocystis Boekhout)

form clade B (Fic. 2), while the M. cognata complex (A5 clade, = sect. Cognatae)

nests with all species in subg. Urticocystis to form clade A (Fic. 2). Therefore,

subg. Melanoleuca as traditionally circumscribed (Bon 1991) is polyphyletic

and subg. Urticocystis is paraphyletic when sect. Cognatae is excluded. It would

appear that even cystidial shape is not a reliable character for tracing higher

phylogenetic relationships.

Our analysis therefore implies that only two subgenera (clades A and B)

should be recognized in Melanoleuca:

Melanoleuca Pat. subg. Melanoleuca, emend. Fontenla, Para & Vizzini

Clade B (the autonymous subgenus, including M. melaleuca), characterized by

basidiomata with non-septate macrocystidia, or rarely without cystidia.

Melanoleuca subg. Urticocystis Boekhout, Persoonia 13(4): 400 (1988),

emend. Fontenla, Para & Vizzini

Clade A (type species: M. grammopodia (Bull.) Murrill), comprising taxa mainly with

urticocystidia but also with macrocystidia and brightly coloured pilei (sect. Cognatae),

or lacking cystidia.

ITS sequence analysis of Melanoleuca ... 371

0.84/56 ———— MCVE 08384 M. sp. 13 A-S3

'_____——— ANC M0217 M. grammopodia1 U2-S3

ous [LT ANC M0218 M. grammopodia 2 U1U2-S2

ANC M0219 M. grammopodia3 U1-S2 A1.1

100 | ANC M0216 M. grammopodia f. macrocarpa 2. U1-S3

GC 08310 M. privernensis A-S3

ANC M0215 M. grammopodia f. macrocarpa’1_U1U2-S3

0.84/50 “ MCVE 04410 M. grammopodia f. macrocarpa 3 U1-S3

11100-— MCVE 04574 M. brevipes 1 U1-S2

1/99 [L MCVE 04505 M. brevipes 2 U1-S2

— MCVE 03316 M. sp. 14 U1-S2

0.57). [0-62 199 —— ANC M0220 M. rasilis U1-S3

L MCVE 19627 M. sp.15 U1-S2

195 —————_————— MCVE 24095 M. sp. 16 U1-S2

oT. '_— ANC M0223 M. sublanipes 3 U1-S2

08757 ANC M0222 M. sublanipes 2 U1-S1

1086. MCVE 01681 M. sp. 17 U1-S1

— ANC M0221 M. sublanipes1 U1-S1

-— ANC M0212 M. exscissa5 U2-S3

+ ANC M0213 M. exscissa6 U2-S2

ANC M0210 M. exscissa3 U1U2-S3

0.99/61 | |}——— ANC M0211 M. iris U2-S2

[|/ ANC M0210B M. exscissa 4 U1U2-S3 A2.1

0.54-| - ANC M0209 M. substrictipes var. sarcophylla U2-S2

oss;— ANC M0207 M. exscissa1 U2-S1 A2

9895 |"! ANC M0206 M. diverticulata U2-S2

"™L ANC M0208 M. exscissa 2 U2-S2

196| [-— ANC M0214 M. substrictipes 1 U2-S3 A2.2

oast MCVE 13934 M. substrictipes 2. U1U2-S3 | clade A

x 1100 ; M. verrucipes DQ490642 (U) . .

———L MCVE 09962 M. verrucipes U2-S3 A2.3 su bg enus Urt icocyst. 1S

0.55/--— ANC M0202 M. striimarginata A-S2

0.87/70/-L_ MCVE 12645 M. paratristis A-S1

ose-| “- ANC M0201 M. graminicola A-S1

L180 ANC M0203 M. angelesiana1 A-S1

0.79) 05t/- ANC M0204 M. angelesiana2 A-S1

ral ANC M0198 M. pseudopaedida U1-S1

82} ANC M0197 M. decembris 1 U1-S1

053: | MCVE 01573 M. decembris 4 U1-S1

ss] ANC M0199 M. decembris 2. U1-S1

0.53). mL ANC M0200 M. decembris 3 U1-S2

{___ ANC M0007 M. stridula A-S1

{___ ANC M0205 M. robertiana_A-S1

1198 ANC M0191 M. pseudoluscina1 U1-S1

o.ge6g/L______________ MCVE 14221 M. sp.12 U2-S2

0.65/- ANC M0193 M. pseudoluscina3 U1-S2

061/661 ANC M0192 M. pseudoluscina 2 U1-S2 A4.1

yoo [| MCVE 14273 M. sp.18 A-S2

ANC M0195 M. pseudoluscina5 U2-S1 A4

ANC M0188 M. sp.2 A-S2

LF ANC M0190 M. paedida2 U1-S1

0.834 | oS] + ANC M0189 M. paedida 1 U1-S2 A42

ANC M0187 M. electropoda U1U2-S1

'_ ANC M0196 M. microcephala A-S1

A1.2

0.95/54

400 - ANC M0170 M. cognata2 M2-S2

1199 MCVE 13939 M. cognata1 M1-S3 A5

> ANC M067 M. arcuata M1M2-S2

1/89 og9i97 - MCVE 08389 M. sp. 10 M1M2-S1

0.97/72,— ANC M0178 M. subpulverulenta2 M1M2-S2

0.7975 | ~ ANC M0004 M. subpulverulenta1 M1M2-S3 B1.1

[- ANC M0176 M. melaleuca MM2-S3

L ANC M0177 M. nivea1 M1-S3

197- MCVE 09821 M. sp. 6 M1-S3

comes [| '—— MCVE 09578 M. nivea3_ M1M2-S3 B1.2

L. ANC M0179 M, robusta M2-S3

-—— ANC M0166 ML. lanipes MiM2-S2

ANC M0175 M. heterocystidiosa 2 M1-S3 | B1.3

195-—|L ANC M0174 M. heterocystidiosa 1 M1-S3

ANC M0224 M. sp. 3. M1M2-S2

MCVE 13410 M. sp.5 M1-S2

MCVE 12248 M. sp. 4 M1-S2

MCVE 08432 M. sp. 11 M2-$3 clade B

os) ANC M0182 M. albifolia 1 M2-S2

ANC M0183 M. nivea2 M2-S1

(ANC Mone Me ae aes Bo | SU bgenus Melanoleuca

os3t oat ANC M0181 M.sp.1 A-S1

| ANC M0180 M. atripes M2-S1

ANC M0184 M. albifolia 2 MiM2-S3

1/89 - ANC M0173 M. strictipes 3 M1-S3

ANC M0172 M. strictipes 2 M1M2-S3

joo (182 ANC MO171 M. strictipes 1 M1-3 B3

MCVE 14576 M. evenosa M1-S3

L MCVE 04112 M. subalpina M1-S3

MCVE 07839 M. oreina M1-S3

om MCVE 01687 M. sp.7 M2-S3 B4

a2 MCVE 19223 M. sp.9 M1-S2

|_____ ANC M0185 M. bataillei_ MM2-S2

0.74)-

MCVE 11243 M. cinereifolia2 M2-S2

o.s464 ++ MCVE 01471 M. cinereifolia1 M2-S3 B5

MCVE 20748 M. cinereifolia3 M2-S2

“ ANC M0186 M. friesii M1-S2

iste Limacella glioderma AY 176451

= Amanita phalloides GQ221842

_ Amanita muscaria var. muscaria AB080790

Macrocystidia cucumis DQ490640

11100 _ Melanoleuca subsejuncta FJ596899

Melanoleuca subsejuncta FJ596898

4400. ———————————— Vol vopluteus gloiocephalus HM562209

0.62/- |_____________ Volvopluteus earlei HI562205

11100 Pluteus salicinus HM562174

Pluteus cervinus var. cervinus EU486448

1/100 -—— Pluteus romellii HM562183

Pluteus aurantiorugosus HM562121

Limnoperdon incarnatum DQ097363

0.08

FIGURE 2. Bayesian phylogram obtained from the ITS rDNA sequences of Melanoleuca and other

taxa of the Pluteoid clade. Limnoperdon incarnatum was used as outgroup taxon. Support values

(BPP in bold >0.5; MLB >50%) are given above branches. Minor supported clades discussed in the

text are numbered A1-A5 and B1-B5. Numbers refer to the collections cited in Tas. 2.

Characters: 1) presence/absence of cheilocystidia and type of cheilocystidia — A = taxa

without cheilocystidia, U1 = taxa with brevipes-type urticiform cystidia, U2 = taxa with exscissa-

type urticiform cystidia, U1-U2 = taxa with both types of urticiform cystidia, M1 = taxa with

fusiform macrocystidia, M2 = taxa with lageniform macrocystidia, M1-M2 = taxa with both type

of macrocystidia [cystidial types are coded in red (A), blue (U), and green (M)]; 2) spore shape

(Qm range) - $1 = Qm < 1.40; $2 = 1.40< Qm $1.60; $3 = Qm > 1.60..

372 ... Vizzini & al.

Basidiospore shape (Qm value) seems to be homoplastic, too variable to use for

circumscribing sections and subsections as proposed by Bon (1991). For example,

in subclade A2.1, collections otherwise assignable to M. exscissa (Fr.) Singer show

a great Qm range, as do M. grammopodia (A1.1) and M. pseudoluscina (A4.1)

collections (Fic. 2).

On the contrary, cystidial subtypes may have some value for delimiting small

clades (Fic. 2). Clades Al and A3 are formed mainly by taxa with brevipes-

type urticocystidia and clade A2 by taxa by mainly exscissa-type urticocystidia.

Clade B1 comprises taxa with mainly fusiform macrocystidia.

Pileus colour is not a reliable phylogenetic marker at any taxonomic level:

three collections (ANC M0177, MCVE 09578, ANC M0183) determined

as M. nivea Métrod ex Boekhout, a species traditionally included in sect.

Alboflavidae based on whitish colouration and presence of macrocystidia (Bon

1991, Boekhout 1988, 1999; Deschuyteneer 2008), are not placed with other

species of that section (clade B3) but distributed throughout clade B. ‘The first

collection represents an albinic form of M. melaleuca (B1.1), the second a taxon

close to M. robusta (Bres.) Fontenla et al. (B1.2), and the third probably an

albinic form of M. albifolia (B2).

In conclusion, taxonomically important morphological characters in

Melanoleuca show a high degree of homoplasy. Although these characters

are useful for species delimitation, and in some cases for the circumscription

of sections or groups, they appear insufficient for a phylogenetically correct

infrageneric concept. A clearer picture will emerge as more Melanoleuca

diversity is included in future analyses.

Groupings and species limits

More extensive sampling of Melanoleuca is needed for a revision of the

whole genus. Additional ITS sequencing is necessary to clarify species limits

and names for taxa occurring in both Europe and North America. For the time

being we only make minor comments on some of the infrageneric clades that

were recovered. These numbered clades and subclades are marked in Fie. 2.

Subgenus Urticocystis (Clade A): clades Al-A5

CLADE Al (0.57 BPP, / MLB), cheilocystidia mainly of the brevipes-type or

rarely absent. Stipe striate longitudinally or not.

SUBCLADE A1.1 (1.0 BPP, 100% MLB) (= M. grammopodia complex = subsect.

Grammopodiae). ‘This well supported subclade comprises M. grammopodia

and M. grammopodia f. macrocarpa Boekhout, all with urticocystidia, and

M. privernensis and M. sp. 13 without cystidia. All taxa have a distinctly

longitudinally striate stipe with a pruinose apex. The eight sequences display

a 96.8 P%IV. Melanoleuca grammopodia f. macrocarpa differs from the type

ITS sequence analysis of Melanoleuca ... 373

only by a stipe that is much shorter than pileus diameter (P%IV = 99.4) and

so should be considered only a growth form of M. grammopodia. Melanoleuca

privernensis is diagnosed by non-amyloid spores, unique in Melanoleuca

(Vizzini et al. 2010). The molecular analysis suggests it might represent an

aberrant neotenic form of M. grammopodia lacking cystidia with non-amyloid

spores.

SUBCLADE A1.2 (0.62 BPP, / MLB) includes M. brevipes (Bull.) Pat., M. rasilis

(Fr.) Singer, and two unidentified species (M. sp. 14, M. sp. 15). Melanoleuca

rasilis is related to M. brevipes based on similar cystidia, grey pileus and stipe

colours, and a non-longitudinally striate stipe but differs by smaller basidiomes,

a stipe length equal to the pileus diameter, and broadly ellipsoid coarsely

ornamented spores (Boekhout 1988, 1999). We also observed that M. rasilis

has a uniformly white context while M. brevipes has a white pileus context and

brown stipe context. Molecularly, the two species are clearly different (P%IV =

94.5; Fic. 2).

SUBCLADE A1.3 (1.0 BPP, 96% MLB) comprises M. sublanipes, M. sp. 16, and

M. sp. 17. Melanoleuca sublanipes (formally described below) is characterized

by a tomentose-woolly pileus margin and stipe base. The two M. sp. could be

conspecific with M. sublanipes, but their collections lack of macromorphological

data and descriptions. We here propose the following new diagnosis:

Melanoleuca sublanipes Fontenla, Para & Vizzini, sp. nov.

MycoBank MB 563140

Pileus -10 cm latus, velutinus, obscure griseo-brunneus, ochraceo-brunneus, griseo-

brunneus, a margine pallescens. Lamellae albae, albidae vel pallide griseo-brunneae. Stipes

1.5-7 x 0.3-1.1 cm, lanatus deinde solum inferne lanato-flocculosus, in senectute omnino

pruinoso-flocculosus, argenteus, a basi griseo-brunneus, cum ripercussis cyaneis. Caro in

pileo albida, in stipite brunnea; odor herbaceus; sapor mitis. Sporae 6.2-9.6 x 3.8-6.6

um; in medio 7.56 x 5.54 um; Q = 1.04-1.70; Qm = 1.36, ellipsoideae, conspicuis verrucis

singulis, globulosis et amyloideis exornatae. Basidia tetraspora. Cheilocystidia numerosa,

31-55 x 5-8 um, pili urticae revocantibus, raro fusoidea. Pilei cutis ad marginem ex

hyphis erectis, 7-8 um latis ad instar trichodermatis efformata; hyphae terminales

cylindricae, usque ad 100 um longae. Stipitis cutis pilis clavatis aggregatis et caulocystidiis

multiformibus ornata. Habitat in herbidis locis.

TyPE: Italy, Veneto, Padova, Montegrotto Terme, 07.X1.2008, leg. G. Zecchin (holotype

ANC M0222).

ErymMo.oey: the specific epithet refers to the tomentose-woolly surface of the stipe

base.

CLADE A2 (1.0 BPP, 96% MLB), cheilocystidia mainly of the exscissa-type:

subclades A2.1, A2.2 and A2.3.

SUBCLADE A2.1 (0.54 BPP, / MLB) (M. exscissa complex) comprises M. exscissa,

M. diverticulata, M. iris, and M. substrictipes var. sarcophylla. The nine sequences

display a 98.5 P%IV. This clade reflects an overemphasis on basidiome colour

37A ... Vizzini & al.

and smell, with some anatomical traits having led to the establishment of

some superfluous species. Melanoleuca iris differs from M. exscissa only in

the strong and sweet smell recalling Lepista irina (Fr.) H.E. Bigelow (Kiihner

1956, Klan 1983, Bon 1991, Boekhout 1988, 1999; Krieglsteiner 2001, Fontenla

et al. 2003). Boekhout (1988, 1999) considered it a variety of M. exscissa and

Krieglsteiner (2001) cited it as only an accidental M. exscissa phenotype; our

molecular data (P%IV between M. iris and M. exscissa sequences = 98.7) clearly

support Krieglsteiner. With an intraspecific variability lower than 3% (Nilsson

et al. 2008), M. iris should be considered a form of M. exscissa. Melanoleuca

diverticulata differs from M. exscissa mainly by the presence of elements with

small outgrowths in the pileipellis (Moreno & Bon 1980). The P%IV between

M. diverticulata and M. exscissa ANC M0207 and ANC M0208 equals 97.9.

Our data show that M. substrictipes var. sarcophylla, which Kihner (1978)

distinguished from the typical variety based on its pinkish lamellae (Kithner

1978, Bon 1991), is more closely related to M. exscissa than to M. substrictipes

Kihner (Fic. 2) and should be considered a form of M. exscissa with coloured

lamellae (P%IV = 98.5).

Therefore, we propose the following new combinations:

Melanoleuca exscissa f. iris (Kiihner) Fontenla, Para & Vizzini,

comb. nov., stat. nov.

MycoBank MB 563141

= Melanoleuca iris Kiithner, Bull. Soc. Linn. Lyon 25(7): 181 (1956).

The holotype of M. iris apparently missing from G (herbarium where Kihner’s collections

of Melanoleuca are kept; Fontenla & Para, unpublished data).

Melanoleuca exscissa f. sarcophylla (Kihner) Fontenla, Para & Vizzini,

comb. nov., stat. nov.

MycoBank MB 563142

= Melanoleuca substrictipes var. sarcophylla Kihner,

Bull. Soc. Linn. Lyon 47(1): 52 (1978).

Holotype (G - K 65-24), with morphological features coincident with those of our

sequenced collection (ANC M0209; Fontenla & Para, unpublished data).

Melanoleuca exscissa f. diverticulata (G. Moreno & Bon) Fontenla, Para & Vizzini,

comb. nov., stat. nov.

MycoBANnkK MB 563143

= Melanoleuca diverticulata G. Moreno & Bon, Docum. Mycol. 11(41): 35 (1980).

Holotype (MA - 3662), with morphological features coincident with those of our

sequenced collection (ANC M0206; Fontenla & Para 2007).

SUBCLADE A2.2 (0.95 BPP, 77% MLB) The two M. substrictipes collections

are very similar (P%IV = 99.1). The first (ANC M0214) has only urticoid

exscissa-type cheilocystidia, whereas the second (MCVE 13934) possesses

ITS sequence analysis of Melanoleuca ... 375

both brevipes- and exscissa-type cheilocystidia. Following Bon (1991) the two

molecularly clearly conspecific collections would be referred to two different

subsections: the first in the subsect. Exscissae (as M. pseudoevenosa Bon ex Bon

& G. Moreno) and the second in subsect. Grammopodiae. The holotype (G-K

66-13) is characterized by both types of urticiform cheilocystidia (Fontenla &

Para 2011).

SUBCLADE A2.3 (1.0 BPP, 100 % MLB) The two M. verrucipes (Fr.) Singer

sequences (GenBank DQ490642 and MCVE 09962) are almost identical

(P%IV = 99.8). The black dotted stipe readily diagnoses the species (Bon 1991,

Boekhout 1999, Gasparini 2001, Fontenla et al. 2003).

CLADE A3 (0.53 BPP, / MLB), cheilocystidia absent or urticocystidia mainly of

the brevipes-type. Both pileus and stipe usually (except for M. robertiana)

dark coloured and lamellae white to whitish. This clade encompasses the

two acystidiate species, M. robertiana Bon and M. stridula (Fr.) Singer, and

subclades A3.1 and A3.2.

SUBCLADE A3.1 (0.66 BPP, / MLB) comprises four acystidiate taxa, M. strii-

marginata Métrod ex Bon, M. “paratristis,’ M. graminicola (Velen.) Kihner

& Maire, and M. angelesiana A.H. Sm. ‘The first three taxa (subg. Acystis sect.

Acystis in Bon 1991) are morphologically and molecularly (P%IV = 98.2)

closely related: they share a collybioid habit, white lamellae, and a whitish stipe,

differing mainly in pileus colour and could probably be reduced to only one

species, M. graminicola (which would have nomenclatural priority). Melanoleuca

angelesiana (the 2 sequences with a P%IV = 99.9) is an independent species

(P%IV = 94.3), distinguished by a tricholomatoid habit, grey lamellae, and

greenish-brown stipe. The two Italian collections (ANC M0203, ANC M0204)

show features consistent with the protologue (Smith 1944) and holotype (MICH

14633) of this North American species (Fontenla & Para, unpub. data).

SUBCLADE A3.2 (0.99 BPP, 89 % MLB) encompasses M. decembris Métrod

ex Bon and M. pseudopaedida Bon. Bon (1991) classified these species as

acystidiate (subg. Acystis, sect. Decembres), but Fontenla & Para (2008) found

numerous urticiform cheilocystidia when they studied the type collections.

Both taxa are characterized by a dark (dark brown-grey to blackish) pileus and

grey lamellae (Bon 1991). Molecular analysis supports M. pseudopaedida as

conspecific with M. decembris (P%IV = 98.1).

CLADE A4 (0.54 BPP, / MLB), cheilocystidia absent or both brevipes- and

exscissa-type urticocystidia, comprising the acystidiate M. microcephala

(P. Karst.) Singer and A4.1 and A4.2 subclades. Bon (1991) described

M. microcephala as having urticiform cystidia, but Fontenla and Para

(unpub. data) found Karsten’s type collection (H 1604) to be acystidiate.

376 ... Vizzini & al.

SUBCLADE A4.1 (1.0 BPP, 99 % MLB) contains M. pseudoluscina, M. sp. 12,

and M. sp. 18. The seven sequences show a P%IV of 97.1. The P%IV (98.6)

between M. sp. 18 and M. pseudoluscina sequences indicates that M. sp. 18 is an

acystidiate form of M. pseudoluscina. Our analysis support M. pseudoluscina as

an independent species and not a variety of M. rasilis (subclade A1.2) as stated

by Boekhout (1988, 1999).

SUBCLADE A4.2 (1.0 BPP, 99 % MLB) encompasses M. paedida, M. sp. 2,

and M. electropoda. The two M. paedida collections are consistent with the

protologue and the observations by Fontenla et al. (2003). Melanoleuca sp. 2

is an acystidiate form of M. paedida. Melanoleuca electropoda was reported by

Bon (1991) as a macrocystidiate species (subg. Melanoleuca, sect. Oreineae);

after finding typical urticoid cheilocystidia in the type collection, Fontenla

et al. (2009) considered M. rufipes Bon a later synonym. As M. electropoda

differs from M. paedida only in an orange red (instead of brown) stipe base

and context, we reduce it to a form of M. paedida based on molecular data

(P%IV = 98.9).

Melanoleuca paedida f. electropoda (Maire & Malencon) Fontenla, Para & Vizzini,

comb. nov., stat. nov.

MycoBank MB 563144

= Melanoleuca electropoda Maire & Malencon, Fl. Champ. sup. Maroc 2: 77 (1975).

CLADE As5 (1.0 BPP, 99% MLB) (= sect. Cognatae), macrocystidiate taxa with

bright coloured pilei.

Melanoleuca cognata (Fr.) Konrad & Maubl. and M. arcuata (Bull.) Singer are

two independent species (P%IV = 95.4). Melanoleuca arcuata differs by having

a brown pileus and yellow-ochre lamellae only at maturity. The collection

named M. sp. 10 is probably referable to a new species (P%IV = 93.1), but its

macromorphological data are lacking.

Subgenus Melanoleuca (Clade B): clades B1-B5 + M. friesii (Fr.) Singer (a

species characterized by a dark-brown blackish, slate-grey pileus, whitish

lamellae and mainly lageniform cystidia).

CLADE Bi (1.0 BPP, 95% MLB), macrocystidia both fusiform and lageniform,

pileus mainly grey-brown and lamellae white to grey. It encompasses the

subclades B1.1, B1.2, B1.3 + 4 M. sp. (M. sp. 3, M. sp. 4, M. sp. 5 and M.

sp. 11).

SUBCLADE B1.1 (0.79 BPP, 75 % MLB) comprises M. subpulverulenta

(Pers.) Singer, M. melaleuca and M. nivea 1. The four collections are closely

related (P%IV = 98.6) despite their morphological differences. Melanoleuca

subpulverulenta is characterized by a pruinose matte grey pileus, pruinose stipe,

ITS sequence analysis of Melanoleuca ... 377

and dark context; M. melaleuca has a grey-brown non-pruinose pileus, non-

pruinose stipe, and whitish context sometimes darkening towards the stipe

base. The M. nivea 1 collection (ANC M0177) we originally referred to M. nivea

due to its whitish colours (sect. Alboflavidae) and <5 cm pileus represents only

an albinic form of the M. subpulverulenta/M. melaleuca complex; its features

nonetheless match those of the M. nivea holotype (PC-2434) (Fontenla & Para,

unpublished data).

SUBCLADE B1.2 (0.56 BPP, 45 % MLB) comprises M. sp. 6, M. nivea 3, and

M. robusta, with the latter distinguished by a grey-brown pileus, grey lamellae,

brown context, caespitose growth, and mainly fusiform macrocystidia.

Melanoleuca nivea 3 is very close to an unidentified collection (M. sp. 6)

characterized by a grey pileus.

