Iv ... MYCOTAXON 117

MY COTAXON

VOLUME ONE HUNDRED SEVENTEEN — TABLE OF CONTENTS

COVER SECTION

PREV ICV CHEE sh abi OPTRA cae Ai ck VG ROMT ich RRM ech Teens AEH Ged escod AER es vii

SUDINISSION. BIOCCH UNOS a SE or noc ir ent eA ke te mets oes hE ae Viii

4a) eld {Bk TM OLA) oh, ieee teal Blan Aira Sabie 8 Met) Pipe oP tar RB i de Deh ean! po ix

RESEARCH ARTICLES

Cortinarius callimorphus, a new species from northern California

Dimitar Bojantchev & R. Michael Davis

A new Phylloporus from two relict Fagus grandifolia var. mexicana populations

in a montane cloud forest Leticia Montoya & Victor M. Bandala

Type studies on two species of Phylloporus (Boletaceae, Boletales) described

from southwestern China Nian-Kai Zeng, Li-Ping Tang & Zhu L. Yang

Arthrobotrys latispora, a new nematode-trapping fungus from

southwest China Hongyan Su, Shuoran Liu, Yunxia Li, Yong Hong Cao,

Minghui Chen & Xiaoyan Yang

Additional records of Volvariella dunensis (Basidiomycota, Agaricales):

morphological and molecular characterization

Alfredo Vizzini, Marco Contu & Alfredo Justo

Moelleriella pumatensis, a new entomogenous species from Vietnam

Suchada Mongkolsamrit, Tai Toan Nguyen,

Ngoc Lan Tran & J. Jennifer Luangsa-ard

Lepiota (Agaricales) in northern Thailand - 1. Lepiota section Stenosporae

P. Sysouphanthong, K.D. Hyde, E. Chukeatirote, A.H. Bahkali& E.C. Vellinga

New records of Digitoramispora from China

Yi-Dong Zhang, Jian Ma, Li-Guo Ma & Xiu-Guo Zhang

Seven new records of foliicolous lichens from Vietnam

Thi Thuy Nguyen, Yogesh Joshi, Robert Licking,

Anh Dzung Nguyen, Xin Yu Wang, Young Jin Koh & Jae-Seoun Hur

Dendrothele latenavicularis sp. nov. (Agaricales, Basidiomycota) from the

Patagonian Andes _ Sergio P. Gorjén, Alina G. Greslebin & Mario Rajchenberg

Endocalyx melanoxanthus var. melanoxanthus (Ascomycota): new to Brazil

and three newhosts_ — Nadja Santos Vitoria, Maria Auxiliadora Q. Cavalcanti,

Edna Dora Martins Newman Luz & José Luiz Bezerra

New records of rust fungi on sedges (Cyperaceae) from Pakistan

Malka Saba & Abdul Nasir Khalid

A new species and new records of Endophragmiella from China

Shou-Cai Ren, Jian Ma & Xiu-Guo Zhang

Two new Minimelanolocus species from southern China

Jian Ma, Yi-Dong Zhang, Li-Guo Ma & Xiu-Guo Zhang

Two new species of Passalora and Pseudocercospora from northeastern

Uttar Pradesh, India Raghvendra Singh, Shambhu Kumar & Kamal

Se)

101

123

137

JULY-SEPTEMBER 2011... V

Lecanora wrightiana and Rhizocarpon inimicum, rare lichens new to

Turkey and the Middle East Kenan Yazici, Andre Aptroot & Ali Aslan 145

A new volvate Macrolepiota (Agaricomycetes, Agaricales) from Italy,

with observations on the M. procera complex

Alfredo Vizzini, Marco Contu, Stefano Ghignone & Else Vellinga 149

Studies in Amanita (Amanitaceae) of Central America. I. Three new species

from Costa Rica and Honduras R.E. Tulloss, R.E. Halling & G.M. Mueller 165

Studies on Cephalotrichum from soils in China —twelve new species

and two new combinations Yu-Lan Jiang, Jun-Jie Xu, Yue-Ming Wu,

Yue-Li Zhang, Hui-Mei Liu, Hao-Qin Pan & Tian-Yu Zhang 207

Morphology and phylogeny of Pseudocercospora kamalii sp. nov.,

a foliar pathogen on Terminalia from India

Kunhiraman C. Rajeshkumar, Rahul Sharma, Rahul P. Hepat,

Santosh V. Swami, Paras Nath Singh & Sanjay K. Singh 227

Mycena sect. Longisetae: a new species, a new name, and an addition

Dollymol M. Aravindakshan & P. Manimohan 239

Three new hyphomycetes from southern China

Jian Ma, Li-Guo Ma, Yi-Dong Zhang & Xiu-Guo Zhang 247

A new species of Junghuhnia (Basidiomycota, Meruliaceae) from

tropical China Hai-Sheng Yuan 255

Caloplaca allochroa (lichenized Ascomycetes), a new saxicolous lichen species

from South Korea Yogesh Joshi, Jan Vondrak, Olga Vondrakova,

Thi Thuy Nguyen & Jae-Seoun Hur 261

Four anamorphic fungi (with two new species) from forests

of Western Ghats, India J. Pratibha, D.J. Bhat & S. Raghukumar 269

Taxonomic studies of Endophragmiella from southern China

Li-Guo Ma, Jian Ma, Yi-Dong Zhang & Xiu-Guo Zhang 279

Rhexodenticula zhengii sp. nov. from fallen leaves from China

De-Wei Li, Jingyuan Chen & Yixun Wang 287

Septobasidium atalantiae sp. nov. (Septobasidiaceae)

and S. henningsii new to China Suzhen Chen & Lin Guo 291

Revision of Glomeromycetes with entrophosporoid and glomoid spore formation

with three new genera Fritz Oehl, Gladstone Alves da Silva,

Ivan Sanchez-Castro, Bruno Tomio Goto, Leonor Costa Maia, Helder Elisio

Evangelista Vieira, José-Miguel Barea, Ewald Sieverding & Javier Palenzuela 29%

Puccinia cortusae (Basidiomycota; Uredinales) on Cortusa brotheri

(Primulaceae), new to southern Asia (Fairy Meadows, Pakistan)

A.N. Khalid & M. Saba 317

Gymnopus fuscotramus (Agaricales), a new species from southern China

Armin Me&i¢, Zdenko Tkaléec, Chun- Ying Deng,

Tai-Hui Li, Bruna PleSe & Helena Cetkovié 321

Biogeographical patterns in pyrenomycetous fungi and their taxonomy.

2. Additions to the Grayan disjunction

Larissa N. Vasilyeva & Steven L. Stephenson 331

vI ... MYCOTAXON 117

Lignincola conchicola from palms with a key to the species of Lignincola

J.K. Liu, E.B.G. Jones, E. Chukeatirote, A.H. Bahkali& K.D. Hyde 343

Craspedodidymum and Corynespora spp. nov. and a new anamorph recorded

from southern China Li-Guo Ma, Jian Ma, Yi-Dong Zhang & Xiu-Guo Zhang 351

A new species of Lepiota (Agaricaceae) from southwestern China

Jun F. Liang & Zhu L. Yang 359

Passalora wangii comb nov. from the genus Tandonella

Feng- Yan Zhai, Ying-Lan Guo, Ying-Jie Liu & Yu Li 365

A new species of Terriera (Rhytismatales, Ascomycota) from China

Zhong-Zhou Yang, Ying-Ren Lin & Cheng-Lin Hou 367

New Bulgarian records of fungi associated with glacial relict plants

Cvetomir M. Denchev, Dimitar Y. Stoykov, Ekaterina EF Sameva & Boris Assyov 373

Two new Ceratocystis species associated with mango disease in Brazil

Marelize Van Wyk, Brenda D. Wingfield, Ali O. Al-Adawi,

Carlos Rossetto, Margarida Fumiko Ito & Michael J. Wingfield 381

The genus Cladonia (lichenized Ascomycota, Cladoniaceae) in South Korea

Xin Yu Wang, Yogesh Joshi & Jae-Seoun Hur 405

First record of Resupinatus poriaeformis (Agaricomycetes) from South America

Georgea Santos Nogueira-Melo, Leif Ryvarden & Tatiana Baptista Gibertoni 423

New recombinations in Glomeromycota

Fritz Oehl, Gladstone Alves da Silva, Bruno Tomio Goto & Ewald Sieverding 429

Pouzarella (Agaricales, Entolomataceae) species from New South Wales (Barrington

Tops National Park) and northeastern Queensland, Australia

David L. Largent, Sarah E. Bergemann, Griffin A. Cummings,

Kathryn L. Ryan, Sandra E. Abell-Davis & Skye Moore 435

Amanita vernicoccora sp. nov. — the vernal fruiting “coccora” from California

Dimitar Bojantchev, Shaun R. Pennycook & R. Michael Davis 485

BOOK REVIEWS AND NOTICES Else C. Vellinga (EpiTor) 499

MYCOBIOTAS ONLINE

Abstracts of newly posted annotated regional species lists 508

NOMENCLATURE

Changes to publication requirements made at the XVII International

Botanical Congress in Melbourne —what does e-publication mean for you?

Sandra Knapp, John McNeill & Nicholas Turland 509

NOMENCLATURAL NOVELTIES proposed in MycoTaxon 117 517

PUBLICATION DATE FOR VOLUME ONE HUNDRED SIXTEEN

MYCOTAXON for APRIL-JUNE, VOLUME 116 (I-x1I + 1-521)

was issued on September 9, 2011

JULY-SEPTEMBER 2011...

REVIEWERS — VOLUME ONE HUNDRED SEVENTEEN

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

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

prepared for this volume.

Najm-u-sehar Afshan

Joe Ammirati

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Javier Etayo

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Edit Farkas

Francisco Das Chagas

Oliveira Freire

Eduardo Furrazola

Zai-Wei Ge

Paolo Giordani

Sergio P. Gorjon

Johannes Zacharias

Groenewald

Roy E Halling

Hiroshi Harada

Shuanghui He

Fabien C.C. Hountondji

Carlos Antonio Inacio

Bryce Kendrick

Alexander Khodosovtsev

Roland Kirschner

Tai-Hui Li

Ying-Ren Lin

Shuyan Liu

D. Jean Lodge

Laszlé Lék6s

W.EO. Marasas

Milagro Mata H.

Eric H.C. McKenzie

Vadim A. Melnik

Andrew S. Methven

Andrew M. Minnis

David W. Minter

Karen K. Nakasone

Maria Alice Neves

Lorelei L. Norvell

Shoji Ohga

Nichole O’Neill

E. Javier Palenzuela

Jiménez

Shaun R. Pennycook

Donald H. Pfister

Marcin Piatek

Francois Roets

Amy Y. Rossman

Giampaolo Simonini

Matthew E. Smith

Dartanha Jose Soares

Ulrik Sochting

Renata Gomes de Souza

Chang-Keun Sung

Michal Tomsovsky

R. Greg Thorn

Else C. Vellinga

Zheng Wang

Felipe Wartchow

Zhu L. Yang

Eugene Yurchenko

Ping Zhang

Ruilin Zhao

Wen- Ying Zhuang

VII

vill ... MYCOTAXON 117

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Warm regards, Lorelei Norvell

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11 November 2011

MYCOTAXON

http://dx.doi.org/10.5248/117.1

Volume 117, pp. 1-8 July-September 2011

Cortinarius callimorphus,

a new species from northern California

DIMITAR BOJANTCHEV’ & R. MICHAEL Davis?

'MushroomHobby.com, 345 Shipwatch Lane, Hercules, CA 94547, USA

*Department of Plant Pathology, University of California, Davis, CA 95616, USA

* CORRESPONDENCE TO: dimitar@pontix.com

Asstract — Described is Cortinarius callimorphus, a new species in subgenus Phlegmacium

from the coastal woods of northern California, with Sitka spruce to its apparent primary host

associate. Cortinarius callimorphus, which resembles other members of sect. Multiformes,

stands out because of its citriform spores. Its closest known relative is C. polymorphus from

Europe.

Key Worps — Cortinariaceae, fungal taxonomy, nrITS data

Introduction

The ecosystem of the northernmost coastal region of California (Del Norte,

Humboldt, and Mendocino counties) is influenced by high annual precipitation

averages (CalFire 2011) that are characteristic of the Pacific Northwest temperate

rainforest and which support its dominant conifers —Picea sitchensis (Bong)

Carriére (Sitka spruce) and Tsuga heterophylla (Raf.) Sarg. (western hemlock).

The extensive wet forests of the region support a rich fungal diversity. The

types of approximately two-thirds of the Cortinarius species described from

California have been collected in this region (Smith 1939; Ammirati & Smith

1977; Moser & Ammirati 1997, 2000). Cortinarius callimorphus, described

here, is common under Sitka spruce along the northern Californian coast.

Materials & methods

Methods for morphological studies and DNA extraction, PCR conditions and

primers, PCR product clean up, and sequencing are outlined in Bojantchev & Davis

(2011). Color codes follow Munsell™ soil color charts (Anonymous 2000). Terminology

follows Brandrud et al. (1998). Collections are stored in the private herbarium of the

first author or at the University of California herbarium in Berkeley (UC) where noted.

2 ... Bojantchev & Davis

C. glaucopus AY174785 Europe

C. glaucopus AF325604 Europe

|staucopus

po Rel OL

C. aleuriosmus AY669537 Europe

C. anserinus AY174805 Europe

C. anserinus AY174807 Europe

C. anserinus AY174806 Europe

C. dionysae AY174813 Europe

C. dionysae DQ083782 Europe

C. callimorphus JF742659 California, USA*

C. callimorphus JF742660 California, USA /polymorphus

C. polymorphus AY669545 Europe

C. cf. viridicoeruleus JF742658 California, USA

C. viridocoeruleus AY174788 Europe

C. cf. multiformis JF750423 Oregon, USA

C. multiformis AY669532 Europe /multiformis

C. allutus AY669531 Europe

C. elegantior AY174850 Europe

C. pseudoglaucopus DQ663395 Europe

C. sodagnitus AY174829 Europe

C. calochrous var. coniferarum EU056956 Europe

/anserinus

QaeOonr T0090 !

|Wiidiooor leus

oan —oO

calochroid clade

Fic 1. Phylogenetic tree inferred by maximum parsimony analysis of 20 Cortinarius subg.

Phlegmacium nrITS sequences. The tree shows the position of C. callimorphus relative to its closest

neighboring stirpes and the more distant calochroid clade. Branch lengths represent estimated

nucleotide substitutions. The GenBank accession numbers are listed after the taxon names.

Cortinarius calochrous var. coniferarum was selected as outgroup.

Phylogenetic analysis

During our studies we have downloaded and reviewed all Cortinarius nrDNA

sequences from the public databases GenBank (http://www.ncbi.nlm.nih.gov) and

UNITE (http://unite.ut.ee/). Preliminary phylogenetic analysis (not shown) of 984

Phlegmacium nrITS sequences from the northern hemisphere, including 278 sequences

from our own collections, clarified the closest relatives of C. callimorphus and resolved

its position outside of the calochroid super-clade as defined by Froslev et al. (2007) and

Garnica et al. (2009).

Twenty Phlegmacium taxa were selected for a detailed phylogenetic analysis. Sixteen

sequences were sourced from GenBank and four sequences representing western North

American taxa came from our collections. The breakdown of the selected sequences is

four members of the calochroid super-clade; two sequences of C. callimorphus plus one

of its closest known relatives; 13 sequences of the most representative taxa of the nearest

stirpes to C. callimorphus — /anserinus, /viridicoeruleus, /multiformis and /glaucopus

from both Europe and Western North America. Cortinarius calochrous var. coniferarum

(M.M. Moser) Nezdojm. was selected as an outgroup.

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

Cortinarius callimorphus sp. nov. (California) ... 3

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 search resulted in

nine most parsimonious trees (length = 249) which only differed in the topology of the

terminal nodes. One tree was selected (Fic. 1).

Taxonomy

Cortinarius callimorphus Bojantchev & R.M. Davis, sp. nov. FIGs 2-6

MycoBank MB 561091

Pileo 60-100 mm lato, hemispherico, dein plano-convexo, glutinoso, margine involuto,

ochraceoluteo vel flavobrunneo interdum centro, ferrugineo maculato, cum KOH

roseobrunneo. Lamellis emarginatis, pallide argillaceis, argillaceobrunneis, in statu senili.

Stipite 80-140 mm longo, cylindrico, bulbo submarginato 30-50 mm lato. Velo universale

albido. Velo partiale copioso, albido, cum KOH ope nullo. Carne albida. Sapore miti. Sporis

9-10 x 5-6 um, limoniformibus, verrucosis, basidiis 24-30 x 6-8 um, tetrasporigeris,

fibulis praesentibus.

Type: USA. California: Mendocino County, Caspar, Caspar Cemetery, under Picea

sitchensis, 22 Nov 2009, Bojantchev DBB25774 (Holotype UC 1860824; Genbank nrITS

JF742659).

Erymo.oey: from Greek: kaAoc = beautiful, wopdoc = form, shape.

Fic 2. Cortinarius callimorphus (collection DBB00080) - a typical form.

4 ... Bojantchev & Davis

straight to involute margin, uniformly ochre yellow (10YR 8/6-8/8), sometimes

darker in older basidiomata (10YR 5/6-5/8) with a persistent lighter yellow

zone near the margin (2.5Y 8/6) and rusty (5YR 6/6) spots where bruised;

surface glutinous when wet, glabrous to dull glossy when dry, innately fibrillose,

radially corrugated near the margin at age. LAMELLAE L = 70-110, crowded,

8-15 mm broad, pale clay (2.5Y 8/1-8/3) to grayish-white when young, never

bluish, shading yellow brown to brown (7.5R 6/6-5/6) as the spores mature;

edge even; attachment notched; lamellulae abundant. Stipe 80-140 mm long,

10-20 mm wide, cylindrical with a submarginate 30-50 mm wide bulb, white,

without bluish tints, spotting brown where bruised. UNIVERSAL VEIL white,

leaving volva-like remnants on the bulb. Cortina white, turning rusty brown

due to mature spore drop, leaving an annular zone of dense fibrils on the

stipe, occasionally forming a hairy appendiculate zone on the pileal margin.

CONTEXT white, bruising brown. Opor weak, aromatic to none. TAsTE fungal,

not unpleasant. MACROCHEMICAL REACTIONS 5% KOH carmine-red on pileus,

uniformly pale yellow brown on context, strongest on mature basidiomata,

negative on basal mycelium. UV non-fluorescent. SpoRE Deposit deep rusty

brown.

BASIDIOSPORES (8.5-)9.0-10.0(-10.5) x (5.0—)5.2-5.8(—6.0) um (mean 9.5 x

5.6 um), Q = 1.54-1.77, Q. = 1.72 (N = 217, 11 basidiomata, seven collections),

distinctly citriform, moderately verrucose. BAsip1a 24-30 x 6-8 um, 4-spored,

cylindro-clavate, clamped. GILL EDGE fertile or with clusters of elongated

septate cells. Cystrp1a not observed. PILEIPELLIS an ixocutis, duplex; epicutis

medium thick, 6-14 layers of hyphae 2-7 um diam., hyaline, hypodermium

well developed, composed of interwoven to parallel cylindrical cells, 20-40 x

12-20 um with refractive cytoplasmic pigment. CLAMP CONNECTIONS common

in all parts.

Fic 3. Cortinarius callimorphus.

a) Basidiospores (UC 1860824, holotype); b) Cuticle structure (UC 1860824, holotype).

Cortinarius callimorphus sp. nov. (California) ... 5

30 ym

Fic 4. Cortinarius callimorphus a) Elongated, septate cells on lamellar edge (UC 1860824, holotype);

b) 5% KOH reaction on pileus surface and stipe context (DBB09495); c) Corrugated pileus surface

near the margin (DBB09495).

HABITAT AND DISTRIBUTION — Very common under Sitka spruce in Northern

California, particularly in the Mendocino, Humboldt and Del Norte counties. We

have never seen it south of Mendocino. Its distribution within the Sitka spruce

range is apparently limited to northern California and southern Oregon (where

we have collected it 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. There are no other closely matching North

American collections in the public sequence databases.

Fic 5. Cortinarius callimorphus collections: a) DBB27800; b) DBB25837.

"fa —

Fic 6. Cortinarius callimorphus collections: a) DBB40200; b) UC 1860824 (holotype)

6 ... Bojantchev & Davis

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: MENDOCINO COUNTY,

Caspar, Caspar Cemetery, under Picea sitchensis, 26 Nov 2010, Bojantchev DBB40200;

4 Nov 2007, Bojantchev DBB00080; Jackson State Forest, under Picea sitchensis, 22 Nov

2008, Bojantchev DBB09495; Jughandle State Reserve, under Picea sitchensis, 22 Nov

2009, Bojantchev DBB25837 (UC 1860825, Genbank nrITS JF742660); DEL NorTE

County, Jedediah Smith Redwood State Park, under Picea sitchensis, 7 Nov 2009,

Bojantchev DBB22704; HumBoLpT County, Arcata, HBMS Mushroom Fair, under

Picea sitchensis. 20 Nov 2010, Bojantchev DBB39217; OREGON: CurRY Counry, Samuel

Boardman State Park, under Picea sitchensis, 11 Nov 2009, Bojantchev DBB27800.

Discussion

Traditional morphological analysis based on the overall combination

of stature, a yellowish viscid pileus, and argillaceous lamellae suggests that

C. callimorphus belongs to sect. Multiformes Rob. Henry ex Brandr. & Melot.

Its carmine-red KOH reaction on the pileus of and submarginate bulb suggest

that C. callimorphus might be related to the calochroid cortinarii, but its well-

developed hypodermium (duplex pileipellis) supports placement in sect.

Multiformes.

Within that section, C. callimorphus stands apart with its distinctly

citriform spores, a relatively uncommon feature in the non-calochroid clades.

Cortinarius polymorphus Rob. Henry is the only other species known with

similar macromorphological features (particularly lacking any blue colors) and

similarly shaped citriform spores. Detailed descriptions of C. polymorphus are

available in Henry (1935, 1951, 1985) and Bidaud et al. (2007); the latter includes

quality iconography. Molecular analysis confirms a close relationship between

the two species (Fic. 1). Cortinarius polymorphus has also been assigned to

Multiformes by Henry (1935, 1951, 1985), Moser (1960), and Bidaud et al.

(2007).

Recent phylogenetic studies within Cortinarius (Garnica et al. 2003) and

our own nrITS analysis show that sect. Multiformes is not well supported. Most

taxa traditionally assigned to it belong to various other Phlegmacium clades.

It is not the aim of this work to present a detailed picture of the infrageneric

relationships within subg. Phlegmacium, but as shown in Fie. 1, C. callimorphus

and C. polymorphus form a clade that is a sibling to the stirpes around

C. viridicoeruleus Chevassut & Rob. Henry and C. anserinus (Velen.) Rob.

Henry. A common character of these clades is the distinctly citriform spores.

Cortinarius callimorphus and C. polymorphus are very similar but can be

separated based on differences in geography (with the former found only in

western North America and the latter in Europe), nrITS sequences, and host

associations. Cortinarius polymorphus is apparently associated with broadleaved

trees, mainly members of Fagaceae, while C. callimorphus is associated with

Picea and possibly other conifers. The spores of C. polymorphus also appear

slightly broader with Q. = 1.5 vs. Q., = 1.7 for C. callimorphus. Henry (1935)

Cortinarius callimorphus sp. nov. (California) ... 7

describes the lamellar edges of C. polymorphus as distinctly denticulate, but we

have not observed that feature on the C. callimorphus basidiomata.

Complete iconography of C. callimorphus and a comparative image study is

available on http://www.mushroomhobby.com.

Acknowledgements

We thank 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

Ammirati JF, Smith AH. 1977. Studies in the genus Cortinarius, II: Section Dermocybe, new North

American species. Mycotaxon 5: 381-397.

Anonymous. 2000. Munsell™ soil color charts, revised edition. Munsell Color, New Windsor, NY.

Bidaud A, Moénne-Loccoz P, Reumaux P, Carteret X. 2007. Atlas des Cortinaires Vol. 16. Editions

Fédération mycologique Dauphiné-Savoie. Lomazzo.

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, Lindstr6m H, Marklund H, Melot J, Muskos S. 1989-98. Cortinarius Flora

Photographica Vol. 1-4. Cortinarius HB, Matfords, Sweden.

CalFire 2011. California Department of Forestry and Fire Protection website.

http://frap.cdf.ca.gov/webdata/maps/statewide/rainmap.pdf (accessed on 04/02/2011)

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, Weifs M, Oertel B, Oberwinkler F. 2003. Phylogenetic relationships of European

Phlegmacium species (Cortinarius, Agaricales). Mycologia 95: 1155-1170.

http://dx.doi.org/10.2307/3761917

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.

http://dx.doi.org/10.1186/1471-2148-9-1

Henry R. 1935. Etude de quelques Cortinaires du groupe des Scauri. Deux espéces nouvelles. Bull.

Soc. Mycol. France 51: 93-94.

Henry R. 1951. Les Scauri. Bull. Soc. Mycol. France 67(3): 1-328.

Henry R. 1985. Novitates. Validations, diagnoses latines. Documents mycologiques 16(61): 21-28.

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

Moser MM. 1960. Die Gattung Phlegmacium (Schleimk6pfe). Die Pilze Mitteleuropas, Vol. 4. J.

Klinkhart, Bad Heilbrunn.

Moser MM, Ammirati JF. 1997. Studies on North American Cortinarii IV: New and interesting

Cortinarius species (subgenus Phlegmacium) from oak forests in Northern California. Sydowia

49: 25-48.

Moser MM, Ammirati JF. 2000. Studies in North American Cortinarii VI. New and interesting taxa

in subgenus Phlegmacium from the Pacific States of North America. Mycotaxon 74: 1-36.

8 ... Bojantchev & Davis

Smith AH. 1939. Studies in the genus Cortinarius I. Contributions from the University of Michigan

Herbarium 2: 1-42.

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/10.5248/117.9

Volume 117, pp. 9-18 July-September 2011

A new Phylloporus from two relict Fagus grandifolia

var. mexicana populations in a montane cloud forest

LETICIA MONTOYA* & VICTOR M. BANDALA

Instituto de Ecologia, A.C., Biodiversidad y Sistematica,

PO. Box 63, Xalapa, Veracruz 91000, Mexico

*CORRESPONDENCE TO *: leticia.montoya@inecol.edu.mx

ABSTRACT — A new species, Phylloporus fagicola, has been discovered growing in association

with two relict populations of Fagus grandifolia var. mexicana in the subtropical region of

central Veracruz (eastern Mexico). The species belongs to a taxonomic group characterized

by broadly ellipsoid oblong to ovoid basidiospores, distinct from the subfusoid boletoid type.

The new taxon is described, illustrated, and compared with closely similar species.

KEY worps — ectomycorrhizal fungi, neotropical fungi, gilled Boletales, taxonomy

Introduction

Currently, Phylloporus rhodoxanthus (Schwein.) Bres., P. bellus (Massee)

Corner, P. phaeoxanthus var. simplex Singer & L.D. Gomez, P. foliiporus (Murrill)

Singer, P centroamericanus Singer & L.D. Gémez, and P. guzmanii Montoya &

Bandala have been recorded from Mexico (Singer 1957, Singer 1978, Singer &

Gomez 1984, Montoya et al. 1987, Montoya & Bandala 1991). Those records

represent a low proportion of the 24 Phylloporus taxa known to occur in the

Neotropics (Neves & Halling 2010). A new species described here, P. guzmanii

from Mexico, and P. aurantiacus Halling & G.M. Muell. from Costa Rica (Halling

et al. 1999), form a group of American taxa distinguished by broadly ellipsoid,

oblong, or ovoid shaped basidiospores that differ from the boletoid (subfusoid)

basidiospores typical of other Phylloporus species (Corner 1970, Heinemann

& Rammeloo 1987, Neves & Halling 2010). The Phylloporus species previously

reported from Mexico have been recorded growing with Quercus or Pinus. The

new species described here represents the first Mexican record of a Phylloporus

growing in association with Fagus grandifolia var. mexicana (Martinez) Little.

Materials & methods

Between September 2005 and September 2009 in the State of Veracruz, random

surveys were conducted in two Fagus grandifolia var. mexicana stands in Mesa de la

10 ... Montoya & Bandala

Yerba (Acajete; 19°33'37.2"N 97°01'9.8"W, 1900 m) and Acatlan Volcano (Naolinco-

Acatlan Road, Acatlan; 19°40'43.9"N 96°51'9.8"W, 1840 m).

Macromorphological characters were recorded from fresh sporocarps. Alphanumeric

color codes refer to Kornerup & Wanscher (1967) and Munsell (1994) (e.g. 2.5Y 5/2).

Micromorphological features were recorded in 3% potassium hydroxide water solution.

Basidiospore measurements were determined according to Montoya & Bandala (2003),

with 45-50 spores measured per collection and where X_ = length x width range of

meansand Q_ = the mean length/width ratio for n collections. Line drawings were made

with the aid of a drawing tube. The examined specimens studied are now deposited in

XAL herbarium. Acronyms for the herbaria follow Holmgren & Holmgren (1998).

Taxonomy

Phylloporus fagicola Montoya & Bandala, sp. nov. FIGs. 1-7

MycoBAnk MB 519849

Pileus 10-53 mm latus, primo hemisphaerico, dein late convexo vel applanato, sicco,

velutinosus, subtomentosus, tenuis squamulosus vel glabro ex parte et ad marginem,

aurantioruber, flavus ex parte, cyanescens, cum NH, caerulescent. Lamellae decurrentes,

confertae, septis connexae, furcatae circa stipitem, crassae, flavidae, rufo maculatae,

cyanescens. Stipes 13-40 x 2-7 mm, centralis vel eccentricus, firmus, rufo, aurantioruber,

velutinosus, basi villosa. Contextus flavus ad pileus, brunneo-rufo ad stipes, cyanescens,

cum NH, caerulescent. Basidiosporae 6.5-10.5(—11) x 4-7(-7.5) um, ellipsoidae, oblongae

olivacei-flavae. Pleurocystidia 84-132 x 7.5-13 jum, subcylindrata, subfusiformia,

ventricosa, sinuosis, numerosa; pariete tenui vel incrassatus (0.8-3.5um). Cheilocystidia

26-114 x 4,8-12(-13) um, clavata, subcylindrata, subfusiformia, interdum mucronata,

numerosa; pariete tenui vel incrassatus (-2.5 um). Pileipellis primo cutis instructa dein

intermitto cum numerosis tumulus catenulatis hyphis, cellulae terminales clavatis,

subcylindraceis, ventricosis, subfusiformis (12-55 x 3.5-7 um). Hyphis defibulatis.

Hotorypus: Ad terram in silvis. Mexico. Veracruz, Mpio. Acatlan, Acatlan Volcano,

August 21, 2007, del Moral 82 (XAL).

PitEus 10-53 mm diam., subhemispheric when young, plane-convex to plane,

reddish-orange (7C8-B8, 7D6) with some intense bright yellowish tones,

yellowish-orange (6B-C8, 10YR 8/8), brick orange to brick-red (2.5YR 4/6-

4/8, 3/6), in some areas orange (6B8) and with a yellow ground or towards the

margin (3A7, 10 YR 6/8), this latter at times discolouring, bluing to blackening

when bruised, dry, pruinose to velutinous, subtomentose to faintly squamulose

or subfloccose towards the center, glabrous and smooth in some areas and

towards the margin or faintly pruinose at margin, cracked in some areas.

LAMELLAE decurrent, thick, broad, 5-8 mm broad, close to crowded. Lamellulae

of 3-5 different lengths, at times furcate towards the stipe attachment, with

interparietal veins, joined in “H appearance’, egg yellow when young to yellow

with orange tinges (5A7-8, 5B7, 6A6-B7) or in age yellow with red (8D8)

irregular stains or almost totally red mainly towards the stipe attachment,

staining green-blue, bluish-brown (25D6) to grayish-black or blackish, margin

Phylloporus fagicola sp. nov. (Mexico) ... 11

Fic. 1. Phylloporus fagicola (Garay 215).

Bar = 10 mm.

irregular. Stipe 13-40 x 2-7 mm, subcylindrical, attenuated towards the base,

velutinous to finely furfuraceous towards the apex, central to excentric, dry,

firm, apex sulcate by the decurrence of the gills, reddish-orange (8C7), reddish

(10C8), vinaceous-brown (7E8) or brick-orange (2.5 4/8), reddish-orange (6B8)

at apex, yellow towards the base (4A6-5) but frequently stained dark grayish to

blackish; base at times villose, yellowish, with rhizomorphs. CoNTExT bright

yellow (3A6, 7B7) to egg yellow (4A7-8) at pileus; vinaceous-brown to reddish-

orange (7D6-D7) at stipe; staining deep blue (23D7-D8) mainly on stipe area.

Opor agreeable, fruity (recalling citrics). Machrochemical reactions: NH4OH

stains greyish-black, greenish on pileus; blue, greenish-blue on context. KOH

stains yellowish-brown on pileus and lamellae and orange-brown on context.

BASIDIOSPORES 6.5-10.5(-11) x 4—-7(-7.5) um, X_ = 7.0-9.0 x 4.2-5.1 um,

Q_= 1.53-1.78, ellipsoid to oblong, obtuse, yellowish-green, smooth, thin-

walled. Bastp1a 46-60 x 6-7 um, clavate, subcylindrical, sinuous, attenuated

towards the base, bisporic or tetrasporic, sterigma up to 8.5 um long, with

yellow contents. PLEUROCySTIDIA 84-132 x 7.5-13 um, subcylindrical,

subfusoid, sinuous, ventricose, frequently thin-walled but other with wall

0.8-3.5um thick, projected from belowthe hymenium, abundant, yellowin KOH.

CHEILOCYSTIDIA 26-114 x 5-12(-13) um, clavate, subcylindrical, mucronate,

12 ... Montoya & Bandala

Fic. 2. Phylloporus fagicola.

a. Basidiospores (del Moral 117). b. Pleurocystidia (del Moral 82, holotype).

Bars: a = 10 um, b = 20 um.

Phylloporus fagicola sp. nov. (Mexico) ... 13

Fic. 3. Phylloporus fagicola.

a. Cheilocystidia. b. Basidia. c. Terminal elements of pileipellis.

(a—b = del Moral 82, holotype; c = del Moral 292). Bars = 20 um.

14 ... Montoya & Bandala

Fic. 4. Phylloporus fagicola. Pleurocystidia.

(a-c: Garay 215; d-f: del Moral 82, holotype). Bars = 20 um.

subfusoid, thin-walled, other thick-walled up to 2.5 um, or with incrustations,

yellow in KOH, the measurements include mucronate subcylindrical or

moderately ventricose sterile elements, 26-45 x 5-8 um present at lamellae

edges. PILEIPELLIS a cutis of loosely intermixed hyphae when young, becoming

as an interrupted layer of frequent mounds of chains of elements disposed in

anticlinal orientation, loosely intermixed, with terminal cells clavate ventricose,

subfusoid, subcylindrical, 12-55 x 3.5-7 um, intercalary elements 12-38 x

Phylloporus fagicola sp. nov. (Mexico) ... 15

y r . oo

ee . . ¢ es _ i Sa

. a oe IF ae we le ~—" * “te

we teat CPi ~ ee LKS : Ay Fe ie, —

— _* Pat) ; wt

Fic. 5. Phylloporus fagicola. a. Basidiospores (Garay 215). b. Pileipellis (Montoya 4721).

Bars: a = 10 um, b = 20 um.

3.5-6 um, the mounds are variable in height and in some basidiomes arranged

in a dense disposition, the hyphae appear more loosely arranged towards the

context, all the elements yellow-amber in KOH, thin-walled. ConTExt hyphae

yellowish in KOH, hyphae loosely arranged 7-12 um broad. HYMENOPHORAL

TRAMA hyphae 3-12 um broad, thin-walled, some with thick wall up to 2.5 um

16 ... Montoya & Bandala

Fic. 6. Phylloporus fagicola, pileipellis (a: del Moral 82, holotype; b: del Moral 66).

Bars = 20 um.

Phylloporus fagicola sp. nov. (Mexico) ... 17

or with incrustations, other with dense, refractive contents. The tissues exude

an intense yellow sap when mounted in KOH.

HasBitaT — Gregarious in naked soil with dead leaves or with mosses in a

Fagus grandifolia var. mexicana forest at 1840 m.

ADDITIONAL SPECIMENS EXAMINED (all conserved at XAL): MEXICO. VERACRUZ:

Mpio. ACATLAN, Acatlan Volcano, August 14, 2007, del Moral 66; September 18, 2007,

del Moral 117; September 10, 2008, del Moral 292; September 29, 2009, Garay 215.

Mpio. ACAJETE, Mesa de la Yerba, August 8, 2006, Montoya 4606; October 22, 2008,

Montoya 4721.

Discussion

Phylloporus fagicola is distinguished by the following features: a short,

medium-sized basidiome, brightly colored (red-orange with yellow tones with

egg-yellow to orange-yellow lamellae) but with context and surfaces bruising

blue, crowded lamellae, thick-walled cystidia, and pileipellis at first a cutis and

developing frequent mounds of anticlinal chains of elements when mature.

Among other Phylloporus taxa with ellipsoid to oblong basidiospores,

P. guzmanii is distinguished by a vinaceous-red to dark purple pileus and stipe,

gills that are more distant and lighter yellow, slightly narrower basidiospores

[(6.4—)7.2-8.8(-10.4) x (3.2-)4.0-4.8(-5.6) um], thin-walled and somewhat

shorter cystidia [pleurocystidia (41.6—)45.0-105.6(-116.8) x 5.6-11.0(-12.0)

um; cheilocystidia 72.0-80.4 x 6.4-11.2 um], and a trichodermial pileipellis

with larger terminal elements [(20-)22-78(-80) x 5.5-10 um]. Phylloporus

guzmanii was recorded from Central and SW Mexico, growing in mesophytic

Quercus and Pinus forests (Montoya & Bandala 1991).

Phylloporus aurantiacus, which possesses oblong basidiospores that are

shorter (5.6—7.7 x 3.5-5 um, X_ = 6.8 x 3.5 um) and more ovoid in shape, is also

distinguished by unchanging flesh when exposed, lamellae lacking red stains,

shorter (60-90 x 7-10 um) thin-walled cystidia, a trichodermial pileipellis, and

an association with Quercus (Halling et al. 1999).

Phylloporus coccineus Corner from Singapore, also with broadly ellipsoid

basidiospores that are somewhat shorter [7.5-9(-10) x 6.5-7.5(-8) um] and

more globose, differs from P fagicola in subdistant gills, thin-walled larger

cystidia [pleurocystidia 200 x 10-16 um; cheilocystidia 70-120 x 10-18 um],

and a pileipellis comprising a compact palisade of terminal elements (25-40 x

5-10 um) with reddish-orange sap and slightly thickened pink walls (Corner

1970).

During repeated surveys of both sites, the P fagicola specimens fruited

infrequently compared with other species recorded in the area. The new species

appears to associate with Fagus grandifolia var. mexicana and has not yet been

found in other montane cloud forest fragments of surrounding the area or in

other regions explored by us.

18 ... Montoya & Bandala

Acknowledgments

We are grateful to Dr. Giampaolo Simonini (Associazione Micologica Bresadola,

Editorial Committee of Rivista di Micologia), Dr. Milagro Mata (INBio, Costa Rica)

and Drs. Lorelei Norvell and Shaun Pennycook (Mycotaxon editors) for the critical

revision of the paper. We appreciate the collaboration in the monitoring in the field

and the assistance in the lab by Biols. Pavel del Moral, David Ramos and MC Edith

Garay (Laboratorio de Biodiversidad y Sistematica de Hongos, Instituto de Ecologia,

A.C., Xalapa).

Literature cited

Corner EJH. 1970. Phylloporus Quél. and Paxillus Fr. in Malaya and Borneo. Nova Hedwigia 20:

793-822.

Halling R, Mueller GM, Dallwitz MD. 1999. A new Phylloporus (Basidiomycetes, Boletaceae) with a

key to species in Colombia and Costa Rica. Mycotaxon 73: 63-67.

Heinemann P, Rammeloo J. 1987. Phylloporus (Boletinae). Flore illustreé champignons Afrique

Centrale 13: 277-309.

Holmgren PK, Holmgren NH. 1998 [continuously updated]. Index Herbariorum: A global

directory of public herbaria and associated staff. New York Botanical Garden's Virtual

Herbarium. http://sweetgum.nybg.org/ih/

Kornerup A, Wanscher JH. 1967. Methuen handbook of colour. 2™ edn. Methuen, London. 243 p.,

30 pl.

Montoya L, Bandala VM, Guzman G. 1987. Nuevos registros de hongos del Estado de Veracruz, IV.

Agaricales (parte II). Rev. Mex. Mic. 3: 83-107.

Montoya L, Bandala VM. 1991. Studies on the genus Phylloporus in Mexico, I. Discussion of the

known species and description of a new species and a new record. Mycotaxon 41: 471-482.

Montoya L, Bandala VM. 2003. Studies on Lactarius a new combination and two new species from

Mexico. Mycotaxon 85: 393-407.

Munsell. 1994. Munsell soil colour charts. Macbeth, New Windsor. 10 p., 9 pl.

Neves MA, Halling R. 2010. Study on species of Phylloporus 1: Neotropics and North America.

Mycologia, 102: 923-943. http://dx.doi.org/10.3852/09-215

Singer R. 1957. Fungi Mexicani, series prima. Agaricales. Sydowia 11: 354-374.

Singer R. 1978. Notes on bolete taxonomy II. Persoonia 9: 421-438.

Singer R, Gdmez LD. 1984. The Basidiomycetes of Costa Rica, IH. The genus Phylloporus

(Boletaceae). Brenesia 22: 163-181.

MYCOTAXON

http://dx.doi.org/10.5248/117.19

Volume 117, pp. 19-28 July-September 2011

Type studies on two species of Phylloporus (Boletaceae, Boletales)

described from southwestern China

NIAN-KaAlI ZENG?“, L1-PING TANG*?4 & ZHU L. YANG'*

'Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany,

Chinese Academy of Sciences, Kunming 650204, China

*Department of Pharmacy, Hainan Medical University, Haikou 571101, China

*Kunming Medical College, Kunming 650021, China

‘Graduate University of Chinese Academy of Sciences, Beijing 100039, China

*CORRESPONDENCE TO: fungi@mail.kib.ac.cn

ABSTRACT —Type specimens of two Phylloporus species originally described from

southwestern China were studied. The concept of P luxiensis is refined with additional recently

collected materials, while P. scabrosus is revealed to be a member of the genus Lentinus.

Key worps — morphology, revision, taxonomy

Introduction

Phylloporus Quél. is a diverse genus in the Boletaceae, the hymenophore of

which is predominantly lamellate instead of poroid (Neves & Halling 2010).

Thirteen species and one variety have been reported from China (Teng 1963;

Zang & Zeng 1978; Li et al. 1992; Bi et al. 1993, 1994, 1997; Dai & Li 1994;

Zang et al. 1996). However, some remain poorly known, especially P. luxiensis

and P. scabrosus, both originally described from the southwestern region of

the country (Zang & Zeng 1978). Only scanty information on microstructures

was provided when the two species were first described with excellent vividly

colored plates of basidiomata (Zang & Zeng 1978); although both taxa were

later cited (Zang et al. 1996, Li & Song 2003), no additional morphological

information was given. Because many Phylloporus collections have recently

been made in the region, it is necessary to re-examine the types of the two

species to characterize them in more detail. The species are described below

based on critical examination of the holotypes and additional collections.

Materials & methods

The P. luxiensis and P. scabrosus holotypes and other P luxiensis collections are

deposited in the Herbarium of Cryptogams, Kunming Institute of Botany, Chinese

20 ... Zeng, Tang & Yang

Academy of Sciences (HKAS). For comparison, the holotype of P sulcatus (Pat.)

E.-J. Gilbert deposited in the Farlow Herbarium, Harvard University (FH) was also

examined. 5% KOH was used as a mounting medium for microscopic studies. The

notations “basidiospores (n/m/p)” indicate that the measurements were made on n

basidiospores from m basidiomata of p collections. Dimensions of basidiospores are

given using the notation (a—)b-c(-d); where the range b-c represents a minimum of

90% of the measured values, and extreme values (a and d) are given in parentheses.

Q refers to the length/breadth ratio of basidiospores; Q. refers to the average Q of

basidiospores + sample standard deviation. All line drawings of microstructures were

made from rehydrated material. For the convenience of the readers, the original Latin

protologue of the two taxa are provided.

Taxonomy

Phylloporus luxiensis M. Zang, Acta Microbiol. Sin. 18: 283, Pl. II: 1-4, 1978.

Figs. 1-3

LATIN PROTOLOGUE — “Pileus centralis, 5-6.5 cm latus, convexus demum plano-convexus

vel concavus, siccus, griseo-umbrinus, subtestaceus vel griseo-fuscus. Stipes 1-1.5 x 5-6.5

cm, cylindricus vel basim versus attenuatus, concolor, spongioso-cavus, myceliis flavido-

albis. Lamellae distantes, irregulares, decurrentes, flavidae. Basidiosporae ellipsoideae,

subfuscoideae, 4-5 x 8.9-12.5 um, laeves, hyalino-flavidae, 1-pleures guttulatae. Basidia

4-sporigera. Pleurocystidia numerosa, 12-16 x 25-35 um, fusiformia, cylindracea vel late

ellipticae, asperata, verruculosa. Cheilocystidia similia. Odor [as “Oder”] gratus et sapor

subnullus.” (Zang & Zeng 1978).

TyPE stupy — The holotype of P luxiensis consists of a basidioma and a spore

print wrapped in a paper possessing the following characters: PILEUS 3.3 cm

in diameter, convex; surface dry, yellowish brown, tomentose, margin inrolled.

HYMENOPHORE lamellate, decurrent. LAMELLAE distant, yellow, about 0.3 cm

in height. LAMELLULAE common, concolorous with lamellae. STrPE central, 4.5

x 0.5 cm, cylindric, base slightly enlarged 1.2 cm; surface dry, with very small,

reddish brown to yellowish brown, grayish brown squamules; the upper ribbed

by the decurrent lines of the lamellae. BasaL MycELium yellow. ANNULUS

absent. SPORE PRINT pale yellowish brown.

Basipiospores [40/1/1] 10-12 x (4-)4.5-5 um, Q = 2.00-2.44(-2.56),

Q = 2.26 + 0.15, 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 (Fics.1 c—d, 2c). PLEUROCYSTIDIA

51-67 x 12-17 um, ventricose or subclavate, thin- to slightly thick-walled (up to

1 um), colorless to yellowish in KOH (Fic. 2e). HYMENOPHORAL TRAMA hyphae

4-15 um in diameter, thin-walled, colorless to yellowish in KOH. Basrp1a,

CHEILOCYSTIDIA and PILEIPELLIS not recovered. CLAMP CONNECTIONS not

seen.

STUDY OF ADDITIONAL COLLECTIONS — BASIDIOMATA medium-sized (Fic.

la—b, 2a). PrLEus (2-)4-8 cm in diameter, convex, then applanate, center

Phylloporus type studies (China) ... 21

S , = rs)

r en i

| 0 = "4 * # ‘Ge

Fic. 1. Phylloporus luxiensis (a from HKAS 57048; b from HKAS 52242; c-d from HKAS 57036).

a—b. Basidiomata. c-d. SEM from dried specimen detailing bacillate ornamentation of basidiospores.

Bars: a-b = 2 cm; c-d = 5 um.

slightly depressed when old, but not infundibuliform; surface dry, brown,

densely tomentose, covered with brown, cinnamon-brown to grayish brown

squamules, margin inrolled; context white, unchanging in color when injured.

HYMENOPHORE lamellate, decurrent. LAMELLAE moderately distant, up to 0.6

cm in height, occasionally anastomosing, yellow, dull yellow to ochre yellow,

unchanging in color when injured. LAMELLULAE attenuate, concolorous with

lamellae. Stipe central, 2-6 x 0.3-1 cm, cylindric, attenuate downwards,

sometimes slightly enlarged at base (up to 1.4 cm), solid, firm; surface dry,

upper half ribbed by the decurrent lines of the lamellae, with minute, reddish

brown or purplish red squamules, lower half with yellowish brown, brown or

grayish brown squamules; context white, unchanging in color when injured.

BASAL MYCELIUM yellow. ANNULUS absent. Opor indistinct.

BASIDIOSPORES [300/15/4] (8-)9.5-12.5(-14) x (4-)4.5-5(-5.5) um, Q =

(1.60-)1.90-2.70(-3.11), Q. = 2.21 + 0.25, yellowish to yellowish brown in

KOH, smooth, subfusiform to ellipsoid, slightly thick-walled (up to 0.7 um)

(Fic. 2c). BAsIDIA 33-44 x 9-10 um, clavate, think-walled, 4-spored, hyaline to

yellowish in KOH; sterigmata 4-5 um long (Fic. 2d). HYMENOPHORAL TRAMA

composed of thin-walled hyphae 3-14 um in width, colorless to yellowish in

22 ... Zeng, Tang & Yang

Fic. 2. Macroscopic and microscopic features of Phylloporus luxiensis (a from HKAS 57048;

b, e from HKAS 40150, holotype; c-d, f-h from HKAS 57036). a. Basidiomata. b-c. Basidiospores.

d. Basidia and pleurocystidium. e. Pleurocystidia. f. Cheilocystidia. g. Pleurocystidia. h. Pileipellis.

Bars: a = 2 cm; b-h = 20 um.

Phylloporus type studies (China) ... 23

Fic. 3. Stipitipellis of Phylloporus luxiensis (HKAS 57036). a. From the apex of the stipe. b. From the

middle part of the stipe. c. From the basal part of the stipe. Bars= 20 um.

KOH. CHEILOCYSTIDIA 36-65 x 10-19 um, abundant, ventricose, subfusiform

or subclavate, thin- to slightly thick-walled (up to 1 um), colorless to yellowish

in KOH, no encrustations (Fic. 2f). PLEUROCysTIDIA 42-105 x 10-19 um,

abundant, ventricose, subfusiform or subclavate, thin- to slightly thick-walled

(up to 1 um), colorless to yellowish in KOH, no encrustations (Fic. 2d,g).

PILEIPELLIS a slightly interwoven trichoderm composed of yellowish, 5-13 um

wide, thin- to slightly thick-walled (up to 1 um), occasionally branching hyphae

usually with yellowish granular encrustations, and clavate or subcylindrical

terminal cells (20-62 x 5-12 um) with attenuated or obtuse apex (Fic. 2h).

PILEAL TRAMA composed of 4-12 um wide, thin-walled, colorless to yellowish,

interwoven hyphae. STIPITIPELLIsS a trichoderm-like structure; at the apex of

the stipe composed of thin- to slightly thick-walled (up to 1 um) hyphae with

24 ... Zeng, Tang & Yang

narrowly or broadly clavate terminal cells (30-46 x 8-18 tm), and occasionally

with clavate, 4-spored basidia (33-43 x 9-11 um) (Fic. 3a); at the middle part

of the stipe composed of thin- to slightly thick-walled (up to 1 um) hyphae

with clavate or subfusiform terminal cells (28-50 x 8-18 um) (Fic. 3b); at the

basal part of the stipe composed of slightly thick-walled (up to 1.5 um) hyphae

with broadly clavate or subfusiform terminal cells (30-71 x 14-20 um), and

many 3-20 um wide, slightly thick-walled (up to 1.5 um), emergent hyphae

with clavate or subcylindrical terminal cells (33-50 x 5-9 um) (Fic. 3c).

STIPE TRAMA composed of 4-10 um wide, cylindrical, thin- to slightly thick-

walled (up to 0.5 um), colorless to yellowish in KOH, parallel hyphae. CLamp

CONNECTIONS absent in all tissues.

KNOWN DISTRIBUTION — Southwestern China.

SPECIMENS EXAMINED — Phylloporus luxiensis: CHINA. YUNNAN PROVINCE: MANG

City (previously called “Luxi County”), Chengguan Town, Xiadong, in the forest

dominated by Pinus and Quercus, 3 July 1977, X.J. Li 90-HK2919 (HKAS 40150,

holotype); CHUXIONG YI AUTONOMOUS PREFECTURE, bought from market, 26 August

2007, Z.L. Yang 4925 (HKAS 52242); NANHUA Country, bought from market, 2 August

2009, L.P Tang 1079 (HKAS 57036); bought from market, 2 August 2009, L.P Tang 1080

(HKAS 57037); bought from market, 3 August 2009, L.P Tang 1091 (HKAS 57048).

Phylloporus sulcatus: VIETNAM. Tonkin: Vallée du Da-Pounian, 1907, Eberhardt LBA

210 (FH 3725, holotype of Phylloporus sulcatus), det. N.T. Patouillard.

ComMENTs — In the protologue, the outer surface of the cell wall of cystidia

was described as “verrucous. However, our observations differ from the

original description.

Phylloporus luxiensis, originally described from Luxi County (now called

“Mang City”) in southwestern Yunnan, is well characterized by its brown,

cinnamon-brown to grayish brown pileal surface, the reddish brown to

yellowish brown, brown, or grayish brown stipe surface, the absence of staining

of lamellae and context, and the non-inflated pileipellis hyphae.

Four collections cited above (HKAS 52242, 57036, 57037, 57048) matched

well with the re-examined holotype and original illustrations. Our nuc-LSU

and ITS sequence analyses (unpublished data) cluster all four collections

together with a high statistic support. Unfortunately, no DNA sequences were

successfully generated from the holotype.

Phylloporus sulcatus, a relatively unknown taxon described originally from

Vietnam, shares some common characters with P. luxiensis (Patouillard 1909,

Perreau & Joly 1964, Corner 1970). One of us (ZLY) studied the P sulcatus

holotype (FH-sheet 3725; Harvard University, Farlow Herbarium), which

consists of the single collection, Eberhardt LBA 210. In the protologue of P.

sulcatus, Patouillard (1909) cited a single locality, which corresponds exactly

with the locality of Eberhardt LBA 210, which Singer (1945, 1978) therefore

designated as the (holo)type. Eberhardt LBA 210 is comprises a single

Phylloporus type studies (China) ... 25

000

(

Fic. 4. Microscopic features of Phylloporus sulcatus (FH 3725, holotype). a. Basidiospores.

b. Pleurocystidia. Bars = 20 um.

basidioma. The dried cap is about 1.3 cm in diameter, convex, dark brown;

the lamellae are decurrent, dark brown to blackish; the stipe is 2 x 0.2-0.3 cm,

subcylindrical, dark, and its base is slightly enlarged; the basidiospores are

[20/1/1] 10.5-12.5(-13) x (4.5-)5-5.5(-6) um, Q = 2.0-2.36(-2.78), Q. = 2.18

+ 0.19, yellowish to brown yellowish in KOH, smooth, ellipsoid (Fic. 4a); the

basidia are 30-40 x 9-12 um, clavate, 4-spored; the pleurocystidia are 55-70

x 9-12 um, fairly abundant, subfusiform or narrowly clavate, with yellowish

to brown yellowish vacuolar pigment, thin-walled, without encrustations (Fic.

4b). Based on our type study, P. sulcatus appears to differ from P luxiensis in

its wider basidiospores and narrower pleurocystidia. Furthermore, the stipe

surface of P sulcatus has no reddish tinge (Patouillard 1909, Perreau & Joly

1964, Corner 1970).

Phylloporus scabrosus M. Zang, Acta Microbiol. Sin. 18: 284, Pl. II: 5-8, 1978. Fie. 5

LATIN PROTOLOGUE — “Pileus centralis, 5-8 cm diam. convexus demum plano-convexus,

siccus, rufobrunneus, chryseus vel subtestaceus. Stipes 1-1.5 x 8-10 cm, subcylindricus vel

subclavatus, aequalis, sordide concolor, flexus, apice striatus, sursum scabroso-furfuraceus,

basim versus squamuloso-furfuraceus. Lamellae distantes, decurrentes, flavido-albidae vel

flavidae, myceliis flavido-testaceis. Basidiosporae subfuscoideae laeves, 5-5.5 x 11-12.5

um, hyalino-flavidae. Basidia 4-sporigera. Pleurocystidia 10-15 x 25-35 yum, lageniformia

vel fusiformia. Cheilocystidia non vidi. spongioso-carnosus. Odor [as “Oder”] gratus et

sapor saccharatus. (Zang & Zeng 1978).

Type stupy — The P. scabrosus holotype comprises five basidiomata possessing

the following characters: PiLEUs 3-4 cm in diameter, convex to applanate;

surface dry, yellowish brown to chocolate brown, glabrous. HYMENOPHORE

lamellate, decurrent. LAMELLAE yellowish brown to dark brown, subdistant.

STIPE central, 7.5-12 x 0.4-0.65 cm, cylindric, base slightly enlarged 0.9-1.2

cm, solid, firm; surface dry, concolorous with the pileus, velutinate, and also

covered with disrupted squamules. ANNULUS absent. TEXTURE very hard.

26 ... Zeng, Tang & Yang

Fic. 5. Microscopic features of Phylloporus scabrosus (HKAS 28981, holotype). a. Basidiospores.

b. Basidium. c. Cystidia. d. Generative hyphae. e. Skeletal hyphae. Bars = 20 um.

BasIDIOSPORES [20/1/1] 6-8.5 x 3-4 um, Q = 1.71-2.33, Q. = 1.97 + 0.20,

yellowish brown in KOH, cylindric, smooth, thin-walled (Fic. 5a). Basrp1a

25-27 x 7-8 um, clavate, yellowish brown in KOH, 4-spored (Fig. 5b). CysTIDIA

25-40 x 5-9 um, finely abundant, subfusiform, cylindric or subclavate, thin-

to slightly thick-walled (up to 1 um), yellowish brown in KOH (Fic. 5c).

GENERATIVE HYPHAE 3-5 um in diameter, not inflated, yellowish to yellowish

brown in KOH, frequently branching (Fic. 5d). SKELETAL HYPHAE 5-13 um in

diameter, cylindric, yellowish brown in KOH, with a thickened wall (usually

2-3 um thick, sometimes up to 5 um thick), frequently branching (Fic. 5e).

CLAMP CONNECTIONS common in every part of the basidioma. PILEIPELLIS

and STIPITIPELLIS not recovered.

SPECIMEN EXAMINED — CHINA. YUNNAN PROVINCE: XISHUANGBANNA DAI

AUTONOMOUS PREFECTURE, Mengla County, Shangyong, Xinanshan, on fallen trunk,

alt. 600 m, 24 September 1974, M. Zang 1640-HK1640 (HKAS 28981, holotype).

CoMMENTS — ‘The five basidiomata of the holotype were probably not fully

mature before being dried; only a few basidia with sterigmata and basidiospores

were observed. The original protologue describes the basidiospores as measuring

Phylloporus type studies (China) ... 27

11-12.5 x 5-5.5 um (Zang & Zeng 1978), but our measurements differed. The

holotype's basidiospores, basidia, and cystidia are filled with a yellowish brown

necropigment, as observed in many other higher fungi (Locquin 1953; Yang

1997; Albee-Scott 2007; Petersen 2008a,b; Alvarado et al. 2010; Cripps et al.

2010).

Judging by the illustrations, texture, observed microscopic features, and

lignicolous habit, this fungus apparently represents the genus Lentinus (see

Corner 1981, Pegler 1983, Karunarathna et al. 2011).

Acknowledgments

The authors are very grateful to the following persons: Prof. Dr. T.H. Li (Guangdong

Institute of Microbiology, China) and Dr. M.A. Neves (Universidade Federal de Santa

Catarina, Departamento de Botanica, Brazil) for serving as reviewers; Prof. Dr. D.H.

Pfister (Farlow Herbarium, Harvard University, USA) for allowing the third author to

access the specimens in the Herbarium and examine them in his laboratory; and Dr. Z.W.

Ge and Dr. X.H. Wang (Kunming Institute of Botany, Chinese Academy of Sciences) for

their suggestions. This study was supported by the Hundred Talents Program of the

Chinese Academy of Sciences and the Ministry of Science and Technology of China

(2008FY110300-03-1).

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MY COTAXON

http://dx.doi.org/10.5248/117.29

Volume 117, pp. 29-36 July-September 2011

Arthrobotrys latispora, anew nematode-trapping fungus

from southwest China

HONGYAN Su’, SHUORAN LIv’, YUNXIA LI',} YONGHONG CAO’,

MINGHUI CHEN? & XIAOYAN YANG"

‘College of Agriculture and Biology, Dali University, Dali 671003, P.R. China

*Laboratory for Conservation and Utilization of Bio-resources, Yunnan University,

Kunming 650091, P.R. China

*CORRESPONDENCE TO: yangxy.dlu@gmail.com

ABSTRACT — Using morphology and molecular phylogenetic analyses, we report a new

nematophagous hyphomycete species, Arthrobotrys latispora, which produces erect, branched

or unbranched conidiophores with conspicuous nodes at the tip; the conidia are nonseptate

(41%) or uniseptate (59%) and broadly ovoid to oval. Three-dimensional adhesive networks

formed in the presence of nematodes.

Key worps — teleomorph-anamorph connection, Orbilia

Introduction

Nematode-trapping fungi have been the subject of decades-long research,

including their ecology, distribution and systematics, due in part to their

potential as biological control agents to manage plant and animal nematode

diseases (Liu et al. 2002, Liu & Zhang 2003, Dong et al. 2004). These fungi are

able to trap and consume nematodes with modified hyphae that include stalked

and sessile adhesive knobs, adhesive nets, and constricting or non-constricting

rings. Species forming adhesive nets are assigned to the anamorphic genus

Arthrobotrys Corda, which currently comprises about 50 species (Scholler et

al. 1999).

Pfister (1994) was the first to connect an Arthrobotrys species to the

ascomycetous genus Orbilia on the basis of cultural studies. Up to now, six

species of Arthrobotrys have been linked to related Orbilia teleomorphs. Orbilia

auricolor (A. Bloxam) Sacc., seemingly a species complex, is known to have four

anamorphs: A. oligospora Fresen., A. cladodes Drechsler (Pfister & Liftik 1995),

A. yunnanensis M.H. Mo & K.Q. Zhang (Mo et al. 2005), and A. psychrophila

(Drechsler) M. Scholler et al. (Rubner 1996). Of the remaining two species, O.

30 ... Su & al.

fimicola Jeng & J.C. Krug pairs with A. superba Corda (Pfister 1994) and an

undescribed Orbilia sp. with A. nonseptata Z.F. Yu et al. (Li et al. 2009).

A survey of Orbilia species and their anamorphs revealed one Arthrobotrys

species that had not yet been characterized. As neither morphological

comparison nor phylogenetic analysis of the internal transcribed spacer (ITS)

regions of rRNA genes could assign this isolate to a previously named taxon, it

is reported here as a new species, Arthrobotrys latispora.

Materials & methods

Collection, isolation, and characterization of teleomorph fungi and their anamorphs

Fresh specimens of an Orbilia species were collected on decaying bark of Castanopsis

orthacantha Franch. (Fagaceae), located in the northwest part of the YongPing County

(25°7'18"-25°13'44"N 99°27'26"-99°36'07"E), Yunnan Province, China where the

dominant plant species are Castanopsis orthacantha, Lithocarpus variolosus Chun,

Cyclobalanopsis glauca Oerst., and Cinnamomum glanduliferum (Wall.) Meisn.

Specimens are preserved in the herbarium of the Biology and Chemistry College of Dali

University, China.