SUBCLADE B1.3 (0.62 BPP, 73 % MLB) consists of two molecularly closely

related taxa (P%IV = 98.8), M. “lanipes” and M. heterocystidiosa (Beller &

Bon) Bon that have both lageniform and fusiform macrocystidia. Melanoleuca

lanipes, which has dark grey-brown lamellae, strongly longitudinally striate

dark grey stipe covered with woolly flocci, and blackish stipe base, resembles

M. cognata but with a tomentose-wooly stipe.

CLADE B2 (0.52 BPP, / MLB), mainly lageniform cystidia, longitudinally striate

and brown to dark brown stipe.

CLADE B2 is formed by M. albifolia, M. atripes, M. nivea 2, M. sp. 1 and

M. sp. 8. All six sequences are closely related (P%IV = 97.6). Morphologically,

M. albifolia is well characterized by a rather small size (2.5-5 cm diam), dark

sepia-brown pileus, white lamellae, grey-brown stipe, and lageniform cystidia

(Boekhout 1988, 1999; Bon 1991); M. atripes has a hygrophanous blackish

brown pileus, dark brown stipe, yellowish beige lamellae, and mainly fusiform

cystidia (Boekhout 1988, 1999; Bon 1991). In contrast, our M. atripes collection

(ANC M0180) shows mainly lageniform cystidia.

The sequence analysis did not support any independent species in the

clade. M. nivea 2 is probably an albinic form of the M. albifolia/M. atripes

complex while M. sp. 1 is the only acystidiate collection nested within subgen.

Melanoleuca (clade B).

CLADE B3 (1.0 BPP, 92% MLB), macrocystidia mainly fusiform, basidiome

white to cream-whitish (sect. Alboflavidae).

CLADE B3 comprises M. strictipes (P. Karst.) Jul. Schaff., M. evenosa (Sacc.)

Konrad, and M. subalpina (Britzelm.) Bresinsky & Stangl. The five samples are

very closely related (P%IV = 98.4) and have characters consistent with those of the

original collections. Bon (1991) circumscribed the species in sect. Alboflavidae

mainly based on Qm value, odour, pileus size, and stipe ornamentation. We

378 ... Vizzini & al.

have now examined morphologically the holotype of M. strictipes (H 2432,

Fontenla & Para, unpubl.), lectotype of M. subalpina (Fontenla & Para 2008),

and eight original Tricholoma cnista sensu Bresadola (= M. evenosa) collections

(TR, BPI). This, combined with our thorough study of the original diagnoses

and phylogenetic analysis, lead us to conclude that M. strictipes, M. evenosa,

and M. subalpina are conspecific, with the name M. strictipes having priority.

CLADE Bq (0.52 BPP, / MLB), cystidia fusiform or lageniform, pileus small (up

to 5 cm broad) and not dark-coloured, lamellae whitish-cream, context

white.

CLADE B4 comprises M. oreina (Fr.) Kihner & Maire, M. bataillei Malencon,

M. sp. 7, and M. sp. 9. The first two species, which are phenetically very similar,

differ mainly on cystidial shape. Bon (1991) placed M. oreina (with fusiform

cystidia) into sect. Oreinae and M. bataillei (with mainly lageniform cystidia)

into sect. Melanoleuca. Molecularly, they are quite distinct (P%IV = 92.5).

CLADE Bs5 (0.84 BPP, 54% MLB) consists of the three M. cinereifolia (Bon) Bon

sequences (P%IV = 100).

Melanoleuca cinereifolia is distinguished by a habit reminiscent of Clitocybe

nebularis (Batsch) P. Kumm., always grey lamellae, short stipe, strictly

lageniform (M2) cystidia, and growth in sand dunes.

Acknowledgements

We are grateful to G. Robich (Venice) for having sent us the Melanoleuca samples

from MCVE DNA bar-coded by Dr. M. Garbelotto (University of California, Berkeley).

Dr. Matteo Garbelotto and Dr. Todd Osmundson (University of California, Berkeley)

kindly provided us with pre-submission Melanoleuca sequences. We also thank Dr.

Vladimir Antonin (Brno, Czech Republic) and Dr. Marco Contu (Olbia, Italy) for their

pre-submission reviews and Dr. Shaun Pennycook (Auckland, New Zealand) for the

nomenclatural review.

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MYCOTAXON

http://dx.doi.org/10.5248/118.383

Volume 118, pp. 383-392 October-December 2011

Geastrum species from the Amazon Forest, Brazil

ANILEIDE GOMES LEITE*', HANNAH KATHREN DE ASSIS’,

BIANCA DENISE BARBOSA DA SILVA?, HELEN MARIA PONTES SOTAO?

& IURI GOULART BASEIA’

’ Universidade Federal do Rio Grande do Norte, Depto. Botanica, Ecologia e Zoologia,

Campus Universitario, CEP: 59072-970, Natal, RN, Brazil

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

Depto. Micologia, Centro de Ciéncias Bioldgicas,

Av. Nelson Chaves s/n, 50670-420, Recife, PE, Brazil

° Ministério da Ciéncia e Tecnologia, Museu Paraense Emilio Goeldi, Departamento de Botanica.

Caixa Postal 399, AV: Perimetral 1901 MARCO, 66040-170 - Belém, PA, Brazil

CORRESPONDENCE TO *: anileidecb@yahoo.com.br

ABSTRACT—Six Geastrum species are reported from the Amazon forest: G. entomophilum,

G. fimbriatum, G. javanicum, G. lageniforme, G. lilloi, and G. saccatum. Geastrum javanicum

and G. lilloi represent first records from Brazil. Descriptions and illustrations of the species

and SEM images of microstructures are given.

Key worps— Basidiomycota, Geastraceae, gasteromycetes, taxonomy

Introduction

The family Geastraceae was first described by Corda (1842) as the Geastrideae

and subsequently classified in the Lycoperdales (Lloyd 1902, Ponce de Leon

1968, Kriiger & Chagas 2008), the Phallales (Hibbett et al. 1997) and, based on

molecular data, recently in the Geastrales (Hosaka et al. 2006). With 43 species

reported, the Geastraceae is the second-most represented gasteromycete

family in Brazil (Trierveiler-Pereira & Baseia 2009). The Geastraceae comprise

eight genera: Geastrum, Myriostoma, Trichaster, Geasteropsis, Phialastrum,

Pyrenogaster, Radiigera, and Terrostella, with Geastrum being the largest

(Sunhede 1989) with 50 species recognized worldwide (Kirk et al. 2008).

Tropical regions contain most of the fungi still unknown to science. Tropical

Amazonian rainforests account for 25% of the remaining forests worldwide

and cover nearly half of Brazil, representing enormous strategic value (Braga-

Neto et al. 2008). The Amazon biome, which covers 4,871,000 km? (INPE

384 ... Leite & al.

2004), extends from the Atlantic Ocean to the eastern slopes of the Andes up

to approximately 600 m altitude, encompassing parts of nine South American

countries and with 69% falling within the boundaries of Brazil (AbSaber

1977).

The mycobiota of the Brazilian Amazon has been poorly studied and needs

to be more thoroughly investigated. However, the following existing studies are

noteworthy: Singer & Araujo (1979), Aguiar (1984), Singer (1984), Singer &

Aguiar (1986), Capelari & Maziero (1988), and Bononi (1992). Several studies

on gasteromycetes in the Amazon region have been conducted: Berkeley

& Cooke (1876), Hennings (1904), Capelari & Maziero (1988), Trierveiler-

Pereira et al. (2009). The purpose of this paper is to contribute to the existing

knowledge of this mycobiota, with special reference to the family Geastraceae.

Material & methods

Material was collected from 1995 to 2008 at the Reserva Nacional de Caxiuana, in the

municipality of Melgaco, Para state, Amazonas, Brazil, and deposited in the herbaria of

the Museu Paraense Emilio Goeldi (MG), Belém, Para, and UFRN, Universidade Federal

do Rio Grande do Norte, Natal. These collections are considered reference depositories

for the study of Amazonian biodiversity (Braga-Neto et al. 2008). Additional Brazilian

specimens held in UFRN were also studied. Macro- and microscopic characters were

studied according to Miller & Miller (1988), Sunhede (1989), Ponce de Ledn (1968),

Soto & Wright (2000), Baseia & Milanez (2002), Calonge et al. (2000), Calonge & Mata

(2004), Baseia & Calonge (2006), Leite & Baseia (2007), and Leite et al. (2007). Color

citations were based on Kornerup & Wanscher (1978). Microstructures were analyzed

in sections mounted on slides, using 5% KOH, and SEM was used to study basidiospore

ornamentation.

Results

Geastrum entomophilum Fazolino, Calonge & Baseia, Mycotaxon 104: 450 (2008).

FiG=.t

Expanded basidiome epigeous, 7.3 cm broad. Exoperidium semi-hygroscopic,

arched, split into 6 rays, recurved under the exoperidial disc; mycelial layer

light orange (6A5), covered with adhering humus, leaf and sand; fibrous layer

light orange (6A4); pseudoparenchymatous layer brown (6F4). Endoperidium

sessile, globose to subglobose, 2 cm diam., ornamentation formed by fascicles

of worm-shaped surface hyphae (Fic. 1b), greyish brown (5D3). Peristome

fibrillose, concolorous with rest of endoperidium, indistinctly delimited. Gleba

brownish grey (5F2). Basidiospores globose, ornamentation more or less

columnar, 3.5-4 um diam. Capillitial hyphae with verrucose surface, 3-3.1 um

diam.

SUBSTRATE & DISTRIBUTION— Sandy soil. Known only from South America,

this is the first record of this species from the Amazon rainforest.

Geastrum in the Amazon (Brazil) ... 385

Fic. 1. Geastrum entomophilum. a. Basidiospores; b. Endoperidial hyphal fascicle.

SPECIMEN EXAMINED: BRAZIL. PARA: Melgaco, Floresta Nacional de Caxiuana, Estacao

Cientifica Ferreira Penna, 20.11.1997, col. H. Sotao & al. (MG 199685).

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO NorTE: Natal,

Parque EstapuaL Dunas do Natal, 15.VII.2006, col. E.P. Fazolino & A.G. Leite. (UFRN-

Fungos 354).

TAXONOMIC REMARKS—Geastrum entomophilum is recognized by its sessile

endoperidium, verrucose surface with vermiform hyphae in fascicles, and

fibrillose and lacerated peristome, non-delimited when older. In the original

description (Fazolino et al. 2008) the small beetles (Coleoptera) observed

inside the gleba led to the name ‘entomophilum: Beetles were not observed by

Trierveiler-Pereira & Baseia (2010).

Geastrum fimbriatum Fr., Syst. mycol. 3(1): 16 (1829). Fic. 2

Basidiomata epigeous when young, globose to depressed-globose, 1.7 cm

diam. x 2 cm high, epigeous at maturity, 2-2.5 cm broad, 0.4-0.7 cm high.

Exoperidium non-hygroscopic, saccate, splitting into 4-6 rays; mycelial

layer dark blond (5D4); fibrous layer adherent, greyish yellow (4B3);

pseudoparenchymatous layer brown (5F5). Endoperidium sessile, globose to

subglobose, 0.8-2 cm diam., brown (6E4). Peristome absent to fibrillose. Gleba

dark brown (6F5); columella present. Basidiospores 3-3.5 um diam., globose,

ornamentation columnar. Capillitial hyphae thick-walled, with surface debris

and verrucose, 3.5-5 um diam.

SUBSTRATE & DISTRIBUTION— Sandy soil. Known from South America,

Central America, North America, Europe, and Australasia.

SPECIMEN EXAMINED: BRAZIL. PARA: Melgaco, Floresta Nacional de Caxiuana, Estacao

Cientifica Ferreira Penna, 30.V.1997, col. H. Sotao & al. (MG-199687, UFRN-fungos

1465).

aranee SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO NORTE: Parque

Estadual Dunas do Natal, 02.V1.2007, col. E.P. Fazolino & B.D.B. Silva (UFRN-Fungos

337).

386 ... Leite & al.

Fic. 2. Geastrum fimbriatum. a. Basidiospore; b. Capillitial hypha.

TAXONOMIC REMARKS—Geastrum fimbriatum strongly resembles G. saccatum,

mainly with respect to its basidiospore ornamentation, but it also differs in

having smaller spores, with those of G. saccatum 4-6 um diam. (Soto & Wright

2000). A marked characteristic in this species observed by both Sunhede (1989)

and us is the interspersed hyphal formation in the mycelial layer.

This is the second record of C. fimbriatum from the Amazon rainforest; the

first was made by Trierveiler-Pereira et al. (2009).

Geastrum javanicum Lév., Annls Sci. Nat., Bot., sér. 3, 5: 161 (1846). Fie. 3-4

Basidiomata epigeous when young, 0.7—1.6 cm broad, 0.5—1.5 cm high,

caespitose on a subiculum, white yellowish (4A1-4A2); expanded basidiome

epigeous. Exoperidium hygroscopic, 1.5-1.9 cm diam. when open, split into

5-8 recurving rays; mycelial layer velvety, separates at maturity, greyish brown

(5D3); fibrous layer light yellowish (5C4); pseudoparenchymatous layer

brownish orange (5C4). Endoperidium brown (6F4) to brownish grey (7E2),

sessile, globose, 1-1.2 cm diam. Peristome delimited, fibrillose, concolorous

with rest of endoperidium. Gleba greyish brown (6F3); columella present.

Fic. 3. Geastrum javanicum. a. Basidiospore; b. Capillitial hypha.

Geastrum in the Amazon (Brazil) ... 387

Fic. 4. Geastrum javanicum. a. Immature basidioma; b. Mature basidioma.

Basidiospores globose, 2.5-3 um diam., with irregular warts. Capillitial hyphae,

3-4 um diam., surface rugose, septate and branched.

SUBSTRATE & DISTRIBUTION— Decaying wood. Known from South America,

Central America, North America, Africa, Australasia, Asia.

SPECIMEN EXAMINED: BRAZIL. PARA: Melgaco, Floresta Nacional de Caxiuana, Estacao

Cientifica Ferreira Penna, 12.IV.1995, col. H. Sotao & al. (MG n° 0149528).

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. RIO GRANDE DO NORTE: Parnamirim,

Mata do Jiqui, 22.V1.2007, col. E.P. Fazolino & al. (UFRN-Fungos 550).

TAXONOMIC REMARKS—Geastrum javanicum is generally found growing in

clusters, forming whitish subiculum on dead wood, leaf litter, and occasionally

in sand impregnated with decomposing organic matter (Ponce de Leon 1968).

A diagnostic character is the velvety exoperidium that detaches itself from the

mycelial layer when the basidioma is mature. Ponce de Leén (1968) reviewed

the holotypes of G. velutinum Morgan and G. javanicum and considered them

to be synonyms. However, on the basis of priority, G. javanicum is the correct

name to use. Sunhede (1989) did not study the type of material of G. javanicum,

and therefore did not confirm its synonymy with G. velutinum.

This species has been reported from Brazil by Sobestiansky (2005), de Meijer

(2006), and Sydow & Sydow (1907) as G. velutinum, but this is the first record

from the Amazon rainforest.

Geastrum lageniforme Vittad., Monograph. Lyc.: 16 (1842). Fic. 5

Mature basidiomata 1-2.4 cm broad, 0.8-1.4 cm high. Exoperidium non-

hygroscopic, saccate with 4-6, long, slender-tipped rays, extended or recurved

388 ... Leite & al.

Fic. 5. Geastrum lageniforme. a. Basidiospore; b. Capillitial hypha.

under the exoperidial disc and the rays are markedly tapered and pointed;

mycelial layer blond (4C4), with longitudinal fissures; fibrous layer pale orange

(5A3); pseudoparenchymatous layer yellowish brown (5F8). Endoperidium

dark blond (5D4), sessile, globose to subglobose, 0.8-1.2 cm diam. Peristome

fibrillose, delimited, conical to mammiform. Gleba pale orange (5A3); columella

present. Basidiospores globose 4.5-5 um diam., ornamentation columnar.

Capillitial hyphae thick-walled, 2-2.8 um diam.

SUBSTRATE & DISTRIBUTION—Sandy soil. Known from South America,

Central America, North America, and Europe; this is the first record of this

species from the Amazon rainforest.

SPECIMEN EXAMINED: BRAZIL. PARA: Melgaco, Floresta Nacional de Caxiuana, Estacao

Cientifica Ferreira Penna, 19.11.1997, col. H. Sotao & al. (MG 199683).

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO NortTE: Natal,

Parque Estadual Dunas do Natal, 29.VII.2006, col. E.P. Fazolino & al. (UFRN-fungos

332).

TAXONOMIC REMARKS— This species is characterized by a saccate exoperidium,

sessile endoperidium and fibrillose, well-delimited peristome. It is very similar

to and often confused with G. saccatum and G. triplex Jungh. (Soto & Wright

2000). Fries (1829) mentions nothing about grooves in the mycelial layer in

G. saccatum. In G. lageniforme, after dehiscence of the exoperidium, the rays

are markedly longitudinally cracked. According to Sunhede (1989), G. triplex

exhibits an endoperidial body of 7.5-19.5 mm diam., while in G. lageniforme it

reaches 11-54 mm.

Geastrum lilloi L.S. Dominguez, Mycologia 88: 858 (1996). Fics. 6-7

Young basidiomata epigeous, 0.8 cm broad, 0.4 cm high, surface irregular,

yellowish white pale (2A2), caespitose, growing on a pale yellow (4A3)

subiculum; mature basidiomata 1.1-1.4 cm diam. Open exoperidium saccate,

splitting into 5—6 rays, hygroscopic; mycelial layer with irregular surface, pale

yellowish white (2A2); fibrous layer white (6D4); pseudoparenchymatous layer

Geastrum in the Amazon (Brazil) ... 389

Fic. 6. Geastrum lilloi. a. Basidiospores; b. Mycosclereid.

brown (5F7). Endoperidium brown (6E4), sessile, globose, 0.6-0.8 cm diam.

Peristome fibrillose, mammiform, greyish brown (5D3), indistinctly delimited.

Gleba dark brown (6F3); mycosclereids present, up to 10.2 x 15.9 um diam.;

columella present. Basidiospores globose, 2.8-3.5 um diam., ornamentation

irregularly columnar. Capillitial hyphae 1.8-2 um. diam., covered with

encrusting surface debris.

SUBSTRATE & DISTRIBUTION— Woody debris. Known from South America

(Argentina, Brazil).

SPECIMEN EXAMINED: BRAZIL. Para: Melgaco, Floresta Nacional de Caxiuana, Estacao

Cientifica Ferreira Penna, 29.V.1997, col. H. Sotao & al. (MG 199686).

TAXONOMIC REMARKS—Distinguishing characteristics of G. lilloi are its small

size, gregarious and xylophilous habitat, velvety mycelial layer, the presence of

mycosclereids in the gleba, the endoperidium with a fibrillose, mammiform

pore, and epigeal development (Dominguez de Toledo 1996). Geastrum

schweinitzii (Berk. & M. A. Curtis) Zeller can easily be confused with G. Jilloi

because it also is small-sized and grows on a subiculum on wood. However,

10 mm

Fic. 7. Geastrum lilloi. a. Immature basidioma; b. Mature basidioma.

390 ... Leite & al.

these two species differ in terms of the non-hygroscopic exoperidium, obovate

fruit body, peristome without a ring, and smooth to velvety mycelial layer in

G. schweinitzii.

This is the first record of G. Jilloi, first described from Argentina, from

Brazil.

Geastrum saccatum Fr., Syst. Mycol. 3(1): 16 (1829). Fic. 8

Mature basidiomata 0.5-2 cm diam. Exoperidium non-hygroscopic,

saccate, splitting into 5-7 rays; mycelial layer encrusted with debris of soil

and leaf litter, light orange (6A4); pseudoparenchymatous layer brown (6E4);

fibrous layer adhered to pseudoparenchymatous layer, pale orange (6A3).

Endoperidium brownish beige (6E3), sessile, globose, 0.8-1.2 cm diam.

Peristome well delimited, fibrillose, conical to mammiform, concolorous

with rest of endoperidium. Gleba greyish brown (6F3); columella present.

Basidiospores globose to subglobose, 3.6—4.5 um diam., ornamentation rather

irregular columnar. Capillitial hyphae 3-3.5 um diam, covered with small wart-

like surface outgrowths.

SUBSTRATE & DISTRIBUTION— Sandy soil. Known from South America,

Central America, North America, Africa, Asia, Australasia, and Europe.

Fic. 8. Geastrum saccatum. a. Basidiospore; b. Capillitial hypha.

SPECIMEN EXAMINED: BRAZIL. PARA: Melgaco, Floresta Nacional de Caxiuana, Estacao

Cientifica Ferreira Penna, 19.11.1997, col. H. Sotaéo & al. (MG-199684).

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO Norte: Natal,

Parque Estadual Dunas do Natal, 15.VII.2006, col. E.P. Fazolino & al. (UFRN-Fungos

334).

TAXONOMIC REMARKS—Geastrum saccatum is characterized mainly by its well-

delimited, fibrillose peristome and sac-like, sessile endoperidium. It resembles

G. lageniforme but differs in its smaller and wider rays and a mycelial layer lacking

longitudinal striations (Soto & Wright 2000). Macroscopically it resembles

young G. triplex specimens, but at maturity the pseudoparenchymatous layer of

Geastrum in the Amazon (Brazil) ... 391

G. triplex detaches from the fibrous layer. Another marked difference between

these two species is the much smaller basidioma of G. saccatum.

This is the second record of G. saccatum, first recorded by Hennings (1904),

from the Amazon rainforest.

Acknowledgments

The authors thank the reviewers, Johan C. Coetzee and Admir Giachini, for

their valuable suggestions. We express our gratitude to the Conselho Nacional de

Desenvolvimento Cientifico e Tecnoldgico (CNPq) for partial financial support; to

CTPETRO-INFR, FINEP/LIEM for collaboration with the SEM and MPEG and to

Estacao Cientifica Ferreira Penna for fieldwork logistical support.

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MY COTAXON

http://dx.doi.org/10.5248/118.393

Volume 118, pp. 393-401 October-December 2011

Hypoderma siculum sp. nov. from Italy

ANGELA LANTIERI’ , PETER R. JOHNSTON”, DUCKCHUL PARK’,

HENRIK LANTZ? & GIANFRANCO MEDARDI*

‘Dipartimento di Biologia “Marcello La Greca”, Universita di Catania,

Via Antonino Longo 19, I-95125 Catania, Italy

*Landcare Research, Private Bag 92170, Auckland 1142, New Zealand

*Swedish University of Agricultural Sciences, Department of Microbiology,

Box 7025, SE-75007, Uppsala, Sweden

°Via Giuseppe Mazzini 21, I-25086 Rezzato (Brescia), Italy

CORRESPONDENCE TO *: angelalantieri@gmail.com

ABSTRACT — Hypoderma siculum is described and illustrated as a new species from

southeast Sicily (Italy) that occurs on remnants of Ferula communis (Apiaceae). Its ecology

and taxonomic and phylogenetic relationships are discussed.

KEY worps — morphology, taxonomy, ITS phylogeny

Introduction

Field investigation of the Ascomycetes of Sicily occurring on decaying

remnants of Ferula communis (Lantieri 2009) revealed a new Hypoderma

species, described here as Hypoderma siculum.

Materials & methods

Collections of the new species were made between 2007 and 2010 at an elevation

of 700 m a.s.l. in southeast Sicily. Morphological and microscopic examinations were

carried out on fresh material and on dried specimens rehydrated in water. Observations

and measurements were made in water and Melzer’s reagent. Ascus and ascospore size

ranges from the holotype were based on 50 measurements, using an Optika optical

microscope (model BK 1301), with 40x or 100x (oil immersion) objectives. All voucher

specimens were deposited in the fungarium of the Royal Botanic Gardens, Kew K(M)

and in the fungal reference collection of Landcare Research in New Zealand (PDD).

DNA was extracted separately from two sets of four fruiting bodies taken from

each of two separate blackened areas on leaf pieces within the isotype specimen (PDD

99894), using REDExtract-N-Amp Plant PCR Kits (Sigma, USA). The perithecia were

394 ... Lantieri & al.

TABLE 1. Specimens used in phylogenetic analysis

COUNTRY OF ORIGIN, COLLECTION GENBANK

SPECIES

HOST VOUCHER ACC. NO.

CeO ES ATEN. ICMP 16772 EF191241

australis Desfontainia spinosa

Hypoderma Sweden, Hanson 2006-451 (UPS) JF690769 *

commune ?Euphorbia sp.

H. cordylines New Zealand, ICMP 17344 JF683421 *

Cordyline australis (ex-holotype culture)

New Zealand, Phormium sp. ICMP 17359 JF683420 *

H. hederae Scotland, Hedera helix Lantz & Minter 421 (UPS) JF690770 *

H. rubi China, host unknown unknown GU138735

China, host unknown unknown GU138736

China, host unknown unknown GU138738

China, host unknown unknown GU138739

China, host unknown unknown GU138741

China, host unknown unknown GU138743

China, host unknown unknown GU138745

China, host unknown unknown GU138746

China, host unknown unknown GU138750

China, host unknown unknown GU138751

China, host unknown unknown GU367895

China, host unknown unknown GU367898

New Zealand, Coprosma sp. ICMP 18768 JF683417 *

New Zealand, ICMP 18325 JE 683418 *

Melicytus ramiflorus

New Zealand, Pseudopanax sp. ICMP 17349 JF683416 *

New Zealand, Rubus cissoides ICMP 17339 JE 683419 *

Sweden, Rubus lindebergii Hanson 2002-230 (UPS) JF690771 *

H. siculum Italy, Ferula communis PDD 99894 (isotype) JF683424 *

H. stephanandrae China, Stephanandra chinensis unknown GU138753

H. vincetoxici Sweden, Lantz 405 (UPS) JF690772 *

Vincetoxicum hirundinaria

Lophodermium New Zealand, ICMP 18327 JF683423 *

agathidis Agathis australis

New Zealand, ICMP 17345 JF683422 *

Metrosideros fulgens

L. eucalypti New Zealand, ICMP 16796 EF191235

Leptospermum scoparium

L. gamundiae Argentina, ICMP 16797 EF191239

Nothofagus dombeyi

* = newly generated for this study).

UPS = Uppsala University; ICMP = International Collection of Microorganisms

from Plants (maintained by Landcare Research)

ground in extraction buffer with a plastic pestle in the Eppendorf tube. Following this,

DNA extraction and PCR were carried out following manufacturer's instructions. ITS

sequences were obtained from each extract following the methods of Johnston & Park

(2005).

Hypoderma siculum sp. nov. (Italy) ... 395

Using ClustalW (Thompson et al. 1994), our newly generated sequences were

aligned with sequences deposited in Genbank as Hypoderma rubi or those that closely

matched H. rubi. Because of the paucity of available data, ITS sequences were generated

from several additional Hypoderma spp. from New Zealand, with DNA extracted from

cultures grown from germinating ascospores. Other taxa included in the analysis were

members Lophodermium eucalypti/Coccomyces tumidus clade recognized by Lantz et al.

(2011) that formed a sister relationship to the core Hypoderma clade, and Lophodermium

agathidis as the outgroup.

The taxa included are listed in TABLE 1, along with the Genbank accession numbers

of our newly generated sequences. A Bayesian phylogenetic analysis was performed

using MrBayes 3.1.2 (Huelsenbeck & Ronquist 2001; Ronquist & Huelsenbeck 2003)

with gaps treated as missing data, using the K80+I+G model which was selected using

the AIC method in MrModelTest 2.3 (Nylander 2004). The data set was run with 2 chains

for 10 million generations, trees sampled every 1000 generations with a burn-in of 25%.

Bayesian posterior probabilities were obtained from 50% majority rule consensus trees.

Taxonomy

Hypoderma siculum Lantieri, P.R. Johnst. & Medardi sp. nov. PLATEs 1-2

MycoBANnkK MB 563145

Ascomata erumpentia inter amplas variiformes nigras areas, haud compositas, supra plantae

superficiem, haud consociata cum zonae lineis. Partes nigrae ex 1-2 stratis hypharum

parietibus obscuris et inflatis, formantibus amplum stratum, 5-8 um latum sub plantae

cuticula, constitutae. Aspectus in superficie: ascomata 1-1.5 x 0.5-0.8 mm, ellipsoidea

vel leviter fusiformia vel navicularia si secta, extremitatibus repente rotundatis vel acutis,

paene mersa in contextu plantae. Ascomata clausa parietibus nigrescentibus, sed plus

minusve obscure griseis ad margines prominentes fissurae. Ascomata aperta per fissuram

longitudinalem, se extendentem in totam longitudinem. Labra dilute griseola adsunt.

Hymenium griseum caesio colore suffusum. Labri cellulae hyalinae ex hyphis inconstanter

cylindricis, inflatis in nonnullis partibus, pariete crassa, cellulis 1 (raro 2) -septatis, 20-25

x 2 um, mersis in involucro gelatino, constitutae. In recta media parte stroma tegens

crassum usque ad 50 um prope ascomatum centrum, sed tenuior ad margines atque tenens

stroma basis ex strato inferiore et obscure brunneo texturae epidermoideae constitutum.

Externa pars parietum cellularum stromatis tegentis obtecta materia densa, nigra, plus

minusve granosa, ex particulis sine structura cellularum manifesta, sed nonnullis cellulis

obscure polygonalibus, efformata. Excipulum abest. Subhymenium 20-40 um crassum,

ex textura intricata, hyphis 3-4 ym latis, parietibus tenuibus et hyalinis constitutum.

Stroma basis 5-15 um crassum, cum 3-4 stratis hypharum hyalinarum, 3-4 um

latarum, parietibus obscure brunneis et leviter inflatis, formantibus texturam intricatam.

Ascosporae fusiformes vel cylindrico-fusiformes, nonnullae leviter curvae, 27-30 x 3-3.5

um, laeves, hyalinae, haud septatae, guttulis haud compositis praeditae, saepe absentibus

prope centrum, cinctae involucro gelatino, laxo, circiter 3 um crasso. Asci clavato-stipitati,

(85—)90-100 x 9-10 um, apice rotundato, haud amyloidei, 8—sporigeri; sporae dispositae

in parte superiore. Paraphyses numerosae, filiformes, 1-2 um latae, emergentes usque ad

30 um supra ascos, curvae, crispatae vel valde plicatae ad apicem, nonnullis parvis guttulis

sparsis per totam longitudinem, sine septis. Specimina cum conidiis haud notata. Habitat:

supra reliqua putrescentia Ferulae communis; inventum solum hieme.

396 ... Lantieri & al.

TyPE: Italy. Sicily, vic. Vittoria (Ragusa), Riserva Naturale Orientata “Pino d’Aleppo’,

on decaying remnants of Ferula communis, 30/12/2007, Leg. Angela Lantieri (holotype,

K(M) 167515; isotype, PDD 99894, GenBank JF683424).

CONIDIOMATA not observed. Ascomata develop among large, irregularly

shaped blackened areas on the plant surface, not associated with zone lines. The

blackened areas have 1-2 layers of hyphae with walls dark and thick forming a

5-8 um wide layer beneath the plant cuticle. In surface view ascomata 1-1.5 x

0.5-0.8 mm, elliptical to slightly fusiform or navicular in outline, with sharply

rounded to acute ends, semi-immersed in the plant tissues. Closed ascomata

with blackish walls, but more or less dark greyish near the prominent edges of

the slit. Ascomata opening by a longitudinal slit, which extends along the whole

length. Lips present, pale greyish. HYMENIUM grey with bluish reflexes. Lip

CELLS hyaline, composed of irregularly cylindrical, in some points enlarged,

thick-walled, 1 (rarely 2) -septate, cells, 20-25 x 2 um embedded in a gelatinous

sheath. IN MEDIAN VERTICAL, covering stroma up to 50 um thick near the

centre of the ascomata, becoming thinner towards the edges, extending to the

basal stroma, consisting of a inner layer of dark brown textura epidermoidea.

OUTER PART of the walls of the cells in the covering stroma with dense, more

or less granulose black matter, composed of small particles with no visible

cellular structure, but some of them obscurely polygonal. ExcipuLuM absent.

SUBHYMENIUM 20-40 um thick, composed of textura intricata, hyphae 1.5-2

um diam. with hyaline, thin walls. BasAL sTroMA 5-15 um thick, comprises

3-4 layers of hyphae 3-4 um diam. with walls dark brown and slightly

thickened, forming a textura intricata. Ascospores fusiform or cylindrical-

fusiform, some slightly curved, 27-30 x 3-3.5 um, smooth, hyaline, aseptate,

containing irregular oil drops often lacking near the middle, surrounded by

loose gelatinous sheath about 3 um thick. Asci clavate-stipitate, (85—)90-100 x

9-10 um, apex rounded, not bluing in iodine, 8-spored, spores confined to the

upper part. PARAPHYSEs abundant, filiform, 1-2 um diam., protruding up to 30

um beyond the asci, curved, curled or remarkably bent at the apex, with some

small oil-drops scattered along the length, without septa.

Hasitat: On decaying remnants of Ferula communis, found only in winter.

ADDITIONAL SPECIMENS EXAMINED: ITALY. SIcILy, vic. Vittoria (Ragusa), Riserva

Naturale Orientata “Pino d’Aleppo’, 13/03/2008, [Leg. A. Lantieri] (K(M) 167516);

06/02/2009, [Leg. A. Lantieri] (K(M) 167517); 13/02/2010, [Leg. A. Lantieri] (K(M)

167518).

Discussion

Hypoderma siculum falls within the Hypoderma sensu stricto clade of Lantz

et al. (2011). Genetic relations within this clade are poorly resolved in the ITS

gene tree (Fic. 3). Despite this, H. siculum is genetically distinct from other

named isolates within the clade, and this genetic isolation and its distinct

Hypoderma siculum sp. nov. (Italy) ... 397

PLATE 1. Hypoderma siculum (Holotype, K(M) 167515). A. fresh ascomata in situ; B. stromatic

tissue surrounding ascomata, vertical section; C. upper wall of ascoma, vertical section; D. detail

of lip cells, vertical section; E. detail of lower wall of ascoma, vertical section; F. detail of lower

wall of ascoma, squash mount; G. asci and ascospores; H. ascospores in KOH; I. ascospores in

water showing gelatinous sheath. Scale bars: A = 1 mm; B, D-H = 20 um; C = 50 um.

398 ... Lantieri & al.

PLATE 2. Hypoderma siculum (Holotype, K(M) 167515).

A. margin of ascoma in vertical section; B. stromatal cells; C. section of stroma;

D. granulose matter; E. asci and ascospores; F. ascospores.

biology supports recognizing the fungus as an independent species. Other

morphologically distinct taxa within this clade, such as Hypoderma cordylines,

are also poorly resolved. More intensive, multi-gene phylogenies of Australasian

species have shown that ITS alone does not adequately resolve relationships

within this clade (unpublished data). The weak quality of the DNA extracted

from the H. siculum dried specimens did not permit reliable sequencing of

single copy genes.

Hypoderma siculum sp. nov. (Italy) ... 399

GU367898 H. rubi China

GU367895 H. rubi China

GU138738 H. rubi China

GU138736 H. rubi China

GU138739 H. rubi China

GU138745 H. rubi China

GU138746 H. rubi China

GU138743 H. rubi China

GU138741 H. rubi China

JF683424 Hypoderma siculum

GU138753 H. stephanandrae

JF690771 H. rubi Sweden on Rubus

GU138750 H. rubi China

GU138751 H. rubi China

JF683416 H. rubi NZ on Pseudopanax

JF683417 H. rubi NZ on Coprosma

JF683418 H. rubi NZ on Melicytus

JF690769 H. commune Sweden

JF690772 H. vincetoxici Sweden

JF690770 H. hederae Sweden

JF683420 H. cordylines NZ

JF683421 H. cordylines NZ

1.00 JF683419 H. rubi NZ on Rubus

EF 191239 Lophodermium gamundiae

EF191235 Lophodermium eucalypti

EF191241 Coccomyces australis

1.00 JF683422 Lophodermium agathidis

JF683423 Lophodermium agathidis

0.99

FIGURE 3. 50% majority-rule consensus phylogenetic tree based on Bayesian analysis of Hypoderma

siculum and related taxa derived from ITS gene sequences. Bayesian posterior probabilities greater

than 90% are shown above the edges. Sclerotinia sclerotiorum was selected as the outgroup.

Ferula communis L. (Apiaceae), a typical herbaceous plant of the

Mediterranean area, is commonly associated with two Hypoderma species,

H. ferulae Lantieri (Lantieri 2009) and H. siculum. Early reports of Hypoderma

rubi (Pers.) DC. and H. commune (Fr.) Duby on F. communis (Duby 1862;

Petrak 1943) are difficult to assess, as neither report is supported by specimens.

It is uncertain in what sense the authors were using these names and impossible

to check whether the specimens they reported match our Ferula-specialised

species.

AOO ... Lantieri & al.

Extensive collecting in Sicily revealed only H. ferulae and H. siculum on

Ferula. Scalia (1900: 37) reported H. commune from Sicily ona species in another

genus of Apiaceae, Thapsia garganica L.; the report included no description of

the fungus, and again there is no material available to verify the record. There

is no evidence from collections made in Sicily that H. siculum occurs on other

genera of Apiaceae (unpubl. data).

Hypoderma siculum is distinguished from H. ferulae by ascospore size and

septation (21-24 x 2.5-3 um and often 1-septate in H. ferulae), by the presence

of an extended area of stromatic subcuticular tissue surrounding the ascomata,

the colour of the hymenium when fresh (yellow-honey with greenish tinges in

H. ferulae), and the lack of an excipular layer.

Hypoderma siculum resembles H. rubi morphologically, but as noted in

the phylogenetic section, the application of this name by different authors is

uncertain, and genetic relationships amongst specimens identified as H. rubi

are poorly resolved. Hypoderma rubi sensu Powell (1974), Johnston (1990), and

Medardi(2006) differs slightly from H. siculum in its ascospore size (20-—25(-26.5)

x 3-4 um) and the fusiform-navicular ascospore shape. Hypoderma commune

differs in having ascomata with yellow-greenish hymenium and smaller, non-

septate spores (17-20 x 3-4 um; Ellis & Ellis 1988).

Acknowledgments

The authors sincerely thank to Prof. G. Consiglio for the Latin diagnosis, Prof. D. Minter

(UK), and Dott. C. Losi (Italy) for critically reviewing the manuscript and G. Cacialli for

significant contributions to the bibliography.

Literature cited

Ellis MB, Ellis JP. 1988. Microfungi on miscellaneous substrates. Croom Helm, London & Sydney.

Huelsenbeck JP, Ronquist F. 2001. MRBAYES: Bayesian inference of phylogeny. Bioinformatics 17:

754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

Johnston PR. 1990. Rhytismataceae in New Zealand 3. The genus Hypoderma. New Zealand J. Bot.

28: 159-283.

Johnston PR, Park D. 2005. Chlorociboria (Fungi, Helotiales) in New Zealand. New Zealand Journal

of Botany 43: 679-719. http://dx.doi.org/10.1080/0028825X.2005.9512985

Lantieri A. 2009. A new species of Hypoderma (Ascomycota) from Italy. Sydowia 61(2): 267-272.

Lantz H, Johnston PR, Park D, Minter DW. 2011. Molecular phylogeny reveals a core clade of

Rhytismatales. Mycologia 103: 57-74. http://dx.doi.org/10.3852/10-060

Medardi G. 2006. Atlante fotografico degli Ascomiceti d'Italia. A.M.B. Centro Studi Micologici,

Vicenza (Italy).

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology

Centre, Uppsala University.

Petrak FE 1943. Fungi. Denkschriften der Akademie der Wissenschaften Mathematische-

Naturwissenschaftliche Klasse 105(2): 9-26.

Powell PE. 1974. Taxonomic studies in the genus Hypoderma (Rhytismataceae). Unpublished PhD

thesis, Cornell University.

Hypoderma siculum sp. nov. (Italy) ... 401

Ronquist F, Huelsenbeck JP. 2003. MRBAYES 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. http://dx.doi.org/10.1093/bioinformatics/btg180

Scalia G. 1900. I funghi della Sicilia orientale e principalmente della regione Etnea (Prima serie).

Atti dell’ Accademia Gioenia di Scienze Naturali di Catania, 4. ser., 13: 1-55.

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http://dx.doi.org/10.1093/nar/22.22.4673

MY COTAXON

http://dx.doi.org/10.5248/118.403

Volume 118, pp. 403-410 October-December 2011

Tuber sinoalbidum and T. polyspermum — new species from China

Li FAN’ CHENG-LIN Hou! & JIN-ZHONG Cao?

' College of Life Science, Capital Normal University,

Xisanhuanbeilu 105, Haidian, Beijing 100048, China

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

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

ABSTRACT — Two new Tuber species from China are described and illustrated. Tuber

sinoalbidum is recognized by its white mid to large sized ascocarps and spinulose-reticulate

ascospores, while T: polyspermum is characterized by Tuber lyonii-like ascospores and smaller

brown ascomata. A molecular study based on the ITS sequences supports the erection of the

two new species.

Key worps — Ascomycota, Tuberaceae, truffle

Introduction

Tuberis a genus of ascomycetous fungi that form ectomycorrhizae with shrubs

and trees. Species of the genus have been extensively studied because many

form edible hypogeous ascocarps — the truffles. Some of these are renowned

for their flavor and are commercially important, with the two European species,

T; magnatum Picco and T: melanosporum Vittad., commanding the highest

prices. Other species harvested include T! aestivum Vittad., T: borchii Vittad.,

T: gibbosum Harkn., and T. indicum Cooke & Massee (Hall et al. 2007). Over the

past two decades the harvest and sale of the Chinese black truffle, T. indicum,

has increased dramatically and has become a significant source of income for

the local farmers who live in the mountainous truffle-producing regions. The

two new Tuber species we describe here are based on specimens collected by

these local farmers.

Materials and methods

Morphological studies

Fresh fruiting bodies were collected from Kunming, Yunnan and Panzhihua City,

Sichuan and deposited in BJTC (Herbarium Biology Department, Capital Normal

404 ... Fan, Hou & Cao

TABLE 1. Specimens used in molecular studies and GenBank accession numbers.

SPECIES VOUCHER SPECIMEN ITS

Tuber aestivum AF516788

AY226042

Tuber huidongense BJTC FAN104 JF9261163

DQ486032

Tuber lyonii EU394704

EU268568

Tuber polyspermum BJTC FAN131 JF9261165

Tuber rufum Picco DQ329375

FM205665

Tuber sinoalbidum BJTC FAN105 JF9261164

Tuber spinoreticulatum Uecker & Burds. FJ80988

FJ74891

Tuber taiyuanense B. Liu GU979033

DQ478662

Tuber umbilicatum Juan Chen & P.G. Liu FJ797880

FJ797879

GU979031

University). The macroscopic characteristics were described from both fresh and

rehydrated specimens. The microscopic characteristics are described from razor-blade

sections of the specimens mounted in 3% KOH, Melzer’s reagent, or cotton blue. For

scanning electron microscopy (SEM), ascospores were scraped from dried gleba of the

fruit bodies 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.

Molecular methods

Herbarium samples were crushed by shaking for 3 min at 30 Hz (Mixer Mill MM 301,

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

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

DNA kit following the manufacturer's protocol. The ITS region was amplified with

PCR using the primers ITS1/ITS4 (White et al. 1990). PCR was performed in 50 ul

reactions containing DNA template 2 ul, primer (10 uM/L) 2 ul each, 2x Master Mix

(Tiangen Biotech (Beijing) Co. Ltd.) 25 ul. PCR reactions were run as follows: an initial

denaturation at 95 °C for 3 min, followed by 30 cycles at 95 °C for 2 min, 55 °C for 25 s,

72 °C for 2 min, and a final extension at 72 °C for 10 min. The PCR products were sent

to Invitrogen Biotechnology Co. Ltd. (Beijing, China) for purifying, sequencing, and

editing. The other sequence data of ITS rDNA included in this study were downloaded

from GenBank. GenBank numbers are shown in TABLE 1.

Phylogenetic analyses

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

was manually adjusted with Se-Al v.2.03a (Rambaut 2000). The aligned dataset was

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

Tuber sinoalbidum e& T. polyspermum spp. nov. (China) ... 405

Tuber umbilicatum FJ797880

97 Tuber umbilicatum FJ797879

100 Tuber umbilicatum GU979031

Tuber taiyuanense GU979033

Tuber taiyuanense DQ478662

Tuber lyonii EU394704

100

Tuber lyonii EU268568

ii6 Tuber huidongense DQ486032

Tuber huidongense JF921163

Tuber sinoalbidum JF921164

97 Tuber rufum DQ329375

Tuber rufum FM205665

100 hé Tuber spinoreticulatum FJ809884

Tuber spinoreticulatum FJ748914

Tuber polyspermum JF921165

Tuber aestivum AF516788

Tuber aestivum AY226042

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

Tuber species with spiny and spinulose-reticulate ornamentation on the ascospore surface, using

T. aestivum as outgroups. Bootstrap values of more than 70% from 1000 replications are shown

above the respective branches.

Maximum parsimony analysis was conducted using heuristic searches with 1000

replicates of random-addition sequence, tree bisection reconnection (TBR) branch

swapping algorithm. All characters were equally weighted and unordered. Gaps were

treated as missing data to minimize homology assumptions. A bootstrap (BS) analysis

was performed with 1000 replicates, each with 10 random taxon addition sequences.

TBR branch swapping was employed.

406 ... Fan, Hou & Cao

Results

Molecular phylogenetics

The maximum parsimony analysis of sequences resulted in one most

parsimonious tree (Fic. 1) with a length (TL) = 1082 steps, consistency index

(CI) = 0.7126, retention index (RI) = 0.7347, homoplasy index (HI) = 0.2874,

and rescaled consistency index (RC) = 0.5731.

Phylogenetic analyses of ITS sequences revealed that all cited species

with light colored ascomata and spiny and spinulose-reticulate ascospore

ornamentations grouped together with a bootstrap support value of 100%.

Tuber polyspermum was placed as a distinct clade in the genus. The sequence

of T: sinoalbidum and two sequences of T: huidongense were grouped in a clade

but with a lower BS support.

Taxonomy

Tuber polyspermum L. Fan & C.L. Hou, sp. nov. FIGs. 2-7

MycoBank MB 519840

Ascomata brunnea vel griseo-brunnea, 0.5-1.5 cm diam, glabra vel subglabra, basi

umbilicata. Peridium 150-200 um crassum, strato exteriore pseudoparenchymato et strato

interiore ex hyphis intricatis instructum. Gleba solida, brunnea vel purpureo-brunnea,

venis albis. Asci 65-85 x 45-60 um, 1-4(-5) spori. Ascosporae ellipsoideae, brunneae vel

flavo-brunneae, 25-37.5(-45)x 20-25(-30) um, spineae vel spineo-reticulatae.

Type: China. Yunnan Province, Kunming, hypogeous, under soil of Pinus yunnanensis

Franch., 20 Dec. 2002, Jin-zhong Cao 2002112001 (Holotype, BJTC FAN131).

EryMo.oecy: polyspermum (Lat.) refers to the large number of ascospores in the

ascomata.

AscoMATa subglobose, 0.5-1.5 cm in diam., usually with an umbilicate

depression at the base, brown to gray-brown when fresh, surface smooth. Odor

pungent when fresh. PERrp1uM 150-200 um in thickness, composed of two

layers; outer layer 50-100 um thick, pseudoparenchymatous, composed of

subglobose or subangular, light brown cells 7.5-15(-20) um in diam.; inner

layer 100-150 um thick, textura intricate, composed of interwoven hyphae with

hyaline, thin-walled cells 2.5-5 um broad. GLEBA brown or purple brown at

maturity, veins white, large and rare, containing a great number of spores. ASCI

globose to subglobose, 65-85 x 45-60 um, sessile or with a short stalk, 1-4

(-5) spored. Ascospores ellipsoid, hyaline at first, brown or yellow brown at

maturity, short spiny or spinulose-reticulate, 25-37.5 (-45) x 20-25 (-30) um

excluding the ornamentation, ornamentations up to 2.5-3 um high, meshes

closed or unclosed, regular to irregular, 5-8 across the spore width.

ComMMENTs — Tuber polyspermum is almost indistinguishable from the

American species T: lyonii Butters in both macro- and micro-morphological

characteristics except for small differences in the shape of the ascomata (more

Tuber sinoalbidum & T. polyspermum spp. nov. (China) ... 407

Fics 2-7. Tuber polyspermum (BJTC FAN131, holotype). 2. Ascocarps. 3. Ascospore observed

under the light microscope. 4-5. Asci and ascospores observed under the light microscope.

6-7. Ascospore observed under the scanning electronic microscope.

or less umbilicate in T’ polyspermum) and the color of the gleba (darker in

T. polyspermum). It is clear that these differences alone are insufficient to erect a

new species. However, the molecular analysis showed that the Chinese material

did not group in the same clade with T. lyonii, and instead was in a clade of its

own (Fic. 1). Because of this and the clear geographic separation between Asia

408 ... Fan, Hou & Cao

and North America, we here treat the Chinese material as a new species that is

similar to, but distinct from, T°: lyonii.