To cultivate the fungus, four apothecia were fixed to the lid of a Petri dish with

hymenia positioned downward in order to collect discharged ascospores on the surface

of CMA media (20 g corn meal, 18 g agar, 40 mg streptomycin, and 30 mg ampicillin

in 1000 ml distilled water). These Petri dishes were placed at room temperature for

4-6 days until the ascospores germinated. Agar blocks with deposited ascospores

were transferred to another CMA plate to avoid contamination and incubated at 25°C.

After 10-15 days, the cultures were examined with an Olympus BX51 microscope

with differential interference contrast. Trapping organs were induced by adding ~100

nematodes (Panagrellus redivivus Goodey) to a square slot (1 x 1 cm) created by

removing the agar at the colony edge.

DNA extraction, PCR, and sequencing

Genomic DNA was extracted from the mycelia grown on cellophane membrane on

PDA according to Jeewon et al. (2002). Primer pairs ITS5 and ITS4 (White et al. 1990)

were used to amplify the complete ITS + 5.88 region. PCR amplification parameters were:

1 minute initial denaturation at 95°C, followed by 30 cycles of 1 minute denaturation

at 94°C, 1 minute primer annealing at 50°C, 1.5 minutes extension at 72°C, and a

final extension period of 10 minutes at 72°C. The purified PCR products were directly

sequenced in forward and reverse directions using the same PCR primers (BGI Co.,

Ltd., China).

Phylogenetic analysis

Complete ITS1+5.8S+ITS2 rDNA sequences were aligned with 20 sequences

(obtained from GenBank, NCBI [http://www.ncbi.nlm.nih.gov/]) by MUSCLE (MUltiple

Sequence Comparison by Log-Expectation) at the EBI (European Bioinformatics

Institute) web server [http://www.ebi.ac.uk/Tools/muscle/index.html]. ITS4 and ITS5

sequences were excluded from the analysis of all taxa. The DNA matrix of 21 taxa and

496 nucleotides was adjusted manually to improve the alignment. Phylogenetic analysis

Arthrobotrys latispora sp. nov. (China) ... 31

was conducted with MrBayes v3.1.2 (Ronquist & Huelsenbeck 2003). Alignment gaps

were treated as missing data; the sequence file set as interleaved; and GTR model was

used for the substitution model. Markov chains were run for 1,000,000 generations, and

trees were sampled every 10th generation resulting in 100,000 trees. About 0.003 average

standard deviation of split frequency was achieved. The first 25,000 burn-in phase trees

were discarded while the remaining 75,000 trees were used for calculating posterior

probabilities in the consensus tree. The consensus tree was analyzed by the program

FigTree v1.3.1. (Rambaut & Drummond 2010). The GenBank accession numbers of all

the analyzed sequences were indicated after names of the species on the phylogenetic

tree.

Results

Phylogenetic analysis

The Bayesian tree generated from the ITS1+5.8S+ITS2 rDNA sequences

of Arthrobotrys latispora and its relative species (shown in PLATE 1) indicates

that Arthrobotrys and Drechslerella species group into two different clades, with

Dactylellina species basal to both. The Bayesian posterior probabilities from

the ITS genotyping resulting in these three major groups correspond to the

different trapping device types and are consistent with the results of Scholler et

100% Arthrobotrys latispora, HQ735415

Arthrobotrys botryospora, U51955

5% Arthrobotrys superba, U51949

Arthrobotrys sinensis, AY773445

Arthrobotrys yunnanensis, AY509930

Arthrobotrys vermicola, AY773454

99% Arthrobotrys oligospora, AB374287

100%|

Arthrobotrys amerospora, EF 192178

99% Arthrobotrys anomala, AY773451

Arthrobotrys eudermatum, AF 106528

Arthrobotrys nonseptata, FJ185261

Dactylellina haptospora, DQ999820

96%

100% Drechslerella anchonia, AY965753

100% Drechslerella brochopaga, U51950

100° Drechslerella polybrochum, U51973

99% Dactylellina phymatopaga, MPU51970

Dactylellina appendiculata, AF106531

99%

Dactylellina parvicollis, AY773472

Dactylellina ellipsospora, AY965759

Dactylellina candida, AY965749

Fusarium oxysporum, EF 192187 (outgroup)

0.04

PLATE 1. Bayesian tree derived from complete ITS1, ITS2 and 5.85 rDNA sequences of nematode-

trapping fungi with Fusarium oxysporum set as outgroup. Bayesian posterior probabilities over 95%

are shown on the branches, and scale bar at the bottom shows the expected changes per site.

32 ... Su & al.

TABLE 1. Morphology of Arthrobotrys species with non-septate conidia.

CONIDIA

a — — COnIdIoGEN. CONIDIOPHORE PREDACIOUS

SPECIES CAPR SIZE SEPTA DENTICLE (height pm) ORGANS

(um) (#)

15-31 x 0 adhesive

A. amerospora obovoid 10-20 short 75-250 at aes

cylindric 13-22 0 adhesive

A. anomala to long 3-7 (-1) short 20-80 branches &

ellipsoid networks

A. arthro- broadly ,

botryoides sensu _cylindric- Nae (a) short 300-450 iene

ear 10-16 1 networks

Drechsler ellipsoid

adhesive

A. botryospora ellipsoid ieee Q short 250-450 hyphae &

11-15 (-1)

networks

broadly 14.8-21.5 see

A. latispora ovoid- x 0-1 short 60-250 LL

oval 10.1-16.3

elongate 11-16.8 adhesive

A. nonseptata ellipsoid o5 86 0 short 40-120 Le

I ,

ape ge adhesive

A, yunnanensis ca as x 0(-1) long 60-200

cylindric, 275-75 networks

or clavate ; |

al. (1999). The trapping devices of Arthrobotrys, Dactylellina, and Drechslerella

species form, respectively, adhesive networks, adhesive knobs, and constricting

rings. Arthrobotrys latispora is situated within the adhesive networks clade

and appears closely related to A. botryospora G.L. Barron. Morphological

comparisons are shown in TABLE 1. Because our isolate differs from previously

described Arthobotrys species, we propose it here as a new species.

Arthrobotrys latispora H.Y. Su & X.Y Yang, sp. nov. PLATE 2

Coloniae in CMA effusae, ad 6 cm diam. post 8 dies 25°C. Mycelium sparsum, effusum,

hyalinum, septatum, romosum, 2-4 um laum. Conidiophora hyalina, simplicia, erecta,

septata, non ramosa, plerumque 60-210 um alta, basi 2-5 um crassa, apice 1.8-4 um

crasso, efferenti 4-12 conidia sola de conidiogenis loci in perspicuis dendriculis in apicie

aut prope apicem. Conidia hyalina, elongato ellipsoideo-cylindrica vel clavata, non- vel

uniseptata, 14.8-21.5 x 10.1-16.3 (x = 18.99 x 13.5) um. Reticula tenacia quae vermiculos

nematodeos capiunt evolventibus. Chlamydosporae globosae vel ellipsoideae, catenulatae.

HotoryPe: Dried agar plate of a culture isolated from a collection of an undescribed

Orbilia teleomorph [see below], (holotype, YMF 1.03168; ex-type culture, YMF 1.03168;

isotype and ex-type culture, College of Agriculture and Biology, Dali University).

EryMo_oey: ‘latispora’ refers to the broadly ovoid conidia.

Arthrobotrys latispora sp. nov. (China) ... 33

PLaTE 2. Arthrobotrys latispora:

A-E. Conidiophores with spore. F-G. Conidia. H. Adhesive nets. Bars: A-H =10 um.

34 ... Su & al.

ASSUMED TELEOMORPH: an unidentified Orbilia sp., YMFT 1.03168, collected on

decaying bark of Castanopsis orthacantha (Fagaceae), PR CHINA, Yunnan Province,

YongPing County, Jinguangsi Virgin Forest, alt. 2820 m, 20 september 2008, coll. X.J. Su,

Z.L. Luo & H.Y. Su. (Duplicate material deposited at College of Agriculture and Biology,

Dali University.)

Colonies grew rapidly on CMA medium, attaining 6 cm diam. in 8 days at

25°C. Mycelium spreading, vegetative hyphae hyaline, septate and branched,

mostly 2-4 um wide. Conidiophores colorless, erect, simple, septate frequently,

60-210 um high, 2-5 um wide at the base and 1.8-4 um at the tip, producing

4-12 conidia singly from conidiogenous loci on conspicuous, partly superposed

nodes at and near the apex. Conidia colorless, broadly ovoid-oval, broadly

rounded at the tip, rounded truncate at the narrowed base, uniseptate or

nonseptate at the center, 14.8-21.5 x 10.1-16.3 (x = 18.3 x 13.5) um (living

state). Approximately 41% of the conidia were nonseptate and 59% uniseptate.

Chlamydospores spherical to ellipsoidal. Nematodes are trapped by three-

dimensional adhesive networks.

Discussion

There are six known species of Arthrobotrys with nonseptate conidia, some

of which form both uni- and nonseptate conidia. Conidia of A. amerospora

(Schenck et al. 1977) and A. nonseptata (Li et al. 2009) are consistently

nonseptate while those of A. anomala (Barron & Davidson 1972), A. botryospora

(Barron 1979), and A. yunnanensis (Mo et al. 2005) are occasionally uniseptate.

A. arthrobotryoides sensu Drechsler (1944), which consistently forms uniseptate

conidia at 20°C, also forms many nonseptate ones at 28-32°C. Van Oorschot

(1995) has noted that the identity of A. arthrobotryoides (Berl.) Lindau in

its original sense cannot be determined from the protologue and cannot be

resolved because of the absence of type material.

Arthrobotrys latispora is characterized by its broad, ovoid or slightly oval,

partly nonseptate conidia, which are borne on nodes. Based on conidial shape,

A. latispora most closely resembles A. arthrobotryoides sensu Drechsler and

A. amerospora. The conidia of all three species are ovoid but differ in conidial

size and septation. Differences between A. latispora and the other six known

species that form non-septate conidia are summarized in TABLE 1.

Three additional collections also made on 20 September 2008 in the same

region were associated with the same teleomorph but produced a very different

anamorph referred to the genus Anguillospora. The question arises as to whether

A. latispora is a contaminant, a conclusion supported by the fact that a similar,

closely related teleomorph, O. luteorubella (Nyl.) P. Karst., also produced a

similar Anguillospora anamorph (Pfister 1997).

Despite the nearly identical conidial morphology of A. latispora and

A. botryospora, our molecular phylogenetic analysis pointed outa clear difference

Arthrobotrys latispora sp. nov. (China) ... 35

between these two species: the ITS1+5.8S+ITS2 rDNA sequence similarity

between A. latispora and a non-type isolate of A. botryospora is 93.7%. Thus

A. latispora and A. botryospora can be considered two distinguishable species.

The phylogenetic analysis also showed that Dactylellina species were basal to

both Drechslerella and Arthrobotrys. These results support the hypothesis of Li

et al. (2005) that the trapping devices of Drechslerella species (with constricting

rings) and Arthrobotrys species (with adhesive networks) developed

independently from adhesive knobs (ancestral types of trapping devices

specific to Dactylellina). Moreover, the analysis indicates that Arthrobotrys

species may have evolved from Dac. haptospora relatives, whereas Drechslerella

species evolved from Dac. phymatopaga relatives. More studies are needed to

investigate the evolutionary process of these predacious fungi.

Acknowledgements

This research was supported by the National Natural Science Foundation Program

of PR China (30960017, 31060019), Reasearch Foundation from Yunnan Education

Department (09y0360). We sincerely thank Dr. Wen Xiao, XiaoMei Gao, XingNeng

Zhao, JiaGuo Yan and XiJun Su for collecting the specimens. Special thanks to

Dr. MingHe Mo and Matthew Scott for their suggestions on the manuscript. We also

thank Hans-Otto Baral, Donald Pfister, Shaun Pennycook, and ChangKeun Sung for

reviewing the manuscript.

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Mo MH, Huang XW, Zhou W, Huang Y, Hao YE, Zhang KQ. 2005. Arthrobotrys yunnanensis sp.

nov., the fourth anamorph of Orbilia auricolor. Fungal Diversity 18: 107-115.

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684-688. http://dx.doi.org/10.2307/3760812

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Rambaut A, Drummond A. 2010. FigTree v1.3.1. Institute of Evolutionary Biology, University of

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MY COTAXON

http://dx.doi.org/10.5248/117.37

Volume 117, pp. 37-43 July-September 2011

Additional records of Volvariella dunensis

(Basidiomycota, Agaricales):

morphological and molecular characterization

ALFREDO VIZZINI*, MARCO CONTU? & ALFREDO JUSTO?

‘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

*Biology Department, Clark University, 950 Main St., Worcester, MA 01610 USA

*CORRESPONDENCE TO: alfredo. vizzini@unito. it

ABSTRACT — Collections morphologically assignable to Volvariella dunensis from Sardinia

and the Atlantic coast of Spain were revised and compared with the original collections.

Molecular data supporting all examined collections as V. dunensis expand its known

geographic distribution. A revised morphological characterization and a phylogenetic

analysis of all Volvariella species sequenced to date are provided.

Key worps — Agaricomycetes, ITS, phylogeny, Volvariella volvacea

Introduction

The genus Volvariella Speg., which is composed of saprotrophic or

mycotrophic agarics, has been historically considered a member of the

family Pluteaceae Kotl. & Pouzar (Singer 1986). The genus is characterized

macroscopically by a pink spore-print, free lamellae, and a universal veil

that forms a saccate volva at the base of the stipe and microscopically by the

inverse hymenophoral trama (Singer 1986). Results from earlier molecular

studies (Moncalvo et al. 2002, Matheny et al. 2006) led to questions about

its monophyly and phylogenetic position in the Pluteaceae. More recent

research focused on the Pluteaceae (Justo et al. 2011) showed that Volvariella is

polyphyletic with two major clades. The large-spored, viscid species belonging

to the V. gloiocephala complex were resolved as the sister group of Pluteus

Fr. and accommodated under the new generic name Volvopluteus Vizzini et

al. Volvariella, the remaining clade, is therefore restricted to the dry, small-

spored species. Volvariella falls outside the pluteoid clade and based on data

from ribosomal loci (nSSU, 5.88, nLSU) shows afhnities to some hygrophoroid

38 ... Vizzini, Contu & Justo

genera (Cuphophyllus (Donk) Bon, Cantharocybe H.E. Bigelow & A.H. Sm.),

but its phylogenetic position within the Agaricales is still unclear.

Volvariella dunensis has been the subject of a detailed taxonomic study

based on the re-examination of the original collections from the Mediterranean

coast of Spain (Justo & Castro 2010). Very recently, a collection assignable to

V. dunensis was reported from the Atlantic coast of Spain (Fernandez-Sasia 2010

as “Volvariella nigrovolvacea var. dunensis”). Also, the identity of the Sardinian

collection reported by Contu & La Rocca (1999) as Volvariella nigrovolvacea

Kosina was recently questioned (Justo & Castro 2010), and it comes close to the

morphological concept of V. dunensis.

The purpose of this study was to determine whether the collections reported

by Fernandez-Sasia (2010) and Contu & La Rocca (1999) represent V. dunensis

using both morphological and molecular characters (ITS region).

Materials & methods

Morphology

Descriptive terminology follows Vellinga (1988) and Boekhout (1990). Microscopical

preparations from dried material were mounted in Congo Red and 5% KOH. The

following abbreviations are used in text: avl for average length, avw for average width,

Q for quotient of length and width and avQ for average quotient. Extreme measurements

are indicated within parentheses. Herbarium acronyms follow Thiers (2011) except that

“SCM” is used for the “Societat Catalana de Micologia” herbarium and RFS refers to the

personal herbarium of Roberto Fernandez-Sasia.

Molecular analysis

Standard methods for DNA extraction, PCR amplification and DNA sequencing

were applied (e.g. Justo et al. 2011). Primer pairs ITS1F and ITS4 (Gardes & Bruns 1993)

were used for both PCR and sequencing. Complete information about the collections

of Volvariella that were sequenced is given in Tas. 1. The remaining sequences used

in the analysis were retrieved from GenBank and come from the studies of Menolli &

Capelari (2008), Li et al. (2009) and Justo et al. (2011). GenBank accession numbers are

given in Fic. 2. No suitable outgroup for Volvariella was found using BLAST searches

(http://blast.ncbi.nlm.nih.gov/Blast.cgi). Therefore, after running a preliminary analysis

using Coprinus comatus (O.F. Mill.) Pers. as an outgroup for all Volvariella sequences

and taking into consideration the results from the combined nLSU-5.8S-nSSU analyses

of Justo et al. (2011), V. bombycina (Schaeff.) Singer and V. volvacea (Bull.) Singer were

used as outgroup taxa in the final dataset. These two species appear to be the sister-

group to all other species of Volvariella sequenced to date. Sequences were aligned with

MAFFT (Katoh et al. 2002) using the Q-INS-i strategy. The alignment was examined

and manually corrected in MacClade 4.05 (Maddison & Maddison 2002). It has been

deposited in TreeBASE (http://purl.org/phylo/treebase/phylows/study/TB2:S11441).

Pairwise similarities were calculated using Jalview (Waterhouse et al. 2009). A Maximum

Likelihood analysis was run in RAxML servers (Stamatakis et al. 2008) with 100 rapid

bootstrap replicates.

TABLE 1. Volvariella samples newly sequenced in this study.

SPECIES

V. dunensis

(Vila et al. 1999,

Justo & Castro 2010)

V. dunensis

(Fernandez-Sasia

2010)

V. dunensis

(Contu & La Rocca 1999

as V. nigrovolvacea)

V. hypopithys

(Fr.) Shaffer

V. pusilla

(Pers.) Singer

V. pusilla

V. strangulata (Romagn.)

Vizzini & Contu

V. terrea

Volvariella sp.

Taxonomy

CouUNTRY—

LOCATION

Spain—

Barcelona, El Prat de

Llobregat, El Pinar

Spain—

Bizkaia,

Muskiz

Italy—

Sardinia,

prov. Cagliari, Chia

Italy—

Stazzo Montesu,

Olbia-Tempio

Spain—

Asturias,

Endriga

Italy—

Oschiri,

Olbia-Tempio

Italy—

Emilia Romagna

France—

Rixheim

Spain—

Madrid,

Parque del Oeste

Slovenia—

Tolminski,

Tolmin

COLLECTOR;

SAMPLING DATE

[herb. number]

F. Angel;

08.11.1997

[SCM 3513]

R. Fernandez-Sasia;

10.11.2007

[RFS 070310-03]

M. Contu;

26.X11.1997

[TO-AV140]

M. Contu;

26.XI.2002

[TO-AV137]

R. Picon;

5.X.2005

[AJ51 (LOU)]

M. Contu;

11.XI.2000

[TO-AV139]

L. Setti;

[TO-AV141]

E. Musumeci;

01.X.2005

[LUG 11010,

holotypus]

L.A. Parra;

6.VIII.2005

[LOU 18294]

J. Kavcic;

12.VII.2007

[TO-AV 143]

Volvariella dunensis ... 39

HABITAT

Open dunes

Dunes with Tamarix gallica

e& Carpobrotus edulis

Sand dune in Juniperus

phoenicea debris;

among Pistacia lentiscus,

J. phoenicea, J. oxycedrus

Under Quercus suber

Under Carpinus

betulus near Agaricus

xanthodermus

Among Cedrus

atlantica remnants

Under Fagus sylvatica

and Fraxinus sp.

ITS sequences from the collections TO-AV140 (Sardinia) and RFS 070310-

03 (Atlantic coast of Spain) are 99.5% and 100% identical to the ITS of SCM

3513 (Mediterranean coast of Spain), respectively. All collections are therefore

considered to represent V. dunensis.

Macromorphological differences between the Sardinian and Atlantic

collections with respect to the original collections of V. dunensis are subtle, but

they concern characters commonly used to delimitate species in Volvariella such

as pileus diameter, stipe colour and volva morphology and colour. Variation in

AO ... Vizzini, Contu & Justo

Figure 1. Volvariella dunensis. Basidiomes (Sardinia, Italy, TO AV140). Bar = 50 mm.

microscopical characters is also subtle and mainly concerns size and shape of

basidiospores and cystidia. Here we provide a revised description of V. dunensis

based on all the collections known so far, highlighting only those characters

that differ from the description provided in Justo & Castro (2010).

Volvariella dunensis (Vila, Angel & Llimona) Justo & M.L. Castro,

Mycotaxon 112: 262. 2010. Fic. 1

= Volvariella nigrovolvacea var. dunensis Vila, Angel & Llimona,

Revista Catalana de Micologia 22: 131. 1999.

PitEus diameter 15-100 mm. StT1PE surface white to grey (similar to pileus

colour). VoLvA white to externally dark sepia to brownish-black, entire or

lobate. BASIDIOSPORES (6-)6.5-9.0 x 4.5-6(-6.7) um, avl x avw = 7.5-7.9 x

5.1-5.5 um, Q = (1.15-)1.3-1.7(-1.8), avQ = 1.4-1.55, ellipsoid to oblong,

rarely broadly ellipsoid. PLEuRocystTip1A (34-)40-95(-112) x 13.5-45(-50)

um, clavate, (narrowly) utriform, obovoid, more rarely broadly lageniform or

fusiform; colourless; with thin, smooth walls, a few with a slightly thickened

wall (up to 0.8 um thick). CHEILOCYsTIDIA 20-80(-120) x 15-60 um, clavate,

utriform, fusiform or lanceolate, without apical appendages, colourless; with

thin, smooth walls, a few with a slightly thickened wall (up to 0.8 um thick).

CAULOCYSTIDIA only observed in the original collections, absent in TO-AV140

and RFS 070310-03.

HABIT, HABITAT AND DISTRIBUTION — Gregarious, usually with most of the

basidiomes growing deeply buried in the sand. In coastal dunes. Known from

the Mediterranean basin (NE Spain, Sardinia) and the Atlantic coast of Spain.

Volvariella dunensis ... 41

Trophic strategy

Litter saprotroph

Lignicolous

Mycotrophic on Clitocybe nebularis ®

Mycotrophic (?) on Agaricus xanthodermus (Musumeci & Riva 2007) ¥¥

Pileus color

White O Volvariella sp. LOU18924 Spain JF415139 O @

Grey or Grey-Brown @ 0

V. hypopithys TOAV137 Sardinia HM246492 O

90 [Lv pusilla TOAV139 Sardinia HM246494 ml CO

95} | V. pusilla AJ51 Spain JF415137 mt O

0.07 V. strangulata TOAV414 Italy HM246493 ill OC

ss V. surrecta AJ55 Spain HM562213 xO

97 V. terrea LUG11010 France JF415141 wv &

V. dunensis RFS07031003 Spain JF415138 mi @

79 100 V. dunensis SCM3513 Spain JF415140 ml @

V. dunensis TOAV140 Sardinia JF415136 mi @

V. caesiotincta MA54717 Spain HM562211 C1 &

Volvariella sp. TOAV143 Slovenia HM246500 mi @

V. taylorii AJ54 Portugal HM562210 mi @

V. nivea China FJ749127 ml C)

100 V. lepiotospora AJ155 USA HM562214 O @

V. nullicystidiata Brazil EU920671 mi @

V. bombycina AJ244 Spain HM562212 C1 OC

100

V. volvacea FJ379274 a @

FiGuRE 2. Best tree from the Maximum Likelihood analysis of ITS sequences of Volvariella.

Bootstrap values above 70% are indicated on the branches. The root length has been reduced to

facilitate graphical representation. Scale bar value indicates nucleotide substitutions/site.

42... Vizzini, Contu & Justo

PHENOLOGY — Volvariella dunensis seems to have a relatively narrow and

particular time of fructification during the winter, spanning from the last days

of December (Sardinia) to early March (Atlantic coast of Spain).

ComMENTS— The arenicolous habitat, winter fructification, and cystidial size

and morphology are the key characters for separating V. dunensis from other

species with grey-fibrillose pilei, especially V. volvacea. Courtecuisse (1984)

reports that V. volvacea could be collected “au niveau des dunes” in the Atlantic

coast of France, and Bon & Gehu (1973) signal this species as typical of the

Ammophiletea communities. In the UK (Orton 1986), Netherlands (Boekhout

1990), and Spain (Justo & Castro 2010), V. volvacea has been consistently

reported as growing inside greenhouses or outside on organic rich substrates

(leaves, compost, sawdust). Given the external resemblance of both species it is

very likely, in our opinion, that at least some coastal records of V. volvacea from

France may, in fact, represent V. dunensis.

In the phylogenetic analysis, V. dunensis appears as sister to the clade

containing V. terrea Musumeci & A. Riva and the V. pusilla group (Fic. 2).

Apart from this relationship, the internal nodes of the tree receive generally low

support and many species appear on relatively long branches. This is probably a

consequence of the high level of divergence in the ITS sequences of Volvariella.

For instance, the relatively low 62% pairwise similarity between V. bombycina

and V. nivea T.H. Li & Xiang L. Chen could indicate either a high rate of

molecular change in the genus or a relatively ancient speciation event.

The number of Volvariella taxa and collections sampled for molecular analyses

is still insufficient to make broad generalizations about the morphological

species-rank taxonomy. However, it seems that many characters traditionally

used for grouping species in the genus such as pileus colour (grey-brown vs.

white) or trophic strategy (litter saprotroph, lignicolous, mycotrophic) have

undergone many transitions during the evolution of Volvariella species (see

character states on Fic. 2). Additional research using both morphological and

molecular data is badly needed to gain further insights in the taxonomy and

evolution of the genus.

Acknowledgements

We would like to thank Dr. A.M. Minnis (Beltsville, USA) and Prof. M. Capelari

(Sao Paulo, Brazil) for their pre-submission reviews. Our sincere thanks also to Roberto

Fernandez-Sasia (RFS), Antoni Sanchez (SCM), and Enzo Musumeci for sending

collections of V. dunensis and V. terrea.

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http://dx.doi.org/10.1093/bioinformatics/btp033

MY COTAXON

http://dx.doi.org/10.5248/117.45

Volume 117, pp. 45-51 July-September 2011

Moelleriella pumatensis, anew entomogenous species

from Vietnam

SUCHADA MONGKOLSAMRIT*, TAI TOAN NGUYEN’,

Nooc LAN TRAN? & J. JENNIFER LUANGSA-ARD’

"BIOTEC, NSTDA Science Park, 113 Paholyothin Road,

Klong 1, Klong Luang, Pathum Thani, Thailand

Faculty of Agriculture, Forestry and Fisheries, Vinh University,

182 Le Duan Street, Vinh, Nghe An, Vietnam

*CORRESPONDENCE TO: suchada@biotec.or.th

AxBstRAcTt — Moelleriella pumatensis, a fungal pathogen infecting scale insect nymphs

(Hemiptera), is described and illustrated as a new species from Pu Mat National Park in

Vietnam. This species is unique in producing a golden yellow spore mass surrounding the

stroma. In surveys throughout the year in Vietnam, only the anamorphic state has been

found in the natural forest. Morphological characters and phylogenetic analysis of translation

elongation factor 1-a (tefl) reveals this species as an anamorph of Moelleriella.

Key worps — morphology, phylogenetics, taxonomy

Introduction

The genus Moelleriella Bres. (Ascomycota, Hypocreales, Clavicipitaceae), a

fungus pathogenic to scale insects and white flies, was recently segregated from

the genus Hypocrella Sacc. together with Samuelsia P. Chaverri & K.T. Hodge

(Chaverri et al. 2008). Moelleriella was described based on molecular data and

morphology: its ascospores disarticulate inside the ascus, whereas Hypocrella

and Samuelsia ascospores do not. The anamorphic state of Moelleriella

is Aschersonia-like, i.e., similar to Aschersonia sensu stricto (teleomorph

Hypocrella sensu lato; Chaverri et al. 2008). Aschersonia sensu lato species

are differentiated mostly on the shape and color of the stromata that cover

the hosts, pycnidium-like conidiomata, phialides, and presence or absence

of paraphyses. These characters have been useful in distinguishing between

subgenera of Aschersonia (Petch 1921, 1925, Mains 1959, Chaverri et al. 2008).

All species of Hypocrella, Moelleriella, and Samuelsia are pathogenic to scale

insects (Coccidae) or white flies (Aleyrodidae) that feed on living leaves and

branches of monocotyledonous and dicotyledonous plants.

46 ... Mongkolsamrit & al.

Exploration of entomopathogenic fungi in Vietnam was initiated under

the program of biodiversity studies in Southeast Asia, a collaborative project

between the Faculty of Agriculture, Forestry and Fisheries, Vinh University,

Vietnam, and the National Center for Genetic Engineering and Biotechnology

(BIOTEC), Thailand. One study site, Pu Mat National Park, is located in the

southwest of Nghe An Province, an area known to have a high diversity of

plants and mammal species. During the surveys in October 2009, an interesting

Aschersonia-like specimen that produced a yellow mass of conidia covering the

stroma was collected from the underside of a leaf of a dicotyledonous plant.

The objectives of this study were to 1) evaluate the taxonomic position of the

isolates derived from the anamorph state, using the sequences of translation

elongation factor 1-a (tefl), and 2) describe this specimen as a new species

from Vietnam.

Materials & methods

Collection and Isolation

Surveys and collections were made during the rainy season in the Khe Moi trail

in Pu Mat National Park (18°46'-19°12'N 104°24'-104°56'E). Material was examined

and isolated into pure culture from the anamorphic state following Mongkolsamrit et al

(2009). Free-hand longitudinal sections of tissues and conidia were mounted in cotton

blue in lactophenol (Heritage et al. 1996) and measured using a light microscope. The

color of fresh specimens and cultures were compared with the colors from standard

code from Kornerup & Wanscher (1962). Specimens were identified using specialized

literature (Petch 1921, Mains 1959, Chaverri et al. 2008). A voucher specimen and

culture were deposited in BIOTEC Bangkok Herbarium (BBH) and BIOTEC Culture

Collection, Thailand.

DNA extraction, amplification and sequencing

Small pieces of tissue (2-3 mm) of each sample were taken from potato-dextrose-

agar (PDA) plates and placed in 50 ml of potato-dextrose-broth (PDB) in a 250 ml

flask. Flasks were incubated in a shaker at 200 rpm for 15-20 d at 20°C in dark. The

mycelial mass was harvested by filtration (Whatman No.1), washed with sterile distilled

water, lyophilized for 1 d, and crushed in liquid nitrogen using a mortar and pestle. Total

DNA of each sample was extracted using Cetyltrimethyl-ammonium bromide (CTAB)

following the procedure described in Mackill & Bonman (1995). PCR amplification

was done in 50 ul volume consisting of 1x PCR buffer, 200 uM of each of the four

dNTPs, 2.5 mM MgCL, 1 U Taq DNA polymerase (Promega, Madison, Wisconsin) and

0.5 uM of each primer. The primers for tefl were 983f (Carbone & Kohn 1999) and

2218r (Rehner 2001). Amplifications were performed using a MJ Research DNA Engine

ALD1244 thermal cycler following the procedure described in Sung et al. (2001). PCR

products were purified using a QIAquick PCR Purification Kit (Qiagen GmbH, Hilden,

Germany), following the manufacturer's instructions. Purified PCR products were sent

to Macrogen Inc. Korea for sequencing.

Moelleriella pumatensis sp. nov. (Vietnam) ... 47

TABLE 1. List of fungi used in this study.

SPECIES VOUCHER/ISOLATE GENBANK

Balansia henningsiana GAM 16112 AY489610

Epichloe elymi C. Schardl 760 AY986951

Hypocrella disciformis M.L.202i = ARSEF 7695 AY986939

H. disciformis P.C.655 = CUP 067861 EU392643

H. disciformis P.C.676 = CUP 067840 EU392645

H. viridans 189-490 = IMI 346739 EU392649

H. viridans P.C.632 = CUP 067849 EU392650

Moelleriella mollii 193-901a = ARSEF 7660 EU392667

M. mollii 193-901c = ARSEF 7667 EU392668

M. ochracea TE1308 = P.C.726 EU392669

M. ochracea P.C.535 = CUP 067777 AY986926

M. ochracea P.C.626 = CUP 067778 EU392670

M. ochracea P.C.648 = CUP 067779 EU392671

M. pumatensis BBH 29281 = BCC 41004 HQ722026*

M. pumatensis BBH 29281 = BCC 41006 HQ722027*

Samuelsia chalalensis P.C.560 = CUP 067856 EU392691

S. geonomis P.C.614 = CUP 067857 EU392692

S. rufobrunnea P.C.613 = CUP 067858 AY986944

*sequences generated in this study.