A very interesting characteristic of T: polyspermum is the very large number

of ascospores in the mature ascomata in the type specimen, especially in the

dried specimens, when compared with other Tuber species. ‘The specific epithet

“polyspermum’ refers to this.

Tuber sinoalbidum L. Fan & J.Z. Cao, sp. nov. Fics. 8-12

MycoBank MB 519841

Ascomata albida, 2-4.5 cm diam., subglobosa vel globosa. Peridium 200-250 um crassum,

strato exteriore pseudoparenchymato et strato interiore ex hyphis intricatis instructum.

Gleba solida, griseo-albida, veins albis. Asci 45-60 x 60-80 um, 1-4(-5) spori. Ascosporae

ellipsoideae, brunneae vel flavo-brunneae, 25-37.5(-45) x 17.5-25(-30) um, spineo-

reticulatae.

Type: China. Sichuan Province, Panzhihua City, hypogeous, under soil in forest, 31 Dec.

2007, De-fu Liu (Holotype, BJTC FAN105).

EryMo toy: sinoalbidum (Lat.) refers to a whitish Tuber species from China.

ASCOMATA hypogeous, globose or subglobose, 2-4.5 cm in diam., white

to whitish when fresh, surface smooth to very fine verrucose. Odor light.

PERIDIUM 200-250 um in thickness, composed of two layers; outer layer

50-100 um thick, pseudoparenchymatous, composed of subglobose or subangular

light brown colored cells 7.5-12.5 um in diam.; inner layer 150-200 um thick,

textura intricate, composed of interwoven hyphae with hyaline, thin-walled

cells 2.5-5 um broad. GLEBA white, pale white or grey white at maturity, veins

white at first and light brown at maturity, narrow and numerous. Asci globose

or subglobose, 45-60 x 60-80 tum excluding the stalk, stalk 8-20 x 3-5 um long,

1-4(-5) spored. Ascosporgs ellipsoid to broad ellipsoid, hyaline at first, brown

or yellow brown at maturity, spinulose-reticulate, 25-37.5(-45) x 17.5-25(-30)

um excluding the ornamentation, ornamentations up to 3.5-5(-7.5) um high,

meshes 4-6 across the spore width.

ADDITIONAL SPECIMEN EXAMINED: CHINA. YUNNAN PROVINCE, Kunming, from

market, 17 Dec. 2005, Jin-zhong Cao (BJTC FAN101).

ComMENtTs —Tuber huidongense Y. Wang, a common endemic species in

Sichuan and Yunnan (Deng et al. 2009), is similar to T. sinoalbidum in ascospore

ornamentation, but differs in having brown ascomata when fresh, and a

blackish colored gleba at maturity. Also T. huidongense ascomata are normally

small, rather than medium to large as in T! sinoalbidum. The phylogenetic

analysis (Fic. 1) also shows that while T: huidongense groups in a clade with

T. sinoalbidum, the bootstrap support value is low. This indicates they are

closely related but clearly separate. Therefore, we conclude that T: sinoalbidum

is a distinct species.

Tuber sinoalbidum e& T. polyspermum spp. nov. (China) ... 409

Fics 8-12. Tuber sinoalbidum (BJTC FAN105, holotype). 8. Ascocarps. 9-10. Asci and

ascospores observed under the light microscope. 11-12. Ascospore observed under the

scanning electronic microscope.

410 ... Fan, Hou & Cao

The color of T’ sinoalbidum when fresh is similar to that of T’ magnatum

and T: latisporum Juan Chen & P.G. Liu, but they can be separated easily from

the new species by the typical reticulate ascospore ornamentations in both

T. magnatum and T. latisporum (Riousset et al. 2001; Chen & Liu 2007).

Acknowledgments

The study was supported by NSFC (No. 30770005, 30870008). We are grateful to

Prof. Wen-Ying Zhuang and Dr. Ian R. Hall for serving as the pre-submission reviewers.

We also thank Prof. Ying-Ren Lin for critically correcting the Latin diagnoses.

Literature cited

Chen J, Liu PG. 2007. Tuber latisporum sp. nov. and related taxa, based on morphology and DNA

sequence data. Mycologia 99: 475-481. http://dx.doi.org/10.3852/mycologia.99.3.475

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

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

Deng XJ, Chen J, Yu FQ, Liu PG. 2009. Notes on Tuber huidongense (Tuberaceae, Ascomycota), an

endemic species from China. Mycotaxon 109: 189-199. http://dx.doi.org/10.5248/109.189

Hall IR, Brown G, Zambonelli A. 2007. Taming the truffle: the history, lore, and science of the

ultimate mushroom. Timber Press, Portland. 304 p.

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

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

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

Riousset L, Riousset G, Chevalier G, Bardet MC. 2001. Truffles d'Europe et de Chine. Institut

National de la Recherche Agronomique, Paris. 181 p.

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

4. Sunderland, MA, USA, Sinauer Associates.

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

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

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

Trappe JM, Jumpponen AM, Cazares E. 1996. NATS truffle and truffle-like fungi 5: Tuber lyonii

(= T. texense), with a key to the spiny-spored Tuber species groups. Mycotaxon 60: 365-372.

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR Protocols: a Guide to

Methods and Applications. Academic Press, San Diego.

MY COTAXON

http://dx.doi.org/10.5248/118.411

Volume 118, pp. 411-422 October-December 2011

Hymenochaete in China. 2. A new species and three new records

from Yunnan Province

SHUANG-HuI HE” & HalI-Jiao LI

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

CORRESPONDENCE TO *: heshh1981@yahoo.cn

ABSTRACT — A new species and three new Chinese records of Hymenochaete are reported:

H. yunnanensis sp. nov., H. fulva, H. spathulata, and H. sphaerospora. They were all collected

from Caiyanghe Nature Reserve, Yunnan Province, southwestern China. Hymenochaete

yunnanensis (in sect. Hymenochaete) is characterized by setal hyphae and heavily encrusted

hyphae and hymenial cells. Complete descriptions with illustrations are provided for the four

species.

Key worps — Hymenochaetaceae, taxonomy, wood-inhabiting fungi

Introduction

Because of the abundant woody plants and the complex topography, the

diversity of wood-inhabiting fungi is extremely high in Yunnan Province and

adjacent areas of southwestern China. During the last decade, the porioid wood-

inhabiting fungi in these areas have been intensively investigated and many

papers were published (Cui et al. 2011; Dai & Zhou 2000, Dai et al. 2007, Dai

2010, 2011a; Niemela et al. 2001; Yu et al. 2008; Yuan & Dai 2008a,b). However,

the corticioid wood-inhabiting fungi in these areas have not been sufficiently

studied. The corticioid genus, Hymenochaete Lév., is one of the most important

genera in the Hymenochaetaceae. Although it has been studied by several

mycologists (Dai 2010, 2011b; Maekawa & Zang 1995; Xu et al. 2003; Zhang &

Dai 2005), the genus has not yet been systematically studied in southwestern

China.

Caiyanghe Natural Reserve, located in southern Yunnan Province, is one of

the famous nature reserves in China because it is a transitional area from the

tropical to the subtropical zone with the altitude ranging from 980-1707 m.

The main vegetation type is a monsoon evergreen broadleaf forest, comprising

412 ... He & Li

mainly species of Castanopsis, Lithocarpus and Schima (Jing & Chen 2007). In

the summer of 2011, an intensive survey of wood-inhabiting fungi in Caiyanghe

and Xishuangbannan Natural Reserve, southern Yunnan Province, was carried

out and more than 130 Hymenochaete specimens were collected. Among

these specimens, Hymenochaete yunnanensis is identified as a new species. In

addition, H. fulva, H. spathulata and H. sphaerospora are found in China for the

first time. Illustrated descriptions of these species are provided in this paper.

Materials & methods

Voucher specimens are deposited in the herbarium of Beijing Forestry University

(BJFC), and the microscopic procedure follows He (2010). In the text the following

abbreviations are 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

between the specimens studied (quotient of the mean spore length and the mean spore

width of each specimen), n = the number of spores measured from given number of

specimens. In presenting the size range of spores, 5% of the measurements were excluded

from each end of the range, and the measurements were given in parentheses. IKI stands

for Melzer’s reagent, KOH for 5% potassium hydroxide, and CB is the abbreviation of

Cotton Blue. IKI- = inamyloid and nondextrinoid, CB- = acyanophilous. Special color

terms follow Petersen (1996).

Taxonomy

Hymenochaete yunnanensis S.H. He & Hai J. Li, sp. nov. Fics. 1-2

MycoBANnkK MB 563149

Carpophorum effusum vel effuso-reflexum, laxe adnatum. Cortex, tomentum et stratum

hypharum adsunt. Hymeniis et hyphae cum resinaceus granulum encrustatae. Setosum

hyphae adsunt. Setae 40-75(-80) x 6-7(-9) ums, porae ellipsoideae, 5-6.5(-6.8) x (2.8-

)3-3.5(-3.8) um.

Type: China. Yunnan Prov., Puer, Caiyanghe Nat. Res., alt ca. 1200 m, on fallen

angiosperm branch, 9 VI 2011 He 709 (holotype, BJFC).

ETYMOLOGY: yunnanensis, refers to its type locality of Yunnan Province, China.

FruiTBopy: Annual, effused or effused-reflexed with slightly elevated margins,

loosely adnate, easily detached, coriaceous, brittle when dry, first as small

colonies, later confluent, resupinate part up to 8 cm long or more in longest

dimension, reflexed part projecting up to 0.3 cm, 180-300 um thick in section.

Pileal surface rust-brown to dark brown, silky, tomentose, curving down when

dry. Hymenophore smooth or with scattered tubercles, grayish brown to clay-

buff, usually covered with many yellowish resinous matters, not cracked or with

few deep crevices when dry; resupinate margin thinning out, distinct, silky,

fimbriate, yellowish to yellowish brown when juvenile, becoming indistinct or

slightly elevated, concolorous with hymenium when mature.

Hymenochaete yunnanensis sp. nov. (China) ... 413

Fic. 1. Basidiocarps of Hymenochaete yunnanensis (He 709, holotype).

HyYPHAL STRUCTURE: Hyphal system subdimitic; generative hyphae without

clamp connections; tissue darkening but otherwise unchanged in KOH.

SUBICULUM: Cortex, tomentum and hyphal layer present. Cortex composed

of strongly agglutinated hyphae, 20-50 um thick. Generative hyphae hyaline

to yellowish brown, thin- to thick-walled, septate, moderately branched,

sometimes collapsed, more or less interwoven and agglutinated, usually heavily

encrusted with resinous matters, 2-4 um in diam. Skeletoids with distinctly

thickened walls, reddish brown, rarely septate and branched. Setal hyphae

(embedded setae) frequently present, up to 200 um long.

STRATIFIED HYMENIUM: Hyphae in this layer similar to those in subiculum,

hyaline, thick-walled, agglutinated, uprightly arranged, 2.5-4.5 um in diam.

Setal layer thickening, composed of several rows of overlapping setae. Setae

numerous, subulate or fusiform, reddish brown, with an acute tip, projecting

up to 45 um above the hymenium, 40-75(-80) x 6-7(-9) um. Hymenial cells

usually heavily encrusted with resinous matters. Cystidia and hyphidia absent.

Basidia clavate, with four sterigmata and a simple septum at base, 12-20 x

3.8-5 um; basidioles in shape similar to basidia, but slightly smaller.

Basipiosporss ellipsoid, some with tapering apex, hyaline, thin-walled,

smooth, IKI-, CB-, 5-6.5(-6.8) x (2.8-)3-3.5(-3.8) um, L = 5.79 um, W = 3.16

um, Q= 1.79-1.87 (n = 60/2).

414 ... He & Li

Fic. 2. Microscopic structures of Hymenochaete yunnanensis (drawn from the holotype).

a: Basidiospores. b: Basidia and basidioles. c: Setae. d: Setal hyphae. E: Hyphae from subiculum.

Hymenochaete yunnanensis sp. nov. (China) ... 415

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PRov., Puer, Caiyanghe Nat.

Res., alt ca. 1200 m, on fallen angiosperm twig, 9 VI 2011 He 690 (BJFC).

REMARKS: Hymenochaete yunnanensis belongs to sect. Hymenochaete (presence

of cortex, hyphal layer and setal layer; Léger 1998) and is characterized by the

presence of setal hyphae and the heavily encrusted hyphae and hymenial cells.

Usually the hymenial cells are so heavily encrusted that the resinous compounds

form numerous yellowish dots on the hymenophore surface.

Microscopically, the new species is very close to H. ulmicola Corfixen &

Parmasto; however, H. ulmicola differs in growing in the bark fissures of old

living Ulmus trees and in having thicker, harder and smaller basidiocarps and

slightly larger basidiospores (5.5-7.5 x 3-4 um; Corfixen & Parmasto 2005).

Another similar species, H. colliculosa (Sacc.) Parmasto, can be distinguished

from the new species by its larger setae (80-110 x 7.5-11 um) and basidiospores

(5.5-7.5 x 3.6-4.8 um; Parmasto 2005). Hymenochaete fulva also has encrusted

hyphae, but differs from H. yunnanensis in the yellowish brown basidiocarps,

broadly ellipsoid basidiospores (5-6 x 3.2-4 um), and absence of setal hyphae

(Parmasto 2001).

Hymenochaete fulva Burt, Ann. Mo. Bot. Gard. 5: 354, 1918 Fics. 3-4

Fruitsopy: Annual, effused, closely adnate, coriaceous, first as small

colonies, later confluent up to 8 cm or more in longest dimension, 150-300

um thick. Hymenophore smooth, cinnamon-brown, yellowish brown or clay-

Fic. 3. A basidiocarp of Hymenochaete fulva (He 620).

416... He & Li

ieee

5 yum

ile

10 pm

OE PEN,

10 pm

DD ASS

Fic. 4. Microscopic structures of Hymenochaete fulva (drawn from He 620).

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

Hymenochaete yunnanensis sp. nov. (China) ... 417

buff, azonate, not cracked or sometimes with numerous deep crevices; margin

thinning out, distinct, paler than hymenophore surface, yellowish brown.

HyPHAL STRUCTURE: Hyphal system monomitic; generative hyphae without

clamp connections; tissue darkening but otherwise unchanged in KOH.

SUBICULUM: Tomentum absent; cortex and hyphal layer present. Cortex

composed of strongly agglutinated hyphae, 10-20 um thick. Generative hyphae

hyaline to yellowish brown, thick-walled with a wide lumen, some lightly or

heavily encrusted with yellowish brown resinous granules, frequently septate,

moderately branched, loosely interwoven, 2.5-5 um in diam.

STRATIFIED HYMENIUM: Hyphae in this layer similar to those in subiculum,

yellowish to yellowish brown, thick-walled, more or less agglutinated,

interwoven, 2-4.8 um in diam. Crystals occasionally present in the hymenium.

Setal layer composed of 1-3 rows of overlapping setae. Setae numerous, reddish

brown, subulate, sometimes enmeshed with a hyphal sheath, with an acute tip,

projecting up to 80 um above the hymenium, 60-100(-110) x 7-10(-11) um.

Hyphidia absent, encrusted cystidia-like hyphal ends present. Basidia clavate,

with four sterigmata and a simple septum at base, 15-22 x 4-5 um; basidioles

in shape similar to basidia, but slightly smaller.

Spores: Basidiospores broadly ellipsoid, hyaline, thin-walled, smooth,

usually bearing a large guttule, IKI-, CB-, (4.8-)5-6 x 3.5-4 um, L = 5.43 um,

W = 3.82 um, Q = 1.40-1.44 (n = 60/2).

SPECIMENS EXAMINED: CHINA. YUNNAN PRov., Puer, Caiyanghe Nat. Res., alt ca. 1400

m, on fallen angiosperm twig, 6 VI 2011 He 620 & 640.

REMARKS: Hymenochaete fulva belongs to sect. Hymenochaete and _ is

characterized by its yellowish brown basidiocarps, encrusted hyphae, and

broadly ellipsoid basidiospores. The “cystidia” of the species cited by Burt

(1918) and Léger (1998) are actually encrusted hyphal ends (Parmasto 2001).

Hymenochaete fulva has also been reported in Mexico and Jamaica (Parmasto

2001). The species is close to H. rhododendricola S.H. He & Hai J. Li and H.

rhabarbarina (Berk.) Cooke; however, both species differ from H. fulva in their

encrusted setae and absence of cortices (He & Li 2011a; Parmasto 2001).

Hymenochaete spathulata J.C. Léger, Bull. Soc. Mycol. Fr. 96: 409,

1981 [“1980"] FIGs. 5-6

Fruitsopy: Annual, effused, closely adnate, coriaceous, first as small

colonies, later confluent up to 20 cm or more in longest dimension, 180-280 um

thick. Hymenophore smooth, pale mouse-gray to light vinaceous gray, azonate,

not cracked; margin thinning out, distinct, whitish, fimbriate when juvenile,

becoming indistinct, concolorous with hymenophore surface when mature.

HyYPHAL STRUCTURE: Hyphal system monomitic; generative hyphae without

clamp connections; tissue darkening but otherwise unchanged in KOH.

A418 ... He & Li

Fic. 5. Basidiocarps of Hymenochaete spathulata (He 685).

SUBICULUM: Tomentum and hyphal layer absent. Cortex locally present,

thin, composed of strongly agglutinated hyphae.

STRATIFIED HYMENIUM: Generative hyphae hyaline to yellowish brown,

thin- to thick-walled with a wide lumen, densely interwoven, 1.8-3.6 um in

diam. Setal layer thickening, composed of several rows of overlapping setae.

Setae scattered, reddish brown, spathulate, with an obtuse tip, usually lightly

encrusted with crystals at the tip, projecting up to 50 um above the hymenium,

65-100 x 8-13 um. Cystidia and hyphidia absent. Basidia clavate, with four

sterigmata and a simple septum at base, 12-16 x 3.5-4 um; basidioles in shape

similar to basidia, but slightly smaller.

Sporss: Basidiospores cylindrical to allantoid, hyaline, thin-walled, smooth,

IKI-, CB-, 6-7.1(-7.5) x 1.8-2.1 pm, L = 6.65 um, W = 1.98 um, Q = 3.36

(n= 30/1).

SPECIMENS EXAMINED: CHINA. YUNNAN PRov., Puer, Caiyanghe Nat. Res., alt ca. 1200

m, on fallen angiosperm twig, 9 VI 2011 He 685 & 704.

REMARKS: Hymenochaete spathulata is distinguished in the genus by its gray

basidiocarps, spathulate setae, and cylindrical to allantoid basidiospores (Léger

1998). Another species with round-tipped setae is H. ryvardenii Parmasto,

which differs from H. spathulata by thicker (< 800 um) basidiocarps and

larger ellipsoid basidiospores (6.5-8 x 3-3.6 um; Parmasto 2000). Previously

Hymenochaete yunnanensis sp. nov. (China) ... 419

1006

G00 2

) 0,0 ¢

10 pm

| ah iechoemmate

10 ym

Fic. 6. Microscopic structures of Hymenochaete spathulata (drawn from the He 685).

a: Basidiospores. b: Basidia and basidioles. c: Setae.

H. spathulata was known only from Gabon and the Central African Republic

(Léger 1998).

Hymenochaete sphaerospora J.C. Léger & Langq., Bull. Soc. Mycol.

Fr. 103: 48, 1987 Figs. 7-8

FruiTBopy: Perennial, effused, closely adnate, woody hard when dry,

confluent up to 20 cm or more in longest dimension, 250-400 um thick.

Hymenophore smooth, pale mouse-gray to grayish brown, azonate, not cracked

or finely cracked when dry; margin thinning out, distinct, fawn to cinnamon,

up to 2 mm wide.

HyPHAL STRUCTURE: Hyphal system monomitic; generative hyphae without

clamp connections; tissue darkening but otherwise unchanged in KOH.

SuBICULUM: Tomentum absent. Cortex and hyphal layer sometimes

present.

A420 ... He & Li

ore

Ot Te

Fic. 7. A basidiocarp of Hymenochaete sphaerospora (He 691).

STRATIFIED HYMENIUM: Generative hyphae hyaline to yellowish brown,

thick-walled, agglutinated, densely interwoven, 2-3 um in diam. Setal layer

thickening, composed of several rows of overlapping setae. Setae numerous,

reddish brown, subulate, encrusted with fine crystals in the upper part, with

an acute tip, projecting up to 40 um above the hymenium, 65-110 x 9-16(-19)

um. Cystidia absent. Simple hyphidia present. Basidia clavate or cylindrical,

with four sterigmata and a simple septum at base, 23-34 x 4.5-6 um; basidioles

in shape similar to basidia, but distinctly smaller.

Spores: Basidiospores broadly ellipsoid or subglobose, hyaline, thin-walled,

smooth, usually bearing a large guttule, IKI-, CB-, 5-6 x 4-5 um, L = 5.52 um,

W = 4.33 um, Q = 1.27 (n = 30/1).

SPECIMENS EXAMINED: CHINA. YUNNAN PRov., Puer, Caiyanghe Nat. Res., alt ca. 1200

m, on fallen angiosperm twig, 9 VI 2011 He 691 & 715.

REMARKS: Léger (1998) and Parmasto (2005) previously reported H. sphaerospora

from Africa. It is diagnosed by the gray basidiocarps, large encrusted setae,

and broadly ellipsoid or subglobose basidiospores. Hymenochaete macrospora

Y.C. Dai, which has similar basidiospores, differs from H. sphaerospora in the

brownish annual basidiocarps and smooth setae (Dai et al. 2000). The also

similar H. megaspora S.H. He & Hai J. Liis distinguished from H. sphaerospora

by its larger spores (7.5-10 x 5-7) and effused-reflexed basidiocarps (He & Li

2011b).

Hymenochaete yunnanensis sp. nov. (China) ... 421

5 ym

Pe aN

10 um

Fic. 8. Microscopic structures of Hymenochaete sphaerospora (drawn from He 691).

a: Basidiospores. b: Basidia and basidioles. c: Setae.

To date, 19 Hymenochaete species (including the four reported here) have been

reported from Yunnan Province (Maekawa & Zang 1995; Xu et al. 2003; Zhang

& Dai 2005).

Acknowledgments

The authors would like to express their deep thanks to Prof. Yu-Cheng Dai (Institute

of Applied Ecology, Chinese Academy of Sciences) and Dr Masoomeh Ghobad-Nejhad

(Botanical Museum, University of Helsinki) for serving as pre-submission reviewers.

This study was supported by the National Natural Science Foundation of China (No.

31000006), the Fundamental Research Funds for the Central Universities (No. YX2010-

22) and the Beijing Forestry University Young Scientist Fund (No. BLX2009023).

422 ... He & Li

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MY COTAXON

http://dx.doi.org/10.5248/118.423

Volume 118, pp. 423-431 October-December 2011

Lichenological notes 3: Sarcogyne plicata in California

KERRY KNUDSEN’ & JANA KOCOURKOVA?

"The Herbarium, Department of Botany and Plant Sciences, University of California,

Riverside, CA 92521-0124, U.S.A.

*Department of Ecology, Faculty of Environmental Sciences, Czech University of Life Sciences,

Prague, Kamycka 129, Praha 6 - Suchdol, CZ-165 21, Czech Republic

CORRESPONDENCE TO *: Knudsen@ucr.edu, kocourkovaj@fzp.czu.cz

ABSTRACT — Sarcogyne plicata, a California endemic, is revised and discussed. A taxon

referred to in the literature as Sarcogyne “privigna,” which looks similar to S. plicata and is

sympatric in California, is discussed. The name is considered invalid for the taxon.

KEY worps — Acarosporaceae, caliciphile, Joshua Tree National Park, nomenclature

Introduction

The type of Sarcogyne Flot. is S. corrugata Flot. (= S. clavus (DC.) Kremp.)

(Jorgensen & Santesson 1993). The genus currently comprises species with

lecideine apothecia with an excipulum of melanized (or even carbonized)

hyphae that lack any epihymenial accretions as are found in species currently

included in Polysporina Vézda. The exact number of species in the genus is

unknown, partly because the current generic circumscription is paraphyletic

(Reeb et al. 2004; Westberg, pers. comm.) and partly because many species

need revision or are known only from lost types (Knudsen & Standley 2007;

Lendemer et al. 2009b).

In North America we currently recognize 15 described Sarcogyne species.

Twelve species are known from collections made in the last 15 years: S. arenosa

(Herre) K. Knudsen & S.M. Standl. (Knudsen & Standley 2007; Lendemer et

al. 2009a), S. clavus (Magnusson 1935a, b; Harris & Ladd 2005; Knudsen &

Standley 2007), S. crustacea K. Knudsen & Kocourk. (Knudsen & Kocourkova

2010), S. dakotensis H. Magn. (Magnusson 1935b; Knudsen & Standley 2007),

S. desolata (H. Magn.) K. Knudsen & S.M. Standl. (Knudsen & Standley

2007), S. novomexicana H. Magn. (Knudsen & Lendemer 2005; Knudsen &

Standley 2007), S. plicata (Magnusson 1935b; Knudsen & Kocourkova 2009),

424 ... Knudsen & Kocourkova

S. “privigna” (Magnusson 1935a, b; Harris & Ladd 2005; Knudsen & Standley

2007), S. reebiae K. Knudsen (Knudsen & Standley 2007; Knudsen et al. 2011),

S. regularis Korb. (Knudsen 2007; Harris & Ladd 2005), S. similis H. Magn.

(Magnusson 1935b; Harris & Ladd 2005; Knudsen & Standley 2007), and

S. sphaerospora J. Steiner (Lendemer et al. 2009b). Three species are known

only from lost type collections: S. athroocarpa H. Magn., S. integra B. de Lesd.

ex H. Magn., and S. magnussonii B. de Lesd. (Magnusson 1935b; Knudsen &

Standley 2007). We have not seen any taxa fitting the descriptions of these three

species that could be used for neotypification.

Weare currently researching the biodiversity and taxonomy of the lichenized,

lichenicolous, and rock-inhabiting fungi of Joshua Tree National Park. In the

park, Sarcogyne plicata, originally described from California (Magnusson

1935b), is sympatric with S. “privigna.” Knudsen & Standley (2007) previously

treated S. plicata as a synonym of S. “privigna” because it was unclear how to

delimit many specimens from S. “privigna” as then interpreted (see discussion

below). Unpublished molecular work by Martin Westberg (Westberg, pers.

comm.) supported S. plicata as distinct from S. “privigna,” and we accepted both

species although we were still unclear how to differentiate the two (Knudsen &

Kocourkova 2009). In this paper we revise Sarcogyne plicata.

Materials & methods

Specimens were examined from COLO, FH, NY, UCR, and UPS. Specimens were

studied in hand sections using standard microscopy. Structures were examined in water

and KOH and measured in water. Amyloid reactions were tested with IKI. Hymenial

height does not include the height of the subhymenium, a valuable secondary character

in distinguishing some species. Thin-layer chromatography was performed by J.C.

Lendemer on Sarcogyne plicata, Knudsen 1230 (NY) (Culberson & Kristinsson 1970).

Images were captured using an Olympus DP20 digital camera with Microsuite Special

Edition. The illustrations were prepared using Adobe Photoshop. All measurements are

based on water mounts of hand cut sections unless otherwise indicated.

The species

Sarcogyne plicata H. Magn., Ann. Crypt. Exot. 7: 134 (1935). FIG. 1

Type: U.S.A. CALIFORNIA. SAN BERNARDINO Co., Upland. On plaster of wall. 17.11.1917,

I.M. Johnston s.n. (Holotype, FH!).

DESCRIPTION — Thallus endolithic to chasmolithic, sometimes visible between

granules of substrate or beneath apothecia as white cushion of plectenchyma.

Vegetative hyphae mostly 1-2 um in diam., cells mostly 4-5 um long, thin-

walled, I-, mixed with crystals of substrate, forming a gelatinized mass binding

the substrate, continuous with the hypothecium and excipulum hyphae, not

forming a distinct medulla. Algal layer usually scattered in substrate or not

observable, algae chlorococcoid 10-15 um in diameter.

Sarcogyne plicata in California (USA) ... 425

Fic. 1. Sarcogyne plicata (Knudsen 1230).

A. Apothecia. Scale = 1mm; B. Apothecia. Scale = 0,5 mm.

426 ... Knudsen & Kocourkova

Apothecia usually contiguous and apparently not dividing vegetatively,

dull black, epruinose, black when wetted, round, angular, to long and narrow

(then vaguely resembling an Opegrapha), the disc black, visible or obscure, not

turning red when wetted, often less than 0.5 um in diameter or in length, up to

1 mm. Surface of disc without epihymenial accretions as in Polysporina species

but rarely with an umbo, which is a remnant of the ontogeny of the apothecium.

Excipulum distinct, melanized, up to 100 um thick, outer layer dark (hyphae

not visible), inner layer to 40 um thick, golden yellow to red-brown, raising

above the disc and even hiding it when dry, not melanized beneath the disc,

sometimes swollen like labia, or in segments fused often at angles to each

other through compression. Hymenium 100-140 um tall, hyaline, I+ blue;

epihymenium golden yellow, to 15 um tall, thick, paraphyses septate, mostly

1.5-2.0 um tall, with oil drops, with moderate branching, upper three cells

sometimes becoming wider to 4 um, apices usually expanded up to 4 um, often

in pigment cap. Asci 70-85 x (17-)20-25 um, over 100 ascospores per ascus.

Ascospores hyaline, simple, 3-5 x 1-2 um, variable in size. Subhymenium

golden yellow, to 50-80 thick, I+ blue. Hypothecium continuous with attaching

hyphae, forming the base of apothecia, to 300 um thick, without algal layer.

Conidiomata not seen. No lichen substances found with TLC.

SELECTED SPECIMENS EXAMINED — U.S.A. CALIFORNIA. ORANGE Co., Santa Ana

Mountains, Fremont Canyon, on granite, 268 m, Sept. 22, 2008, K. Knudsen 10329

(UCR); on granite rocks eroded out of sandstone, 343 m, Dec. 6, 2007, K. Knudsen 9354

(UCR); 200 m, on conglomerate rock, Oct. 5, 3007, K. Knudsen 9004.2 (UCR); 956 m,

Sept. 18, 2008, K. Knudsen 10310 (UCR); RiveRsIDE Co., Wildomar, Menifee Hills, on

granite wall along seasonal stream, 558 m, Jan. 2, 2004, K. Knudsen 2124 (UCR); 635

m, granite drainages, Dec. 28, 2005, K. Knudsen 4837 (UCR); 543 m, crumbling granite

along stream, Oct. 14, 2009, K. Knudsen 11782 (UCR); Joshua Tree National Park:

Pleasant Valley, wash crossing Geology Tour Road, 1049 m, on flat granite rock, Dec.

3, 2010, K. Knudsen 12689 (UCR); Malapai Hill, south-facing base on granite rubble,

1165 m, Dec. 2, 201, K. Knudsen 12636.1 & 12636.2 (UCR); Squaw Tank, on dissected

decaying quartz dike on monzogranite dome, 1092 m, Dec. 4, 2010, K. Knudsen 12748

(UCR); desert between Skull Rock & Jumbo Rocks, 1347 m, on monzogranite, Dec.

19, 2010, K. Knudsen 13150 (UCR); San Jacinto Mountains, along Forbes Ranch Road,

on granite on pebble plain, 1651 m, March 26, 2006, K. Knudsen 5698 (UCR); SAN

BERNARDINO Co., Granite Mountains, above Granite Cove, along wash, 1335 m, on

granite slab, June 6, 2008, K. Knudsen 9687 (UCR); Joshua Tree National Park, Key’s

Ranch, 1267 m, on granite in washes, May 26, 2005, K. Knudsen 3041 w/ T. LaDoux

(UCR); San Gabriel Mountains, San Antonio Creek, in wash behind San Antonio Dam,

761 m, on hard granite boulders in flood plain, June 4, 2004, K. Knudsen 1230 (ASU,

MIN, NY, UCR); Santa Ana River Valley, base of San Bernardino Mountains, along

Greenspot Road, on granite boulders in floodplain, 533 m, May 1, 2006, K. Knudsen

5936 w/ M. Knudsen (UCR); Woolly Star Preserve, 327 m, on granite boulder, Feb. 7,

2011, K. Knudsen et al. 13570 (UCR); VENTURA Co., Santa Monica Mountains, Tri-

Peaks near Backbone Trail, 829 m, on Conejo volcanics, May 13, 2007, K. Knudsen

8397 (UCR); Yerba Buena Canyon area, 409 m, March 22, 2006, K. Knudsen 5623 w/ D.

Magney (UCR).

Sarcogyne plicata in California (USA) ... 427

ECOLOGY & DISTRIBUTION — Sarcogyne plicata is frequent in southern

California, often locally abundant, usually occurring on granite in drainages,

washes and flood plains, often a solitary pioneer in full sun, at elevations from

200 to 1651 meters. A few collections from the Santa Monica Mountains are

on Conejo volcanics. No populations have been found on calcareous rock.

It is currently considered endemic to California, occurring in the Mojave

Desert (Granite Mountains and Joshua Tree National Park) and in the coastal

mountains and foothills of southern California in the Peninsular Ranges

(Menifee Hills, Santa Ana Mountains, San Jacinto Mountains) and in the

Transverse Ranges (San Bernardino Mountains, San Gabriel Mountains, and

Santa Monica Mountains).

Discussion — A typical S. plicata specimen (Fic. 1) is quite distinctive from

S. “privigna” (Fic. 2), but many specimens can be quite similar. For instance, a

S. plicata thallus with many round apothecia may resemble S. “privigna,” while

a S. “privigna” thallus with many compressed apothecia may be identified as

S. plicata. While S. “privigna” usually has discs that appear red even when

dry, some thalli have mostly blackish apothecia. Fortunately S. “privigna”

“= Ss eo sas

o oleae

2 &

Fic. 2. Sarcogyne ‘privigna (Harris 42777).

Apothecia. Scale = 250 um.

428 ... Knudsen & Kocourkova

discs always turn red when dampened with water while S. plicata discs

remain black. Otherwise S. plicata has a taller hymenium (100-140 um) and

deeper subhymenium (50-80 um) than S. “privigna” (60-85 um and 30-40

um, respectively). One site where both species occurred was in Joshua Tree

National Park at Key's Ranch. The site has extensive monzogranite boulders

forming a maze of small alkaline washes, which commonly contain rock

crevices and fissures rich in calcium deposited during seasonal flooding and

subsequent evaporation. Sarcogyne “privigna” was common in these calcareous

microhabitats. Sarcogyne plicata was frequent in the more acidic microhabitats

found on the tops of monzogranite boulders that are not inundated during

flooding and thus lack calcium deposits. In areas of the Mojave Desert like

Cactus Flats (San Bernardino Mountains) or in the Clark Mountains, where

limestone predominates, S. “privigna” was abundant and no S. plicata was

collected.

The problem of Sarcogyne “privigna” auct., non (Ach.) A. Massal. Fia. 2

Lichenologists generally agree as to which taxon should be referred to

Sarcogyne “privigna” (for instance: Magnusson 1935a, b; Clauzade & Roux

1985; Golubkova 1988; Knudsen & Standley 2007; Seppelt et al. 1998; Fletcher

& Hawksworth 2009). However, the Acharius type of the basionym Lecidea

privigna Ach. 1803 (and thus Massalongo’s 1854 combination in Sarcogyne)

represents Polysporina simplex (Taylor) Vézda (Magnusson 1935a). The

Acharius types of the other possible basionym, Lecanora milvina var. privigna

Ach. 1810, appear to represent Polysporina simplex and/or an Acarospora. For

descriptions see Clauzade & Roux (1985), Magnusson (1935a, b), Knudsen &

Standley (2007), and Fletcher & Hawksworth (2009). Thus the S. “privigna”

now recognized by many authors does not have a validly published name

(Magnusson 1935a; Knudsen & Standley 2007).

Sarcogyne plicata is restricted to a small area within the range of the more

cosmopolitan S. “privigna”. As noted above S. “privigna,” can generally be

separated from the often similar to S. plicata based on its shorter hymenium

and shallower subhymenium. The species also sometimes retains a vestige

of apothecial development in the form of an umbo in the center of the disc

that usually eventually disappears. Also as noted above, the easiest way to

differentiate the two species is that S. “privigna” discs (usually red or red-orange

when dry) are brighter when wet, while the black S. plicata discs remain black

even when wet. The S. “privigna” pycnidia we observed were ca. 0.1-0.2 mm

diam., while conidia were simple, hyaline, and 1.0-2.0 x 0.5-1.0 um.

SELECTED SPECIMENS EXAMINED — CZECH REPUBLIC. Bouemia, Jeseniky, 500 m,

May 9, 1961, A. Vézda, Lichenes Selecti Exsiccati 95 (COLO); GREECE. RETHIMNO,

Mt. Psiloritis, on calcareous rock, 800 m, March 31, 1993, A. Nordin 3129 (UPS); Samos:

west end of the island, 156 m, on limestone, D.J. Hill s.n. (UCR); ITALY. LomBarpy,

Sarcogyne plicata in California (USA) ... 429

Bormio, 1250 m, on stone fence, July 24, 1927, A.H. Magnusson 10675 (UPS); NORWAY.

Troms, Storfjord Par., rocky slope, 60 m, on horizontal calcareous rock, Aug. 6, 2003,

A. Nordin 5633 (UPS); SLOVAKIA. Bratislavsky Svaty Jur, in vineyard, July 20, 1922,

A.H. Magnusson 6825 (UPS); SWEDEN. GoTLanD, Faré par., Gotska Sand6én, on

human bone, May 14, 1921, T. Vestergren (UPS); LycKsELLE LAPPMARK, Tarna par.,

520 m, Aug. 30, 1963, G.E. Du Rietz 675D (UPS); SODERMANLAND, Ostra Vingaker

par., on calcareous rocks, Aug. 11, 1913, A.H. Magnusson s.n. (UPS) VASTERGOTLAND,

Lerdala pars., Sparresater, on rocks in wood, July 12, 1934, A.LH. Magnusson 14261

(UPS); Meddelplana par., Hallekis, on gneiss, July 12, 1942, A.H. Magnusson 18214

(UPS); U.S.A. CALIFORNIA. Los ANGELES Co., Santa Monica Mountains, Hennessey,

south of Castro Crest, 445 m, on calcium carbonate accretions on sandstone, May 31,

2009, K. Knudsen 11169 (UCR); ORANGE Co., Santa Ana Mountains, Fremont Canyon,

587 m, on sandstone, Sept. 18, 2008, K. Knudsen 10303 (UCR); RIVERSIDE Co., Joshua

Tree National Park, Hexie Mountains, Stirrup Tank, 1040 m, on granite, Dec. 26, 2010,

K. Knudsen 13313 (UCR); base of Hexie Mountains at head of Fried Liver Wash, 983

m, rare on granite rocks along wash, Dec. 12, 2010, K. Knudsen 13013 (UCR); Ryan

Mountain, 1568 m, on small granite rock in drainage, Dec. 6, 2010, K. Knudsen 12842

(UCR); Sheep’s Pass, west-facing slope covered with rubble and pebbles, 1369 m, on

decaying granite, Dec. 20, 2010, K. Knudsen 13218 (UCR); Upper Juniper Flats, on soft

granite, 1497 m, Dec. 18, 2010, K. Knudsen 13107 (UCR); SAN BERNARDINO Co., Clark

Mountains, 1535 m, on limestone, Oct. 11, 2009, K. Knudsen 11768 w/ N. Pietrasiak

(UCR); Granite Mountains, canyon above Yucca Bajada camp, 1219 m, Oct. 20, 2007,

K. Knudsen 9391 (UCR); Joshua Tree National Park, Key’s Ranch, on monzogranite

in alkaline wash, 1283 m, Apr. 1, 2005, K. Knudsen et al. 2622 (UCR); San Bernardino

Mountains, Cactus Flats, 1905 m, common on calcareous rock, June 7, 2005, K. Knudsen

3284 (UCR); Bear Mountain, 2652 m, on dolomite, Aug. 25, 2005, K. Knudsen 1609 w/

C.L. Wagner (ASU, UCR); pebble plain along Polique Canyon Road, on granite rock,

2282 m, Aug. 25, 2011, Knudsen 13674 w/ S. Eliason (UCR); SAN DiEGo Co., Anza

Borrego, Sentenac Canyon, 677 m, on granite in drainage, Dec. 23, 2005, K. Knudsen

4832 et al. (UCR); Laguna Mountains, The Point, 1825 m, on vertical granite walls, June

2, 2004, K. Knudsen 1223 (ASU, UCR); SANTA BARBARA Co., Santa Rosa Island, bottom

of Dry Canyon, 96 m, on sandstone along seasonal stream, Oct. 24, 2008, K. Knudsen

10538 (UCR); GEorGIA. PUTNAM Co., Oconee National Forest, on sandstone, Sept.

16, 1996, W.R. Buck 30477 (NY); KANSAS. RUSSELL Co., west of Bunker Hill, 479 m,

on sandstone, June 25, 2007, C.A. Morse 15459A (KANU, UCR); KENTUCKY. HARLAN

Co., Profile Rock, 763 m, on sandstone, Sept. 16, 1991, R.C. Harris 27175 (NY); MAINE.

KENNEBEC Co., Mud Pond, disturbed area under power lines, Sept. 19, 1987, on granite,

R.C. Harris 20896 (NY). MARYLAND. ANNE ARINDEL Co., On Wishing Rock, Raritan

formation, Nov. 25, 1987, Clyde F. Reed (NY); FREDERICK Co., Sugar Loft Mountain,

on rocks along Mt. Ephram Rd., Apr. 19, 1962, Clyde E Reed (NY); Missourt.

CaRTER Co., bluff on e-side of Current River, on top of granite boulder, R.C. Harris

25629 (NY); Mapison Co., Castor River Shut-Ins Natural Area, on granite bedrock,

Oct. 21, 2001, R.C. Harris 45075, 45069-A (NY). NEw MExIco. SAN JUAN Co., Chaco

Canyon National Monument, 1090 m, on sandstone, Aug. 6, 1979, T.H. Nash III, 16362

(ASU); NoRTH CAROLINA. JACKSON Co., Cedar Cliff Mountains, Oct. 11, 1998, on

schist and gneiss, R.C. Harris 42777 (NY); OHIO. GALLIA Co., Wayne National Forest,

Symes Creek Natural Area, 215 m, May 21, 2006, on sandstone, W.R. Buck 50330 (NY);

PENNSLYVANNIA. BLAIR Co., Tussey Mountain, on sandstone talus, Apr. 22, 2008, R.C.

Harris 54291 (NY); FRANKLIN Co., Michaux State Park, n-slope of Rocky mountain,

c. 350 m, June 1, 2009, J.C. Lendemer 18182 (NY); LycoMiNG Co., Tioga State Forest,

430 ... Knudsen & Kocourkova

Algerine Swamp Natural Area, c. 548 m, May 14, 2009, on rock, J.C. Lendemer 16992

(NY); VERMONT. LAMOILLE Co., Smuggler’s Notch, rocks at base of a cliff, Sept. 21, 1985,

R.C. Harris 18222 (NY); WEsT VIRGINIA. GRANT Co., Shroud Ridge, on sandstone, 425

m, W.R. Buck 38252 (NY).

ECOLOGY & DISTRIBUTION — Sarcogyne “privigna” occurs on calcareous

rock as well as non-calcareous rock such as granite, gneiss, sandstone and

volcanic rock in drainages, washes, and flood plains potentially rich in calcium

carbonate deposited in the fissures of rocks during inundation and subsequent

evaporation. The species is widespread and found in Africa, Asia, Europe, North

America, Antarctica, and Australia (Fletcher & Hawksworth 2009; Golubkova

1988; Seppelt et al. 1998).

Acknowledgments

We thank our reviewers, T. Wheeler and M.G. Halici (Erciyes University, Turkey).

We thank J.C. Lendemer for photographs and TLC. The work of Kerry Knudsen was

supported by a co-operative agreement between UCR and the Joshua Tree National

Park. His visit to New York Botanical Garden was supported by an Academic Fellowship

from the Santa Monica Mountains Fund. The work of Jana Kocourkova was supported

financially by the KONTAKT II, Program of International Cooperation in Research

and Development for scientific cooperation between the CR and USA, LH 11057 from

Ministry of Education, Youth and Sports.

Literature cited

Clauzade G, Roux C. 1985. Likenoj de Okcidenta Europo. Ilustrita Determinlibro. Bulletin de la

Societe Botanique du Centre-Ouest, Nouvelle Serie, Numero Special 7. Royan, France. 893 pp.

Culberson CF, Kristinsson H. 1970. A standardized method for the indentification of lichen products.

Journal of Chromatography 46: 85-93. http://dx.doi.org/10.1016/S0021-9673(00)83967-9

Fletcher A, Hawksworth DL. 2009. Sarcogyne A. Massal. (1851). 829-830, in: CW Smith et al.

(eds.): The lichens of Great Britain and Ireland. British Lichen Society, Natural History Museum

Pub., UK.

Golubkova NS. 1988. The lichen family Acarosporaceae in the U.S.S.R. Komarov Botanical Institute,

Academy of Sciences of the U.S.S.R. (‘Nauka’), Leningrad. 136 pp.

Harris RC, Ladd D. 2005. Preliminary draft: Ozark lichens. Available at:

http://www.duke.edu/~bph8/Harris_and_Ladd_2005_Ozark_Lichens.pdf

Jorgensen PM, Santesson R. 1993. Nine proposals to conserve generic names of lichenized fungi.

Taxon 42(4): 881-887. http://dx.doi.org/10.2307/1223274

Knudsen K, Kocourkova J. 2009. Lichens and lichenicolous fungi of the northwestern Santa Ana

Mountains. Crossosoma 35(2): 66-81.

Knudsen K, Kocourkova J. 2010. Lichenological notes 1: Acarosporaceae. Mycotaxon 112: 361-366.

http://dx.doi.org/10.5248/112.361

Knudsen K, Lendemer JC. 2005. Changes and additions to the checklist of North American lichens

— III. Mycotaxon 93, 277-281.

Knudsen K, Standley SM. 2007. Sarcogyne. 289-296, in: TH Nash III et al. (eds.): Lichen flora of

the Greater Sonoran Desert Region. Volume 3. Lichens Unlimited, Arizona State University,

Tempe.

Sarcogyne plicata in California (USA) ... 431

Knudsen K, Lendemer JC, Harris RC. 2011. Studies in lichens and lichenicolous fungi - no. 15:

miscellaneous notes on species eastern North America. Opuscula Philolichenum 9: 45-75.

Lendemer JC, Kocourkova J, Knudsen K. 2009a. Studies in lichens and lichenicolous fungi: more

notes on taxa from North America. Mycotaxon 110: 373-378.

http://dx.doi.org/10.5248/110.373

Lendemer JC, Kocourkova J, Knudsen K. 2009b. Studies in lichens and lichenicolous fungi: more

notes on some taxa from North America. Mycotaxon 108: 491-497.

http://dx.doi.org/10.5248/108.491

Magnusson AH. 1935a. Acarosporaceae, Thelocarpaceae. 1-318, in: A Zahlbruckner (ed.):

Rabenhorst’s Kryptogamen-Flora von Deutschland, Osterreich, und der Schweiz, 2nd ed. Band

9, Abt. 5(1). Borntraeger, Leipzig.

Magnusson AH. 1935b. On the species of Biatorella and Sarcogyne in America. Annales de

Cryptogamie Exotique 7: 115-145.

Reeb V, Lutzoni FE, Roux C. 2004. Contribution of RPB2 to multilocus phylogenetic studies of

the euascomycetes (Pezizomycotina, Fungi) with special emphasis on the lichen-forming

Acarosporaceae and evolution of polyspory. Molecular Phylogenetics and Evolution 32:

1036-1060. http://dx.doi.org/10.1016/j.ympev.2004.04.012

Seppelt RD, Nimis PL, Castello M. 1998. The genus Sarcogyne (Acarosporaceae) in Antarctica.

Lichenologist 30(3): 249-258. http://dx.doi.org/10.1006/lich.1998.0135

MY COTAXON

http://dx.doi.org/10.5248/118.433

Volume 118, pp. 433-440 October-December 2011

Homolaphlyctis polyrhiza gen. et sp. nov.,

a species in the Rhizophydiales (Chytridiomycetes)

with multiple rhizoidal axes

Joyce E. LONGCORE™, PETER M. LETCHER’ & TIMOTHY Y. JAMES?

'School of Biology & Ecology, University of Maine

5722 Deering Hall, Orono, ME 04469-5722, USA

*Department of Biological Sciences, The University of Alabama

Box 870344, Tuscaloosa, AL 35487 USA

*Department of Ecology & Evolutionary Biology, University of Michigan

830 North University, Ann Arbor, MI 48109-1048 USA

* CORRESPONDENCE TO: longcore@maine.edu

AxsstrRactT — An undescribed cellulosic chytrid with multiple rhizoidal axes, JEL142,

has grouped in molecular hypotheses with Batrachochytrium dendrobatidis, the chytrid

pathogen of amphibians, and thus is of interest for genetic and physiological comparisons.