Phylogenetic analysis

Sequences were proofed manually, assembled using BioEdit v. 6.0.7 (Hall 2004), and

submitted to Genbank (TABLE 1). Sequences were aligned using ClustalW incorporated

in BioEdit and alignments were refined manually by direct examination. Maximum

parsimony analysis was performed using random addition sequence (10 replications)

and gaps were treated as missing data. Bootstrap analysis was performed using maximum

parsimony criterion in 1000 replication samples.

Results

Molecular analysis

Amplification and sequencing of tefl were successful for 16 strains comprising

two unidentified Aschersonia-like specimens. Fourteen selected closely related

taxa were obtained from GenBank for this study. Sequences of Epichloe elymi

and Balansia henningsiana were used as outgroup taxa. Of the 899-character

alignment in the tefl data set, 188 characters were parsimony informative.

Maximum parsimony analyses of this data set yielded one parsimonious tree

(tree length 436; CI = 0.651, RI = 0.795, RC = 0.518, HI = 0.349) as shown in

Fic 1. The phylogenic tree suggests that the two anamorphic isolates with yellow

conidial masses (BCC41004 and BCC41006) belong to the genus Moelleriella

with a strong bootstrap support of 99%.

48 ... Mongkolsamrit & al.

= H. disciformis M.L.202i

Hypocrella 400 |H. disciformis P.C.655

= H. disciformis P.C.676

at a H. viridans P.C.632

H. viridans \89-490

64 M. mollii 193-901a

400| MM. mollii 193-901c

M. ochracea P.C.535

400| | M. ochracea P.C.626

M. ochracea P.C.648

M. ochracea |IE1308

100 | M. pumatensis BCC41006

M. pumatensis BCC41004

Samuelsia _99[— S- chalalensis P.C.560

S. rufobrunnea P.C.613

S. geonomis P.C.614

Epichloe elymi C. Schardl 760

Balansia henningsiana GAM16112

78 Moelleriella 7%

100

—— 10 changes

Fic. 1. Phylogenetic relationship of Moelleriella pumatensis and related species based on maximum

parsimony analysis of the TEF1 gene. Numbers above each branch represent bootstrap support

from 1000 replicates.

Taxonomy

Moelleriella pumatensis T.T. Nguyen & N.L. Tran, sp. nov. FIG. 2

MycoBank MB 561074

Stromata discoidea vel pulvinata, ad 2.5 mm diam, 1.5 mm alta, sursum rotundata,

aureus, Conidiomata pycnidialia, immerse, disperses, 250-280 um alta, 90-100 um diam.

Phialides cylindricae, ad 25 um longae. Paraphyses pycnidiales praesentes, filiformes,

flexuosae, ad 180 um longae, 1.5 um latae. Conidia fusoidea, utrinque acutata 12-15 x

2-2.5 um.

Type — Vietnam: Nghe An Province, Khe Moi trail, Pu Mat National Park, on scale

insect nymph (Hemiptera) on the underside of dicotyledonous leaf, 18 Oct. 2009, J.J.

Luangsa-ard, S. Mongkolsamrit, T.T. Nguyen, R. Ridkaew & N.L. Tran (BBH 29281,

holotype; ex-type culture BCC41006).

ETymMo.ocy — referring to Pu Mat National Park, the collection location.

ANAMORPE: Aschersonia-like

Moelleriella pumatensis sp. nov. (Vietnam) ... 49

Fic. 2. Moelleriella pumatensis A, B, stroma on host showing conidia mass (white arrow);

C, pycnidium and sporulating structure; D, E, conidiogenous cells and paraphyses; F, conidia;

G, colony on PDA at 20°C after 4 wk (sporulation present). Scale bars: A, B, G = 1 mm; C = 100

um; D, E, F = 20 um.

STROMATA discoid to pulvinate up to 2.5 mm diam and 1 mm high, upper

surface round, golden yellow (Kornerup & Wanscher 1962: 5B7). Conidiomata

pycnidial, embedded, scattered in stroma, ovoid, 250-280 um high, 90-100 um

diam. Conidiogenous cells phialidic, cylindrical, up to 25 um long. Pycnidial

paraphyses present, linear, filiform, flexuous, up to 180 um long, 1.5 um wide.

Conidia narrowly fusiform, with acute ends, 12-15 x 2-2.5 um.

50 ... Mongkolsamrit & al.

CULTURAL CHARACTERISTICS: Conidia germinating within 24 h on PDA.

Colonies on PDA slow-growing, attaining 5 mm diam in 4 wk. Optimal

temperature 20-25°C, with no growth at <5°C and >35°C. Stromatic colonies

yellow, forming moderately compact stromata. Conidial masses golden yellow

(Kornerup & Wanscher 1962: 5B7), appearing as abundant slimy masses from

immersed pycnidia scattered over the surface.

CoMMENTs: A teleomorphic state of this species was not found in the field

although several attempts to find it were made throughout the year. However,

the teleomorph name is used for this new species because both phylogenetic

analysis and morphology support the placement of this species in Moelleriella.

In this case, as well as for Moelleriella madidiensis from Bolivia also with only

an anamorphic state, the teleomorphic name is used (Chaverri et al. 2008).

The anamorphic state of M. pumatensis is similar to Aschersonia aleyrodis

(teleomorph: M. libera) reported from Brazil, Florida, and Venezuela by

Petch (1921) based on pulvinate stroma, yellow mass of extruded spores, and

fusiform conidia. The conidia and paraphyses of M. pumatensis are somewhat

longer; the conidia are 12-15 x 2-2.5 um and paraphyses up to 180 um long.

In contrast, A. aleyrodis has conidia 8-14 x 1.5-2 um, paraphyses 50-110 um

long. In addition, M. pumatensis was compared morphologically with several

species collected from the Neotropics (Chaverri et al. 2008), China (Qiu et al.

2009, Qiu & Guan 2010), and Thailand (Luangsa-ard et al. 2007, 2008, 2010,

Mongkolsamrit et al. 2009). The Vietnamese specimens are mostly similar

to Aschersonia andropogonis (teleomorph: M. ochracea) in having pulvinate

stroma, yellow, yellowish orange to orange spore masses, fusiform conidia

(8-14 x 1.5-2 um), which is commonly found in the Neotropics. However, M.

pumatensis differs from A. andropogonis in the color of the stroma. Moelleriella

pumatensis has a golden yellow stroma while that of A. andropogonis is white

to pale yellow. The DNA sequence analysis of translation elongation factor l-a

(tefl) also supports M. pumatensis as a new species.

Acknowledgments

The authors would like to thank Dr. Amy Rossman (Systematic Mycology &

Microbiology Laboratory, Beltsville, USA) and Dr. Priscila Chaverri (Department

of Plant Sciences and Landscape Architecture, Maryland University, USA) for their

comments and suggestion to improve the manuscript for Mycotaxon. We also would like

to thank Dr. Nigel Hywel-Jones for his guidance in specimen identification. This research

was supported by the project “Research collaboration in identifying entomopathogenic

fungi and selecting specific species with high bioactive compound served as medicine”

between National Center for Genetic Engineering and Biotechnology, Thailand, and

Vinh University, Vietnam, number: 04/2009/HD-NDT. We also would like to thank

Prof. Ngoc Hoi Nguyen, Prof. Morakot Tanticharoen, and Dr. Kanyawim Kirtikara

for their support of the program ‘Biodiversity studies of entomopathogenic fungi in

Moelleriella pumatensis sp. nov. (Vietnam) ... 51

Southeast Asia.

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rDNA andbeta-tubulin phylogeny of Thaispecies of Hypocrellawithnon-fragmenting ascospores.

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Qiu JZ, Ma HF, Wang YY, Guan X. 2009, Two Aschersonia species from Fujian new to China.

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Rehner SA. 2001. Primers for Elongation Factor 1-alpha (EF1-alpha). Available from:

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MY COTAXON

http://dx.doi.org/10.5248/117.53

Volume 117, pp. 53-85 July-September 2011

Lepiota (Agaricales) in northern Thailand — 1.

L. section Stenosporae

P. SYSOUPHANTHONG"” , K.D. HypgE??4, E. CHUKEATIROTE’,

A.H. BAHKALI‘ & E.C. VELLINGA?”

'School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand

*Department of Plant and Microbial Biology, University of California at Berkeley,

Berkeley, CA 94720-3102 U.S.A.

*International Fungal Research and Development Centre,

Research Institute of Insect Resources, Chinese Academy of Forests,

Bailongsi, Kunming, Yunnan Province 650224, P.R. China

“Botany and Microbiology Department, College of Science, King Saud University,

PO. Box: 2455, Riyadh 1145, Saudi Arabia

*CORRESPONDENCE TO: phongeunsysou@gmail.com

ABSTRACT —Eleven species of Lepiota section Stenosporae, reported as new for ‘Thailand, are

fully described and illustrated. Five are compared with European and North American collections

based on nrITS sequence data. Two species possess a cutis-like pileipellis (L. aureofulvella,

L. sp. 2) and nine are characterized by a trichodermium (L. alopochroa, L. castanea, L. citrophylla,

L. erythrosticta, L. griseovirens, L. infelix, L. papillata, L. poliochloodes, L. sp. 1). New to science

are Lepiota aureofulvella (close to L. boudieri) and L. papillata, which is characterized by small

basidiomata covered with gray-brown to olive brown squamules, an orange-white to brownish-

orange context, spurred basidiospores with straight or outgrown base, clavate to cylindrical

cheilocystidia, and a trichodermal pileus covering. A key to Lepiota sect. Stenosporae in northern

Thailand is provided.

KEY worps — Agaricaceae, biodiversity, taxonomy, saprotroph

Introduction

Members of the genus Lepiota (Pers.) Gray (Agaricaceae) are in general

saprotrophic forest floor dwellers (Singer 1986, Vellinga 2004b). They occur

worldwide in tropical and temperate regions, with only a few species reported

from desert and arctic-alpine areas (Vellinga 2004b). Sequence analyses

of nrITS and LSU data in the Agaricaceae have shown that Lepiota forms a

monophyletic clade together with Cystolepiota, Echinoderma, Melanophyllum,

and Pulverolepiota (Vellinga 2003ab, 2004a).

54 ... Sysouphanthong & al.

Lepiota was traditionally divided on morphological grounds into sections

based on spore shape and (secondarily) pileus covering structure. Section

Stenosporae (J.E. Lange) Kihner, typified by L. pseudofelina J.E. Lange, is

characterized by spores with a distinct outgrowth (spur) at the base, and

a trichodermal or cutis-like pileus covering (e.g., Singer 1986, Bon 1993).

Molecular-phylogenetic analyses have since reorganized the morphologically

defined sections (Vellinga 2003b). Vellinga (2003b) noted that species with

spurred spores and a hymenidermal pileus covering have been found to relate

closely to species with ellipsoid spores and a hymenidermal pileus covering,

while species with spores that lack a distinct spur, are slender, and have a

trichoderm (e.g., L. cortinarius J.E. Lange) group with species with fusiform

spores. Vellinga (2003b) also reported that species with ellipsoid spores and

a trichoderm without basal short elements form a monophyletic clade with

Lepiota sect. Stenosporae, but that three subclades can be recognized — two

with spurred spores plus one with ellipsoid spores. Here we focus on the two

spurred-spored clades.

Among the few Lepiota species previously reported from Thailand are

L. clypeolaria (Bull.) P. Kumm., L. cortinarius, L. cristata (Bolton) P. Kumm.,

and L. pseudohelveola Hora (Soytong 1994, Chandrasrikul 1996, Chandrasrikul

et al. 2008), none of which belongs to Lepiota sect. Stenosporae. However, the

following species in Lepiota sect. Stenosporae are known from other parts of

tropical Asia: L. alopochroa, L. citrophylla, L. erythrosticta, L. leontoderes (Berk.

& Broome) Sacc., and L. pyrrhaes (Berk. & Broome) Sacc. from Sri Lanka

(Pegler 1972, 1986; Petch & Bisby 1950); L. castanea, L. erythrosticta, and

L. griseovirens from southern India (Kumar & Manimohan 2009); and L. aurora

E. Horak, L. castanea, L. crepusculata E. Horak, L. erythrosticta, L. infelix,

L. luteocastanea E. Horak, and L. squamatula E. Horak from Papua New Guinea

(Horak 1981).

Lepiotaceous fungi have been studied in East Asia in recent years (Ge &

Yang 2006; Ge et al. 2010; Liang et al. 2009, 2010b; Vellinga et al. 2011). The

present study focuses on Lepiota sect. Stenosporae in northern Thailand, as part

of our ongoing efforts to document the diversity of Agaricaceae from this area

(Zhao et al. 2010, Vellinga et al. 2011). We used morphological and molecular

methods to identify and circumscribe our taxa.

Materials & methods

Collecting and examination methods

Collecting sites in Chiang Mai and Chiang Rai provinces, Thailand, were surveyed

during the rainy seasons from April to October, 2007-2010. The sites, located within the

coordinates 16°06.16'N 98°29.64'E and 19°48.60'N 99°54.60'E at altitudes of 596-1700

m, comprise different vegetation types. Detailed notes, concerning location, vegetation

type, soil and substrate were taken in the field. Most specimens were also photographed

in the field. All material is deposited in the herbarium of Mae Fah Luang University

Lepiota section Stenosporae (Thailand) ... 55

(MFLU). Macromorphological characters were noted in the laboratory based on the

fresh materials, using standard procedures and terms (Vellinga & Noordeloos 2001).

Color annotations in the macroscopical descriptions are from Kornerup & Wanscher

(1978), or if preceded by “Mu: from Munsell (1975). Microscopical characters were

studied from dry specimens and drawn using a drawing tube attached to an Olympus

CX-4lresearch compound microscope. Characters were observed in Congo red in

ammonia, water, or in 2.5-10% of KOH. Chemical reactions in Melzer’s reagent, Cotton

blue and Cresyl blue were noted. At least 20 spores per collection were measured in side

view. The notation [180,7,6] indicates that measurements were made on 180 spores in

seven samples in six collections. The following abbreviations are used: ‘L for lamellae, ‘T

for lamellulae, ‘av!’ for average length, ‘avw’ for average width, ‘Q’ for quotient of length

and width and ‘avQ’ for average quotient.

Molecular and phylogenetic methods

DNA EXTRACTION—DNA was extracted from herbarium collections (TaBLE 1)

according to the instructions of the Biospin Fungus Genomic DNA Extraction Kit

(Bioer Technology Co., Ltd., Hangzhou, P.R. China).

TABLE 1. Lepiota species for the ITS sequences used in the phylogenetic analyses in

this study.

ITS GENBANK

SPECIES COUNTRY COLLECTION & HERBARIUM

ACCESSION NUMBER

L. alopochroa Thailand MFLU090178 (MFLU) HQ647294***

L. andegavensis France P.D.H. Roux 2121 (herb. Roux) AY176461*

L. aureofulvella Thailand MFLU090183 HQ647293***

L. boudieri The Netherlands E.C. Vellinga 1180 (L) AF391025*

Italy MCVE: 474 FJ998388*

L. cf. boudieri US.A E.C. Vellinga 2601 (UC) AY 176479**

L. castanea The Netherlands N.J. Dam 97020 (herb. Dam) AY 176463*

The Netherlands H.A. Huijser (herb. Huijser) AF391026*

L. cingulum Germany M. Enderle (L) AY176359*

L. citrophylla Thailand MFLU090172 HQ647295***

L. cristata US.A P.B. Matheny 1958 (WTU) AF391051**

L. grangei Belgium H.A. Huijser (herb. Huijser) AY176471*

Italy MCVE:4666 FJ998399*

L. griseovirens Italy MCVE:13747 FJ998403*

L. ignicolor The Netherlands H.A. Huijser (herb. Huijser) AY176472*

L. cf. ignipes Italy MCVE: 480 FJ998390**

L. pilodes The Netherlands H.A. Huijser (herb. Huijser) AY176476*

US.A E.C. Vellinga 3234 (UC) EF080865**

L. poliochloodes Thailand MFLU081272 HQ647296***

L. pseudofelina Italy MCVE: 3553 FJ998398*

L. rhodophylla US.A E.C. Vellinga 2610 (UC) AY176480**

US.A E.C. Vellinga 3026 (UC) EF080864**

L. subalba The Netherlands E.C. Vellinga 2242 (L) AY 176489*

L. tomentella The Netherlands H.A.Huijser (L) EF080868*

Lepiota sp. Thailand E.C. Vellinga 3881 (MFLU) HQ647297***

US.A E.C. Vellinga 3327 (UC) EF080867**

US.A E.C. Vellinga 2574 (UC) AY176484**

US.A E.C. Vellinga 2603 (UCB) AY176481**

U.S.A E.C. Vellinga 3014 (UC) EF080866**

“= sequence from Europe, “= sequence from U.S.A, ™ = new sequence from Thailand

56 ... Sysouphanthong & al.

PCR AND SEQUENCING—Primers ITS1-E ITS 1, and ITS4 were used for the nrITS1,

5.88 and nrITS2 regions, and PCR conditions followed Gardes & Bruns (1993). PCR

amplified products were cleaned up by Shanghai Sangon Biological Engineering

Technology & Services Co. Ltd.). Sequencing was performed using Big Dye chemistry,

with the same primers as for PCR, and an ABI PRISM 3100 Genetic Analyzer (both

from Applied Biosystems, Foster City, CA, USA). All new sequences were deposited in

GenBank.

DaTA ANALYsIS—Sequences were edited and contigs assembled using Sequencher

4.2.2 (Gene Codes Corporation, Ann Arbor, MI, USA). The sequences of all taxa

belonging to Lepiota sect. Stenosporae present in GenBank were downloaded, and

the complete data set aligned using MAFFT (Katoh et al. 2002, Katoh & Toh 2008)

and minimally manually adjusted. Heuristic search of ITS was performed using the

maximum parsimony (MP) option of the program PAUP* 4.0 b10 (Swofford 2004),

using 1000 heuristic searches, employing TBR branch swapping and random sequence

addition. Other setting were as follows: gaps were treated as missing data; all characters

are of type unordered and equally weighted; multistate taxa interpreted as uncertainty;

starting trees were obtained via stepwise addition; one tree was held at each step during

stepwise addition; the steepest descent option was not in effect, branches were collapsed

(creating polytomies) if minimum branch length was zero, and MulTrees option was

in effect. Bootstrap supports were evaluated using 1000 bootstrap replicates with 10

heuristic searches per replicate, random sequence addition and TBR branch swapping.

A Maximum Likelihood analysis was performed with the on line program RAxML

(Stamatakis et al. 2008). All free model parameters were estimated by RAxML using

a general time-reversible (GTR) substitution matrix and a proportion of invariable

sites estimate. One hundred rapid ML bootstraps were performed. The ML tree was

visualized with the program Figtree v. 1.3.1 (Rambaut 2009).

Results

Nine named and two unnamed species of Lepiota sect. Stenosporae were

recognized in this study. A key to the eleven taxa is provided and each taxon

is provided with a full description, colored photographs (Frcure 1), and

illustrations. TABLE 2 summarizes the morphological characters of the Thai

species. Two species (L. aureofulvella, L. sp. 2) have a cutis pileus covering and

most (except L. papillata) have dextrinoid spores.

ITS sequence data for five species from Thailand were compared with those

from 23 European and U.S.A. specimens; Lepiota cristata with spurred spores

and a hymeniform pileus covering is used as outgroup (TABLE 1). The topology

of a tree based on Maximum Likelihood analysis is identical to the one presented

here (FIGURE 2) and two clades are present in the tree. Clade 1 comprises

species with spurred spores and a trichodermal pileus covering; the Thai species

L. citrophylla and L. poliochloodes group together and are sister to L. grangei,

L. griseovirens, and L. pseudofelina from Europe with low bootstrap support;

L. alopochroa groups with L. castanea and forms a clade with L. cf. ignipes (from

North America) with high bootstrap support. Clade 2, comprising species

Lepiota section Stenosporae (Thailand) ... 57

FIGURE 1. Basidiomata of species of Lepiota section Stenosporae in the field: a. L. aureofulvella;

b. L. citrophylla; c. L. castanea; d. L. alopochroa; e. L. poliochloodes; f. L. infelix; g. L. papillata;

h. L. sp. 2; i. L. griseovirens; j. L. erythrosticta; k. L. sp.1.

with spurred spores and a cutis-like pileus covering, includes L. andegavensis

and L. boudieri (Europe), L. cf. boudieri and L. rhodophylla (U.S.A.), and

L. aureofulvella (Thailand); L. aureofulvella groups with L. boudieri with low

bootstrap support (FIGURE 2).

58 ... Sysouphanthong & al.

ll 100 ra

ig (7 L. sp.1 HQ647297

L. citrophylla HQ647295

EL. poliochloodes HQ647296

x * I. grangei FJ998399

100 «| T grangei AY176471

og -L- griseovirens FJ998403

100 ET. pseudofelina F3998398

15 [--L pilodes AY176476

9 EF pilodes EFO80865

L. sp EFO80867

L. sp AY176484

L. sp EFO80866

L. cingulum AY176359

13 [~ L. castanea AY176463

? TL. castanea AF391026

11 L alopochroa HQ647294

100 |_18 __1F cf ignipes FI998390

3 [OL subalba AY 176489

10 =} _1F. ignicolor AY176472

L. sp AY176481

L. tomenteila EFO80868

10 «| L- rhodophylla AY176480

10017 rhodophyila EFOS0864

48 33 7

10 $_ TF cf boudieri AY176479

LT. boudierti AF391025 | Clade 2

100 3 fF pboudieri FI998388

L. aureofilvella HQ647293

L. andegavensis AY 176461

L. cristata AF391051

—— 10 changes

Clade 1

FIGURE 2. Phylogenetic relationships of Lepiota section Stenosporae based on a Maximum

Parsimony analysis of nrITS sequences. Bootstrap values 260 % are indicated below the

branches. GenBank accession numbers are given for each collection. The topology of a tree

based on Maximum Likelihood analysis is identical. Two clades are present in the tree, clade

1 is composed of species with spurred spores and a trichodermal pileus covering; clade 2 is

composed of species with spurred spores and a cutis-like pileus covering. Lepiota cristata with

spurred spores and a hymeniform pileus covering is used as outgroup.

Lepiota section Stenosporae (Thailand) ... 59

TABLE 2. Selected Lepiota basidiospore and pileus covering structure characters

in collections from Thailand.*

SPECIES SPORE SIZE (\tM) AVL X AVW Q AVQ_DEXT* ~—s PC?

L. alopochroa 7.5-9.3 x 3.8-4.0 8.5 x 3.9 2.0-2.3 2.2 + Trichoderm

L. aureofulva 5.8-7.5 x 3.0-4.8 7.2 x 3.8 1.6-2.1 1.9 + Cutis

L. castanea 9.5-13.0 x 3.8-5.2 11.5 x 4.5 2.4-2.6 2.5 + Trichoderm

L. citrophylla 6.0-8.5 x 3.8-4.3 7.2 x 4.0 1.5-2.0 1.8 + Trichoderm

L. erythrosticta 7.8-11.3 x 3.8-4.8 9.8 x 4.2 2.2-2.8 2.3 + Trichoderm

L. griseovirens 6.2-7.0 x 3.5-4.2 6.7 x 3.8 1.5-1.8 1.7 + Trichoderm

L. infelix 7.5-9.0 x 4.0-4.2 8.3 x 4.0 1.7-2.2 2.0 + Trichoderm

L. papillata 7.2-11.8 x 4.0-4.8 8.2 x 4.1 1.8-2.5 2.0 - Trichoderm

L. poliochloodes 6.5-8.2 x 3.0-3.8 7.2 x 3.5 1.6-2.4 2.0 + Trichoderm

Lepiota sp. 1 7.1-8.4 x 2.8-4.2 7.8 x 3.2 1.9-2.8 2.4 + Trichoderm

Lepiota sp. 2 6.2-8.0 x 3.2-4.0 7.3 X 3.7 1.6-2.5 1.9 + Cutis

* No taxa have metachromatic basidiospores; all have cheilocystidia.

'dext = dextrinoid, * pc = pileus covering.

Taxonomy

Key to species of Lepiota section Stenosporae identified in this study

. Pileus covering a cutis made up of cylindrical elements....................0.06. 2

Pileus covering a trichoderm made up of erect cylindrical to narrowly

GlAV Ate PICTITOM EG, Ata 3'F ted eae D ama Ot. Ot tas hen hots Path eg kaks Vaekols Pa 3

. Pileus covering yellowish brown, golden-brown to brown; pileus covering elements

cylindrical, septate and not branched ....................004. L. aureofulvella

. Pileus covering grayish yellow, grayish brown, or slightly reddish brown;

pileus covering elements cylindrical, septate and sometimes branched... L. sp. 2

3. Basidiomata pale yellow to:light-yellow . 0... .cnjS Gabe Ge ee eee t a L. citrophylla

8. ASG Olata OLIVE OW shits Rae ala Wesnacale Ride des lia Rd Rdg wt Wibeecalt Rides 4

4, Basidiomata pinkish; pileus with reddish white to pinkish squamules. . . L. erythrosticta

4, Basidiomata not pinkish; pileus without reddish white to pinkish squamules,

but with gray,-brown, greenish to-dark Colors. 0 iisic.6 ou sicier ge leiaie evade date aie dea 5

5. Pileus covered with gray to grayish brown, dark gray or olive brown squamules. . . .6

5. Pileus covered with brown to dark brown or reddish brown squamules ........... 8

6. Pileus umbonate with high umbo or papilla, covered with brownish gray to

Olive: brownSopuarile ss te Ph eh cries x went g manent soieene L. papillata

. Pileus umbonate with wide umbo, covered with gray to grayish brown,

sometimes cliyacedus:squamules; .: 4.0. sven siheestheos teens tease kes the 5 7

. Pileus with gray to grayish brown small plush-like squamules or floccules;

smell mild; basidiospores 6.5-8.2 x 3.0-3.8 um. ...........006- L. poliochloodes

. Pileus with small plush-like squamules or floccules, grayish brown to

black, sometimes with a hue of green to olive-brown; smell fruity;

basidiospores 6.2-7.0 x 3.5-4.2 UM... . cece eee eee eee L. griseovirens

60 ... Sysouphanthong & al.

8. Pileus covered with reddish brown small scales or squamules............... L. sp. 1

8. Pileus covered with light brown to dark brown squamules....................0.. 9

9; Pileus'convex without-or with I6w-uimbor.s.¢ eine: gies laos Maes Meee L. infelix

o Pileussunibonate Or withtdistincttmbo. Sainte te keh dad che tees 10

10. Pileus covered with dark brown to dark at center (umbo), with brown squamules

around center toward margin; smell spicy; basidiospores 9.5-13 x 3.8-5.2 um

Bn Pi cheats epg het Tk ec Maton cate Re ES Se GEM na Reo xl aire Noun De iia yar a L. castanea

10. Pileus umbonate, covered with brown squamules toward margin; smell fruity;

basidiospores: 4529. 3-xe3-8=4-07n.. 73 Fleet ent htt L. alopochroa

Lepiota alopochroa (Berk. & Broome) Sacc., Syll. Fung. 5: 63. 1887. FIG. 3

= Agaricus alopochrous Berk. & Broome, J. Linn. Soc., Bot. 11: 510. 1871.

= Lepiotula alopochroa (Berk. & Broome) E. Horak, N.Z. J. Bot. 18: 185. 1980.

PiLEus 14 mm, umbonate, expanding to plano-concave with low umbo, with

inflexed margin, brown (7E6-7) at umbo, rough or with crowded squamules,

with surface breaking up around umbo, with brown (7E6-7) fibrillose

squamules toward margin, on light brown (6D5-6) fibrillose background, with

peeling surface at margin and white background, with white fibrillose remnants

of partial veil and margin exceeding lamellae. LAMELLAE free, crowded,

ventricose, 1 mm wide, white or pale yellow to orange-white (4A3, 5A2), with

white serrulate edge. STIPE 35 x 2.5-4 mm, cylindrical or slightly tapering to

apex and wider at base, with grayish orange to brownish orange (5B4-5, 6C5)

background, white fibrillose at annular zone, with brown (7E6-—7) squamules or

fibrillose squamules at middle zone downward base, with white rhizomorphs at

base, hollow. ANNULUS as an annular zone. CONTEXT in pileus white, 0.5 mm

wide; in stipe concolorous with surface. SMELL fruity. TasTE unknown. SPORE

PRINT white.

BASIDIOSPORES [25,1,1] 7.5-9.3 x 3.8-4.0 um, avl x avw = 8.5 x 3.9 um, Q =

2.0-2.3, avQ = 2.2, in side view oblong to cylindrical, spurred, truncate or with

outgrown base, with acute apex, in frontal view fusiform to oblong, thick-walled,

hyaline, dextrinoid, congophilous, cyanophilous, not metachromatic in Cresyl

blue. Basrp1A 13-17 x 6.0-8.0 um, clavate, narrowly clavate, rarely subclavate,

4-spored, thick-walled, hyaline. LAMELLA EDGE sterile. CHEILOCYSTIDIA 25-37

x 5-7 um, narrowly clavate to cylindrical, with rounded apex, rarely utriform,

thin-walled, hyaline. PLEUROCysTIDIA absent. PILEUS COVERING a trichoderm

made up of narrowly clavate to cylindrical elements, 50-113 x 3.5-18 um,

brown and thick-walled, with parietal pale brown pigment; underlayer with

hyaline to pale brown hyphae, 2.5-4 um wide. CLAMP CONNECTIONS present

in all tissues.

HABITAT AND DISTRIBUTION—Solitary, saprotrophic and terrestrial on

soil, on dead leaves in bamboo forest. Known from one locality in northern

Thailand.

Lepiota section Stenosporae (Thailand) ... 61

e le d

FiGuRE 3. Lepiota alopochroa (MFLU090178).

a. basidiomata; b. basidiospores; c. basidia; d. cheilocystidia; e. pileus covering.

Scale bars: a = 10 mm; b-d = 10 um; e = 20 um.

MATERIAL EXAMINED: THAILAND, CHIANG Mal PRov., MAE TaenG Distr., Mok

Fah Waterfall, 20°02'43.1"N, 99°52'35.0"E, 596 m alt., 6.VIII.2008, P. Sysouphanthong,

MFLU090178.

DiscussIon—As a representative of the Lepiota castanea group, L. alopochroa is

characterized by relatively short spores (averaging 8.5 x 3.9 um) and relatively

short elements in the pileus covering; the cheilocystidia are narrowly clavate

to cylindrical. Despite these short spores and short pileus covering elements

it differs from L. ignicolor Bres. in nrITS sequences (FIGURE 2). The Thai

collections of L. castanea have darker basidiomata, longer spores, and longer

elements in the pileus covering.

62 ... Sysouphanthong & al.

Although the collection reasonably well fits L. alopochroa as described by

Pegler (1972, 1986) and Horak (1980, as Lepiotula alopochroa), there are no

available molecular data to support this conclusion.

Lepiota aureofulvella Sysouphanthong, K. D. Hyde, Chukeatirote & Vellinga,

sp. nov. FIG. 4

MycoBank MB 519671

Pileus 15-40 mm, convexus vel campanulatus, umbonatus, plano-concavus, fibrillosus,

flavidus-brunneus vel auratus. Lamellae liberae, albae vel pallide luteae, 5-6 mm. Stipes

25-45 x 2.5-5 mm, cylindricus vel subclavatus, fibrillosus, albus dein laete brunneus.