To describe this member of the Rhizophydiales, we examined its zoospore ultrastructure and

developmental morphology. Based on a reanalysis of rDNA data plus ultrastructural and

morphological characters, we name this fungus Homolaphlyctis polyrhiza gen. et sp. nov.

Key worps — Chytridiomycota, phylogeny, Rhizophlyctis, TEM

Introduction

A chytrid referred to by its isolate number, JEL142, has been in phylogenies

since the first molecular hypotheses of the Chytridiomycota (James et al.

2000, 2006; Letcher et al. 2008b) but has not been identified to species or

received a formal name. This isolate is of interest because it groups as sister

to Batrachochytrium dendrobatidis Longcore et al. (Longcore et al. 1999), the

pathogen of amphibians, and because it is one of the few members of the

Rhizophydiales Letcher (Letcher et al. 2006) that have thalli with multiple

rhizoidal axes. Although the phylogenetic distance based on branch lengths

in phylograms is large and support values for the relationship have been low

(James et al. 2006; Letcher et al. 2008b), JEL142 seems to be the closest relative

to B. dendrobatidis now in culture. In an effort to identify genes that enable

434 ... Longcore, Letcher & James

B. dendrobatidis to be pathogenic, the genome of JEL142 has been sequenced

for comparison (Joneson et al. 2011). In this paper, we reanalyze the phylogenetic

placement of JEL142 and provide a description of the ultrastructure of its

zoospore and its developmental morphology. These analyses place isolate JEL142

in the Rhizophydiales, but outside of described families or genera. Therefore we

describe a new genus and species to accommodate isolate JEL142.

Materials & methods

We first observed this chytrid on onionskin bait (outer, dry bulb scales of white

onions) placed in a gross culture containing the aquatic plant Eriocaulon aquaticum,

associated debris and lake water in a deep Petri dish. The collection site was Mud Pond, a

small (1.7 ha), acidic (pH 4.6), oligotrophic and fishless lake in Hancock County, Maine

(Davis et al. 1994, Rhodes & Davis 1995). With standard methods (Barr 1986) we isolated

JEL142 into pure culture on modified PmTG nutrient agar (Longcore 1992). Since initial

isolation in 1994 this chytrid has grown on nutrient agar slants with transfers at three-

month intervals and storage at ~4 °C after growth. We collected zoospores from growth

on 20 mPmTG plates and fixed them for TEM with s-collidine buffered glutaraldehyde

followed by osmium tetroxide. Zoospores were rinsed, added to agar and en bloc stained

with uranyl acetate before embedment in epon araldite plastic (Barr 1981, Longcore

1992). Sections were examined at 60 kV. We photographed developmental morphology

of thalli grown on mPmTG nutrient agar with a Spot RT digital camera. Phylogenetic

analysis of Rhizophydiales was conducted using the initial alignments of 18S, 28S, and

5.88 rRNA subunits produced by James et al. (2006). Additional sequences were added

from Letcher et al. (2004, 2008b), and the alignment manually adjusted in MacClade

4.08 (Maddison & Maddison 1992). The phylogeny was estimated using maximum

likelihood in RAXML-7.0.4 (Stamatakis 2006) with 100 heuristic searches beginning

from random trees. Support for nodes was established by bootstrapping in RAxML (1000

pseudo-replicates) and Bayesian posterior probabilities in MrBayes v3.1.2 (Huelsenbeck

& Ronquist 2001).

Results

The phylogenetic analysis of Rhizophydiales based on combined 18S, 28S,

and 5.8S subunits provides strong support, 77 bootstrap percentage and

0.99 posterior probability, for a sister relationship between H. polyrhiza and

B. dendrobatidis (Fic. 1). These two species group with a strain identified as

Entophlyctis helioformis (JEL326), and this clade is sister to the other members

of the Rhizophydiales. As seen in previous studies, the phylogenetic distance

between H. polyrhiza and B. dendrobatidis is rather large as suggested by

branch lengths, and the percentage difference between rRNA sequences of the

two species is ~10%.

Homolaphlyctis polyrhiza gen. et sp. nov. ... 435

100/1.0 »MP8 Globomycetaceae clade

85/0.99 --., ARGO68 Globomyces pollinis-pini

ARG046 Pateramyces corrientinensis

JEL136 Rhizophydium brooksianum

JEL316 Rhizophydium clade

JEL223 Operculomyces laminatus

PL163L Coralloidiomyces digitatus

100/1.0--4. JEL171 Rhizophlyctis harderi

2 PLAUS21 Boothiomyces macroporosum

PLO8 Terramyces clade

97/1.0 ARGO008 Angulomyces argentinensis

61/0.99 ARG018 Aquamyces chlorogonii

ARGO71 Protrudomyces lateralis

PL98 Kappamyces laurelensis

a ARGO025 Alphamyces chaetiferum

100/1.0 JEL299 Gorgonomyces clade

83/1.0 ARG026 Gorgonomyces haynaldii

89/1.0 JEL174 Entophlyctis sp.

7710.99 JEL197 Batrachochytrium dendrobatidis

JEL142 Homolaphlyctis polyrhiza

/ JEL326 Entophlyctis helioformis

Si goue Perry UCB-91-10 Gaertneriomyces semiglobifer

62/0.96 ----J96/1.0 ATCC48900 Spizellomyces punctatus

JEL318 Rhizophlyctis rosea

JEL127 Nowakowskiella sp.

100/1.0 JELO6 Rhizoclosmatium globosum

ee 100/1.0 JEL30 Podochytrium dentatum

JEL129 Entophlyctis luteolus

JEL109 Polychytrium aggregatum

0.03

FiGurRE 1. Phylogeny of Rhizophydiales showing sister relationship between Homolaphlyctis

polyrhiza and Batrachochytrium dendrobatidis. The phylogeny was based on a combined

analysis of 18S, 28S and 5.88 ribosomal RNA subunits and was estimated using maximum

likelihood under a GTR+I+I model of evolution with RAxML-7.0.4 (Stamatakis 2006). Values

at nodes indicate support as estimated by maximum likelihood from 1,000 bootstrap pseudo-

replicates followed by Bayesian posterior probabilities. Only nodes receiving >60% bootstrap

support are labeled. Further strain details and GenBank accession numbers can be found in

James et al. (2006) and Letcher et al. (2004, 2008b).

Taxonomy

Homolaphlyctis Longcore, Letcher & T.Y. James, gen. nov.

MycoBank MB561579

Zoosporarum ultrastructura Rhizophydialium characteristica, constans ex microtubulis

separatis duo vel tres e kinetosomate ad rumposoma in globulo solitario extensorum;

microcorpus in latere interiore globuli lipoidei. Calcar kinetosomale et zona vesiculata

436 ... Longcore, Letcher & James

circum kinetosoma absunt. Mitochondrion lobatum. Vesiculae corde denso per cytoplasma

dispersae.

TYPE SPECIES: Homolaphlyctis. polyrhiza Longcore, Letcher & T.Y. James

EryMoLocy: Named for R.L. Homola in whose laboratory this work began, and A.D.

Homola, who helped collect the sample.

Zoospore ultrastructure typical for members of the Rhizophydiales, consisting

of stack of 2-3 separate microtubules leading from the kinetosome to a

rumposome on a single lipid globule; microbody on inner side of lipid.

Lacking kinetosome-associated spur and vesiculated area around kinetosome.

Mitochondrion lobed. Dense-cored vesicles throughout cytoplasm.

Homolaphlyctis polyrhiza Longcore, Letcher & T.Y. James, sp. nov. FIGS. 2, 3

MycoBank MB561580

Thallus axibus rhizoidalibus tres vel pluris extense dispersis. Papillae evacuationis

tubulosae numerosae, breves usque ad longae.

Type: Fic. 2a—-h (holotype) photographed from pure culture JEL142 isolated from

cellulosic substrate placed with a collection of Eriocaulon aquaticum collected from

Mud Pond, (44°38'00.26"N 68°05'15.81"W) Hancock County, Maine on 15 April 1994.

[GenBank DNA sequences: AH009051 (SSU rDNA); EF634247 (LSU rDNA); EF634249

(ITS1-5.8S-ITS2).]

Erymo.oey: The epithet refers to the multiple rhizoidal axes.

Thallus with three or more widely distributed rhizoidal axes. Multiple, short to

long tubular discharge papillae.

EXPANDED DESCRIPTION — ‘The generation time for H. polyrhiza on

mPmTG nutrient agar is 2-3 days at 23 °C. In motion, zoospores are spherical

(Fic. 2h, arrowhead) and contain a single lipid globule near the base of the

flagellum (Fic. 2a). They encyst and within 24 h form a thallus with multiple

rhizoidal axes (Fic. 2b). At maturity rhizoids can extend to more than twice

the diameter of the zoosporangium (Fic. 2c); distally rhizoids are fine (<0.5

um) and isodiametric, but near the thallus they are slightly swollen (Fic. 2d).

Zoosporangia form discharge papillae (Fics. 2e, f) within 52 hr, with the

number of papillae increasing with size of the zoosporangia. Papillae may be

barely raised (Fic. 2f), short tubes (Fic. 2e) or, in crowded conditions, long

tubes (Fic. 2g). Zoospores are elongate when discharged (arrows, Fic. 2h) but

soon become spherical (dia 3.5-4.5 um with flagellum up to 28 um).

ULTRASTRUCTURAL CHARACTERS — ‘The ultrastructure (Fics 3a-e) of

the H. polyrhiza zoospore is nearly identical to Barr's illustration of a typical

Rhizophydium zoospore (Barr & Hadland-Hartmann 1978). A membrane-

bound aggregation of ribosomes is in the center of the zoospore, outside of

which are the nucleus, a single lobed mitochondrion, and a single lipid globule

(Fics. 3a, b). A lobe of a mitochondrion is often anterior to the kinetosome

Homolaphlyctis polyrhiza gen. et sp. nov. ... 437

FiGuRE 2. Developmental morphology of Homolaphlyctis polyrhiza in pure culture on mPmTG

nutrient agar. a. Zoospores with single lipid globule, aggregated ribosomes and single posteriorly

directed flagellum. b. Young thallus with multiple rhizoidal axes (arrows). c. Nearly mature thallus

showing the extent of rhizoids. d. Rhizoidal bases (arrow) are enlarged. e. Papilla (arrowhead) on

nearly mature zoosporangium. f. Short, evenly spaced discharge papillae (arrowheads). g. Long

discharge papilla, nearly ready to release zoospores. Note breakdown of apical, hyaline papillar

material. h. Zoospores (arrows) elongate during discharge through papillar opening (arrowheads)

but become spherical shortly after release (large arrowhead). Scale bar in a, for all figures except c.

(Fic. 3a). The microbody is appressed to the side of the lipid globule toward the

center of the zoospore (Fic. 3b), and a rumposome (= fenestrated cisterna of

Letcher et al. 2004) is appressed to the lipid near the surface of the cell (Fic. 3b).

The non-flagellated centriole is parallel to the kinetosome and joined to it by a

bridge of partly overlapping fibers centrally located between the two structures

(Fras. 3c, d; arrow); the fibrils form a wide (>0.075 um) zone of convergence. The

non-flagellated centriole is shorter than its width (Fras. 3c, e). A microtubular

root, composed of separate, parallel microtubules, extends from the side of the

kinetosome to the rumposome (Fic. 3e). A terminal plate is in the lumen of

438 ... Longcore, Letcher & James

“aaa Sra h 4

tie ee ca $ “”

5 os OL =

Ficure 3. Ultrastructural features of Homolaphlyctis polyrhiza zoospores. a. Longitudinal section

of zoospore with nucleus (N); aggregated ribosomes (R); lobed mitochondrion (M); kinetosome

(K); non-flagellated centriole (nfc). b. Transverse section with lipid globule (L); microbody (mb);

rumposome (Ru); vacuole (Va); and vesicles (arrowheads). c. Longitudinal section of kinetosomal

region with kinetosomal props (P) and terminal plate (T). d. Cross section of kinetosome and

non-flagellated centriole with fibrillar connection and wide zone of convergence (arrow) between

the two structures. Microtubule root (mt) extends from the kinetosome. e. Longitudinal section

through the kinetosome and non-flagellated centriole; microtubules extend from the kinetosome

to the rumposome, shown in face view. Scale bar in b also for a; scale bar in c also for d-e.

the axoneme (Fic. 3c). Typical for members of the Rhizophydiales (Letcher et

al. 2006), a flagellar plug is absent from the transition region of the flagellum.

In the peripheral cytoplasm outside the central core are numerous, large

Homolaphlyctis polyrhiza gen. et sp. nov. ... 439

FiGurE 4. Summary diagram of zoospore ultrastructure of Homolaphlyctis polyrhiza.

Kinetosome (K); lipid globule (L); mitochondrion (M); microbody (mb); microtubule root

(mt); nucleus (N); nonflagellated centriole (nfc); prop (P); vacuole (Va); vesicle (Ve).

(~200 nm diameter) cored vesicles (Fic. 3b; arrowheads); a large vacuole is

often present. Ultrastructural features are summarized in Fic. 4.

ComMENTS — We found this saprobic organism only once, from an oligotrophic,

acidic lake. Based on descriptions of genera of the Chytridiales sensu Sparrow

(1960), it would have been placed in the genus Rhizophlyctis A. Fisch. because

it develops endogenously and has multiple rhizoidal axes. Molecular and

TEM evidence have altered our concept of the importance of morphological

characters (e.g., James et al. 2000, 2006; Mozley-Standridge et al. 2009) and

chytrids with endogenous development and multiple rhizoidal axes occur not

only in the Rhizophlyctidales (Letcher et al. 2008a) but also other orders of the

Chytridiomycetes, now including the Rhizophydiales.

Acknowledgments

This work was supported by NSF: PEET grant DEB-0529694, AFTOL grant DEB-

0732599 and REVSYS grant DEB-0516173. We thank Dr. James Pringle of the Royal

Botanical Garden, Ontario for translating the Latin description.

440 ... Longcore, Letcher & James

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17: 754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

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336-350. http://dx.doi.org/10.1139/cjb-78-3-336

James TY, Letcher PM, Longcore JE, Mozley-Standridge SE, Porter D, Powell MJ, Griffith GW,

Vilgalys R. 2006. A molecular phylogeny of the flagellated fungi (Chytridiomycota) and

description of a new phylum (Blastocladiomycota). Mycologia 98: 860-871.

http://dx.doi.org/10.3852/mycologia.98.6.860

Joneson S, Stajich JE, Shiu S-H, Rosenblum EB. 2011. Genomic transition to pathogenicity in

chytrid fungi. PLoS Pathog 7(11): e1002338. http://dx.doi.org/ 10.1371/journal.ppat. 1002338

Letcher PM, Powell MJ, Chambers JG, Holznagel WE. 2004. Phylogenetic relationships among

Rhizophydium isolates from North America and Australia. Mycologia 96: 1339-1351.

http://dx.doi.org/10.2307/3762150

Letcher PM, Powell MJ, Churchill PE Chambers JG. 2006. Ultrastructural and molecular

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898-915. http://dx.doi.org/10.1016/j.mycres.2006.06.011

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— anew order in Chytridiomycota. Mycol Res 112: 1031-1048.

http://dx.doi.org/10.1016/j.mycres.2008.03.007

Letcher PM, Powell MJ, Viusent MC. 2008b. Rediscovery of an unusual chytridiaceous fungus new

to the order Rhizophydiales. Mycologia 100: 325-334.

http://dx.doi.org/10.3852/mycologia.100.2.325

Longcore JE. 1992. Morphology, occurrence, and zoospore ultrastructure of Podochytrium

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http://dx.doi.org/10.1093/bioinformatics/btl446

MY COTAXON

http://dx.doi.org/10.5248/118.441

Volume 118, pp. 441-446 October-December 2011

Eutypella phaeospora, a new species on Chenopodiaceae

JACQUES FOURNIER’ & CHRISTIAN LECHAT”

’ Las Muros, F-09420 Rimont, France

? 64 route de Chizé, F-79360 Villiers en Bois, France

*CORRESPONDENCE TO: jacques.fournier@club-internet.fr

ABSTRACT — The authors describe a new species of Eutypella (Diatrypaceae, Xylariales) based

on several collections on Suaeda vera in France and one on Salsola vermiculata in Spain. The

Libertella-like asexual state was obtained in culture.

KEY worpDs — Ascomycota, taxonomy, salt marsh, disjunct distribution, semi-arid land

Introduction

An ascomycete referable to Eutypella (Nitschke) Sacc. was repeatedly

collected on Suaeda vera J.F. Gmel. (Chenopodiaceae), a woody shrub occurring

in salt marshes in Ile de Ré (France), and once on Salsola vermiculata L.

(Chenopodiaceae) in a semi-arid environment in northern Spain. Salt marshes

are known to be peculiar ecosystems in which halophytes associate with both

terrestrial and marine mycota (Barata 2002). Previous investigations have been

carried out mainly on monocots such as Juncus roemerianus (Kohlmeyer et

al. 1995), Spartina sp. (Gessner & Kohlmeyer 1976), S. alterniflora (Gessner

1977), and S. maritima (Barata 2002), but little attention has been paid to

woody shrubs.

Based on extensive collecting of ascomycetes in Ile de Ré, this Eutypella

appears to be somewhat common in salt marshes but was never encountered

on the island out of this biotope. However, its occurrence on S. vermiculata

in a very different environment in Spain suggests a stronger relationship to

Chenopodiaceae than to a specific ecological niche as first assumed.

As it is also remarkable in possessing ellipsoid brown ascospores, we decided

to describe it as new. A typically diatrypaceous anamorph obtained in culture

confirmed its placement in Diatrypaceae and a living culture was deposited in

CBS, allowing further phylogenetic investigations in the future.

442 ... Fournier & Lechat

Materials & methods

Macro and microscopic observations were carried out following Rappaz’s (1987)

methodology, especially by testing the reaction of subapical ring to Melzer’s reagent

after pre-treatment in 3% KOH and by making measurements of asci and ascospores in

heated lactic cotton blue. Micrographs of asci were taken in Lugol's solution (IKI), those

of conidiogenous structures and conidia were taken in dilute Waterman blue ink and

water. Cultures were made from single ascospores which were isolated on malt extract

agar (Difco™ Malt Extract Agar) according to Rossman’s (1999) method. The material

collected, along with duplicates of holotype, is kept in the the authors’ private herbaria

(JF and CLL numbers). The holotype is deposited in LIP herbarium (Lille) and cultures

at CBS.

Taxonomy

Eutypella phaeospora J. Fourn. & Lechat, sp. nov. PLATE 1

MyYCOBANK 561649

Ab alteribus speciebus generis Eutypella differt combinatione crescentiae in

Chenopodiaceae, ascosporarum ellipsoidearum et brunneorum et apiculi annuli leviter

amyloidei.

TyPeE: France, Charente Maritime: Ile de Ré, Loix, Le Feneau, edge of a salt marsh, on

dead or dying stems of Suaeda vera, 16 Apr. 2008, leg. MH, CLL & JF (Holotype, LIP

JF08078; ex-type culture, CBS129148).

Erymo oey: The epithet is derived from the unusual brown colour of ascospores.

STROMATA corticolous or lignicolous, pustulate, pustules 0.8-1.8 mm diam,

comprising 2-8 perithecia, irregularly scattered, separate to sometimes

confluent; in bark, stromata irregularly rounded, hardly raising the periderm,

under a conspicuous dorsal black line which is absent when the periderm

is tightly adherent, cortical tissue hardly altered, without differentiated

entostroma; in wood, stromata fully immersed to slightly raising wood surface,

elongated in the grain of wood, substrate not altered but surface more or less

blackened. OsTIOLEs emerging collectively as small bunches 0.5-1 mm diam,

shiny black, rectangular with cruciform apices and deeply sulcate sides, (80-)

150-840 um high, 210-290 um diam. PerirHeciA subglobose, 380-420

um diam, in contact, with short to long convergent necks. Ascr unitunicate,

thin-walled, long-pedicellate, the spore-bearing parts cylindrical to fusiform

PLATE 1. Eutypella phaeospora (holotype). A: Surface view of host bark showing bunches of ostioles

erumpent through the periderm. B: Cross section through a stroma in bark showing convergent

ostiolar necks. C-D: close-ups showing the stout cruciform ostioles. E: Cross section through a

stroma in wood showing convergent ostiolar necks. F: Mature and immature asci mounted in

Lugol’s solution showing the very faint reaction of apical ring. G: Ascospores mounted in lactic

cotton blue. H: Conidiophores, conidiogenous cells and conidia of the Libertella-like anamorph

mounted in dilute Waterman blue ink. I: Culture in Petri dish. J: Conidia in water.

Scale bars: A = 5 mm; B-C,E = 1 mm; D = 0.5 mm; F-H,J = 10 um; I= 1 cm.

Eutypella phaeospora sp. nov. ... 443

444 ... Fournier & Lechat

with a truncate apex, 35-40 um long x 7.5-8.5 um broad with a minute, very

inconspicuous subapical ring bluing in Lugol's solution or in Melzer’s reagent

after pre-treatment with 3% KOH, the stipes 30-35 um long, filiform, very

fragile. Paraphyses copious, persistent, hyphae-like, hyaline, septate, 4-5 tm at

base, tapering above asci. ASCOSPORES 6-7.5 x 3.4—4 um, oblong to ellipsoid-

equilateral with broadly rounded ends, brown, one-celled, smooth, obliquely

uniseriate to irregularly biseriate in the ascus.

ADDITIONAL SPECIMENS EXAMINED: FRANCE, CHARENTE MARITIME: Ars en Ré, edge

of a salt marsh, on dead or dying stems of Suaeda vera, 07 May 2007, leg. C. Lechat,

CLL7129. SPAIN, Navarra: Arguedas, 80 km south of Pamplona, on roadside in the

semi-desert of Bardenas Reales, 310 alt., on a dead corticated stem of Salsola vermiculata,

26 Sep. 2010, leg. Jean-Paul Priou, CLL10033.

ANAMORPH in culture: After two months on MEA, colony wrinkled, 2.5-3 cm

diam., 0.3-0.5 cm high, producing a Libertella-like anamorph, white at first,

then covered by greyish-white hyphal elements, white at margin, diffusing a

brown to blackish coloration in the medium; reverse side greyish-yellow in

the center, blackish at margin. Conidiophores branched, conidiogenous cells

cylindrical, producing holoblastic conidia 16-23(-—26) x 1.1-1.3 um, hyaline,

white en masse, cylindrical, tapered at ends, irregularly curved, e.g. curved on

upper third of length or regularly and strongly curved, produced in whitish

clusters on the median part of the colony.

Discussion

This ascomycete meets all the key features of the family Diatrypaceae Nitschke

in macromorphology, ascal morphology, and Libertella-like anamorph. It

only deviates in possessing ascospores that are ellipsoid and brown instead

of allantoid and yellowish as encountered in typical taxa. However, several

species in Anthostoma Nitschke, Cryptosphaeria Ces. & De Not., Diatrype

Fr. and Eutypella are known to have brown ascospores (Rappaz 1987, 1992),

and several of them —A. decipiens (DC.) Nitschke, C. subcutanea (Wahlenb.)

Rappaz, D. whitmanensis J. D. Rogers & Glawe, E. grandis (Nitschke) Sacc.,

E. corynostomoides (Rehm) Rappaz— produce such ellipsoid ascospores along

with broadly allantoid ones (Rappaz 1987).

The new taxon appears best accommodated in Eutypella as defined by

Rappaz (1987) based on its pustulate stromata with stout, converging ostiolar

necks. Although the recent ITS sequence analysis of Diatrypaceae by Acero et al.

(2004) showed that Eutypella as currently conceived is most likely polyphyletic,

those authors did not propose any formal taxonomic changes. We therefore

refer the present fungus to Eutypella and await further molecular studies that

might shed some light on its affinities within Diatrypaceae. A case was made

to reconsider its placement following Carmaran et al. (2006), who reinstated

Peroneutypa Berl. for taxa differing from Eutypella in ascal morphology.

Eutypella phaeospora sp. nov. ... 445

Although we did not use fluorescence microscopy to study the morphology of

asci in E. phaeospora, the ascal shape and subapical ring clearly point towards

type I as defined by the authors and therefore suggest a closer affinity with

Eutypella as currently accepted (Rappaz 1987).

Eutypella phaeospora can be readily distinguished from other Eutypella

species through its consistently ellipsoid brown ascospores and occurrence on

members of Chenopodiaceae. The usually uniseriate ascospore arrangement

in the ascus correlated with the cylindrical shape of its spore-bearing part is

likewise a distinctive feature contrasting with the spindle-shaped ascus most

often encountered in Diatrypaceae. The fungus seems to be a common saprobe

occurring on Suaeda vera in France and on Salsola vermiculata in Spain.