Basidiosporae 5.8-7.5 x 3.0-4.8 um, calcaratae. Basidia 16. -21.5 x 5.5-6.3 um,

clavata. Cheilocystidia 17.5-27.5 x 5-10 ym, clavata, anguste clavata. Epicutis ex cellulis

cylindraceis vel anguste clavatis. Fibulae adsunt.

Hotorypus: Thailand, Chiang Mai Prov., Mae Taeng Distr., Mok Fah Waterfall National

Park, 20°02'43.1"N, 99°52'35.0"E, 596 m. alt., 8.VIII.2008, P. Sysouphanthong PS110

(MFLU090183, holotype).

Erymo oey: ‘aureofulvella’ means golden-brown and refers to a close relationship with

L. boudieri (syn. L. fulvella Rea)

PitEus 15-40 mm, convex to umbonate, expanding to plane or plano-

concave, with straight margin, with crowded brown (6E7-8) fibrils at umbo,

with concentrically crowded fibrillose squamules from umbo toward margin,

yellowish brown to golden brown (5D5-7), on white background, when mature

surface peeling from background; margin fringed and exceeding lamellae.

LAMELLAE free, crowded, ventricose, 5—6 mm wide, white, with white serrulate

edge. Stipe 25-45 x 2.5-5 mm, cylindrical to subclavate, with white to light

brown (6D4-5) background, white fibrillose in upper zone, at base with slightly

brown (6E4-8) fibrillose squamules, with white rhizomorphs at base, hollow.

ANNULUS an annular zone, with white fibrils. CONTEXT in pileus white and

4.5-5 mm wide; in stipe concolorous with surface. SMELL fruity. TasTE

unknown. SPORE PRINT white.

BasIpiosPores [60,3,3] 5.8-7.5 x 3.0-4.8 um, avl x avw = 7.2 x 3.8 um,

Q = 1.6-2.1, Qav = 1.9, cylindrical to oblong, with truncate to spurred base,

triangular or with curved abaxial side, in frontal view cylindrical, thick-walled,

hyaline, dextrinoid, congophilous, cyanophilous, not metachromatic in Cresyl

blue. Basrp1A 16.3-21.5 x 5.5-6.3 um, clavate, slightly thick-walled, 4-spored.

Lamella edge sterile. CHEILOCYsTIDIA 17.5-27.5 x 5-10 um, clavate, narrowly

clavate, utriform, cylindrical, hyaline, slightly thick-walled. PLEUROCYSTIDIA

absent. PILEUS COVERING a cutis made up of cylindrical elements, sometimes

with narrowly clavate terminal elements, 40-130 x 5.5-14 um, thick-walled,

with brown parietal pigment; underlayer with hyaline to pale brown hyphae,

3-4 septate, 5.0-13 um wide. CLAMP CONNECTIONS present in all tissues.

Lepiota section Stenosporae (Thailand) ... 63

b () f ff

mea

need

4. {.

pads

ray

Jit

ran

% 7,

Bet

Gna ae Bp

h :

qe.

Tay

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coe RECA ger oe

aS ae ae

CR ne para rarces

FiGuRE 4. Lepiota aureofulvella (MFLU090183).

a. basidioma and a section; b. basidiospores; c. basidia;

d. cheilocystidia; e. elements of pileus covering.

Scale bars: a = 10 mm, b-e =10 um.

HABITAT AND DISTRIBUTION—growing in a small to large group;

saprotrophic on decayed wood, soil rich in humus, and found in forest with

dominant bamboo; in one locality in northern Thailand.

64 ... Sysouphanthong & al.

ADDITIONAL MATERIAL EXAMINED: THAILAND, CHIANG Mar PRov., MAE TAENG

Distr., Mok Fah Waterfall National Park, 20°02'43.1"N, 99°52'35.0"E, 596 m. alt.,

24.V1.2010, P. Sysouphanthong, MFLU 100592; ibidem, 9.VI.2010, P. Sysouphanthong,

MFLU100621.

Discusston—Lepiota aureofulvella is characterized by the yellowish brown or

light brown to golden brown color of the pileus covering, clavate to utriform

cheilocystidia, and cutis-like pileus covering.

The only similar species, L. alopochroa, resembles L. aureofulvella in color

and general stature but differs in the trichodermal pileus covering.

Lepiota squamatula from Papua New Guinea, which is discussed and

illustrated as having a cutis-like pileus covering but diagnosed in Latin as

having a trichoderm, differs in having orange-brown squamules and fibrils on

the pileus, longer (8.0-9.0 um) and narrower (2.5-3.5 um) basidiospores, and

a pileus covering “composed of cylindric interwoven hyphae forming a cutis or

trichoderm” (Horak 1980).

Some north temperate species with a cutis-like pileus covering and spurred

basidiospores are similar to L. aureofulvella. Lepiota boudieri Bres. is similar

in morphology but has somewhat darker and less golden colors and shorter

(5.8-7.5 um) basidiospores (Vellinga 2001); the western North American

L. rhodophylla Vellinga is distinguished by a pinkish-brownish pileus covering

with a pinkish brown margin and pink to pale brownish lamellae (Vellinga

2006); and the French species, L. andegavensis Mornand (Mornand 1983), has

very dark pileus colors.

The nrITS based phylogeny (FiGuRE 2) clearly separates L. aureofulvella

from all the other species in clade 2 with a cutis-like pileus covering and for

which sequence data are available.

Lepiota castanea Quél., C.r. Ass. Frang. Av. Sci. (Reims 1880) 9: 661. 1881. Fic. 5

PitEus 15-20 mm, umbonate to campanulate with wide umbo, with

inflexed margin, with crowded fibrils at umbo, brown (7E6-8) and changing to

dark brown to dark when dry (7F6-8), with rough umbo, with rough surface

around umbo to margin, brown (6E5-6) and paler at margin (6D5-7), with

surface soon breaking up and becoming squamules or patch-like squames

with upcurved tips, on white background, at margin white fibrillose and with

light brown fibrillose squamules. LAMELLAE free, crowded, ventricose, 3 mm

wide, white, with smooth edge. StrpE 40-58 x 1.8-3 mm, cylindrical or slightly

tapering to apex, white from apex to 1/3 of length, white or pale orange to light

orange from annular zone to base with squamule-like, light brown to brown

(6D5-7, 6E6) patches and bands, hollow, with white rhizomorphs at base.

ANNULUS an annular zone, with squamules or fibrillose partial veil. CONTEXT

in pileus white and dull, 2-2.5 mm wide, in stipe white. SMELL spicy. TASTE

unknown. SPORE PRINT white.

Lepiota section Stenosporae (Thailand) ... 65

SON

FiGuRE 5. Lepiota castanea (MFLU090106).

a. basidioma and a section; b. basidiospores; c. basidia; d. cheilocystidia; e. pileus covering.

Scale bars: a = 10 mm; b-e = 10 um..

BASIDIOSPORES [75,2,2] in side view 9.5-13 x 3.8-5.2 um, avl x avw = 11.5

x 4.5 um, Q = 2.4-2.6, Qav = 2.5, in side view cylindrical, with truncate to

distinctly spurred base, mostly with outgrown broad basal spur and long hilar

appendage, some with a straight base, in frontal view oblong to cylindrical,

hyaline, slightly thick-walled, dextrinoid, congophilous, cyanophilous, not

metachromatic in Cresyl blue. Basip1a 18-23 x 9.0-10 um, clavate, 4-spored,

with or without mucilaginous contents. LAMELLA EDGE sterile. CHEILOCYSTIDIA

66 ... Sysouphanthong & al.

18-36 x 5.5-12.5 um, usually clavate to narrowly clavate, with acute or rounded

apex, often utriform, thin-walled. PLEUROCysTIDIA absent. PILEUS COVERING

and stipe covering a trichoderm made up of narrowly clavate to cylindrical

elements, wider at middle and narrowing to apex and base, 40-375 x 6.5-35

um, with slightly thickened walls, pale brown to brown-walled, with parietal

brown pigment, with rare short clavate elements; under layer with cylindrical

hyphae, hyaline to pale brown-walled, 2.5-5 um wide. CLAMP CONNECTIONS

present in all tissues.

HABITAT AND DISTRIBUTION—growing solitary or in a small group,

saprotrophic and terrestrial on forest floor with high amount of humus, only

found in deciduous forest at high elevation (Doi Inthanon National Park, Chiang

Mai Province, Thailand). Widely distributed in Europe and North America and

temperate parts of Asia (Vellinga 2001), also recorded from India and Nepal

(Manandhar & Adhikari 1994), and from Papua New Guinea (Horak 1981).

This is a new record for Thailand.

MATERIAL EXAMINED: THAILAND, CHIANG Mal PRov., CHom Tuone Distr., Doi

Inthanon National Park, Junction of Highway 1009 and Road to Mae Cheam, 19°31.58'N,

98°29.64'E, 1700 m alt., 20.V1.2008, P. Sysouphanthong MFLU090106; 15.VII.2008, P.

Sysouphanthong, MFLU090154.

Discuss1oNn—The L. castanea complex is in need of a combined morphological-

molecular approach to resolve the species borders. Vellinga (2001), who took a

pragmatic view, considered L. castanea a very variable species, variable in terms

of spore length and length of pileus covering elements. Migliozzi & Zecchin

(1997, 2000) recognized several species in the complex: L. rufidula Bres. for

the long-spored taxon, commonly (also by us) named L. castanea; L. ignipes

Bon is considered a synonym of L. rufidula; L. castanea s. str. for one short-

spored species and L. ignicolor for a taxon with small basidiomata and spores

tending toward ovoid. A collection referred to L. ignicolor is sister to L. subalba

P.D. Orton in the phylogenetic analyses based on nrITS sequences (FIGURE 2);

L. subalba has short elements in the pileus covering but whitish basidiomata.

The characters of the Italian collection named L. ignipes in GenBank (accession

number FJ998390) are not known, but the molecular data indicates that it

differs from the other taxa in this complex.

Lepiota cingulum Kelderman, which resembles L. castanea, has predominantly

pinkish brown basidiomata and a pileus covering with terminal elements that

tend to taper towards the tips (Kelderman 1994).

The record of L. castanea from Nepal (Manandhar & Adhikari 1994-95)

probably refers to another species, as the spores are described as ellipsoid.

Lepiota castanea is rare in northern Thailand and was found only at one

locality at 1700 m asl on Thailand’s highest mountain early in the rainy season.

It represents the temperate Eurasian element of the Thai Lepiota mycota.

Lepiota section Stenosporae (Thailand) ... 67

aa?)

Lee avd

Wot

FiGure 6. Lepiota citrophylla (MFLU090101).

a. basidiomata; b. cheilocystidia; c. basidiospores; d. basidia; e. pileus and stipe covering.

Scale bars a = 10 mm; b-e = 10 um.

Lepiota citrophylla (Berk. & Broome) Sacc., Syll. Fung. 5: 57. 1887. FIG. 6

= Agaricus citrophyllus Berk. & Broome, J. Linn. Soc., Bot. 11: 509. 1871.

ExcLuDED: Lepiota citrophylla sensu Rea (1922), Akers & Sundberg (1998), Liang et al.

(2010a).

PitEus 15-35 mm, subglobose to conical, expanding to campanulate, with

moderately wide umbo, with inflexed margin, at umbo covered with fine

68 ... Sysouphanthong & al.

squamules to floccules, brown (6D7) at umbo and with light brown to

yellowish brown (5D6-8) squamules toward margin on pale yellow to light

yellow (4A3-4) background; margin with distant squamules, sometimes with

partial veil. LAMELLAE free, crowded, ventricose, 2.5-3.8 mm wide, pale yellow

to light yellow (3A4-4A4), with concolorous serrulate edge. STIPE 60-65

x 2-3 mm, cylindrical, slightly wider at annular zone; surface covered with

squamules from annular zone to middle and with small light brown to brown

(5D6-6D7) squamules at middle to base, on light yellow (4D4) background;

hollow. ANNULUS an annular zone or made up of squamules. CONTEXT in

pileus 1.8-2 mm wide and pale yellow (3A3), in stipe light yellow (4D4). SMELL

as burnt rubber. TasTE unknown. SPORE PRINT white.

BASIDIOSPORES [100, 5, 5] in side view 6.0-8.5 x 3.8-4.3 um, avl x avw = 7.2

x 4.0 um, Q = 1.5-2.0, avQ = 1.8, in side view with straight or outgrown spur

at base, ellipsoidal to cylindrical with rounded or more acute apex, in frontal

view oval, cylindrical, hyaline, thick-walled, strongly dextrinoid, congophilous,

cyanophilous, not metachromatic in Cresyl Blue. BAsip1a 19-37 x 6.5-8.5 um,

clavate to slightly narrowly clavate, 4-spored, hyaline, with thin wall. LAMELLA

EDGE Sterile. CHEILOCYSTIDIA 15-60 x 5.0-10 um, narrowly clavate, sometimes

clavate, utriform, rarely narrowly fusiform, or wider at center. Pleurocystidia

absent. PILEUS COVERING a trichoderm made up of erect narrowly clavate

elements, often narrowed into pedicel, 50-160 x 8.5-17 um, with thin brown

wall, with parietal and intracellular brown pigment, with an under layer of

cylindrical hyphae, 2-6.5 um wide, with thin hyaline to slightly pale brown

walls, with intracellular pigment. Stipe covering with same structure as pileus

covering. CLAMP CONNECTIONS present in all tissues.

HABITAT AND DISTRIBUTION—Solitary or in small groups, terrestrial in soil

with decaying leaves and wood, found in deciduous forest and pine forests of

northern Thailand. Widely distributed in the tropics: Sri Lanka (Pegler 1972),

Trinidad (Dennis 1952), Kenya and East Africa (Pegler 1977), and India

(Natarajan & Manjula 1983). This is the first record from Thailand.

MATERIAL EXAMINED: THAILAND, CHIANG Mal PrRov., Mak TaENG Distr., Hot

Spring National Park, 16°06'16.1"N, 99°43'07.9"E, 780-805 m alt., 30.VII.2008,

P. Sysouphanthong, MFLU090172; 8.VIII.2010, MFLU090185; Mae Sae Village,

19°07'13.7"N, 98°43'52.9"E, 962 m alt., 14.V1.2008, P. Sysouphanthong, MFLU090101;

CHIANG Ral PrRov., Mak Fa Luanc Distr., Doi Tung National Park, 10.V1.2009, P.

Sysouphanthong, MFLU 100405; W1ANG CHIANG RunG DistTr., Hui Mae Sack Waterfall

National Park, 26.VHI.2009, P. Sysouphanthong, MFLU 100424.

DiscusstoN—This species is found during the whole rainy season and is

widespread in northern Thailand. It is quite variable in basidiome size and

pileus covering color. Lepiota citrophylla is the only species of sect. Stenosporae

in northern Thailand characterized by predominantly yellow basidiome colors.

The Thai material is very similar to collections from Sri Lanka described by

Lepiota section Stenosporae (Thailand) ... 69

Pegler (1972, 1986), but he did not recover cheilocystidia in the material

from Sri Lanka and his described shorter pileus covering elements are shorter

(30-80 x 12-17 um). There can be two reasons for this: the material he studied

is young, or the longer elements were degraded, broken, or otherwise not easily

found and measured.

The name L. citrophylla has been applied to both a European species (e.g.

Rea 1922; Kiihner 1934) and an American species (Akers & Sundberg 1998),

but those have non-spurred spores; the European species was subsequently

described as L. xanthophylla P.D. Orton (Orton 1960).

Lepiota luteocastanea is very close to L. citrophylla in yellow basidiome

colors, but L. luteocastanea is distinctly purple tinged on the pileus and has

fusoid-cylindric elements in the pileus squamules (Horak 1981). Lepiota

subcitrophylla Hongo (1956) from Japan also comes close to L. citrophylla but

turns blue when bruised or damaged. Verrucospora vulgaris Pegler, which

resembles L. citrophylla in the brown squamules, yellow tinged basidiomata,

and persistent annulus, does not have free lamellae and produces angular

basidiospores (Pegler 1977).

Lepiota citrophylla has previously been known only from tropical parts of

Asia and is reported from Thailand here for the first time. A recent report of

this species from China refers to a different species representing Lepiota sect.

Lepiota, as the spores are not spurred (Liang et al. 2010a).

Lepiota erythrosticta (Berk. & Broome) Sacc., Syll. Fung. 5: 62. 1887. FIG. 7

= Agaricus erythrostictus Berk. & Broome, J. Linn. Soc., Bot. 11: 508. 1871.

Piteus 14 mm, parabolic, with straight margin, with crowded reddish or

pinkish (8A2-3) squamules at center and toward margin, sometimes with

fibrillose squamules, on white to pale red (8A3) background; margin sulcate,

with reddish white (8A2-3) fibrillose squamules and partial veil remnants.

LAMELLAE free, slightly crowded, broadly ventricose, white to pale orange

(6A3), with white floccose edge. STIPE 32 x 2-2.5 mm, cylindrical, with, 3-3.5

mm wide basal bulb, white (6A2) at apex, covered with light brown to reddish

or pinkish (6D4, 8A2) fibrillose squamules on orange fibrillose background

from annular zone downward, hollow. ANNULUS an annular zone, with fibrils

and fibrillose squamules similar to those on pileus. CONTEXT in pileus white,

turning orange white to pale orange (5A2-3), 1 mm wide; in stipe orange white

(5A2) in apical zone, grayish red to reddish brown (8C5-8D5) from midway

downward. TasTE unknown. SMELL unknown. SPORE PRINT white.

BASIDIOSPORES [25,1,1] 7.8-11.3 x 3.8-4.8 um, on average 9.8 x 4.2 um,

Q = 2.2-2.8, avQ = 2.3, in side-view cylindrical with truncate to spurred base,

narrowly triangular, in frontal view cylindrical, thick-walled, hyaline, not

dextrinoid, congophilous, cyanophilous, metachromatic in Cresyl blue. BAsIDIA

70 ... Sysouphanthong & al.

FiGurRE 7. Lepiota erythrosticta (MFLU090034).

a. basidioma; b. basidiospores; c. basidia; d. cheilocystidia; e. pileus covering.

Scale bars: a = 1 mm; b-d = 10um; e = 20 um.

16-28 x 5-8 um, clavate, 4-spored, rarely 2-spored. LAMELLA EDGE Sterile.

CHEILOCYSTIDIA 19-32 x 8-13 um, irregularly clavate or sphaeropedunculate,

utriform, often narrowly fusiform, thick-walled, hyaline. PLEUROCYSTIDIA

absent. PILEUS COVERING a trichoderm made up of long cylindrical elements,

70-170 x 4.5-12.0 um, hyaline to pale pink-walled, with parietal and

intracellular pink pigment, with underlayer made up of hyaline hyphae, 4-10

Lepiota section Stenosporae (Thailand) ... 71

um wide. STIPE COVERING a trichoderm similar to pileus covering. CLAMP

CONNECTIONS present in all tissues.

HABITAT AND DISTRIBUTION—Growing solitary, on humus-rich soil with

decayed leaves and wood, in forest with dominant bamboo; found in one

locality in northern Thailand. Distributed in Sri Lanka (Pegler 1972), Brazil

(Wartchow et al. 2008), and Trinidad (Dennis 1952). This is a new record for

Thailand.

MATERIAL EXAMINED: THAILAND, CHIANG Mal PRov., Mak TAaENG Distr., Hot

Spring National Park, 16°06'16.1"N, 99°43'07.9"E, 780-805 m alt., 4.VIII.2007, P.

Sysouphanthong, MFLU090034.

Discusston—Lepiota erythrosticta is recognized by the reddish or pinkish

squamules on pileus and stipe and pinkish lamellae. The non-dextrinoid spores

and metachromatic reaction of the spore wall in Cresyl blue distinguishes

L. erythrosticta from all other species in sect. Stenosporae.

The sole Thai collection consisted of young basidiomata. Wartchow et al.

(2008) recorded longer pileus and stipe covering elements for material from

Brazil; Dennis (1952), who recorded this species from Trinidad, noted smaller

basidiospores than those found in material from Thailand. Pegler (1972), who

studied the Sri Lankan type material but did not report the reaction of spore

walls in Cresyl blue, also cited slightly smaller basidiospores. It is possible that

the South American reports represent a different taxon.

This species is rare in Thailand; it was only found once (August 2007) in

the middle of rainy season at Hot Spring National Park but not the other three

years the survey was conducted. Like L. citrophylla, L. erythrosticta is a species

with a tropical distribution pattern.

Lepiota griseovirens Maire, Bull. trimest. Soc. mycol. Fr. 44:37. 1928. — F1aG.8

MISAPPLIED NAME: Lepiota pseudofelina sensu Bon (1993), Candusso & Lanzoni (1990).

EXCLUDED: Lepiota griseovirens sensu Reid (1972), Bon (1993).

PriLEus 8-13 mm, conico-campanulate, expanding to umbonate, plano-convex

with broad umbo, with straight margin, at centre covered with tomentose tufts,

grayish brown (7F3, 7E3, 7D3) to black, sometimes with a hue of green to

olive-brown (4F4-5), toward margin cracked into small squamules, paler than

at center, on orange-white to pale orange (5A2-3) background; margin slightly

fringed, exceeding lamellae in mature specimen. LAMELLAE free, slightly

crowded, ventricose, and rounded near stipe, 2 mm wide, orange-white to pale

orange (5A2), with white eroded to flocculose edge. StrpE 16-20 x 1.5-2.0

mm, with 2.0-2.5 mm wide base, cylindrical, slightly wider at base, white to

orange-white (5A2) innately fibrillose above annular zone and below annular

zone downward toward base, with scattered grayish brown (7F3, 7E3, 7D3)

to black squamules as on pileus, with brownish-orange (6C4-5) background,

72 ... Sysouphanthong & al.

d e

FiGurReE 8. Lepiota griseovirens (MFLU100554).

a. basidiomata and a section; b. basidiospores; c. basidia; d. pileus covering; e. cheilocystidia.

Scale bars: a = 10 mm; b-e = 10 um.

with white mycelium cords, hollow. CONTEXT in pileus white, 1.5-2 mm wide,

in stipe concolorous with surface. SMELL fruity. TASTE unknown. SPORE PRINT

white.

BasIpiIosPpores [50,2,1] in side view 6.2-7.0 x 3.5-4.2 um, avl x avw =

6.7 x 3.8 um, Q = 1.5-1.8, avQ = 1.7, oblong, with truncate base, with hilar

Lepiota section Stenosporae (Thailand) ... 73

appendage, thick-walled, strongly dextrinoid, congophilous, cyanophilous, not

metachromatic in Cresyl blue. Bastp1a 17-20 x 5.5-7 um, clavate, 4-spored,

often with 2 sterigmata. LAMELLA EDGE Sterile. CHEILOCYSTIDIA 22-30 x 6-8

um, clavate, narrowly clavate, narrowly utriform. Pleurocystidia absent. PILEUS

COVERING a trichoderm made up of cylindrical to narrowly fusiform elements,

without short clavate elements at base, 40-140 x 7.0-13 um, thin-walled,

with brown parietal pigment; basal hyphae cylindrical and hyaline, 7.5-12.5

um wide. Stipe covering (squamules) a trichoderm as pileus covering. CLAMP

CONNECTIONS present in all tissues.

HABITAT AND DISTRIBUTION—growing in a small group with few

basidiomata, terrestrial, and saprotrophic on ground of rain forest with

dominant pine. This is the first record for Thailand. Also known from Europe

(Vellinga 2001, Bon 1993).

MATERIAL EXAMINED: THAILAND, CHIANG Ral PRov., MAE Fa Luanc Distr., Doi

Tung National Park, 17.VII.2010, P. Sysouphanthong, MFLU100554.

Discuss1on—Lepiota griseovirens is rare in Thailand and was only found once,

in Doi Tung National Park. The Thai collection is characterized by grayish brown

to black pileus squamules that sometimes have a green to olive-brown hue. Its

spores fall into the lower part of the range recorded for European collections

(Vellinga 2001), but all other characters (including shape of the cheilocystidia)

fit well with the European species concept.

Vellinga & Huijser (1993) discussed the naming of this species, which was

then mainly known as L. pseudofelina. The two nrITS collections labeled as

L. pseudofelina in FiGuRE 2 previously were referred to two different names:

L. pseudofelina and L. griseovirens.

Lepiota infelix E. Horak, Sydowia 33: 133. (1981). FIG. 9

PiLEus 19-23 mm, when young convex or with low umbo, expanding to

plano-concave with low umbo, with straight or slightly inflexed margin, when

young dark brown (7F5-6), soon breaking open into squamules, crowded

and dark brown (7F5-6) at centre and umbo, with fibrillose to fibrillose light

brown to brown (6D7-8) squamules around centre toward margin, on white

fibrillose background; margin split, fringed, exceeding lamellae when mature.

LAMELLAE free, slightly crowded, ventricose, 3 mm wide, white, fragile, with

white eroded edge. Stipe 20-25 x 1.2-1.5 mm, cylindrical, slightly wider at

base; background white at apical zone, darker down towards base, light brown

(6D7) at base, when mature turning grayish orange to brownish orange (6B4,

6C4-6), with brown (6E5-7) squamules from centre down towards base,

hollow. ANNULUS as annular zone, with squamules. CONTEXT in pileus white to

pale yellow (3A2), 1 mm wide; in stipe concolorous with surface. SMELL mild.

TASTE unknown. SPORE PRINT white.

7A ... Sysouphanthong & al.

FiGure 9. Lepiota infelix (MFLU090014).

a. basidioma; b. basidiospores; c. basidia; d. pileus covering; e. cheilocystidia.

Scale bars: a = 20 mm; b-e = 10 um.

Lepiota section Stenosporae (Thailand) ... 75

BASIDIOSPORES [25,2,1] 7.5-9.0 x 4.0-4.2 um, avl x avw = 8.3 x 4.0 um, Q

= 1.7-2.2, avQ = 2.0, oblong to cylindrical, with rounded apex, with distinct

spur at base, with or without lateral hilar appendage, in frontal view oblong,

hyaline, slightly thick - walled, dextrinoid, congophilous, cyanophilous, not

metachromatic in Cresyl blue. Basrp1a 15-18 x 6.0-8.0 um, clavate, 4-spored.

LAMELLA EDGE sterile. CHEILOCYSTIDIA 16-25 x 5-7 um, clavate to narrowly

clavate, often utriform, colorless. PLEUROCYSTIDIA absent. PILEUS COVERING

a trichoderm made up of cylindrical to narrowly clavate elements, 60-150 x

6-15 um, slightly thick-walled, with pale brown parietal pigment, with under

layer made up of hyaline to pale brown-walled, 3-6 um wide, hyphae. STIPE

COVERING a trichoderm, similar to pileus covering. CLAMP CONNECTIONS

present in all tissues.

HABITAT AND DISTRIBUTION—growing in a small group, saprotrophic

and terrestrial on nutrient-rich soil, in deciduous rain forest with dominant

Lithocarpus spp. In Thailand known from one locality. Found in Papua New

Guinea (Horak 1981), and reported from India (Natarajan & Manjula 1983).

This is a new record for Thailand.

MATERIAL EXAMINED: THAILAND, CHIANG Mat PRov., MAE TAENG Distr., Pha

Deng Village, 19°07'13.7"N, 98°43'52.9"E, 905 m alt., 11.VII.2007, P. Sysouphanthong,

MFLU090014.

Discusston—Lepiota infelix, originally described from Papua New Guinea by

Horak (1981), is characterized by reddish brown pileus and stipe squamules.

The species resembles L. ignipes in pileus color, but the slender basidiospore

shape in L. infelix separates the two taxa.

The also similar L. alopochroa, described from Sri Lanka and recorded

from New Zealand (as Lepiotula alopochroa, Horak 1980), Papua New Guinea

(Horak 1981), and now northern Thailand, differs in the color of the pileus

squamules.

Lepiota papillata Sysouphanthong, K. D. Hyde, Chukeatirote & Vellinga, sp. nov.

MycoBANK MB 519794 FIG. 10

Pileus 9-15 mm, conico-convexus dein campanulatus, umbonatus, vel papillatus,

squamuloso-fibrillosus, brunneo-griseus vel olivaceo-brunneus. Lamellae liberae, albae

dein pallide luteae. Stipes 25-30 x 1.5-2 mm, cylindricus vel subclavatus, squamuloso-

fibrillosus. Basidiosporae 7.2-11.8 x 4.0-4.8 ym, calcaratae. Cheilocystidia 30-52 x 6-29

um, cylindracea, clavata. Epicutis ex cellulis substrictis fusoideo-cylindraceis. Fibulae

adsunt.

Hotorypus: Thailand, Chiang Mai Prov., Mae Taeng Distr., Hot Spring National Park,

16°06'16.1"N, 99°43'07.9"E, 780-805 m alt., 9.VII.2007, P. Sysouphanthong PNG041

(MFLU090041, holotype).

EryMo_oey: ‘papillata’ means with a papilla, referring to the shape of the pileus and of

the pileus covering elements.

76 ... Sysouphanthong & al.

mnt Soule

FIGURE 10. Lepiota papillata (MFLU090041).

a. basidioma; b. basidiospores; c. basidia; d. cheilocystidia; e. pileus covering.

Scale bars: a = 10 mm; b-e = 10 um.

PiLEus 9-15 mm, conical at first, expanding to campanulate, umbonate with

umbo or high papilla, with inflexed margin, applanate to slightly plano-concave

with uplifted margin; background brown (5F4), white to yellowish brown

(5E5) on margin, covered with crowded brownish gray to olive brown (4F2-3)

squamules at umbo, with brown (6E6) fibrillose squamules around umbo toward

margin; margin split, slightly appendiculate, exceeding lamellae. LAMELLAE

free, slightly crowded, ventricose,1.5-2 mm wide, when young white, yellowish

Lepiota section Stenosporae (Thailand) ... 77

white to pale yellow (4A2-3) when mature, turning grayish orange (5B5) when

dried, with serrate concolorous edge. Stipe 25-30 x 1.5-2 mm, cylindrical

or slightly tapering upward, with orange-white to grayish red (5A2-7B4)

background at apex, grayish orange to brownish orange (5B4-6C5) at center

to base, turning brown (6D7) when bruised, covered with brownish gray to

olive-brown (4F2-3) squamules from annular zone to base, with orange-white

to brownish orange (5A2-6C5) fibrils between squamules; hollow. ANNULUS

with annular zone, with squamules as stipe covering. CONTEXT in pileus white,

less than 1 mm wide; in stipe grayish orange (5-6B6). SMELL grass-like. TASTE

mild. SPORE PRINT white.

BASIDIOSPORES [50,2,1] 7.2-11.8 x 4.0-4.8 um, avl x avw = 8.2 x 4.1 um,

Q = 1.8-2.5, avQ= 2.0, with straight or outgrown spur, cylindrical to oblong,

with rounded or more acute apex, in frontal view oblong to cylindrical

fusiform, with or without hilar appendage, hyaline, slightly thick-walled,

not strongly dextrinoid in Melzer’s reagent, congophilous, cyanophilous, not

metachromatic in Cresyl blue. Basidia 12-20 x 5.5-7.5 um, clavate, 4-spored,

rarely 2-spored. LAMELLA EDGE Sterile. CHEILOCYSTIDIA 30-52 x 6-29 um,

variable in shape, clavate to narrowly clavate, slightly cylindrical, sometimes

pyriform, spathulate, rarely narrowly clavate with long pedicel, hyaline, thin

— walled. PLEUROCysTIDIA absent. PILEUS COVERING a trichoderm made up

of erect narrowly clavate elements 30-130 x 5.5-23 um, with rounded apex,

often with abrupt apical excrescence, pale brown-walled, with parietal and

intracellular brown pigment, with cylindrical hyphae in under layer, thin-

walled, 2.5-5.0 um wide. STIPE COVERING in squamules a trichoderm similar

to pileus covering. CLAMP CONNECTIONS present in all tissues.