Eutypella kochiana Rehm and E. alsophila (Durieu & Mont.) Berl. (which

occur on Chenopodiaceae in marine environments) are worth notice here, but

their asci have a clearly amyloid subapical ring and their ascospores markedly

differ in being yellowish and allantoid. Eutypella kochiana var. salsolae Urries,

collected on Salsola vermiculata in Spain, resembles E. phaeospora in slightly

larger ascospores (4.8-8 x 1.8-2.2 um —vs. 4.8-6 x 1.5-1.8 um in E. kochiana)

that are sometimes oblong to broadly ellipsoid (Rappaz 1987). However that

variety still differs in having asci with a clearly amyloid subapical ring and

narrower, yellowish ascospores. The above collection of E. kochiana var. salsolae

does suggest that at least two different Eutypella species do occur on Salsola

vermiculata, whereas E. phaeospora is the only Eutypella species known thus

far to occur on Suaeda vera.

Based on many occurrences of E. phaeospora on S. vera in salt marshes

in Ile de Ré and absence inland, it was first assumed that the new species was

restricted to coastal environments. However, the material collected in Spain

from a very different environment —a semi-arid area far inland— points

towards a disjunct distribution not correlated with salt marshes. The fact that

Salsola vermiculata is an invasive shrub that can thrive in both coastal wetlands

and inland semi-arid areas may explain this apparent discrepancy. ‘Therefore,

it could be assumed that Eutypella phaeospora exhibits a strong preference

for Chenopodiaceae species regardless of ecological niche. Occurrence of

E. phaeospora on S. vermiculata in salt marshes is predictable but should be

confirmed by further sampling in coastal areas. Whether E. phaeospora occurs

on other members of Chenopodiaceae or possibly on other shrubs is likewise

still unknown, and it is hoped the morphological characterization given above

will help elucidate its apparently complex ecology.

Acknowledgements

The authors gratefully acknowledge Dr. Frangois Rappaz (Switzerland), Dr. Shaun

Pennycook (New Zealand), and Prof. Jack D. Rogers (USA) for their remarks and

valuable suggestions to improve the text, Michel Hairaud and Jean Paul Priou (France)

446 ... Fournier & Lechat

for help with collecting, and Dr. Marc Stadler (Germany) and Prof. Kevin D. Hyde

(Thailand) for help with literature.

Literature cited

Acero FJ, Gonzalez V, Sanchez-Ballesteros J, Rubio V, Checa J, Bills GF, Salazar O, Platas G, Peldez

EF, 2004. Molecular phylogenetic studies on the Diatrypaceae based on rDNA-ITS sequences.

Mycologia 96(2): 249-259. http://dx.doi.org/10.2307/3762061

Barata M. 2002. Fungi on the halophyte Spartina maritima in salt marshes. 179-193, in: KD Hyde

(ed.). Fungi in Marine Environments. Fungal Diversity Research Series 7.

Carmaran CC, Romero AI, Giussani LM. 2006. An approach towards a new phylogenetic

classification in Diatrypaceae. Fungal Diversity 23: 67-87.

Gessner RV. 1977. Seasonal occurrence and distribution of fungi associated with Spartina alterniflora

from Rhode Island estuary. Mycologia 69: 479-491. http://dx.doi.org/10.2307/3758551

Gessner RV, Kohlmeyer J. 1976. Geographical distribution and taxonomy of fungi from salt marsh

Spartina. Canadian Journal of Botany 54: 2023-2037. http://dx.doi.org/10.1139/b76-216

Kohlmeyer J, Volkmann-Kohlmeyer B, Eriksson O. 1995. Fungi on Juncus roemerianus. 2. New

dictyosporous ascomycetes. Botanica Marina 38: 165-174.

http://dx.doi.org/10.1515/botm.1995.38.1-6.165

Rappaz F. 1987. Taxonomie et nomenclature des Diatrypacées a4 asques octosporés, Mycologia

Helvetica 2(3): 285-648.

Rappaz E. 1992. Anthostoma decipiens et sa position systématique. Mycologia Helvetica 5: 21-32.

Rossman AY, Samuels GJ, Rogerson CT, Lowen R. 1999. Genera of Bionectriaceae, Hypocreaceae

and Nectriaceae (Hypocreales, Ascomycetes). Studies in Mycology 42: 1-248.

MY COTAXON

http://dx.doi.org/10.5248/118.447

Volume 118, pp. 447-453 October-December 2011

New combinations in Lactifluus. 1. L. subgenera Edules,

Lactariopsis, and Russulopsis

A. VERBEKEN’ , J. NUYTINCK' & B. BUYCK?

'Ghent University, Department of Biology, Research Group Mycology,

K.L. Ledeganckstraat 35, B-9000 Gent, Belgium

?Muséum National d’Histoire Naturelle, Dépt. Systématique et Evolution,

CP39, UMR7205, 12 Rue Buffon, F-75005 Paris, France

*CORRESPONDENCE TO: mieke.verbeken@ugent.be

AsBsTRACT — In this first of a series of three papers, new combinations in the genus

Lactifluus are proposed. This paper treats the subgenera Edules, Lactariopsis, and Russulopsis

(all proposed here as new combinations in Lactifluus). In Lactifluus subg. Edules, eight

combinations at species level are proposed. In Lactifluus subg. Lactariopsis, the following

three new combinations are proposed at sectional level: Lactifluus sect. Lactariopsis with

seven newly combined species, L. sect. Chamaeleontini with eight newly combined species,

and L. sect. Albati with four newly combined species plus two species previously combined

in Lactifluus. Finally, in L. subg. Russulopsis, eight new combinations at species level are

proposed.

Key worps — milkcaps, nomenclature

Introduction

Multigene-based phylogenies of the Russulaceae show that Lactarius and

Russula, described by Persoon in 1797 and 1796 respectively, are not supported

as two monophyletic clades (Buyck et al. 2008). Russula appears to be

monophyletic only when a small group of species (R. subsect. Ochricompactae)

is removed. This small group of species, which forms a clade where Lactarius

and Russula are mixed, was described recently as the new genus Multifurca

Buyck & V. Hofst. (Buyck et al. 2008).

Apart from the representatives that belong to Multifurca, Lactarius divides

into two clades, a larger and a smaller one, and splitting the genus seems a

better solution than lumping everything in a giant genus Russula.

In a proposal to conserve Lactarius with a conserved type (Buyck et al.

2010), arguments were given to justify the conservation of Lactarius torminosus

448 ... Verbeken, Nuytinck & Buyck

(Schaeff.: Fr.) Pers. as the type of Lactarius. This has the advantage of retaining

the name Lactarius for the larger clade, so that only the smaller clade (containing

20% of the species) would require new combinations. Since most of the species

in the smaller clade are tropical, almost all temperate milkcaps would remain

in Lactarius.

The subgenera Lactarius subg. Piperites, Lactarius subg. Russularia, and

Lactarius subg. Plinthogalus would constitute the larger genus Lactarius

sensu novo, while the former subgenera Lactarius subg. Lactarius, Lactarius

subg. Lactariopsis, Lactarius subg. Russulopsis, Lactarius subg. Lactifluus, and

Lactarius subg. Gerardii, and the former section Lactarius sect. Edules would

constitute the genus Lactifluus (Pers.) Roussel.

The proposal (Buyck et al. 2010) was supported by the Nomenclatural

Committee for Fungi (Norvell 2011) and has been approved by the General

Committee and accepted by the 2011 International Botanical Congress (Barrie

2011, McNeill et al. 2011). This first paper proposes new combinations in

Lactifluus, recombinations of species from the subgenera Lactariopsis and

Russulopsis, and the section Edules.

Taxonomy

Lactifluus subg. Edules

Molecular phylogenies show that Lactarius sect. Edules is a well-supported,

monophyletic clade (Buyck et al. 2008). It is preferable to treat it at the subgenus

rank. Currently, this group consists of eight species, all endemic for tropical

Africa.

Lactifluus subg. Edules (Verbeken) Verbeken, comb. nov.

MycoBank MB 563158

= Lactarius section Edules Verbeken, Belg. J. Bot. 132: 176. 2000 (“1999”).

TYPE SPECIES: Lactarius edulis Verbeken & Buyck

Lactifluus aureifolius (Verbeken) Verbeken, comb. nov.

MycoBank MB 563150

= Lactarius aureifolius Verbeken, Bull. Jard. Bot. Belg. 65: 198. 1996.

Lactifluus densifolius (Verbeken & Karhula) Verbeken, comb. nov.

MycoBank MB 563151

= Lactarius densifolius Verbeken & Karhula, Bull. Jard. Bot. Belg. 65: 200. 1996.

Lactifluus edulis (Verbeken & Buyck) Buyck, comb. nov.

MycoBank MB 563152

= Lactarius edulis Verbeken & Buyck, Champ. Comest. Ouest Burundi: 103. 1994.

Lactifluus inversus (Gooss.-Font. & R. Heim) Verbeken, comb. nov.

MycoBank MB 563153

= Lactarius inversus Gooss.-Font. & R. Heim, Bull. Jard. Bot. Etat 25: 78. 1955.

Lactifluus combs. nov. and new subgenera ... 449

Lactifluus latifolius (Gooss.-Font. & R. Heim) Verbeken, comb. nov.

MycoBAnk MB 563154

= Lactarius latifolius Gooss.-Font. & R. Heim, Bull. Jard. Bot. Etat 25: 82. 1955.

Lactifluus nodosicystidiosus (Verbeken & Buyck) Buyck, comb. nov.

MycoBank MB 563155

= Lactarius nodosicystidiosus Verbeken & Buyck, Mycol. Res. 111: 794. 2007.

Lactifluus phlebophyllus (R. Heim) Buyck, comb. nov.

MycoBank MB 563156

= Lactarius phlebophyllus R. Heim, Candollea 7: 378. 1938.

Lactifluus roseolus (Verbeken) Verbeken, comb. nov.

MycoBAnk MB 563157

= Lactarius roseolus Verbeken, Bull. Jard. Bot. Belg. 65: 207. 1996.

Lactifluus subg. Lactariopsis

Three sections are recognized in this subgenus. One, L. sect. Lactariopsis, is

characterized by species with a distinct veil resulting from pseudoangiocarpic

development (Heim 1937) and is known only in tropical Africa and South

America. Lactifluus sect. Chamaeleontini also has its major distribution in

tropical Africa but contains some Asian species. Lactifluus sect. Albati is

mainly temperate with representatives from North America and Europe,

but some Asian species are also placed in this group. The molecular analyses

contradict the monophyly of L. subgenus Lactariopsis. Lactifluus sect. Albati is

an evolutionarily distinct group that is not closely related to L. sect. Lactariopsis

and L. sect. Chamaeleontini. The monophyly of both latter sections remains to

be further investigated. We provisionally maintain the traditional division of L.

subg. Lactariopsis in three sections here for practical reasons.

Lactifluus subg. Lactariopsis (Henn.) Verbeken, comb. nov.

MycoBank MB 563214

= Lactariopsis Henn., Bot. Jahrb. Syst. 30: 51. 1901.

= Lactarius sect. Lactariopsis (Henn.) Singer, Ann. Mycol

40: 111. 1942 (as “Lactariopsideae”).

= Lactarius subg. Lactariopsis (Henn.) R. Heim, Prodr.

Fl. Mycologique Madagascar 1: 36. 1938.

TYPE SPECIES: Lactariopsis zenkeri Henn.

Lactifluus sect. Lactariopsis Verbeken, sect. nov.

MycoBank MB 563215

Velum praesens distinctumque, annulum formans. Pileipellis et stipitipellis cum pilis

crassiparietini abundantes. Pseudopleurocystidia abundantia, fusiformia, emergentia.

TYPE SPECIES: Lactariopsis zenkeri Henn.

450 ... Verbeken, Nuytinck & Buyck

Lactifluus annulatoangustifolius (Beeli) Buyck, comb. nov.

MycoBank MB 563170

= Russula annulatoangustifolia Beeli, Bull. Jard. Bot. Etat 14: 87. 1936.

= Lactarius annulatoangustifolius (Beeli) Buyck, Bull. Jard. Bot. Belg. 59: 241. 1989.

Lactifluus annulifer (Singer) Nuytinck, comb. nov.

MycoBank MB 563171

= Lactarius annulifer Singer, Beih. Nova Hedwigia 77: 290. 1983.

Lactifluus heimii (Verbeken) Verbeken, comb. nov.

MycoBank MB 563172

= Lactarius heimii Verbeken, Bull. Jard. Bot. Belg. 65: 201. 1996.

Lactifluus neotropicus (Singer) Nuytinck, comb. nov.

MycoBank MB 563173

= Lactarius neotropicus Singer, Kew Bull. 7: 299. 1952.

Lactifluus pelliculatus (Beeli) Buyck, comb. nov.

MycoBANnkK MB 563174

= Armillaria pelliculata Beeli, Bull. Soc. Roy. Bot. Belgique 59: 111. 1927.

= Lactarius pelliculatus (Beeli) Buyck, Bull. Jard. Bot. Belg. 59: 142. 1989.

Lactifluus velutissimus (Verbeken) Verbeken, comb. nov.

MycoBank MB 563175

= Lactarius velutissimus Verbeken, Bull. Jard. Bot. Belg. 65: 212. 1996.

Lactifluus zenkeri (Henn.) Verbeken, comb. nov.

MycoBank MB 563176

= Lactariopsis zenkeri Henn., Bot. Jahrb. Syst. 30: 51. 1902 (“1901”).

= Lactarius zenkeri (Henn.) Singer, Ann. Mycol. 40: 111. 1942.

Lactifluus sect. Chamaeleontini (Verbeken) Verbeken, comb. nov.

MycoBank MB 563216

= Lactarius section Chamaeleontini Verbeken, Mycotaxon 66: 393. 1998.

TYPE SPECIES: Lactarius chamaeleontinus R. Heim

Lactifluus chamaeleontinus (R. Heim) Verbeken, comb. nov.

MycoBank MB 563217

= Lactarius chamaeleontinus R. Heim, Bull. Jard. Bot. Etat 25: 87. 1955.

Lactifluus emergens (Verbeken) Verbeken, comb. nov.

MycoBank MB 563218

= Lactarius emergens Verbeken, Syst. Geogr. Pl. 70(1): 192. 2000.

Lactifluus indusiatus (Verbeken) Verbeken, comb. nov.

MycoBank MB 563219

= Lactarius indusiatus Verbeken, Bull. Jard. Bot. Belg. 65: 201. 1996.

Lactifluus laevigatus (Verbeken) Verbeken, comb. nov.

MycoBank MB 563220

= Lactarius laevigatus Verbeken, Bull. Jard. Bot. Belg. 65: 203. 1996.

Lactifluus combs. nov. and new subgenera ...

Lactifluus leoninus (Verbeken & E. Horak) Verbeken, comb. nov.

MycoBank MB 563221

= Lactarius leoninus Verbeken & E. Horak, Austr. Syst. Bot. 12: 775. 1999.

Lactifluus madagascariensis (Verbeken & Buyck) Buyck, comb. nov.

MycoBank MB 563222

= Lactarius madagascariensis Verbeken & Buyck, Mycol. Res. 111: 791. 2007.

Lactifluus pruinatus (Verbeken & Buyck) Verbeken, comb. nov.

MycoBank MB 563223

= Lactarius pruinatus Verbeken & Buyck, Mycotaxon 66: 399. 1998.

Lactifluus sesemotani (Beeli) Buyck, comb. nov.

MycoBank MB 563224

= Russula sesemotani Beeli, Bull. Soc. Roy. Bot. Belgique 60(2): 169. 1928.

= Lactarius sesemotani (Beeli) Buyck, Bull. Jard. Bot. Belg. 59: 241. 1989.

Lactifluus sect. Albati (Bataille) Verbeken, comb. nov.

MycoBank MB 563735

= Lactarius [unranked] Albati Bataille, Fl. Monogr. Astéro.: 35. 1908.

= Lactarius subsect. Albati (Bataille) Konrad, Bull. Soc. Mycol. France 51: 188. 1935.

= Lactarius sect. Albati (Bataille) Singer, Ann. Mycol 40: 109. 1942.

TYPE SPECIES: Agaricus vellereus Fr. : Fr.

Lactifluus bertillonii (Neuhoff ex Z. Schaef.) Verbeken, comb. nov.

MycoBank MB 563226

= Lactarius vellereus var. bertillonii Neuhoff ex Z. Schaef., Ceska Mykol. 33: 9. 1979.

= Lactarius bertillonii (Neuhoff ex Z. Schaef.) Bon, Doc.

Mycol. 10(37-38): 92. 1980 (“1979”).

Lactifluus deceptivus (Peck) Kuntze, Revis. Gen. Pl. 2: 856. 1891.

= Lactarius deceptivus Peck, Annual Rep. New York State Mus. 38: 125. 1885.

Lactifluus pilosus (Verbeken, H.T. Le & Lumyong) Verbeken, comb. nov.

MycoBank MB 563227

= Lactarius pilosus Verbeken, H.T. Le & Lumyong, Mycotaxon 102: 287. 2007.

Lactifluus puberulus (H.A. Wen & J.Z. Ying) Nuytinck, comb. nov.

MycoBank MB 563228

= Lactarius puberulus H.A. Wen & J.Z. Ying, Mycosystema 24(2): 155. 2005.

Lactifluus subvellereus (Peck) Nuytinck, comb. nov.

MycoBank MB 563229

= Lactarius subvellereus Peck, Bull. Torrey Bot. Club 25: 369. 1898.

Lactifluus vellereus (Fr. : Fr.) Kuntze, Revis. Gen. Pl. 2: 857. 1891.

= Agaricus vellereus Fr. : Fr., Syst. Mycol. 1: 76. 1821.

= Lactarius vellereus (Fr. : Fr.) Fr., Epicr. Syst. Mycol.: 340. 1838.

451

452 ... Verbeken, Nuytinck & Buyck

Lactifluus subg. Russulopsis

Currently, this group comprises eight species, all endemic for tropical Africa.

All species are recombined in the genus Lactifluus here.

Lactifluus subg. Russulopsis (Verbeken) Verbeken, comb. nov.

MycoBank MB 563736

= Lactarius subg. Russulopsis Verbeken, Mycotaxon 77: 439. 2001.

TYPE SPECIES: Lactarius ruvubuensis Verbeken

Lactifluus sect. Russulopsidei (Verbeken) Verbeken, comb. nov.

MycoBank MB 563737

= Lactarius sect. Russulopsidei Verbeken, Mycotaxon 77: 440. 2001.

TYPE spEcIEs: Lactarius ruvubuensis Verbeken

Lactifluus brachystegiae (Verbeken & C. Sharp) Verbeken, comb. nov.

MycoBank MB 563230

= Lactarius brachystegiae Verbeken & C. Sharp, Syst. Geogr. Pl. 70(1): 187. 2000.

Lactifluus claricolor (R. Heim) Verbeken, comb. nov.

MycoBank MB 563231

= Lactarius claricolor R. Heim, Candollea 7: 381. 1938.

Lactifluus corbula (R. Heim & Gooss.-Font.) Verbeken, comb. nov.

MycoBank MB 563232

= Lactarius corbula R. Heim & Gooss.-Font., Bull. Jard. Bot. Etat 25: 64. 1955.

Lactifluus cyanovirescens (Verbeken) Verbeken, comb. nov.

MycoBank MB 563233

= Lactarius cyanovirescens Verbeken, Bull. Jard. Bot. Belg. 65: 199. 1996.

Lactifluus longipes (Verbeken) Verbeken, comb. nov.

MycoBank MB 563234

= Lactarius longipes Verbeken, Bull. Jard. Bot. Belg. 65: 203. 1996.

Lactifluus pseudotorminosus (R. Heim) Verbeken, comb. nov.

MycoBank MB 563235

= Lactarius pseudotorminosus R. Heim, Candollea 7: 379. 1938.

Lactifluus ruvubuensis (Verbeken) Verbeken, comb. nov.

MycoBank MB 563236

= Lactarius ruvubuensis Verbeken, Bull. Jard. Bot. Belg. 65: 208. 1996.

Lactifluus urens (Verbeken) Verbeken, comb. nov.

MycoBank MB 563237

= Lactarius urens Verbeken, Bull. Jard. Bot. Belg. 65: 211. 1996.

Acknowledgments

The authors acknowledge Lorelei Norvell, Shaun Pennycook, and Scott Redhead for

valuable comments.

Lactifluus combs. noy. and new subgenera ... 453

Literature cited

Barrie FR. 2011. Report of the General Committee: 11. Taxon 60: 1211-1214.

Buyck B, Hofstetter V, Eberhardt U, Verbeken A, Kauff F. 2008. Walking the thin line between

Lactarius and Russula: the dilemma of Russula sect. Ochricompactae. Fungal Diversity 28:

15-40.

Buyck B, Hofstetter V, Verbeken A, Walleyn R. 2010. Proposal to conserve Lactarius nom. cons.

(Basidiomycota) with a conserved type. Taxon 59(1): 295-296.

Heim R. 1937. Observations sur la flore mycologique malgache V. Les Lactario-Russulés a anneau:

ontogénie et phylogénie. Rev. Mycol. (Paris) 2: 4-17, 61-75, 109-117.

McNeill J, Turland NJ, Monro AM, Lepschi BJ. 2011. XVII International Botanical Congress:

Preliminary mail vote and report of Congress action on nomenclature proposals. Taxon 60:

1507-1520.

Norvell LL. 2011. Report of the Nomenclature Committee for Fungi: 16. Taxon 60: 223-226.

MY COTAXON

http://dx.doi.org/10.5248/118.455

Volume 118, pp. 455-458 October-December 2011

Validation of combinations with basionyms published by Fries

in 1861

Scott A. REDHEAD’, JOSEPH F. AMMIRATI’, LORELEI L. NORVELL’,

ALFREDO VIZZINI* & MARCO CONTU®

' National Mycological Herbarium, Eastern Cereal & Oilseed Research Centre C.E.F.,

Agriculture & Agri-Food Canada, Ottawa, Ontario, Canada, K1A 0C6

? Department of Biology, 351330, University of Washington, Seattle, WA 98195 USA

> Pacific Northwest Mycology Service,

6720 NW Skyline Boulevard, Portland, OR 97229-1309 USA

* 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

* CORRESPONDENCE TO: scott.redhead@agr.gc.ca

Axstract — Authors (including some of us) have incorrectly cited as basionyms names

treated by Fries in 1863 that were actually originally published by him in 1861. As these

basionym citation errors mean that the intended new combinations are not validly published,

the following combinations are again proposed as new: Chromosera cyanophylla, Mythicomyces

corneipes, Tephrocybe misera, T. tesquorum. Three other intended combinations are noted as

also not validly published, but the species are currently treated under the different (and validly

published) names Haasiella venustissima, Phaeoclavulina curta, and Rhodonia placenta.

KEY worps — Ceriporiopsis, Gerronema, Ramaria, International Code of Botanical

Nomenclature

Introduction

Fries (1861) published a series of observations on new or little known

hymenomycetes from Sweden. Most of his new taxa were included again under

the same names in his MONOGRAPHIA HYMENOMYCETUM SUECIAE (Fries

1863). In his subsequently published HYMENOMYCETES EuROPAEI (Fries 1874),

Fries referred back only to the 1863 publication for each of the names originally

published in 1861. This has led to a series of citation errors by later authors and

indices, some of which affect the valid publication of proposed combinations.

456 ... Redhead & al.

In 1863, Fries did refer back to the 1861 publication as the source for

each of the names corrected below. Therefore the authors, who proposed the

new combinations after 1 January 1953 and who cited only the 1863 or later

publications, violated Articles 33.4 and 33.7 of the INTERNATIONAL CODE OF

BOTANICAL NOMENCLATURE (McNeill et al. 2006) and did not validly publish

their new combinations.

Under Art. 33.5 & 33.7(a), such citation errors are not correctable. Here we

validate the previously proposed binomials that are now generally accepted and

in current use, despite their current lack of status under the CopE (McNeill

et al. 2006). With one exception in Haasiella, we are unaware of incidental

subsequent validations.

Chromosera cyanophylla (Fr.) Redhead, Ammirati & Norvell, comb. nov.

MycoBank MB 563787

= Agaricus cyanophyllus Fr., Ofvers. K. Svensk. Vetensk.-

Akad. Forhandl. 18(1): 23 (1861).

Redhead et al. (1995) proposed the genus Chromosera Redhead, Ammirati &

Norvell, citing as type “Chromosera cyanophylla” and listing Fries (1863) (and

not the earlier Fries 1861) for the basionym. They did, however, list as obligate

synonyms three validly published names: Agaricus cyanophyllus Fr. (albeit with

the incorrect citation), Omphalia cyanophylla (Fr.) Quél. (Quélet 1872: 99),

and Omphalina cyanophylla (Fr.) Quél. (Quélet 1886: 45). This indication of

the type fulfilled the requirements for valid publication of the generic name

(Art. 37.2), but the incorrect citation did not meet the requirements for valid

publication of the binomial. Notably, the requirements for valid publication of

new combinations prior to 1953 were far more lenient.

Mythicomyces corneipes (Fr.) Redhead & A.H. Sm. comb. nov.

MycoBank MB 563788

= Agaricus corneipes Fr., Ofvers. K. Svensk. Vetensk.-Akad. Férhandl. 18(1): 25 (1861).

Similarly, Redhead & Smith (1986) proposed the genus Mythicomyces Redhead &

A.H. Sm., citing as type “Mythicomyces corneipes (Fries) comb. nov.’ They listed

as obligate synonyms two validly published names: the first, Agaricus corneipes

Fr., incorrectly cited Fries (1863), and the second was Psilocybe corneipes (Fr.)

P. Karst. (Karsten 1879: 504). These actions fulfilled the requirements for valid

publication of the generic name (Art. 37.2) but failed to meet requirements for

valid publication of the binomial, and so we publish it here.

Tephrocybe misera (Fr.) M.M. Moser ex Contu & Vizzini comb. nov.

MycoBank MB 563789

= Agaricus miser Fr., Ofvers. K. Svensk. Vetensk.-Akad. Férhandl. 18(1): 21 (1861).

Validation of Chromosera, Mythicomyces & Tephrocybe spp. ... 457

Tephrocybe tesquorum (Fr.) M.M. Moser ex Contu & Vizzini, comb. nov.

MycoBank MB 563790

= Agaricus tesquorum Fr., Ofvers. K. Svensk. Vetensk.-Akad. Férhandl. 18(1): 22 (1861).

Moser (1967) proposed two combinations in Tephrocybe but cited Fries (1863)

rather than (1861).

Notes on other cases of names not being validly published

Haasiella venustissima (Fr.) Kotl. & Pouzar ex Chiaffi & Surault, Bull. Soc. Mycol.

Fr. 112: 127 (1996).

= Agaricus venustissimus Fr., Ofvers. K. Svensk. Vetensk.-

Akad. Forhandl. 18(1): 21 (1861).

Kotlaba & Pouzar (1966) erected the genus Haasiella Kotl. & Pouzar, typified

by H. splendidissima Kotl. & Pouzar, and included a second species, for which

they proposed the combination “Haasiella venustissimus,” citing Fries (1863)

for the basionym. Their combination was inadvertently validated by Chiafhi &

Surault (1996), who cited the basionym and correct place of publication. The

proposed combination “Gerronema venustissimum” by Singer (1962) is not

validly published, as only Fries (1863) was cited.

Phaeoclavulina curta (Fr.) Giachini, Mycotaxon 115: 190 (2011).

= Clavaria curta Fr., Ofvers. K. Svensk. Vetensk.-Akad. Férhandl. 18(1): 31 (1861).

The intended combination in Ramaria by Schild (1994) was not validly

published because he referred only to Fries (1863) “Monogr. Hym. Suec.,”

which he mistakenly cited as “1857,” the date of the earlier parts of that title.

Giachini & Castellano (2011) correctly cited Fries (1861).

Rhodonia placenta (Fr.) Niemela, K.H. Larss. & Schigel, Karstenia 45(2): 79 (2005).

= Polyporus placenta Fr., Ofvers. K. Svensk. Vetensk.-Akad. Férhandl. 18(1): 30 (1861).

Domanski’s (1963) intended combination in Ceriporiopsis Domanski, where he

cites Fries (1874), was not validly published. Niemela et al. (2005) correctly

cited Fries (1861).

Acknowledgements

We thank Shaun Pennycook and John McNeill for reviewing the manuscript and

confirming that the genus Chromosera was validly published according to the ICBN even

though the earlier combination of Chromosera cyanophylla was not validly published.

Literature cited

Chiafhi M, Surault J-L. 1996. Une espéce rare et remarquable, Haasiella venustissima (Fr.) Kotl. &

Pouz. Bull. Soc. Mycol. France 112: 127-135.

Domaniski S. 1963. Dwa nowe rodzaje grzyboéw z grupy “Poria Pers. ex S.F. Gray”. Acta Soc. Bot.

Poloniae 32: 731-739.

458 ... Redhead & al.

Fries EM. 1861. Hymenomycetes novi vel minus cogniti, in Suecia 1852-1860 observati. Ofvers. K.

VetenskAkad. Forh. 18(1): 19-34.

Fries EM. 1863. Monographia Hymenomycetum Sueciae 2: [147]-355. Uppsala, CA Leffler.

Fries EM. 1874. Hymenomycetes Europaei sive Epicriseos systematis mycologici. Editio altera.

Upsaliae, E. Berling.

Giachini, AJ, Castellano MA. 2011. A new taxonomic classification for species in Gomphus sensu

lato. Mycotaxon 115: 183-201. http://dx.doi.org/10.5248/115.183

Karsten PA. 1879. Rysslands, Finlands och den Skandinaviska half6ns Hattsvampar. Forra Delen:

Skifsvampar. Bidr. Kann. Finl. Nat. Folk 32. XXVIII + 571 p.

Kotlaba F, Pouzar Z. 1966. Haasiella, a new agaric genus and H. splendidissima sp. nov. : Ceska

Mykologie 20: 135-140.

McNeill J, Barrie FF, Burdet HM, Demoulin V, Hawksworth DL, Marhold K, Nicolson DH, Prado

J, Silva PC, Skog JE, Wiersema J, Turland NJ (eds.) 2006. International code of botanical

nomenclature (Vienna Code). Adopted by the Seventeenth International Botanical Congress

Vienna, Austria, July 2005. Reg. Veget. 146: i-xvi, 1-568.

Moser M. 1967. Basidiomyceten II. Teil. Die ROhrlinge und Blatterpilze (Agaricales). Kleine

Kryptogamenflora, ed. 3, IIb/2. 443 p.

Niemela T, Kinnunen J, Larsson K-H, Schigel DD, Larsson E. 2005. Genus revisions and new

combinations of some North European polypores. Karstenia 45: 75-80.

Quélet L. 1872. Les champignons du Jura et des Vosges. Mém. Soc. Emul. Montbéliard, sér. II, 5:

43-332.

Queélet L. 1886. Enchiridion fungorum in Europa media et prasertim in Gallia vigentium. Lutetiae.

a5 2}

Redhead SA, Smith AH. 1986. Two new genera of agarics based on Psilocybe corneipes and

Phaeocollybia perplexa. Canad. Jour. Bot. 64: 643-647. http://dx.doi.org/10.1139/b86-082

Redhead SA, Ammirati JF, Norvell LL. 1995. Omphalina sensu lato in North America 3: Chromosera

gen. nov. Beih. Sydowia 10: 155-167.

Schild E. 1994. Ramaria-Studien Clavaria curta Fries (1857) eine eigene Art. Z. Mykol. 60: 123-130.

Singer R. 1962 (1961). Diagnoses fungorum novorum Agaricalium II. Sydowia 15: 45-83.

MYCOTAXON

http://dx.doi.org/10.5248/118.459

Volume 118, pp. 459-462 October-December 2011

BOOK REVIEWS AND NOTICES

ELSE C. VELLINGA, Book Review Editor*

861 Keeler Avenue, Berkeley CA 94708 U.S.A.

CORRESPONDENCE TO: bookreviews@mycotaxon.com

INTRODUCTION

Three totally different books are reviewed here: a beautifully illustrated

French book on myxomycetes, an Indian book on the smut fungi of the

subcontinent, and a well-written, popular-science introduction to mushroom

forming fungi, succinctly titled ‘Mushroom,

Book announcements include the 2011 publications in the series ‘FUNGI

NON DELINEATI, a very nicely illustrated book on tropical fungi in China, anda

checklist for all fungi and myxomycetes of Rhode Island (U.S.A.).

MYXOMYCETES

Les myxomycetes. By M. Poulain, M. Meyer & J. Bozonnet, 2010. FMBDS, 8,

Avenue de la Plaine, 74000 Annecy, France, <philippecattin74@orange.fr>. 2 vols,

pp. 1119, col. plates 544. ISBN 978-2-9518540-2-4. Price 159.00 €

As anyone who has ever studied myxomycetes knows, the monograph by Martin

and Alexopoulos, published by the University of Iowa Press in 1969, represents

the single most definitive treatment of this group of organisms published to date.

This monograph (THE MyxomyceteEs) has long since been out of print, but it

still remains—more than 40 years after it first appeared—the standard against

which any of the more recently published works on the myxomycetes are judged.

The two volumes that make up Les Myxomycétes undoubtedly represent the

most noteworthy recent contribution to the study of myxomycetes. Although

not totally comprehensive, since some species (particularly tropical examples)

are not included, this work covers virtually any species one is likely to encounter

“Books for consideration for coverage in this column should be mailed to the Book Review Editor

at the address above. All unsigned entries are by the Book Review Editor.

460 ... Vellinga, BOOK REVIEW EDITOR

during a lifetime of collecting. Information is provided on 853 taxa (species

and varieties), and 530 of these are illustrated with color images. These images

represent the most important feature of the work itself. Never before have so

many high-quality (often stunning) macroscopic color images been provided

for so many different myxomycetes. For anyone studying myxomycetes, this

alone is reason enough to obtain a copy of Les Myxomycetes. The first of the

two volumes begins with a short introduction to the biology, ecology and

morphology of myxomycetes. Since the primary language used throughout the

work is French, this information is not readily available to everyone. However,

the remainder of the first volume is devoted to keys and short descriptions

of the species being considered, and this material is presented in both French

and English, which is a major plus for those of us whose knowledge of the

former language is limited. The descriptions include both macroscopic as well

as microscopic characters, and reference is made to the number of the image

provided for a particular species in the second volume. ‘The first volume also

contains a glossary of terms, an index to all of the taxa being considered and a

list of relevant bibliographical references. The nomenclature used throughout

both volumes is that found in the current version of Nomenmyx (www.nomen.

eumycetozoa.com), which is generally followed by most of us who work with

myxomycetes.

The second volume contains the color images of the 530 taxa. The general

approach used throughout is to have each page devoted to a single image that

shows the habit of the species in question, but some pages contain several

images of the same species. Also included are line drawings that show various

microscopic structures (e.g., spores, capillitium and peridium) that are relevant

to that species. Altogether, the images, drawings and descriptions provide

enough information to allow most specimens of myxomycetes to be identified

with a high degree of certainty. Since the authors are especially well known

for their studies of nivicolous (“snowbank”) myxomycetes, there is extensive

coverage of this group. In fact, information on some of the nivicolous taxa

they have included in this work was available previously only in the primary

literature. The majority of the color images are truly outstanding, and the only

complaint that anyone might have is that in a few instances (e.g., Physarum

pusillum) the specimen photographed might not have been absolutely typical

for a given species. I found the images of the various species of Licea particularly

well done and thus potentially very useful when working with members of this

genus.

Overall, Les Myxomycétes is an extraordinary work that is the result of an

enormous amount of time and effort on the part of the three authors. Simply

looking through the color images in the second volume is enjoyable, but

the complementary information provided in the first volume makes this an

MycotTaxon 118 Book Reviews ... 461

exceedingly useful scientific work on the myxomycetes. Anyone with even the

slightest interest in this group of organisms should have a copy of this work.

STEVEN L. STEPHENSON

Department of Biological Sciences, University of Arkansas

Fayetteville, Arkansas 72701, U.S.A.

slsteph@uark.edu

USTILAGINALES

Ustilaginales of India. By R.V. Gandhe, 2011. Bishen Singh Mahendra Pal Singh,

23-A, New Connaught Place, Dehra Dun, 248001 India, <bsmps@vsnl.com>. ISBN

978-81-211-0788-4. Pp. 414, col. pl. 22, figs. Price circa US$ 154

Dedicated to ‘the doyen of mycology in India the late Professor M_J.

Thirumalachar, this book gives an overview of the Ustilaginales (s.l.) species

in India.

The economically important smut fungi of crop plants and the smut fungi

of native plants are treated in this book. It gives descriptions of 343 species in

45 genera, several of them new for science (e.g., three new Sorosporium species

and two new Sphacelotheca species), the genera and species per genus are

treated in alphabetical order. The genus Sporisorium is the largest, represented

by 96 species, followed by Tilletia with 46, and Ustilago with 42.

Besides an introduction to smut fungi and coverage of their economic

importance, diversity, and morphological variations, a useful host index is

provided.

The author, who has spent 2 decades studying smut fungi and especially

their germination patterns, places a strong emphasis on morphotaxonomy and

symptomatology, and although in many cases descriptions are literally the same

as in Vanky (2007), morphological characters have been added to the keys. The

illustrations entail eight colour plates and over 50 drawings of infected plants,

spores, and germinating spores.

The book makes the impression of having been written at the same time as

Vanky’s treatment of the smut fungi of the Indian subcontinent was published,

although that publication is lacking from the list of references. There are no

post-2007 publications cited, not even the phylogenetic paper by Begerow et

al. (2007). The illustrations in Vanky’s 2007 work are of higher quality than the

ones in the present book, but I doubt whether the former is easily available in

India, where it should be used.

Begerow D, Stoll M, Bauer R. 2007 ['2006’] A phylogenetic hypothesis of

Ustilaginomycotina based on multiple gene analyses and morphological data.

Mycologia 98: 906-916.

Vanky, K. 2007. Smut fungi of the Indian Subcontinent. Polish Botanical Studies 26.

462 ... Vellinga, BOOK REVIEW EDITOR

MISCELLANEOUS

Mushroom. By N.P. Money, 2011. Oxford University Press, 198 Madison Avenue,

New York, NY 10016, U.S.A. <www.oup.com>. ISBN 978-0-19-973256-2. Pp. 201,

col. pl. 16, figs. Price US$ 24.95

In eight chapters the reader is given a dazzling, fast-spaced and thorough

introduction into the development of mushrooms (including life cycles), spore

launching, diversity in shape and function, edible mushrooms and conservation,

mushroom cultivation, toxicity, intoxication, and the medicinal mushroom

industry. The text moves back and forth between the author's experiences in

the England of his youth and present-day life, science, and economics and

yet is always anchored safely in the writings and illustrations of the scientists

of the past. Central to all is the biology of the mushroom, as masterpiece of

bioengineering. It is by no means a text book or field guide. Nonetheless,

MUSHROOM gives a huge amount of information in its 200 pages, makes for

excellent reading, and would be a good present to friends, family, and all who

are intrigued by nature.

Book ANNOUNCEMENTS

Characteristic and rare species of gasteromycetes in Eupannonicum. By I. Riméczo.

M. Jeppson & L. Benedek, 2011. FUNGI NON DELINEATI 56-57. Edizioni Candusso, Via

Ottone Primo 90, 17021 Alassio SV, Italy. <maxcandusso@libero.it>. Pp. 230, col. plates

108, figs. Price 28.00 €

Cortinarius Ibero-insulares - 3. By Grupo Iberoinsular de Cortinariologos (GIC), 2011.

FUNGI NON DELINEATI 58-59. Edizioni Candusso, Via Ottone Primo 90, 17021 Alassio

SV, Italy. <maxcandusso@libero.it>. Pp. 236, col. plates 236. Price 33.00 €

Fungi of tropical China. By W. Xingliang et al., 2011. Chinese Corporation for

Promotion of Humanities. ISBN-13: 9787030294708. Pp. 548, col. plates 495. Price circa

US$ 220

Inocybe dai litorali alla zona alpina. By E. Ferrari, 2011. FUNGI NON DELINEATI 54-55.

Edizioni Candusso, Via Ottone Primo 90, 17021 Alassio SV, Italy. <maxcandusso@libero.

it>. Pp. 216, col. plates 106, figs 69. Price 23.00 €

Mycota of Rhode Island: A checklist of the fungi recorded in Rhode Island (including

lichens and myxomycetes). By R.D. Goos, 2010. THE BioTA OF RHODE ISLAND

vol. 4. Rhode Island Natural History Survey, P.O. Box 1858, Kingston, RI 02881,

<programadmin@rinhs.org>. ISBN 1-887771-09-3. Pp. 228. Price US$60.00

Rare and noteworthy species of agarics from the western Caucasus. By E.F. Malysheva

& T. Yu. Svetasheva, 2011. FUNGI NON DELINEATI 60. Edizioni Candusso, Via Ottone

Primo 90, 17021 Alassio SV, Italy. <maxcandusso@libero.it>. Pp. 104, col. plates 45.

Price 12.00 €

MYCOTAXON

http://dx.doi.org/10.5248/118.nom

Volume 118, pp. 463-465 October-December 2011

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 118

Amicodisca castaneae J.G. Han, Hosoya & H.D. Shin, p. 90

Arrasia Bernicchia, Gorjon & Nakasone, p. 258

Arrasia rostrata Bernicchia, Gorjon & Nakasone, p. 258

Aschersonia conica Jun Z. Qiu, Y.B. Su & C.S. Weng, p. 326

Asterophora salvaterrensis Blanco-Dios, p. 84

Chromosera cyanophylla (Fr.) Redhead, Ammirati & Norvell (validated), p. 456

Coccomyces minimus Y.R. Lin, C.L. Hou & GJ. Jia, p. 232

Cortinarius mikedavisii Bojantchev, p. 267

Corynespora carrisae R. Singh & Kamal, p. 124

Corynespora peristrophicola R. Singh & Kamal, p. 126

Diversispora clara Oehl, B. Estrada, G.A. Silva & Palenz., p. 75

Eremiomyces magnisporus G. Moreno, P. Alvarado, Manjon & Sanz p. 105

Eutypella phaeospora J. Fourn. & Lechat, p. 442

Exserohilum neoregeliae Sakoda & Tsukib., p. 214

Exserticlava manglietiae S.C. Ren & X.G. Zhang, p. 350

Fomitiporia apiahyna (Speg.) Vlasak & Kout, p. 161

Glomus cubense Y. Rodr. & Dalpé, p. 339

Hallenbergia Dhingra & Priyanka, p. 289

Hallenbergia singularis Dhingra & Priyanka, p. 291

Homolaphlyctis Longcore, Letcher & T.Y. James, p. 435

Homolaphlyctis polyrhiza Longcore, Letcher & TY. James, p. 436

Hymenochaete yunnanensis S.H. He & Hai J. Li, p. 412

Hypoderma siculum Lantieri, P.R. Johnst. & Medardi, p. 395

Lactifluus subg. Edules (Verbeken) Verbeken, p. 448

Lactifluus subg. Lactariopsis (Henn.) Verbeken, p. 449

Lactifluus subg. Russulopsis (Verbeken) Verbeken, p. 452

Lactifluus sect. Albati (Bataille) Verbeken, p. 451

Lactifluus sect. Chamaeleontini (Verbeken) Verbeken, p. 450

Lactifluus sect. Lactariopsis Verbeken, p. 449

464 ... MYCOTAXON 118

Lactifluus sect. Russulopsidei (Verbeken) Verbeken, p. 452

Lactifluus annulatoangustifolius (Beeli) Buyck, p. 450

Lactifluus annulifer (Singer) Nuytinck, p. 450

Lactifluus aureifolius (Verbeken) Verbeken, p. 448

Lactifluus bertillonii (Neuhoff ex Z. Schaef.) Verbeken, p. 451

Lactifluus brachystegiae (Verbeken & C. Sharp) Verbeken, p. 452

Lactifluus chamaeleontinus (R. Heim) Verbeken, p. 450

Lactifluus claricolor (R. Heim) Verbeken, p. 452

Lactifluus corbula (R. Heim & Gooss.-Font.) Verbeken, p. 452

Lactifluus cyanovirescens (Verbeken) Verbeken, p. 452

Lactifluus densifolius (Verbeken & Karhula) Verbeken, p. 2448

Lactifluus edulis (Verbeken & Buyck) Buyck, p. 448

Lactifluus emergens (Verbeken) Verbeken, p. 450

Lactifluus heimii (Verbeken) Verbeken, p. 450

Lactifluus indusiatus (Verbeken) Verbeken, p. 450

Lactifluus inversus (Gooss.-Font. & R. Heim) Verbeken, p. 448

Lactifluus laevigatus (Verbeken) Verbeken, p. 450

Lactifluus latifolius (Gooss.-Font. & R. Heim) Verbeken, p. 449

Lactifluus leoninus (Verbeken & E. Horak) Verbeken, p. 451

Lactifluus longipes (Verbeken) Verbeken, p. 452

Lactifluus madagascariensis (Verbeken & Buyck) Buyck, p. 451

Lactifluus neotropicus (Singer) Nuytinck, p. 450

Lactifluus nodosicystidiosus (Verbeken & Buyck) Buyck, p. 449

Lactifluus pelliculatus (Beeli) Buyck, p. 450

Lactifluus phlebophyllus (R. Heim) Buyck, p. 449

Lactifluus pilosus (Verbeken, H.T. Le & Lumyong) Verbeken, p. 451

Lactifluus pruinatus (Verbeken & Buyck) Verbeken, p. 451

Lactifluus pseudotorminosus (R. Heim) Verbeken, p. 452

Lactifluus puberulus (H.A. Wen & J.Z. Ying) Nuytinck, p. 451

Lactifluus roseolus (Verbeken) Verbeken, p. 449

Lactifluus ruvubuensis (Verbeken) Verbeken, p. 452

Lactifluus sesemotani (Beeli) Buyck, p. 451

Lactifluus subvellereus (Peck) Nuytinck, p. 451

Lactifluus urens (Verbeken) Verbeken, p. 452

Lactifluus velutissimus (Verbeken) Verbeken, p. 450

Lactifluus zenkeri (Henn.) Verbeken, p. 450

Lophodermium sinense Y.R. Lin, C.L. Hou & Jiang L. Chen, p. 225

NOMENCLATURAL NOVELTIES & TYPIFICATIONS ... 46 5

Marasmius tyrius B.E. Lechner & Papinutti, p. 252

Melanoleuca exscissa f. diverticulata (G. Moreno & Bon) Fontenla, Para &

Vizzini, p. 374

Melanoleuca exscissa f. iris (Kithner) Fontenla, Para & Vizzini, p. 374

Melanoleuca exscissa f. sarcophylla (Kihner) Fontenla, Para & Vizzini, p. 374

Melanoleuca paedida f. electropoda (Maire & Malencon) Fontenla, Para &

Vizzini, p. 376

Melanoleuca sublanipes Fontenla, Para & Vizzini, p. 373

Microbotryum coronariae (Liro) Denchev & T. Denchey, p. 54

Microcyclospora rumicis Arzanlou & Bakhshi, p. 182

Mycena guldeniana Aronsen & B.A. Perry, p. 190

Mythicomyces corneipes (Fr.) Redhead & A.H. Sm. (validated), p. 456

Paraphysoderma Boussiba, Zarka & T.Y. James, p. 178

Paraphysoderma sedebokerense Boussiba, Zarka & T.Y. James, p. 178

Peziza paludicola (Boud.) Sacc. & Traverso 1911 (lectotypified, epitypified), p. 117

Peziza paludicola var. kilimanjarensis (J. Moravec) Cacialli, Lantieri & Medardi, p. 120

Pleurotus giganteus (Berk.) Karunarathna & K.D. Hyde, p. 62

= Lentinus giganteus Berk. 1847 (epitypified)

Pseudocercospora epidendri Meir. Silva & O.L. Pereira, p. 2

Septobasidium saurauiae S.Z. Chen & L. Guo, p. 283

Spiropes terminaliae S.C. Ren & X.G. Zhang, p. 351

Subulicystidium curvisporum Gorjon, Gresl. & Rajchenb., p. 48

Tephrocybe misera (Fr.) M.M. Moser ex Contu & Vizzini, p. 456

Tephrocybe tesquorum (Fr.) M.M. Moser ex Contu & Vizzini, p. 457

Terriera camelliae (Teng) Y.R. Lin & Jiang L. Chen, p. 227

Terriera coacervata Y.R. Lin & Q. Zheng, p. 318

Terriera illiciicola (S.J. Wang, Y.F. He & Y.R. Lin) Q. Zheng & Y.R. Lin, p. 321

Tothia fuscella (Sacc.) Bat., in Toth 1960 (epitypified), p. 207

Tuber polyspermum L. Fan & C.L. Hou, p. 406

Tuber sinoalbidum L. Fan & J.Z. Cao, p. 408

Urocystis bolboschoeni Denchev, T. Denchev, Spooner & Legon, p. 249