HABITAT AND DISTRIBUTION—growing in small group, saprotrophic and

terrestrial, on humus-rich, nutrient-rich soil and dead leaves, found during

rainy season in rain forest with dominant bamboo. Only known from one

locality in northern Thailand.

Discusston—Lepiota papillata resembles L. griseovirens but differs especially

in the pyriform, spathuliform to narrowly clavate shape of the cheilocystidia

(which are narrowly utriform, obovoid, to nearly cylindrical cheilocystidia in

L. griseovirens; Vellinga 2001) and in the abrupt apical excrescences often found

on the pileus covering elements.

Lepiota grangei (Eyre) Kihner comes close in morphology but differs in the

dark gray-green, grayish blue, green-brown, or blue-brown to brown pileus, the

cylindrical to utriform cheilocystidia, and the intracellular pigment in the long

pileus covering elements.

This species comes very close to L. tomentella J.E. Lange in pileus color and

basidiospore size, but the latter has longer pileus covering elements that lack

abrupt apical excrescences (Vellinga & Huijser 1993).

78 ... Sysouphanthong & al.

Lepiota pilodes Vellinga & Huijser differs from L. papillata by dull orange-

brown to dark brown and ochraceous brown colors at centre of pileus, and

longer pileus- and stipe-covering elements (60-280(-330) x (6.5-)8.0-21 um;

Vellinga & Huijser 1993, Vellinga 2001).

Lepiota poliochloodes Vellinga & Huijser, Persoonia 15: 229. 1993. FIG. 11

MISAPPLIED NAMES: Lepiota griseovirens sensu Bon (1993), Reid (1972);

Lepiota griseovirens var. griseovirens sensu Bon (1981).

PitEus 12-25 mm, campanulate to umbonate, applanate to plano-concave

with wide umbo, with straight margin, at centre with crowded squamules to

small plush-like squamules or floccules, grayish brown, grayish green brown to

dark brown (Mu. 2.5 Y 4-5/2, 6E3-6, 6F5-6), with squamules or fibrils around

umbo toward margin and distant at marginal zone, on brownish orange, slightly

orange or light brown fibrillose background (5C4, 6D4); margin peeling from

background when mature, exceeding lamellae. LAMELLAE, L = around 40, | =

0-3, crowded to moderately crowded, just free, ventricose, 1.7-3 mm wide, white

to yellowish white, distinctly pinkish cream (4A2, Mu. 10 YR 8/6), with eroded

to floccose, distinctly cystidiose edge. STIPE 25-36 x 1-2.5 mm, cylindrical and

slightly wider at middle, with white or yellowish white to orange-white or pale

pink (4A2, 5A2) fibrillose background, with dark brown to dark gray-green

(6F5-6) squamules from annular zone to base, with white to yellowish white

(4A2) fibrils between squamules, hollow, with white rhizomorphs at base.

ANNULUS an annular zone, with gray-green to dark brown squamules. CONTEXT

in pileus white to pale cream, 1.5-2 mm wide, in stipe white, yellowish white

(4A2) to pale orange. SMELL mild. TasTE unknown. SPORE PRINT WHITE.

BasIp1osPorEs [70,3,3] 6.2-8.2 x 3.0-3.8 um, avl x avw = 7.2 x 3.5 um, Q

= 1.6-2.4, Qav = 2.0, in side-view cylindrical or narrowly triangular with or

without lateral spur, but spur not protruding abaxially, and with rather abrupt

base, in frontal view cylindrical, with rounded apex or tapering toward apex,

with 1 or 2 guttules, congophilous, dextrinoid, cyanophilous, not metachromatic

in Cresyl Blue. Basip1a 15-19 x 6 - 8 um, 4-spored, rarely 2-spored, relatively

short and plump. LAMELLA EDGE sterile. CHEILOCYSTIDIA 18-40 x 3-9 um,

mostly cylindrical, to very narrowly clavate, some narrowly lageniform or

slightly fusiform, strangulate, digitate. Pleurocystidia absent. PILEUS AND STIPE

COVERING a trichoderm or slightly irregular trichoderm made up of erect

cylindrical, narrowly clavate to narrowly lageniform elements, (20-)30-130

x 6.5-15 um, with pale brown walls, and with intracellular brown pigment,

thin-walled, with under layer made up of cylindrical, hyaline, 2.5-9.0 um wide

hyphae. CLAMP CONNECTIONS present in all tissues.

HABITAT AND DISTRIBUTION—growing solitary, saprotrophic and terrestrial

in deciduous forest and bamboo forest. Reported from the Netherlands,

Lepiota section Stenosporae (Thailand) ... 79

FIGURE 11. Lepiota poliochloodes (MFLU090182).

a. basidioma; b. cheilocystidia; c. basidiospores; d. basidia; e. pileus covering.

Scale bars: a = 20 mm; b-d = 10 um; e = 20 um.

Denmark, France, and Great Britain (Vellinga 2001); this is the first record for

Thailand.

MATERIAL EXAMINED: THAILAND, CHIANG Mat Prov., MAE TAENG DiIstTR., Mae

Sae village, 19°07'13.7"N, 98°43'52.9"E, 905 m alt., 8.VIII.2008, P. Sysouphanthong,

80 ... Sysouphanthong & al.

MFLU090182; Mok Fah Waterfall National Park, 20°02'43.1"N, 99°52'35.0"E, 596 m

alt., 5.VII.2008, P. Sysouphanthong, MFLU090118; Mar Rim Distr., Mae Sa Valley,

12.VII.2008, J-K. Liu (collection ecv3877), MFLU081272.

Discusston—Lepiota poliochloodes is rare in Northern ‘Thailand, where it is

known from three localities. The macro- and micromorphology of the Thai

material agree with the description of L. poliochloodes from Europe (Vellinga

& Huijser 1993); unfortunately, there are no European nrITS sequence data

available for comparison.

The relatively small spores and short pileus-covering elements combine with

the pileus with grayish brown to green colors over a slightly orange background

to characterize this species.

Lepiota griseovirens, which has similarly sized basidiospores and

cheilocystidia differs from L. poliochloodes in the much darker basidiomata and

the longer pileipellis elements.

Lepiota sp. 1 Fic. 12

PitEus 15 mm, plano-convex with umbo and slightly inflexed margin, at

umbo not completely closed, with tufted scales; around umbo scales are more

spreading out on yellowish background; scales small and reddish brown (Mu.

2.5 YR3/4-6). LAMELLAE, L = around 35,1= 1-3, free, not very crowded, slightly

ventricose, yellowish (Mu. 10 YR-2.5 Y 8/4) with lighter, slightly cystidiose

edge. StipE 24 x 2 mm, cylindrical, above annular zone innately lengthwise

fibrillose, pale brown, below annular zone with small patches, sometimes spiny,

of same material and color as on pileus, hollow, with white rhizomorphs. SMELL

a bit unpleasant, when old astringent.

BASIDIOSPORES [20,1,1] in side view 7.1-8.4 x 2.8-4.2 um, avl x avw =

7.8 x 3.2 um, Q = 1.9-2.8, avQ = 2.4, with distinct basal spur, in some spores

spur also abaxially bulging, with round to slightly amygdaliform apex, in

frontal view obovoid to subcylindrical, thick-walled, smooth, dextrinoid,

and congophilous. Basidia 15-22 x 6.5-8.0 um, 4-spored, some 2-spored,

with basal clamp connection. LAMELLA EDGE sterile. CHEILOCYSTIDIA

22-28 x 7.0-10.5 um, utriform, fusi-utriform, and narrowly clavate, with basal

clamp-connection. PLEUROCysTIDIA absent. Pileus covering trichodermal,

made up of erect elements, 52-170 x 7.5-12.5 um, cylindrical with rounded

apex, almost always narrowed into pedicel, some very narrowly lageniform,

with intracellular brown pigment. STIPE COVERING similar to pileus covering.

CLAMP CONNECTIONS present in all tissues.

HABITAT AND DISTRIBUTION—Solitary, saprotrophic on humus rich soil in

orchard with bamboo; only collected once close to Chiang Mai.

MATERIAL EXAMINED: THAILAND, CHIANG Mat PRov., MAE Rim Distr., Mae Sa

Valley, 12.VII.2008, leg. J-K. Liu (collection E.C. Vellinga 3881), MFLU08-1259.

Lepiota section Stenosporae (Thailand) ... 81

yaa

FIGURE 12. Lepiota sp. 1 (MFLU081259).

a. basidioma; b. cheilocystidia; c. basidiospores; d. basidia; e. pileus covering elements.

Scale bars: a = 1 cm; b-d = 10 um.

Discusston—Macroscopically, this undescribed taxon resembles L. echinella

Quél. & G.E. Bernard because of the tufted reddish brown pileus scales but

differs in the spurred spores and shorter elements in the pileus covering.

We refrain from describing this taxon as a new species, as the sole collection

contained only one specimen.

Lepiota sp. 2 Fic. 13

PILEus 30 mm, campanulate or umbonate with broad umbo, with straight

margin, covered with crowded grayish brown to slightly reddish brown (8F6-8)

squamules at center and around umbo toward margin and there more radially

fibrillose, on grayish yellow (4B3), white to yellowish white (4A2), radially

82 ... Sysouphanthong & al.

IOOD0

ES eeeiern,

ee SN

WLI

FIGURE 13. Lepiota sp. 2 (MFLU090006).

a. basidioma and context; b. cheilocystidia; c. basidiospores; d. basidia; e. pileus covering.

Scale bars: a = 10 mm; b-d = 10 um; e = 20 um.

fibrillose background; margin exceeding lamellae. LAMELLAE free, crowded,

broadly ventricose, 2.5-3 mm wide, white to yellowish white (3.2), with slightly

wavy edge. STIPE 40 x 3-4 mm, cylindrical, slightly wider at center and tapering

to apex and curved at base; surface with white to pale orange (5A3) fibrils, with

squamules from middle downwards, concolorous with squamules on pileus,

with yellowish brown (5E5) fibrillose squamules at basal zone. CONTEXT in

pileus thick at umbo, 4 mm wide, white to yellowish white (3A2); in stipe pale

orange (6A3), hollow. Smell Termitomyces-like. Taste sweet. Spore print white.

Lepiota section Stenosporae (Thailand) ... 83

BASIDIOSPORES [25,1,1] 6.2-8.0 x 3.2-4.0 um, avl x avw = 7.3 x 3.7 um,

Q = 1.6-2.5, Qav = 1.9, in side-view with truncate to spurred base, oblong to

cylindrical triangular or with curved abaxial side, sometimes with rounded

or acute apex, in frontal view oval or cylindrical, strongly dextrinoid,

congophilous, cyanophilous, not metachromatic in Cresyl blue. BAsip1a 16-21

x 7.0-8.0 um, narrowly clavate, 4—-spored, hyaline, thin-walled. Lamella edge

sterile, with crowded cheilocystidia. CHEmLocystTip1A 12-45 x 5.2-10 um,

cylindrical, sometimes narrowly lageniform, narrowly clavate, with rounded

apex, subcapitate, sometimes slightly swollen at center, utriform, colorless,

thin-walled. PLEUROCysTIDIA absent. PILEUS COVERING a cutis made up

of irregular narrowly clavate hyphae, 25-128 x 10-18 um, usually narrowed

into pedicel, with rounded or subcapitate apex, slightly thick-walled, with or

without septum, with basal clamp connections, with yellowish brown parietal

pigment; under layer made up of cylindrical 3.0-7.0 um wide hyphae, thin-

walled, with yellowish brown walls in upper part, with hyaline walls in lower

part. STIPE COVERING a cutis similar to pileus covering. CLamMP CONNECTIONS

present in all tissues.

HABITAT AND DISTRIBUTION—growing solitary, saprotrophic and terrestrial

on humus-rich soil with dead leaves and wood, in deciduous forest with

Lithocarpus spp. Only found in one locality in northern Thailand.

MATERIAL EXAMINED: THAILAND, CHIANG Mar PRov., Mar TaENnG Distr., Pha

Deng Village, 19°07'13.7"N, 98°43'52.9"E, 905 m alt., 19. VII.2007, P. Sysouphanthong,

MFLU090006.

DiscusstoNn—This undescribed taxon is characterized by grayish brown to

reddish brown squamules on pileus and stipe, truncate to spurred basidiospores,

cylindrical to narrowly clavate cheilocystidia, and a cutis-like pileus covering.

Lepiota griseovirens, which can be confused with this species in the field

because of the color of the pileus squamules, differs in the trichodermal pileus

covering.

Here also the single collection comprised only one basidioma, and we were

unable to obtain nrITS sequence data. We await further collections before

proposing a name.

Acknowledgements

We wish to acknowledge the Mushroom Research Centre in Mae Taeng District,

Chiang Mai Province, Thailand for providing funding, and facilities for collecting and

study. The International Fungal Research and Development Centre, The Research

Institute of Insect Resources (RIRI) is thanked for providing materials and laboratory

facility for the molecular work. Funding of ECV and PS in 2007-2009 by NSF grant DEB

0618293 is gratefully acknowledged. This study was also partially supported by the project

“Value added products from Basidiomycetes: Putting Thailand’s biodiversity to use”

(BRN049/2553). The comments by the two reviewers, Dr R.-L. Zhao and Dr Zai-Wei Ge,

are gratefully acknowledged.

84 ... Sysouphanthong & al.

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http://dx.doi.org/10.1007/s13225-010-0050-4

MY COTAXON

http://dx.doi.org/10.5248/117.87

Volume 117, pp. 87-92 July-September 2011

New records of Digitoramispora from China

YI-DONG ZHANG, JIAN Ma, LIi-Guo Ma & X1U-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

* CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Four species collected from plant debris in natural areas of southern China

are recorded for the first time from China. Digitoramispora caribensis, D. tambdisurlensis,

D. excentrica, and D. lageniformis are described and illustrated, anda key to currently accepted

Digitoramispora species is provided. The specimens are deposited in Herbarium of Shandong

Agricultural University, Plant Pathology (HSAUP), and Mycological Herbarium, Institute of

Microbiology, Chinese Academy of Sciences (HMAS).

KEY worpDs — anamorphic fungi, taxonomy

Introduction

Castaneda & Kendrick (1990) introduced Digitoramispora to accommodate

two species, D. caribensis (type species) and D. excentrica. Two other species,

D. lageniformis and D. tambdisurlensis were described from Thailand and

India, respectively (Somrithipol & Jones 2003, Pratibha et al. 2009). Until now,

Digitoramispora has contained only these four species with no reports from

China. Digitoramispora species produce dictyosporous, digitate or irregular

conidia with peripheral hyaline or colourless short radiating branches or cells,

and irregularly (often percurrently) extending conidiophores (Castafieda &

Kendrick 1990). During investigations of tropical fungi from the forests of

southern China, all four species of Digitoramispora were collected. They are

illustrated and described as new records for China.

Taxonomy

Digitoramispora caribensis R. F. Castafieda & W.B. Kendr., University of Waterloo

Biology Series 33: 20 (1990) Fic. 1

Colonies on the natural substrate effuse, black. Mycelium partly superficial,

partly immersed in the substrate, composed of smooth, brown, branched,

septate hyphae, 1-2 um wide. Conidiophores macronematous, mononematous,

88 ... Zhang & al.

Fig. 1. Digitoramispora caribensis A. Colonies on the natural substratum.

B-D. Conidiophores, conidiogenous cells and conidia. E. Mature conidia.

single, straight to slightly flexuous, unbranched, dark brown, often swollen at

the base, up to 110 um long, 3-5 um thick. Conidiogenous cells monoblastic,

integrated, terminal, cylindrical to lageniform, 6-12 x 2-2.5 um. Conidia

holoblastic, digitate, lobed at the apex, 17-21 x 14-20 um, dark brown at the

base and centre, pale brown to colourless at apical cells and short branches,

seceding schizolytically.

SPECIMEN EXAMINED: CHINA. GUNGDONG PRovINCE: Mountain Danxia, on dead

branches of unidentified plant, 22 Oct. 2010, L.Y. Sun, HSAUP H3273 (duplicate HMAS

146118).

ComMMENTS—Digitoramispora caribensis, the type species of Digitoramispora,

was previously known only from Cuba. Our specimen is similar to the type

species, but it lacks annellidic conidiogenous cells and its conidia are somewhat

larger (17-21 x 14-20 um vs. 15-20 x 10-15 um).

Digitoramispora tambdisurlensis Pratibha, Raghuk. & Bhat, Mycotaxon 107: 383

(2009) Fic. 2

Colonies on the natural substrate effuse, dark brown. Mycelium immersed

in the substrate, composed of smooth, brown, branched, septate hyphae, 2-3

uum wide. Conidiophores macronematous, mononematous, straight to slightly

flexuous, unbranched, dark brown, swollen at the base, 200-230 x 5.5-11 um.

Digitoramispora spp. new to China... 89

20um

Fic. 2. Digitoramispora tambdisurlensis A. Colonies on the natural substratum.

B. Conidiophores and conidiogenous cells. C. Conidiophores with conidia. D. Conidia

Conidiogenous cells monoblastic, integrated, terminal, cylindrical, producing

only one terminal conidium, 9-12.5 x 4.5-6 um. Conidia holoblastic, muriform,

digitate, variable in shape, dark brown at the base and centre with peripheral

light brown cells or branches, 64.5-80 x 38.5-55.5 um.

SPECIMEN EXAMINED: CHINA. HuNAN PROVINCE: forest park of Zhang jiajie, on dead

branches of unidentified plant, 18 Aug. 2010, Y.D. Zhang, HSAUP H3168 (duplicate

HMAS 146119).

CoMMENTS—Digitoramispora tambdisurlensis differs from the other three

species in the genus by its larger conidia and conidiophores. The shape and size

of the conidia in our collection compare well with those described by Pratibha

et al. (2009). The only difference is that the conidia and conidiophores in our

collection are slightly smaller.

Digitoramispora excentrica (B. Sutton) R.E. Castafieda & W.B. Kendr., University of

Waterloo Biology Series 33: 20 (1990) FIG. 3

Colonies on natural substrate effuse, black. Mycelium partly superficial,

partly immersed in the substrate, composed of smooth, brown, branched,

septate hyphae, 1-2 um wide. Conidiophores macronematous, mononematous,

single, straight to slightly flexuous, unbranched, dark brown, often swollen at

the base, up to 140 um long, 3-4 um thick, with up to 5 lageniform or doliiform

90 ... Zhang & al.

an * oe

lim

Fic. 3. Digitoramispora excentrica A. Colonies on the natural substratum. B. Conidiophores and

conidiogenous cells. C. Conidiophores with conidia. D. Conidia.

percurrent proliferations. Conidiogenous cells monoblastic, integrated,

terminal, cylindrical to lageniform, 7-10 x 2-2.5 um. Conidia holoblastic,

muriform or cheiroid, lobed at the apex, variable in shape, with 1-2 transverse

septa and 1-3 longitudinal septa, the septa are partly obscured by a band of wall

which is more deeply pigmented and almost black, apical cell of each branch

colourless, 16.5-21 x 11.5-14 um.

SPECIMEN EXAMINED: CHINA. GUNGDONG PROVINCE: Mountain Danxia, on dead

branches of unidentified plant, 22 Oct. 2010, Y.D. Zhang, HSAUP H3273 (duplicate

HMAS 146120).

CoMMENTS—Digitoramispora excentrica was originally described as Acrodictys

excentrica by Sutton (1969) in Canada. Hughes (1979) transferred it to

Arachnophora (as A. excentrica (B. Sutton) S. Hughes) based on the rhexolytic

conidial secession. Castafieda & Kendrick (1990) provided the combination

Digitoramispora excentrica based on the percurrently proliferating

Digitoramispora spp. new to China... 91

20pm

20m

Fic. 4. Digitoramispora lageniformis A. Colonies on the natural substratum. B. Conidiophores and

conidiogenous cells. C. Conidiophores with conidia. D. Conidia.

conidiophores and dictyosporous, digitate conidia with colourless apical

cells. Our specimen lacks the Selenosporella-like synanamorph at the apex. The

conidial shape is similar to the type specimen, but the conidia are somewhat

longer (16.5-21 um vs. 13-18 tm) than those of the type.

Digitoramispora lageniformis Somrith. & E.B.G. Jones, Nova Hedwigia 77: 374

(2003) Fic. 4

Colonies on natural substrate effuse, black. Mycelium partly superficial,

partly immersed in the substrate, composed of smooth, brown, branched, septate

hyphae, 1.5-3 um wide. Conidiophores macronematous, mononematous,

straight to slightly flexuous, unbranched, dark brown, often swollen at the

base, up to 260 um long, 6-10.5 um thick, with up to 3 lageniform or doliiform

percurrent proliferations. Conidiogenous cells monoblastic, integrated,

terminal, producing only one terminal conidium, cylindrical to lageniform,

12-17 x 4-6 um. Conidia holoblastic, muriform or digitate, variable in shape,

38-57 x 40-51 um, with many short, 1-3 septate, aggregated branches, brown or

dark brown at the base and in the centre, apical cell of each branch colourless.

SPECIMEN EXAMINED: CHINA. GUNGDONG PROVINCE: Mountain Danxia, on dead

branches of unidentified plant, 22 Oct. 2010, L.Y. Sun, HSAUP H7034 (duplicate HMAS

146121).

92 ... Zhang & al.

CoMMENTS—Digitoramispora lageniformis is distinguished from other species

by the lageniform or doliiform percurrently proliferating conidiophores and

numerous short branches on the conidia. Compared with the morphology of

the type specimen described by Somrithipol & Jones (2003), the conidia in our

collection are slightly larger.

Key to Digitoramispora species

ly “Gonidiosenous-cells, without proliterationy. 25:0 .8 tt 1 tote etek bok lek Be RRL es 2

Conidiogenous cells with lageniform or doliiform proliferations ............. 3

2. Conidia 15-20 um long, 10-15 um wide .....................06. D. caribensis

Conidia 50-90 um long, 40-75 um wide ................00. D. tambdisurlensis

3. Conidia 13-18 um long, 12-17 um wide ....................06. D. excentrica

Conidia 37-45 um long, 28-32 um wide ..................0.. D. lageniformis

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. 30499340, 30770015) and the Ministry of Science and Technology of the

People’s Republic of China (Nos. 2006FY120100, 2006FY110500-5).

Literature cited

Castaneda Ruiz RF, Kendrick WB. 1990. Conidial fungi from Cuba: II. Univ. Waterloo Biol. Ser.

33: 1-61.

Hughes SJ. 1979. Relocation of species of Endophragmia auct. with notes on relevant generic names.

New Zealand J. Bot 17: 139-188.

Pratibha J, Raghukumar S, Bhat DJ. 2009. New species of Digitoramispora and Spondylocladiopsis

from the forests of Western Ghats, India. Mycotaxon 107: 383-390.

http://dx.doi.org/10.5248/107.383

Somrithipol S, Jones EBG. 2003. Digitoramispora lageniformis sp. nov., a new graminicolous

hyphomycetes from Thailand. Nova Hedwigia 77: 373-378.

http://dx.doi.org/10.1127/0029-5035/2003/0077-0373.

Sutton BC. 1969. Forest microfungi. II. Additions to Acrodictys. Can. J. Bot. 47: 853-858.

http://dx.doi.org/10.1139/b69-123.

MY COTAXON

http://dx.doi.org/10.5248/117.93

Volume 117, pp. 93-99 July-September 2011

Seven new records of foliicolous lichens from Vietnam

Tor THuy NGUYEN™, YOGESH JOSHI’, ROBERT LUCKING?,

ANH DZUNG NGUYEN*, XIN YU WANG’, YOUNG JIN KOH?"

& JAE-SEOUN Hur™

"Korean Lichen Research Institute, Sunchon National University, Sunchon 540-742, South Korea

*Department of Botany, S.S.J. Campus, Almora 263601, Uttarakhand, India

° Botany Department, The Field Museum, 1400 South Lake Shore Drive,

Chicago IIlinois 60605-2496, USA

‘Plant Biological Department, Faculty of Agriculture, Tay Nguyen University,

Buon Ma Thuot City, Daklak Province, Vietnam

CORRESPONDENCE TO *: ‘pthuydhtn@yahoo.com, *dryogeshcalo@gmail.com,

>rlucking@fieldmuseum.org, ‘nadzung@dng.vnn.vn, & *jshur1@sunchon.ac.kr

ABSTRACT — Seven foliicolous species growing in tropical regions of Vietnam are reported

as new to the country. Described are Arthonia accolens, Calenia aspidota, Calopadia

subcoerulescens, Coenogonium minimum, Fellhanera rhapidophylli, F. semecarpi, and Porina

subnitidula. Among them, Coenogonium minimum and Porina subnitidula are reported for

the first time from the paleotropics.

KEY worps — Ascomycota, geographical distribution, lichen-forming fungi, taxonomy

Introduction

The present paper, which is a further report on the taxonomic study of

foliicolous lichens from Vietnam by Nguyen et al. (2010, 2011), describes

seven species newly recorded from three different localities [Thac Dray Sap (=

Dray Sap waterfall), Vuon quéc gia Bach Ma (= Bach Ma National Park), and

Vuon quéc gia Phong Nha-Ké Bang (= Phong Nha Ke Bang National Park)]

in Vietnam. The discovery in Vietnam of Coenogonium minimum and Porina

subnitidula, previously known only from the Neotropics, provides further

information about their distribution in southeastern Asia. In addition to the

newly recorded species, the authors also collected previously reported (by

Santesson 1952, Vézda 1977, Farkas & Sipman 1993, 1997, Licking & Vézda

1998, Aptroot & Sparrius 2006) the foliicolous lichens Bacidina apiahica (Mill.

Arg.) Vézda, B. pallidocarnea (Mill. Arg.) Vézda, Byssoloma chlorinum (Vain.)

94 ... Nguyen & al.

Zahlbr., Calopadia puiggarii (Mull. Arg.) Vézda, Coenogonium dilucidum

(Kremp.) Kalb & Licking, C. subluteum (Rehm) Kalb & Licking, Echinoplaca

epiphylla Fée, Porina corruscans (Rehm) R. Sant., P. diaphana Vézda, P. nitidula

Mill. Arg., Sporopodium phyllocharis (Mont.) A. Massal., Strigula orbicularis

Fr., S. nitidula Mont., and Trichothelium alboatrum Vain. The most common

and widely distributed species found during the survey were Bacidina apiahica,

Calopadia puiggarii, Coenogonium dilucidum, Porina nitidula, and Strigula

nitidula.

This is the first report of lichens from central Vietnam. Earlier workers

(Santesson 1952, Vézda 1977, Farkas & Sipman 1993, 1997, Liicking & Vézda

1998, Aptroot & Sparrius 2006) have reported Vietnamese lichens mostly from

the northern parts of the country. Surveys of the several provinces still left

unexplored will definitely contribute further to our knowledge of the Vietnamese

lichen flora. Brief taxonomic descriptions, chemical and ecological notes, and

comments are provided below for each of the newly recorded species.

Materials & methods

The lichen samples were collected from three different localities between 22 July 2006

and 08 February 2010. The study area covers the three Vietnamese provinces Daknong,

Quang Binh, and Thua Thien Hue. After drying at room temperature, the lichen samples

were identified using stereo and light microscopes: a NIKON Eclipse E200 dissecting

microscope was used for identifying morphological characters of thallus, reproductive

structures, color, size and shapes, while an OLYMPUS BX 50 compound microscope

was used for studying the anatomy of thalli and fruiting bodies. All measurements were

made from material mounted in water and stained in lactophenol cotton blue (LCB).

An average of ten measurements per structure was recorded for the sizes of thallus,

ascomata, and ascospores and the thicknesses of hymenium, hypothecium, exciple, and

involucrellum. Only free ascospores lying outside the asci were measured. Ascospore

dimensions are generally presented as minimum value observed - maximum value

observed. Spot test reactions were carried out on hand sections of thalli and apothecia

under the compound microscope. Iodine (I) was used to check the color reactions of

the ascus wall and the hymenium. Thin Layer Chromatography (TLC) was performed

in solvent system C (Toluene:Acetic acid 85:15) as described by Orange et al. (2010).

Terminology used in this study follows Liicking (2008). Vouchers have been deposited in

the herbarium of the Lichen & Allied Bioresource Center at the Korean Lichen Research

Institute (KoLRI), Sunchon National University, South Korea.

New records

Arthonia accolens Stirt., Proc. Roy. Phil. Soc. Glasgow 11: 105, 1879

Thallus dispersed into rounded patches, smooth, 2-20 mm across, ecorticate,

brownish green. Photobiont cells rectangular in radiate plates. Apothecia

adnate, rounded, 0.1-0.3 mm diam., dark brown. Hypothecium 2-4 um high,

Folicolous lichens new to Vietnam ... 95

brown. Hymenium pale brown. Asci globose, 15-20 x 13-15 um, 8-spored.

Ascospores obovate, hyaline, 2-septate, 10-12 x 3-4 um, distal cell enlarged.

CHEMISTRY — Spot test reactions: thallus and apothecia K-, C-, KC-, P-,

N-. Epithecium N-. Secondary metabolites: none detected.

DISTRIBUTION & ECOLOGY — Pantropical (Liicking 2008); new to Vietnam.

At the collection site the species was found growing on the understory leaves

in lowland tropical rain forest along with Coenogonium dilucidum and Porina

spp.

SPECIMEN EXAMINED - VIETNAM: Tuva THIEN HUE PROVINCE: Vu6n qu6c gia Bach

Ma, 16°12'55.50"N, 107°51'40.20"E, alt. 685 m, on leaf of Ochna spp., 08 February 2010,

T. Nguyen 100001 (KoLRI).

REMARKS — Arthonia cyanea Mill. Arg., another Arthonia species known from

Vietnam, differs in having bluish to greenish gray, white pruinose apothecia.

The apothecia in A. accolens are light to dark brown colored and never pruinose.

The species can also be confused with A. leptosperma (Mill. Arg.) R. Sant.,

which differs in having smaller, 1-septate ascospores and a continuous thallus

with net-like photobiont. For further descriptions of A. accolens, see Liicking

(2008).

Calenia aspidota (Vain.) Vézda, Folia geobot. phytotax. 19: 195, 1984

Thallus dispersed into rounded patches, 5-13 mm across, greenish gray,

47-80 um thick, cortex formed by rounded cells, strongly inflated with calcium

oxalate crystals, white. Setae not observed. Apothecia immersed, zeorine,

rounded, 0.2-0.5 mm diam. and 70-120 um high; disc whitish gray with white

pruina; margin distinct, irregularly lobulate, white. Hymenium 90-100 um,

hyaline. Epithecium 3-5 um thick, with algal layer. Asci ellipsoid, 70-85 x

20-35 um, single spored. Ascospores hyaline, muriform, 50-80 x 15-30 um.

Hyphophores white, 0.2—0.4 mm high.

CHEMISTRY — Spot test reactions: thallus and apothecia K-, C-, KC-, P-.

Secondary metabolites: none detected.

DISTRIBUTION & ECOLOGY — Pantropical (Liicking 2008); new to Vietnam.

At the collection site the species was found growing on the understory leaves

in lowland tropical rain forest along with Calopadia puiggarii and Porina

nitidula.

SPECIMEN EXAMINED - VIETNAM: DakNONG PROVINCE: Thac Dray Sap,

12°32'21.39"N, 107°53'19.80"E, alt. 399 m, on leaf, 22 July 2009, Y. Joshi & T. Nguyen

090183 (KoLRI).

REMARKS — Calenia thelotremella Vain., another Calenia species known from

Vietnam, differs in having transversely septate ascospores, a verrucose, pale

greenish, or yellowish gray thallus, and epruinose, yellowish gray apothecia.

The species can also be confused with C. bullatinoides Liicking, which differs in

96 ... Nguyen & al.

having 2-4-spored asci and smaller hyphophores with blunt, darkened apices.

For further descriptions of C. aspidota, see Liicking (2008).

Calopadia subcoerulescens (Zahlbr.) Vézda, Sched. Lichenes Selecti Exsiccati

Fascicle 88: 3, no. 2185, 1988

Thallus dispersed into rounded patches, 10-15 mm across, smooth,

pale greenish gray. Apothecia rounded, 0.2-0.4 mm diam., plane, brownish

black; margin thin, prominent, gray. Excipulum 35-40 um broad, hyaline,

paraplectenchymatous. Hymenium 100-120 um, hyaline. Hypothecium 30-40

um high, aeruginous. Apothecial base aeruginous. Epithecium 7-10 um high.

Asci 70-95 x 20-25 um, single spored. Ascospores hyaline, muriform, 70-85 x

15-20 um. Campylidia not observed.

CHEMISTRY — Spot test reactions: thallus and apothecia K-, C-, KC-, P-.

Secondary metabolites: none detected.

DISTRIBUTION & ECOLOGY — Pantropical (Liicking 2008); new to Vietnam.

At the collection site the species was found growing on the understory leaves in

lowland tropical rain forest at an elevation of 136 m.

SPECIMEN EXAMINED - VIETNAM: QUANG BINH PROVINCE: Vudn quéc gia Phong

Nha-Ké Bang, 17°28'51.60"N, 106°18'39.10"E, alt. 136 m, on leaf of Lauraceae tree, 06

February 2010, T. Nguyen 100028b (KoLRI).

REMARKS — Calopadia puiggarii, another Calopadia species known from

Vietnam, differs in having a dark brown hypothecium and grayish brown

apothecia. Calopladia subcoerulescens always has an aeruginous hypothecium

and grayish black to black apothecia. For additional details see Liicking

(2008).

Coenogonium minimum (Mill. Arg.) Licking, Fl. Neotrop. Monogr. 103: 572, 2008

Thallus continuous, smooth, green. Apothecia sessile, rounded, 0.1-0.13

mm diam. Disc concave, yellowish brown, slightly translucent; margin distinct,

prominent, concolorous with disc. Excipulum 5-8 um high, pale yellowish

brown. Hypothecium 5-8 um high, hyaline. Hymenium 40-45 um high,

hyaline. Asci 42-48 x 5-6 um, 8-spored. Ascospores ellipsoid, 1-septate, 12-14

x 3-4 um. Pycnidia not observed.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

DISTRIBUTION & ECOLOGY — Neotropical (Licking 2008); new to Vietnam.

At the collection site the species was found growing along with Coenogonium

dilucidum on the understory leaves in lowland tropical rain forests between

elevations of 136-556 m.

SPECIMENS EXAMINED — VIETNAM: QUANG BINH PROVINCE: Vudn quéc gia Phong

Nha-Ké Bang, 17°33'07.10"N, 106°18'06.10"E, alt. 556 m, on leaf, 05 February 2010, T.

Nguyen 100168h (KoLRI); 17°33'37.10"N, 106°18'06.10"E, alt. 556 m, on leaf of Rubiaceae

Folicolous lichens new to Vietnam ... 97

tree, 06 February 2010, T. Nguyen 100016 (KoLRI); 17°28'51.60"N, 106°18'39.10"E, alt.

136 m, on leaf, 06 February 2010, T. Nguyen 100031, 100066b (KoLRI); 17°33'07.10"N,

106°18'06.10"E, alt. 556 m, on leaf of Fagaceae tree, 06 February 2010, T. Nguyen

100081a (KoLRI).

REMARKS — Coenogonium dilucidum, C. disciforme Papong et al., C. luteum

(Dicks.) Kalb & Licking, and C. subluteum are the other Coenogonium species

known from Vietnam. Coenogonium dilucidum differs in having wax-colored

to pale yellow, slightly concave apothecia; C. luteum and C. subluteum have

larger apothecia (0.2-1.5 mm diam.), while C. disciforme differs in bearing

numerous disc-shaped isidia. For further descriptions of C. minimum, see

Liicking (2008).

Fellhanera rhapidophylli (Rehm) Vézda, Folia geobot. phytotax. 21: 214, 1986

Thallus continuous, farinose, greenish gray to green. Apothecia rounded,

12-14 um diam.; disc plane to slightly convex, grayish brown to dark reddish

brown; margin thin, persistent, pale gray. Excipulum paraplectenchymatous,

20-25 um broad. Hypothecium dark brown. Hymenium colorless, 40-55 um

high. Asci 42-55 x 10-11 um, 8-spored. Ascospores ellipsoid, 3-septate, 13-16

x 3-4 um. Pycnidia not observed.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

DISTRIBUTION & ECOLOGY — Pantropical (Liicking 2008); new to Vietnam.

At the collection site F rhapidophylli was found growing on the understory

leaves in tropical rain forest along with E bouteillei (Desm.) Vézda.

SPECIMEN EXAMINED - VIETNAM: DaKNONG PROVINCE: Thac Dray Sap,

12°32'21.39"N, 107°53'19.80"E, alt. 685 m, on leaf, 22 July 2009, Y. Joshi & T. Nguyen

090094 (KoLRI).

REMARKS — Fellhanera bouteillei, F emarginata Licking, F. microdiscus (Vain.)

Vézda, F sublecanorina (Nyl.) Vézda, and F. semecarpi are the other Fellhanera

species reported from Vietnam. Fellhanera bouteillei and FE. semecarpi differ

in having 1-septate ascospores and light-colored apothecia; E emarginata

has a reduced exciple and broader ascospores with constrictions at the septa;

F. sublecanorina has a bluish gray thallus containing isousnic acid and other

substances and an apothecium with a well-developed whitish margin; F.

microdiscus has 5-septate ascospores that are slightly curved and attenuated

at one end and an apothecium with a thin but distinct margin. For further

descriptions of FE. rhapidophylli, see Liicking (2008).

Fellhanera semecarpi (Vain.) Vézda, Folia geobot. phytotax. 21: 215, 1986

Thallus dispersed into rounded patches, 3-7 mm across, smooth, greenish

gray. Apothecia rounded to irregular in outline, 0.15-0.3 mm diam.; disc

plane, ochraceous yellow to reddish brown; margin thin, pale gray. Excipulum

98 ... Nguyen & al.

paraplectenchymatous. Hypothecium brown. Hymenium 40-50 um high,

hyaline. Asci 30-40 x 8-10 um, 8-spored. Ascospores 1-septate, with

constriction at septum, 12-15 x 4-5 um. Pycnidia not observed.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

DISTRIBUTION & ECOLOGY — Pantropical (Liicking 2008); new to Vietnam.

At the collection site the species was found growing on the understory leaves

in lowland tropical rain forest along with Calopadia puiggarii and Porina

nitidula.

SPECIMENS EXAMINED — VIETNAM: QUANG BINH PROVINCE: Vudn quéc gia Phong

Nha-Ké Bang, 17°33'04.20"N, 106°18'03.60"E, alt. 132 m, on leaf of Cinnamomum, 05

February 2010, T. Nguyen 100168a, 100168c, 100168e (KoLRI).

REMARKS — Fellhanera bouteillei, F. emarginata, FE. microdiscus, E. rhapidophylli

and F. sublecanorina are the other Fellhanera species reported from Vietnam

(Vézda 1977, Nguyen et al. 2010). Fellhanera emarginata, F. microdiscus, F

rhapidophylli, and EF. sublecanorina differ in having 3-5-septate ascospores.

Fellhanera bouteillei has 1-septate ascospores but differs in its farinose to

granulose thallus bearing secondary metabolites (usnic acid, isousnic acid,

zeorin and sometimes asemone). For further descriptions of F semecarpi, see

Liicking (2008).

Porina subnitidula Coliin & A.B. Pefia, Phyton (Horn) 44: 176, 2004

Thallus dispersed into irregular patches, brownish gray. Perithecia black,

sessile, hemispherical with base spreading, 0.4-0.6 mm diam., glabrous,

pure black, slightly shiny. Excipulum 20-30 um thick, brownish black, K+

black. Involucrellum black, K-, exposed. Asci fusiform, 80-100 x 10-12 um.

Ascospores fusiform, 5-septate, without constriction at septa, 15-30 x 5-7 um,

colorless. Pycnidia not observed.

CHEMISTRY — Spot test reactions: thallus K-, C-, KC-, P-. Secondary

metabolites: none detected.

DISTRIBUTION & ECOLOGY — Neotropical (Liicking 2008); new to Vietnam.

At the collection site the species was found growing on the understory leaves in

lowland tropical rain forest at an elevation of 780 m.

SPECIMEN EXAMINED - VIETNAM: QuANG BINH PROVINCE: Vudn quéc gia Phong

Nha-Ké Bang, 17°28'42.10"N, 106°27'35.60"E, alt. 780 m, on leaf of Rhaphidophora, 06

February 2010, T. Nguyen 100080a (KoLRI).

REMARKS — Many Porina species have been reported from Vietnam (Aptroot

& Sparrius 2006). Here we compare P. subnitidula with those having blackish,

brownish or colorless involucrellum (P. atrocoerulea Mill. Arg., P. diaphana,

P. nitidula). Porina diaphana differs in having pale yellowish-white perithecia

and colorless involucrellum; P. nitidula has subglobose perithecia surrounded

Folicolous lichens new to Vietnam ... 99

by white tomentum, while P. atrocoerulea has oblong, 7-septate ascospores. For

further descriptions of P. subnitidula, see Liicking (2008).

Acknowledgments

This work was supported by a grant from the NRF (National Research Foundation)

of Korea (KRF-2208-313-C0081), and the Higher Education Project 2 (HEP2), Vietnam.

The authors are thankful to Drs L. Lékés and E. Farkas for reviewing the manuscript and

providing valuable comments.

Literature cited

Aptroot A, Sparrius LB. 2006. Additions to the lichen flora of Vietnam, with an annotated checklist

and bibliography. Bryologist 109(3): 358-371.

http://dx.doi.org/10.1639/0007-2745(2006)109[358:ATTLFO]2.0.CO;2

Farkas EE, Sipman HJM. 1993. Bibliography and checklist of foliicolous lichenized fungi up to

1992. Tropical Bryology 7: 93-148.

Farkas EE, Sipman HJM. 1997. Checklist of foliicolous lichenized fungi. Abstracta Botanica 21(1):

173-206.

Liicking R. 2008. Foliicolous lichenized fungi. Flora Neotropica Monograph 103: 1-866.

Licking R, Vézda A. 1998. Taxonomic studies in foliicolous species of the genus Porina (lichenized

Ascomycotina: Trichotheliaceae) I. The Porina epiphylla group. Willdenowia 28: 181-225

Nguyen TTT, Joshi Y, Liicking R, Wang XY, Nguyen AD, Koh YJ, Hur J-S. 2010. Notes on some new

records of foliicolous lichens from Vietnam. Taiwania 55(4): 402-406.

Nguyen TT, Joshi Y, Dzung NA, Hur J-S. 2011. First report of fertile specimen of Coenogonium

disciforme: a species new to Vietnam lichen flora. Lichenologist 43: 184-186.

http://dx.doi.org/10.1017/S0024282910000691

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens.

Second edition. British Lichen Society, London, pp. 1-101.

Papong K, Boonpragob K, Liicking R. 2007. New species and new records of foliicolous lichens

from Thailand. Lichenologist 39: 47-56. http://dx.doi.org/10.1017/S0024282906006104

Santesson R. 1952. Foliicolous lichens I. A revision of the taxonomy of the obligately foliicolous,

lichenized fungi. Symbolae Bot. Upsal. 12(1): 1-590.

Vézda A. 1977. Beitrag zur Kenntnis foliikoler Flechten Vietnams. Casopis Slezskeho Muz., Ser. A

26: 21-33.

MY COTAXON

http://dx.doi.org/10.5248/117.101

Volume 117, pp. 101-108 July-September 2011

Dendrothele latenavicularis sp. nov. (Agaricales, Basidiomycota)

from the Patagonian Andes

SERGIO P. GORJON*’', ALINA G. GRESLEBIN’” & MARIO RAJCHENBERG’”

‘Centro de Investigacion y Extension Forestal Andino Patagénico, Area de Proteccion.

CC 14, 9200 Esquel, Chubut, Argentina

?Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET) Argentina

*CORRESPONDENCE TO: spgorjon@usal.es

Asstract — Dendrothele latenavicularis is described as a new species from the Patagonian

Andes forest. It is closely related to other species of Dendrothele with navicular or arachiform

basidiospores —D. arachispora, D. cymbiformis D. navicularis, D. magnenavicularis, and

D. pitrae— but with distinctly broader basidiospores. A key to the previous species is included.

KEY worpDs — corticioid fungi, Patagonia, Saxegothaea conspicua, taxonomy

Introduction

The genus Dendrothele Hohn. & Litsch. comprises about 45 species

(Parmasto et al. 2004), most described and exclusively recorded from the

Northern Hemisphere. Dendrothele includes species with discoid or crustose

basidiomes occurring on bark of living trees and shrubs, with only few

species known from fallen wood. Microscopically, it is characterized by the

abundant dendrohyphidia and crystalline deposits, an adaptation to drought

and exposure, large basidia, and variably shaped basidiospores, in some cases

slightly thick-walled or cyanophilous. There are several worldwide taxonomic

studies of the genus (e.g., Lemke 1964, Boidin et al. 1996), but only a few that

focus on the Southern Hemisphere species (Cunningham 1954, 1963; Greslebin

& Rajchenberg 1998; Nakasone & Burdsall 2011).

Dendrothele is a polyphyletic genus (Goranova et al. 2003, Bodensteiner et

al. 2004, Binder et al. 2005, Larsson 2007), and the generic type, Dendrothele

papillosa Hohn. & Litsch. [= D. griseocana (Bres.) Bourdot & Galzin], is included

in the Lachnellaceae Boud. (Niaceae Jiilich) within the Agaricales Underw.,

closely related to such cyphelloid genera as Lachnella Fr. and Cyphellopsis

Donk. Recently, Nakasone & Burdsall (2011) described the first navicular

102 ... Gorjén, Greslebin & Rajchenberg

basidiospores in Dendrothele for some species from New Zealand. Some genera

of cyphelloid fungi, such as Flagelloscypha Donk and Lachnella, have navicular

basidiospores, which morphologically support their phylogenetic relationship

with Dendrothele.

As a result of some collecting trips in the Valdivian rainforest we have

found growing on bark of living Saxegothaea conspicua Lindl. (Podocarpaceae),

a Dendrothele species with broad navicular basidiospores, different from

previously described species, that is herein proposed as new.

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 establish

cyanophily of basidiospores. Line drawings were made with a camera lucida attachment.

Specimens are deposited in the herbarium of the Centro de Investigacioén y Extensién

Forestal Andino-Patagonico (Esquel, Argentina), BAFC, and SALA.

Taxonomy

Dendrothele latenavicularis Gorjon, sp. nov. PLATES 1-4

MycoBank MB 519378

Basidiomata resupinata, effusa, crustacea, leve vel rimosa, subalbida. Systema hypharum

monomiticum, hyphae fibulatae, crystallis perabundantibus. Dendrophyses copiosae,

graciles, leves vel incrustates. Cystidia desunt. Basidia suburniformia vel subcylindracea,

cum fibula basali, 4-sterigmatibus. Basidiosporae leves, late navicularis, tenuitunicatae,

hyalinae, 16-21 x 8-11 um, inamyloideae, indextrinoideae, forte cyanophileae. Differt

Dendrothele naviculari, D. magnenaviculari et D. cymbiformi basidiosporis latioribus. Ad

corticem arborum coniferarum viventes.

Type: Argentina, Neuquén-Rio Negro: Nahuel Huapi National Park, Puerto Blest

(41°00'05"S 71°49'42"W), 800 m a.s.l., 30 May 2010-on bark of living Saxegothaea

conspicua, leg. S.P.Gorjén, coll. SPG 2963. Holotype, BAFC; isotypes, SALA and

herbarium of Centro de Investigacién y Extension Forestal Andino-Patagonico.

ErymMo oey: latus — Latin for broad, and navicularis — Latin for boat-shaped, referred

to the shape of the basidiospores.

BASIDIOMATA annual, resupinate, adnate, at first orbicular, then coalescent and

effused up to 3-5 cm, smooth, rimose when mature, at first whitish, turning

yellowish cream with age, margin abrupt (PLATE 1). HYPHAL SYSTEM monomitic,

generative hyphae with clamps, hyaline, thin-walled, straight or some tortuose,

1.5-2.5(-3) um diam. DENDROHYPHIDIA variable, some with few branches

and smooth, others richly ramified and covered by a crystalline encrustations,

1-1.5(-3) um in diam, projecting above the basidia. cystT1p1a absent. BASIDIA

at first suburniform, cylindrical when mature, with a median constriction,

thin-walled, (30-)35-45 x 10-14 um, with 4 sterigmata, basally clamped.

BASIDIOSPORES broadly navicular, tapering gradually to the vermiform distal

Dendrothele latenavicularis sp. nov. ... 103

PLATE 1. Dendrothele latenavicularis.

Basidiomes on bark (coll. S.P. Gorjon 2963, holotype)

end, with a blunt and round prominent apiculus, hyaline, smooth, thin-walled,

mostly attached in groups of two or four, (16-)17-20(-21) x (8-)9-10(-11)

um, IKI-, with distinctly and strongly cyanophilic walls (PLATES 2-4).

DISTRIBUTION AND EcoLtocy — Dendrothele latenavicularis is known

only from the Patagonian Andes forests of Argentina (Valdivian rainforest).

It grows on bark of living Saxegothaea conspicua, an endemic southern South

American conifer, in mixed forests with Nothofagus dombeyi (Mirb.) Oerst.,

Luma apiculata (DC.) Burret, and Chusquea culeou E. Desv. If its occurrence

coincides with host distribution, then it should be present in southern Chile

also, although we have surveyed some areas in the closest regions (Region X and

XI) with no success. In many specimens of S. conspicua where D. latenavicularis

grows, it is also frequent to find associated to the bark Globulicium hiemale

(Laurila) Hjortstam, easily to distinguish by the pinkish hymenial surface and

the conspicuous white fibrillose to rhizomorphic margin.

ADDITIONAL SPECIMENS EXAMINED — ARGENTINA. Neuquén-Rio Negro: Nahuel

Huapi National Park, Puerto Blest (41°00'05"S 71°49'42"W), 800 m a.s.1., 29-30 May

2010-on bark of living Saxegothaea conspicua, leg. S.P.Gorjén coll. SPG 2911, 2912,

2914, 2915, 2917, 2918, 2920, 2923, 2926, 2930, 2962. Idem, 12-14 April 2011, coll. SPG

3215, 3242. Neuquén: Lanin National Park, Lago Queni (40°08'35"S 71°43'05"W), 1080

m a.s.l., 18 May 2010-on bark of living S. conspicua, leg. S.P.Gorjon coll. SPG 2831,

2832.

104 ... Gorjén, Greslebin & Rajchenberg

PLATE 2. Dendrothele latenavicularis.

Hymenial elements a) basidiospores, b) basidium, c) dendrohyphidia, d) generative hyphae

(coll. S.P. Gorjén 2963, holotype)

106 ... Gorjén, Greslebin & Rajchenberg

20 um

PiatE 4. Dendrothele latenavicularis.

Basidiosporogenesis (coll. $.P. Gorjén 2963, holotype)

Comments — Dendrothele latenavicularis belongs to the group of Dendrothele

species with navicular basidiospores, a spore feature recently described by

Nakasone & Burdsall (2011) for three species from New Zealand. Dendrothele

navicularis Nakasone & Burds. has irregularly subfusiform to navicular

basidiospores, often medially constricted, sometimes curved, with a small,

distinct apiculus, (11-)14-16(-19) x 5.5-7.2(-8) um. In addition it has rare

suburniform, pyriform, or narrowly clavate to cylindric cystidia. Dendrothele

magnenavicularis Nakasone & Burds. produces long, narrowly navicular

basidiospores (17.3-)18-22(-24) x (5.5-)6-7(-7.8) um, with a distinct median

constriction. Dendrothele cymbiformis Nakasone & Burds. is characterized

by a hymenophore with hyphal pegs and narrowly navicular basidiospores,

(11.5-)14-17.5 x 4-5(-6) um.

Two Dendrothele species with arachiform basidiospores also appear to be

morphologically closely related. Dendrothele pitrae Gresl. & Rajchenb. from

Argentina, has cylindric, slightly or distinctly curved basidiospores with

a median constriction, and distally obtuse or tapered, (11—)12-15(-16) x

5-5.5(-6) um. Dendrothele arachispora Nakasone & Burds., from New Zealand,

has ellipsoid basidiospores, (14—)16-18 x (6.5—)7.8-8.5(-9.5) um, with a slight

median constriction (peanut-shape).

Dendrothele latenavicularis differs from the species discussed above in its

distinctly broader navicular basidiospores that lacks a median constriction and

tapers to a narrow, vermiform distal end (PLATE 3). Moreover, all these species

Dendrothele latenavicularis sp. nov. ... 107

with navicular or arachiform basidiospores have different host preferences.

Because no Dendrothele species with navicular or arachiform basidiospores is

known from the Northern Hemisphere, it is likely that they have a common

origin in the Southern Hemisphere. It would be very interesting to study these

species from a molecular perspective, to determine their genetic relationships

and phylogeographical patterns.

Key to Dendrothele species with navicular and arachiform basidiospores

la. Basidiospores cylindric to ellipsoid with a distinct median constriction .......... 2

Ib Basidiospores Navicular to fistlOrias vgula wp slens 6 ah ena a 4tclennd Plane.» trecsute-gurshee mae 3

2a. Basidiospores usually more than 6.5 um wide .................06. D. arachispora

2b. Basidiospores narrow, usually up to 6 um wide................. 0. eee D. pitrae

3a. Basidiospores (8-)9-11 um wide ............ 0. eee eee eee eee D. latenavicularis

3b. Basidiospores up to 6-7(-8) um wide ........ 0... eee eee eee eee teens 4

4a. Hymenophore with hyphal pegs, basidiospores up to 6 um wide ... D. cymbiformis

4b. Hymenophore smooth, basidiospores 5.5-8 um wide.......... 0... cece eee eee 5

5a. Cystidia present, basidiospores 11-19 um long ................... D. navicularis

5b. Cystidia absent, basidiospores 17-24 um long.............. D. magnenavicularis

Acknowledgments

Annarosa Bernicchia and Karen K. Nakasone reviewed this manuscript. The Consejo

Nacional de Investigaciones Cientificas y Técnicas (CONICET, Argentina) supported

this research by PIP 80101000. Sergio Pérez Gorjon is a postdoctoral research fellow of

the Agencia Espafiola de Cooperacion Internacional (MAEC-AECID). Alina Greslebin

and Mario Rajchenberg are researchers of CONICET.

Literature cited

Binder M, Hibbett DS, Larsson KH, Larsson E, Langer E, Langer G. 2005. The phylogenetic

distribution of resupinate forms across the major clades of mushroom-forming fungi

(Homobasidiomycetes). Systematics and Biodiversity 3: 113-157.

http://dx.doi.org/10.1017/S1477200005001623

Bodensteiner P, Binder M, Agerer R, Moncalvo JM, Hibbett DS. 2004. Phylogenetic relationships

of cyphelloid Homobasidiomycetes. Molecular Phylogenetics and Evolution 33(2): 501-515.

http://dx.doi.org/10.1016/j.ympev.2004.06.007

Boidin J, Lanquetin P, Duhem B. 1996. Contribution a la connaissance du genre Dendrothele

(Basidiomycotina, Aphyllophorales). Bulletin de la Société Mycologique de France 112: 87-126.

Cunningham GH. 1954. Thelephoraceae of New Zealand Part III: The genus Corticium. Transactions

of the Royal Society of New Zealand 82: 271-327.

Cunningham GH. 1963. The Thelephoraceae of Australia and New Zealand. Bulletin New Zealand

Department of Scientific and Industrial Research 145: 1-359.

Goranova G, Binder M, Hibbett DS. 2003. Molecular phylogenetics indicate that the corticioid

genus Dendrothele is highly polyphyletic. Inoculum 54: 22.

Greslebin A, Rajchenberg M. 1998. Corticioid Aphyllophorales (Basidiomycota) from the Patagonian

Andes forests of Argentina. 3. The genus Dendrothele. Mycotaxon 67: 469-486.

108 ... Gorjén, Greslebin & Rajchenberg

Larsson KH. 2007. Re-thinking the classification of corticioid fungi. Mycological Research 111:

1040-1063. http://dx.doi.org/10.1016/j.mycres.2007.08.001

Lemke PA. 1964. The genus Aleurodiscus (sensu lato) in North America. Canadian Journal of

Botany 42(6): 723-768.

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

Parmasto E, Nilsson RH, Larsson KH. 2004. Cortbase version 2. Extensive updates of a

nomenclatural database for corticioid fungi (Hymenomycetes). Phyloinformatics 1: 5.

MY COTAXON

http://dx.doi.org/10.5248/117.109

Volume 117, pp. 109-113 July-September 2011

Endocalyx melanoxanthus var. melanoxanthus (Ascomycota):

new to Brazil and three new hosts

NApjA SANTOS VITORIA?, MARIA AUXILIADORA Q. CAVALCANTI,

EpNA DorA MARTINS NEWMAN Luz?” & JOSE LuIZ BEZERRA?

Universidade Federal de Pernambuco, Departamento de Micologia,

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

?CEPLAC/CEPEC, Km 22, Rod Ilhéus/Itabuna, 45600-970, Ilhéus, Bahia, Brazil

>Universidade Estadual de Santa Cruz, Departamento de Ciéncias Agrarias,

Rodovia Ilhéus-Itabuna, Km 16, 45662-900, Ilhéus, BA, Brazil

* CORRESPONDENCE TO: nadjasv@hotmail.com

AxBstRact — Endocalyx melanoxanthus var. melanoxanthus was recovered during a

survey of microfungi on palms in Atlantic Rainforest areas of Northeast Brazil. Three new

hosts were identified for the fungus, newly reported for Brazil.

KEY worps —anamorph, biodiversity, Brazilian mycota, palm fungi, taxonomy

Introduction

Brazil has one of the richest Arecaceae (palm family) diversity in the world,

hosting at least 266 species (39 genera), of which 111 are regarded as endemic

(Leitman et al. 2010). Although Farr et al. (2011) list about 100 species of

microfungi associated with this family for the country, the palm mycobiota

remains almost unknown. Silva & Minter (1995) cite 38 ascomycete species

on palms in Pernambuco State and only six in Bahia State based on works

by Chaves Batista and collaborators. During palm microfungal surveys of

Pernambuco and Bahia states along the Brazilian northeastern coast (Vitoria

et al. 2008, 2010, 2011; Souza et al. 2008), the fungus Endocalyx melanoxanthus

var. melanoxanthus was found on Acrocomia intumescens Drude, Euterpe edulis

Mart., and E. oleracea Mart. This report of the fungus for the first time for Brazil

on three new hosts is part of an ongoing project to further knowledge of palm

microfungi in the Brazilian Northeast Atlantic Rainforest.

Materials & methods

Dead palm leaves were collected in the municipalities of Igarassu, Recife, Cabo

de Santo Agostinho, and Tamandaré in Pernambuco State and Urucuca in Bahia

110 ... Vitoria & al.

GEORAHIA — Instituto do Meio Ambiente

collections

PiaTE 1. Map of the northeastern Brazilian coast with the collecting sites indicated:

1. Municipality of Igarassu: 07°48'56.7"S 34°57'17.3"W, 49 m; 2. Municipality of Recife:

08°00'36.9"S 34°56'57.2"W, 30 m; 3. Municipality of Cabo de Santo Agostinho: 08°14'01.0"S

35°02'49.1"W, 54 m; 4. Municipality of Tamandaré: 08°43'22.8"S 35°10'40.5"W, 109 m;

5. Municipality of Urucguca: 14°56'132"S 39°16'601" W, 101 m.

State (PLATE 1). The collected specimens were incorporated into CEPEC Herbarium

(Mycological Collection) in Itabuna, Bahia. Observations under the stereomicroscope

preceded study of squash preparations and vertical, free hand sections of the conidiomata.

Morphological features were described, measured and photographed using a Carl Zeiss

microscope. All measurements were made in water preparations. In most cases, the

samples were stained with lacto-glycerol cotton blue.

Taxonomy

Endocalyx melanoxanthus (Berk. & Broome) Petch

var. melanoxanthus PL. 2-8

Conidiomata 0.4-1.0 mm high, up to 0.5 mm diam, yellow, cup-shaped to

cylindrical, scattered. Conidiophores up to 2 um wide, septate, hyaline, thread-

like or hyphoid. Conidiogenous cells 1-1.5 um long, integrated, determinate,

unicellular, hyaline, knob-like. Conidia 10.4-17 x 10-15 um in face view, and

Endocalyx melanoxanthus new to Brazil... 111

Piates 2-8. Endocalyx melanoxanthus var. melanoxanthus: 2-3. Conidiomata on host surface;

4. Section of conidioma; 5-8. Conidia (arrows indicate germ-slits). (Scale bars: 2 = 0.5 mm; 3 = 1 mm;

4= 100 um; 5-8 = 10 um).

7-9 um in side view (mean = 13.5 x 11.5 x 7.3 um, n = 50), 1-celled, smooth-

walled, flattened, round, oval or slightly polygonal in face view, at first pale, dark

brown at maturity, solitary, guttulate, with a longitudinal, straight germ slit.

SPECIMENS EXAMINED — BRAZIL. PERNAMBUCO: IGARAssuU, Refugio Ecolédgico

Charles Darwin, on dead leaf (rachis) of Acrocomia intumescens, 18.V1. 2009, coll. Nadja

Vitoria, det. JL Bezerra (CEPEC 2169); ReciFE, Parque Estadual Dois Irmaos, on dead

leaf (rachis) of Euterpe oleracea, 13. VI. 2009, coll. Nadja Vitoria, det. JL Bezerra (CEPEC

2170); CABO DE SANTO AGOSTINHO, Reserva Ecolégica de Gurjau, on dead leaf (rachis)

of E. oleracea, 09. VI. 2009, coll. Nadja Vitoria, det. JL Bezerra (CEPEC 2171); 16.XI.

2009, coll. Nadja Vitoria, det. JL Bezerra (CEPEC 2172); TAMANDARE, Reserva Biolégica

de Saltinho, on dead leaf (rachis) of E. oleracea, 03. IX. 2010, coll. Nadja Vitoria, det. JL

Bezerra (CEPEC 2173). Banta: URucGucA, EMARC, on dead leaf (rachis) of E. edulis,

18. III. 2010, coll. Nadja Vitoria, det. JL Bezerra (CEPEC 2174).

Hosts — Arecaceae: Acrocomia (this paper), Archontophoenix, Borassus, Cocos, Elaeis,

Euterpe (this paper), Livistona, Oncosperma, Phoenix, Satakentia, Trachycarpus,

Washingtonia; Dipterocarpaceae: Shorea; Smilacaceae: Ripogonum, Smilax.

112... Vitoria & al.

TABLE 1. Synopsis of conidia of Endocalyx species

TAXA

E. amarkantakensis U.S. Patel et al.

E. cinctus Petch

E. collantesis J. Mena & Mercado

E. indicus J.N. Kapoor & Munjal

E. indumentum G. Okada & Tubaki

E. melanoxanthus [var. melanoxanthus]|

E. melanoxanthus var. melanoxanthus

(CEPEC 2169)

E. melanoxanthus var. melanoxanthus

CONIDIA

5-10 x 4-5 um,

smooth walled

10-13 (-17) x 6-12 x 7-8 ym,

verrucose

10-13.5 x 7.5-11.5 x 5-7 um,

smooth

7-10 x 6-8 x 1.5-2 um,

smooth

8-12 um diam,

wall ornamented

14-19 x 12-14 x 6-7 um

10.4-17 x 10-15 x 7-9 um,

smooth

12-17 x 10-14 x 7-9 (-10) um,

SOURCE

Patel et al.

2002

Okada & Tubaki 1984

Mena Portales &

Mercado Sierra 1984

Kapoor &

Munjal 1966

Okada & Tubaki 1984

Petch 1908

This publication

Okada & Tubaki 1984

smooth

9-16 x 9-15 x 6-9 um,

verrucose

17-21 x 16-19 um,

verrucose

E. melanoxanthus var. grossus

G. Okada & Tubaki

E. thwaitesii Berk. & Broome

Okada & Tubaki 1984

DISTRIBUTION — Australia, Brazil (this paper), China, Cuba, Ghana, Hong Kong, India,

Jamaica, Japan, Malaya, New Zealand, Pakistan, Papua New Guinea, Philippines, Sabah,

Sarawak, Seychelles, Sierra Leone, Singapore, Sri Lanka, Taiwan, USA.

CoMMENTS — ‘The genus Endocalyx includes seven species, one of which is

divided into two varieties (TABLE 1). According to Okada & Tubaki (1984),

Endocalyx species colonize palm litter or (rarely) vine and lilies. Our report

represents the first record of E. melanoxanthus for Brazil, and Acrocomia

intumescens, Euterpe edulis, and E. oleracea represent new hosts for the

species.

Acknowledgments

The authors thank Dr. Michelline Silvério, Dr. Jadergudson Pereira and Alisson Neves

for collaboration, CAPES-MEC Brazil, CNPq-Brazil for scholarships and CEPLAC for

facilities and laboratories used to lead the research. Also sincere thanks are given to Drs.

Carlos Inacio, Francisco Freire, and Dartanha Soares for the pre-submission review of

our manuscript.

Literature cited

Farr DF, Rossman AY. 2011. Fungal databases. Systematic Mycology and Microbiology Laboratory,

ARS, USDA. htt:/nt.ars-grin.gov/fungaldatabases/fungushost.cfm/2011 (access20/jan/2011).

Kapoor JN, Munjal RL. 1966. Indian species of Stilbaceae. Indian Phytopathology 19: 348-350.

Leitman P, Henderson A, Noblick L. 2010. Arecaceae. In: Lista de Espécies da Flora do Brasil.

Jardim Botanico do Rio de Janeiro. http://floradobrasil.jbrj.gov.br/2010/FB000053 (access30/

sep/ 2010).

Endocalyx melanoxanthus new to Brazil ... 113

Mena Portales J, Mercado Sierra A. 1984. Nuevas especies de Endocalyx y Stachylidium

(Hyphomycetes, Deuteromycotina) de Cuba. Revista del Jardin Botanico Nacional 5(3): 53-60.

Okada G, Tubai K. 1984. A new species and new variety of Endocalyx (Deuteromycotina) from

Japan. Mycologia 76: 300-313. http://dx.doi.org/10.2307/3793106

Patel US, Pandey AK, Rajak RC. 2002. Two new hyphomycetes. Journal of Mycology and Plant

Pathology 32(1): 70-71.

Petch T. 1908. The genus Endocalyx Berkeley and Broome. Annals of Botany 22: 389-400.

Silva MS, Minter DW. 1995. Fungi from Brazil recorded by Batista and co-workers. Mycological

Papers 169. 585 p.

Souza CAP, Vitoria NS, Bezerra JL, Luz EDMN, Inacio CA, Dianese JC. 2008. Camarotella

brasiliensis sp. nov. (Phyllachoraceae) on Syagrus schizophylla (Arecaceae) from Brazil.

Mycotaxon 103: 313-317.

Vitoria NS, Bezerra JL, Gramacho KP, Luz EDMN. 2008. Camaroetella torrendiella comb. nov. e

C. acrocomiae: agents etiolégicos das lixas do coqueiro. Tropical Plant Pathology 33(4):

295-301. http://dx.doi.org/10.1590/S1982-56762008000400006

Vitoria NS, Bezerra JL, Gramacho KP. 2010. A simplified DNA extraction method for PCR analysis

of Camarotella spp. Brazilian Archives of Biology and Technology 53(2): 249-252.

http://dx.doi.org/10.1590/S1516-89132010000200001

Vitoria NS, Cavalcanti MAQ, Hyde KD, Bezerra JL. 2011. Arecomyces new to Brazil, including

A. attaleae sp. nov. Cryptogamie Mycologie 32(1): 103-108.

MY COTAXON

http://dx.doi.org/10.5248/117.115

Volume 117, pp. 115-122 July-September 2011

New records of rust fungi on sedges (Cyperaceae) from Pakistan

M. SABA'* & A.N. KHALID?

Department of Botany, University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

CORRESPONDENCE TO *: *'rustflora@gmail.com, *drankhalid@gmail.com

ABSTRACT —Puccinia caricis-kouriyamensis on Carex karoi, P. caricis-pocilliformis on Carex

sp., PR extensicola var. linosyridis-caricis on Carex divulsa, and P. conclusa on Cyperus difformis

(all three new to the country) and P. conclusa on Cyperus difformis were recently recorded

during a survey of rust fungi on sedges in Pakistan. These rust species have the potential to

serve as biological agents to control weeds and increase agricultural productivity. Carex karoi

and C. divulsa are newly reported hosts for rust fungi in Pakistan.

Key worps — Kaghan valley, Khyber-Pakhtoonkhwa, Punjab, Uredinales

Introduction

The Cyperaceae is a cosmopolitan family with 106 genera and ca. 5400

species (Govaerts et al. 2007) worldwide with 22 genera and 179 species known

in Pakistan (Kukkonen 2001). Members of the Cyperaceae commonly called

sedges are primarily grass-like herbs with linear leaves and parallel venation.

The most important cyperaceous weeds in terms of their adverse effect on

agriculture include Cyperus difformis, C. esculentus, C. iria, C. rotundus, and

Fimbristylis miliacea (Bryson & Carter 2008). Among the important fungal

pathogens of sedges are rust fungi (Basidiomycota: Uredinales). Previously,

thirteen species of Cyperaceae have been reported as hosts for rust fungi from

Pakistan (Ahmad et al. 1997; Afshan & Khalid 2008, 2009; Afshan et al. 2009).

In the present study, four Puccinia taxa, three of them new records for Pakistan,

are recorded on four cyperaceous hosts.

Materials & methods

During the survey of rust fungi from Pakistan, infected plants were collected from

different areas of Pakistan. Healthy plants were collected along with inflorescences and

fruits for accurate identification. Host plants were identified by comparing them with

specimens in the herbarium of the Department of Botany, University of the Punjab,

Lahore (LAH).

116 ... Saba & Khalid

Free hand sections of infected portions of material and spores were mounted in

lactophenol. Semi-permanent slides were prepared by cementing cover slips with nail

lacquer (Dade & Gunnell 1969). Preparations were observed under a NIKON YS 100

microscope. Drawings of spores were made by using a Camera Lucida (Ernst Leitz,

Wetzlar, Germany). Spores were measured using an ocular micrometer (Zeiss, St Albans,

Hertfordshire, England). At least twenty-five spores were measured for each spore stage.

Measurements include the usual range and the arithmetic means; extremes are given in

parentheses.

Enumeration of Taxa

Fic. 1. Lucida drawings of 1-2 celled teliospores of Puccinia caricis-kouriyamensis.

Scale bar = 15 um.

Puccinia caricis-kouriyamensis Morim., J. Jap. Bot. 47(4): 117 (1972). Fic. 1

SPERMOGONIA, AECIA and UREDINIA not seen. TELIA amphigenous,

naked, black, scattered, compact, rounded, erumpent, pulverulent, 140-280 x

240-440 um. TeLiosporss ellipsoid or oblong, pale brown to chestnut brown,

constricted at septum, attenuate or rounded at base, 19-27 x 39-59(-63) um

(mean 22 x 50.8 um); wall smooth, two-layered, pale brown to chestnut brown,

Sedge rusts new to Pakistan ... 117

1.8-4.7 um; germ pores two, one per cell, apical in upper cell, obscure in lower

cell, most probably adjacent to septum; apex rounded or conical, dark brown

with hyaline region, 4-9 um thick; pedicel persistent, hyaline, 5-10 um wide,

26-118 tm long. Mesospores intermixed, pale brown, sometimes chestnut

brown, clavate to oblong, attenuated below, 14-24 x 31-53 um (mean 18 x

40.8 um); wall pale brown to dark brown, 1-2.4 um, smooth; apex conical,

pale brown to dark brown with hyaline region, 4-10 um thick; pedicel hyaline,

persistent, up to 73 um long.

MATERIAL EXAMINED: PAKISTAN, GILGIT BALTISTAN, on Carex karoi Freyn, 26 July

2010, coll. & det. Malka Saba 01, with III stage (LAH 1140).

Rust fungi previously reported on Carex spp. from Pakistan include Puccinia

caricina DC. and P. caricis-filicinae Barclay on Carex filicina Nees, P. dioicae

Magnus and P. pakistani S. Ahmad on Carex nubigena D. Don, P. bolleyana

Sacc. on Carex flacca Schreb., and P. subepidermalis Afshan et al. on Carex curta

Gooden. (Ahmad et al. 1997, Afshan & Khalid 2009, Afshan et al. 2009).

Puccinia caricis-kouriyamensis, first reported on Carex brunnea Thunb.

(kogome-suge) from Japan (Hiratsuka et al. 1992), is a new record for Pakistan.

Carex karoi is a newly recorded host.

Fic. 2. Lucida drawings of teliospores of Puccinia caricis-pocilliformis.

Scale bar = 10 um.

Puccinia caricis-pocilliformis Morim., J. Jap. Bot. 49(8): 228 (1974). Fic. 2

SPERMOGONIA, AECIA and UREDINIA not seen. TELIA on abaxial side of leaf,

black, erumpent, scattered, first covered by epidermis, soon naked, compact,

80-200 x 100-600 um. TELIOsPoREs clavate, obovate-clavate or oblong-clavate,

yellowish brown to dark brown, constricted at septum, attenuated below,

118 ... Saba & Khalid

(15-)18-24(-28) x (32-)37-60(-74) um (mean 21.2 x 48.4 um); wall smooth,

yellowish brown or paler, two-layered, 1.5-2 um; apex rounded, conical or

obliquely conical, rarely truncate, dark brown, 6-17 um thick; pedicel long,

persistent, hyaline to pale yellow, 4-9.4 x (4-)7-71 um. Mesospores present,

clavate, pale brown, attenuated below, 13-15 x 36-47 um (mean 13.6 x 39 um);

apex conical, 2-9 um thick; pedicel short, up to 12 um long.

MATERIAL EXAMINED: PAKISTAN, GILGIT BALTISTAN, on Carex L., 26 July 2010, coll.

& det. Malka Saba 02, with III stage (LAH 1141).

Puccinia caricis-pocilliformis, first reported on Carex tristachya var. pocilliformis

(Boott) Kuk. (kotsubu-moegisuge) from Japan (Hiratsuka et al. 1992) , isa new

record for Pakistan.

Fic. 3. Lucida drawings of urediniospores of Puccinia conclusa,

showing echinulate ornamentation.

Scale bar = 10 um.

Puccinia conclusa Thiim., J. Sci. math. phys. nat. Lisboa, 1 Ser. 6(24): 237 (1878).

Fics. 3-4

SPERMOGONIA and AECIA unknown. UREDINIA amphigenous, mostly on

abaxial side, brown, scattered, somewhat aggregated, erumpent, naked, 100-250

x 100-400 um. UREDINIOSPORES subglobose, ellipsoid or obovoid, pale brown,

18-26 x 21-35 um (mean 20.9 x 27 um); wall pale brown, echinulate, 1-2(2.5-)

um; germ pores 2, equatorial; paraphyses absent. TELIA amphigenous, black,

loculate, covered with epidermis, aggregated, in groups, 90-110 x 90-170 um.

TELIOSPORES cinnamon brown, clavate or fusiform, constricted at septum,

attenuated below, 16-21 x 43-61 um (mean 18.3 x 50 um); wall cinnamon

brown, smooth, 1-1.4 um; apex rounded or conical, cinnamon brown to dark

brown, 3-6 um thick; pedicel persistent, pale brown, 6-9 x 8-41 um.

MATERIAL EXAMINED: PAKISTAN, PunyjaB, Lahore, at 216 m a.s.l., Cyperus difformis

L., 6 October 2010, coll. & det. Malka Saba 03, with II + III stages (LAH 1142).

Puccinia species previously reported on Cyperus in Pakistan include P. cypericola

S. Ahmad and P. philippinensis Syd. & P. Syd. on C. rotundus L., P. conclusa

[including misdeterminations as P romagnoliana Maire & Sacc.] on C. difformis

and C. rotundus from Chuharkana (Sheikhupura) and Karachi and on C. iria L.

Sedge rusts new to Pakistan ... 119

Fic. 4. Lucida drawings of teliospores of Puccinia conclusa.

Scale bar = 10 um.

from Lahore, and P. cyperi-laevigati Afshan & Khalid on C. laevigatus L. from

Khanspur (NWFP) (Ahmad 1956a,b; Ghaffar & Kafi 1968, Hasnain et al. 1959,

Afshan & Khalid 2008, Afshan et al. 2008).

Although Puccinia conclusa has been previously reported from other

Pakistan localities, the Lahore collection is the first found on Cyperus difformis,

a new host.

Puccinia extensicola var. linosyridis-caricis (E. Fisch.) Zwetko, Biblthca Mycol.

153: 109 (1993) . Fic. 5

SPERMOGONIA and AECIA not seen. UREDINIA amphigenous, scattered in

rows, golden brown, naked, covered with epidermis, 70-100 x 100-300 um.

UREDINIOSPORES sub-globose, ovoid or ellipsoid, pale brown, 16-24 x 20-27

um (mean 20.5 x 23.4 um); wall pale brown, echinulate, 1-2(-2.5) um; germ

pores 2, supraequatorial; pedicel hyaline, deciduous, 6-18 x 18-29 um. TELIA

on abaxial side, dark brown to blackish brown, scattered in rows, naked,

erumpent, 95-150 x 100-300 um. TeLiosporgs ellipsoid or oblong, cinnamon

brown to dark brown, 16-26 x 39-64 um (mean 21.5 x 51.4 um), constricted

at septum, attenuated at base; wall smooth, 1.5-3 um; apex rounded, conical

or sometimes truncated, dark brown, 8-15 um; pedicel hyaline to pale brown,

persistent, 5-13 x 15-41 um.

MATERIAL EXAMINED: PAKISTAN, KHYBER-PAKHTOONKHWA, Kaghan valley, Jalkhud,

3300 ma. s. 1., Carex divulsa Stokes, 16 August 2009, coll. & det. Malka Saba 04, with II

+ III stages (LAH 1143).

120 ... Saba & Khalid

Fic. 5. Lucida drawings of Puccinia extensicola var. linosyridis-caricis.

(A) Urediniospores showing echinulate ornamentation and supraequatorial germ pores.

(B) Teliospores. Scale bar = 10 um.

Puccinia extensicola var. linosyridis-caricis, first reported on Carex humilis

Leyss. from Japan (Hiratsuka et al. 1992), is a new record for Pakistan, and

Carex divulsa is a newly recorded host.

Key to the rust fungi on Cyperaceae in Pakistan

On Carex

JPET Te Freya) 01 fesco0 (010 [Spe earene Miter one Baer D Ribearn AM Uparen S Wireren PW eran Sharan Yiserin © Vora» Screven oo 2,

L Telia Aypaphyllous..rarely amp nisen ous 04.5. af db, whk bey slokbs sloth y ald be oho Abin a

2. Largest teliospore <50 «um long, oblong or oblong to clavate;

teliospore apex 8-14 um thick ...................00.. Puccinia caricis-filicinae

2 Jiareest teliospore > 50st 16mg. faces, 1 oes 1, ats 28 Ree 20 A re UBS og SA 3

Sedge rusts new to Pakistan ... 121

3. Teliospore pedicel short, <10 um; teliospore apex 4-6 um thick;

uredinial paraphyses clavate to capitate.................0000- P. subepidermalis

3. Teliospore pedicel long, >10 um; teliospore apex 4-9 um thick;

uiredinial-paraphyses absent... 24, Aves steesin « slovsle «lems P. caricis-kouriyamensis

4. earpesttelospore-=60 Pini lone txts Plat rl ttas wlettat laGel wlital s Patol Peels Pay 5

Advateestiteliospore->6O ions... «diss, +-a ute oa ditacea Liber dtbter a dike di Baw ano 7

5, Leliosporepedicelslor ty = UO) Mk gb ocgoz opp whee dyes peg dye peg ye pega eee P. pakistani

Steliospere pediceblonig a Or min wets Fate tte ser erty ete as 6

6. Teliospores clavate to oblong, 21-24 um wide; urediniospore germ pores 2,

roa Oy We Eee Se ore ROSE ere eS SR a P. bolleyana

6. Teliospores clavate, 16-24 um wide; urediniospore germ pores 2,

SUpracgatorial ssa ine ests ciety. ge laa cae sane use a eee aha ieee A URY Aansee P. dioicae

7: leliespotespedice 250 [itd IONS. 2, havi, slants atoucdee wlonbe t alivabe- olinbe ot lanibe wtb se enti 8

7. Teliospore pedicel >50 um long; teliospores obovate-clavate or oblong-clavate; apex

eS Ueac gis vol qt ASME NO Ai NARS ELSE aa RO Stn, Witenes Ce Oe P. caricis-pocilliformis

&: Urediiospore sermupores. 3=4;equigtorial:... a. ete ree 5 eed areted Sok Paricina

8. Urediniospore germ pores 2, supraequatorial. ... PR extensicola var. linosyridis-caricis

On Cyperus

i Wredimiospote-erim. pores two, Sq Walton al vs. uot hi a 4% Widnes Wid tale ine wigle Win eae ed Z

1. Urediniospore germ pores >two, scattered; paraphyses clavate;

teliospores 10-18 x (31-) 34-45 um ......... eee eee eee P. cyperi-laevigati

Zaleaeestteliospore > 90 titel One is. tp Notas ans oha an sie stadt ol sarc tsa ob ditt Ae anata 3

2. Largest teliospore <50 um long; teliospore apex <7 um long;

urediniospores 14-18 x 14-19 um ..... eee eee P. philippinensis

3. Teliospore apex <10 um thick; urediniospores 18-26 x 21-35 um....... P. conclusa

3. Teliospore apex >10 um thick; urediniospores 15-20 x 20-25 um...... P. cypericola

On Fimbristylis

Urediniospores 11-18 x 18-23 um, germ pores 3, supraequatorial; teliospores

long ellipsoid or fusoid, 10-18 x 35-55 um ............ P. flavipes Syd. & P. Syd.

On Scirpus

1. Teliospores two-celled; urediniospores 12-24 x 19-32 um............ P. scirpi DC.

1. Teliospores one-celled; urediniospores 18-24 x 24-34 um

ee Se ee See Mee op Oe PRE Uromyces lineolatus (Desm.) J. Schrot.

Acknowledgments

We sincerely thank Dr. Amy Rossman, Systematic Mycology and Microbiology

Laboratory, USDA-ARS, Beltsville and Dr. Marcin Piatek, W. Szafer Institute of Botany,

Polish Academy of Sciences, Poland for their valuable suggestions to improve the

manuscript and acting as presubmission reviewers. We are cordially thankful to Dr.

Shaun Pennycook, nomenclature editor, for reviewing the manuscript critically. We are

122 ... Saba & Khalid

highly obliged to the Pakistan Science Foundation, Islamabad for funding this research

work. We are also thankful to Dr. Najam-ul-Sehar Afshan and Dr. Abdul Rehman Khan

Niazi for accompanying the first author on field trips and assistance in the field.

Literature cited

Afs han NS, Khalid AN. 2008. New rust fungi on noxious weeds from Pakistan. Pak. J. Phytopathol.

20(1): 82-87.

Afshan NS, Khalid AN. 2009. New records of Puccinia and Pucciniastrum from Pakistan. Mycotaxon

108: 137-146. http://dx.doi.org/10.5248/108.137

Afshan NS, Khalid AN, Niazi AR. 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 Fairy Meadows, Northern Pakistan. Mycotaxon 110: 173-182.

http://dx.doi.org/10.5248/110.173

Ahmad S. 1956a. Uredinales of West Pakistan. Biologia 2(1): 29-101.

Ahmad S. 1956b. Fungi of Pakistan. Biological Society of Pakistan, Lahore Monograph 1: 1-126.

Ahmad §, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Nabiza Printing Press, Karachi, Pakistan.

Bryson CT, Carter R. 2008. Sedges: uses, diversity and systematic of the Cyperaceae. Monographs

in Systematic Botany, Missouri Botanical Garden 108: 15-101.

Dade HA, Gunnell J. 1969. Class work with fungi. Commonwealth Mycological Institute Kew.

Ghaffar A, Kafi A. 1968. Fungi of Karachi. Pak. J. Sci. 20: 5-10.

Govaerts R, Simpson DA, Bruhl J, Egorova T, Goetghebeur P, Wilson K. 2007. World checklist of

Cyperaceae sedges. Royal Botanic Gardens, Kew.

Hasnain SZ, Khan A, Zaidi AJ. 1959. Rusts and smuts of Karachi. Bot. Dept. Karachi Univ. Mont.

2. 36 p.

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y,

Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba Shuppankai, Ibaraki.

Kukkonen I. 2001. Cyperaceae. Flora of Pakistan, vol. 206. University of Karachi and Missouri

Botanical Press, Karachi and St. Louis. 277 p.

MY COTAXON

http://dx.doi.org/10.5248/117.123

Volume 117, pp. 123-130 July-September 2011

A new species and new records of Endophragmiella from 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— Four Endophragmiella species are described and illustrated: E. nanlingensis

occurring on dead branches of Melicope triphylla is proposed as new, and E. pulchra,

E. rostrata, and E. resinae found on dead branches of unidentified plants are recorded for the

first time from China. The specimens are deposited in Herbarium of Shandong Agricultural

University, Plant Pathology (HSAUP) and Mycological Herbarium, Institute of Microbiology,

Chinese Academy of Sciences (HMAS).

Key worps—anamorphic fungi, hyphomycete, taxonomy

Sutton (1973) proposed the genus Endophragmiella to accommodate two

species, E. pallescens B. Sutton and E. canadensis (Ellis & Everh.) B. Sutton.

Subsequently, Hughes (1979) emended the genus and gaveavery detailed account

of conidiogenesis and generic concepts. Endophragmiella is characterized by

macronematous, mononematous conidiophores with integrated, percurrently

proliferating conidiogenous cells and acrogenous, solitary, septate conidia

with rhexolytic secession. Presently, more than 80 species are accepted in

Endophragmiella, most of which are reported from rotten wood, dead branches,

and decaying leaves of various plants; a few species have associations with other

fungi (Sutton 1973, Hughes 1979).

During a continuing survey of saprobic fungi in tropical forests of Guangdong

Province, China, four Endophragmiella species were found on dead branches.

Endophragmiella nanlingensis is proposed herein as new, based on its distinctive

conidial characters, and E. pulchra, E. rostrata and E. resinae are recorded from

China for the first time.

Endophragmiella nanlingensis S.C. Ren & X.G. Zhang, sp. nov. Fic. 1

MycoBank MB561001

COLONIAE in substrato naturali effusae, pilosae, fuscae ad usque atrae. Mycelium

immersum, sparsum, ex hyphis septatis, laevibus, pallide brunneis, ramosis, 2-4 um latis

124 ... Ren, Ma & Zhang

Fic. 1. Endophragmiella nanlingensis. A. Conidiophore and conidiogenous cell.

B. Conidiophores, conidiogenous cells and conidia. C-D. Conidia.

compositum. CONIDIOPHORA macronematosa, mononematosa, singularia vel fasciculata,

simplicia, erecta, recta vel leviter flexuosa, laevia, septata, brunnea, apicem versus

pallidiora, 80-160 um alta, 4.5-9 um lata, interdum ad basim inflata, 1-5 proliferationes

percurrentes elongantia. CELLULAE CONIDIOGENAE monoblasticae, integratae, terminales,

percurrentes, cylindricae, ad apicem contractae et truncatae. CONIDIA acrogena, solitaria,

late fusiformia, rostrata, (3-)4(-5)-euseptata, ad septa interdum leviter constricta, cum

cellula centrali brunnea, cellula basali et apicali pallide brunnea, laevia, 55-80 um longa,

10-13 um lata, cum rostro 23-33 um longo, ad basim 1.5-2 um, ad fundamentum distincta

fractam ob partem cellulae conidiogenae superiorem fimbriata.

HoLotyPpe: CHINA. GUNGDONG PROVINCE: tropical forest of Nanling, on dead

branches of Melicope triphylla (Lam.) Merr. (Rutaceae), 10 Dec. 2010, Sh.C. Ren, HSAUP

H8334 (isotype HMAS 146104).

Erymo toey: named for the collection locality.

CoLoNies on natural substratum effuse, hairy, dark blackish brown to

black. Mycelium immersed, sparse, composed of septate, smooth, pale

Endophragmiella nanlingensis sp. nov. (China) ... 125

brown, branched hyphae 2-4 um wide. CONIDIOPHORES macronematous,

mononematous, arising singly or sometimes fasciculate, simple, erect, straight

or slightly flexuous, smooth, septate, brown, paler towards the apex, 80-160

um high, 4.5-9 um wide, sometimes swollen at the base, with 1-5 percurrent

proliferations. CONIDIOGENOUS CELLS monoblastic, integrated, terminal,

percurrent, cylindrical, tapered to a truncate apex. CONIDIA acrogenous,

solitary, broadly fusiform, rostrate, (3—)4(-5)-euseptate, slightly constricted at

the septa, central cells brown, basal cells and apical cells pale brown, smooth,

55-80 um long, 10-13 um wide, rostrum 23-33 um long, base truncate, 1.5-2

um wide, with a distinct basal frill of 2.5-3.5 um long, derived from the apex of

the conidiogenous cell.

Notes: Endophragmiella nanlingensis is closely related to E. fusiformis W.P. Wu

(Wu & Zhuang 2005) in its proliferating conidiophores producing fusiform,

rostrate, multiseptate conidia, except that E. fusiformis produces conidia that

are narrower (7.5-9 um) and have more (6-7) septa. In addition, the conidia

of E. nanlingensis slightly constricted at the septa, while those in E. fusiformis

are not.

Endophragmiella rostrata (Kirk 1985) and E. variabilis R.F. Castafeda

(Castafieda Ruiz 1988) are also morphologically similar to E. nanlingensis.

However, E. rostrata conidia are obclavate, smaller (13-24 x 4-5 um), and

have fewer (3) septa, while E. variabilis produces obclavate or Y-shaped,

smaller conidia (15-23 x 4-5 um, ramuli 9-10 x 3 um) with fewer septa (2-4-

septate).

Endophragmiella pulchra (B. Sutton & Hodges) P.M. Kirk. Trans. Br. Mycol. Soc.

78: 298 (1982) Fic. 2

ANAMORPHIC FUNGI. COLONIES effuse, blackish brown to black, hairy.

Mycelium immersed in the substratum, composed of branched, septate, pale

brown hyphae. CONIDIOPHORES macronematous, mononematous, arising

singly or in a group, erect, straight or slightly flexuous, sometimes swollen

at the base, smooth, brown, paler towards the apex, septate, 120-400 um

long, 4-7 um wide, with up to 3 or more percurrent proliferations at the

apex. CONIDIOGENOUS CELLS monoblastic, integrated, terminal, percurrent,

cylindrical. Conip1A holoblastic, terminal, solitary, dry, smooth, ellipsoid,

rostrate, 3-septate, sometimes slightly constricted at the septa, median cells

brown, basal and apical cell pale brown, conidia 35-40 um long (including

rostrum), 5.5-6.5 um wide, rostrum 8-11 um long, with a small basal frill

derived from the apex of the conidiogenous cell.

SPECIMENS EXAMINED: CHINA. GUNGDONG PROVINCE: tropical forest of Nanling, on

dead branches of unidentified plant, 10 Dec 2010, Sh.C. Ren, HSAUP H8423; Fujian

PROVINCE: forest park of Wuyishan, on dead branches of unidentified plant, 8 Aug.

2010, Y.D. Zhang, HSAUP H3101 (duplicate HMAS 146105).

126 ... Ren, Ma & Zhang

Fic. 2. Endophragmiella pulchra.

A. Conidiophores, conidiogenous cells and conidium. B. Conidia.

Notes: Endophragmiella pulchra was originally described by Sutton & Hodges

(1978) as Chaetendophragmiopsis pulchra. Kirk (1982) subsequently transferred

it to Endophragmiella based on its percurrently proliferating conidiogenous

cells and rhexolytically seceding conidia. The conidial features in our specimen

agreed well with those reported by Kirk. The species differs from others

Endophragmiella species by its ellipsoid, 3-4-septate conidia with a filiform

rostrum.

Endophragmiella rostrata P.M. Kirk, Mycotaxon 23: 325 (1985) Fig. 3

ANAMORPHIC FUNGI. COLONIES effuse, blackish brown to black, hairy.

Mycelium mostly immersed in the substratum, composed of branched, septate,

pale brown to brown, smooth-walled hyphae, 1.5-3 um wide. CONIDIOPHORES

macronematous, mononematous, arising singly or in group, erect, straight

or slightly flexuous, sometimes swollen at the base, smooth, brown, paler

towards the apex, septate, 120-180 um long, 3-4 um wide, with up to 4 or more

percurrent proliferations at the apex. CONIDIOGENOUS CELLS monoblastic,

Endophragmiella nanlingensis sp. nov. (China) ... 127

Fic. 3. Endophragmiella rostrata.

A-B. Conidiophores, conidiogenous cells and conidia. C. Conidia.

integrated, terminal, percurrent, cylindrical. Conip1A holoblastic, terminal,

solitary, dry, smooth, obclavate, rostrate, (3-)4-septate, sometimes slightly

constricted at the septa, pale brown to brown, body of conidium 20-30 um

long, 4-5 um wide, rostrum 25-40 um long, with a small basal frill derived

from the apex of the conidiogenous cell.

SPECIMEN EXAMINED: CHINA. GUNGDONG PROVINCE: tropical forest of Nanling, on

dead branches of unidentified plant, 10 Dec 2010, Sh.C. Ren, HSAUP H8226 (duplicate

HMAS 146106).

Notes: Compared with the type specimen described by Kirk (1985), the conidia

of our collection are slightly shorter and have more septa than those of type

128 ... Ren, Ma & Zhang

material (40-85 um long, 2-3-septate). Despite these minor differences, we

believe they represent the same species. Endophragmiella rostrata bears some

affinities with E. corticola P.M. Kirk (Kirk 1982) in conidial morphology, but

E. corticola can be separated from the former by its narrowly obclavate to

broadly fusiform conidia with more variable septation (1-3(-4)-septate), more

variable conidial dimensions (l-septate conidia 14-23 x 5-6 um, 2-septate

conidia 18-38 x 5.5-6(-6.5) um, 3-septate conidia 25-42 x 5.5-6.5 um), and

shorter apical cell measuring 8-12 um long.

Fic. 4. Endophragmiella resinae.

A-B. Conidiophores. C. Conidiophores, conidiogenous cells and conidia. D. Conidia.

Endophragmiella resinae P.M. Kirk. Trans. Br. Mycol. Soc. 76: 78 (1981) Fic. 4

CoLontgs effuse, hairy, dark blackish brown to black, inconspicuous.

Mycelium immersed in the substratum, composed of septate, smooth, pale

Endophragmiella nanlingensis sp. nov. (China) ... 129

brown, branched hyphae 2-4 um wide. CONIDIOPHORES macronematous,

mononematous, arising singly, unbranched, erect, straight or flexuous, septate,

smooth, dark brown, paler towards the apex, percurrently regenerating,

145-210 um long, 3.5-4.5 um wide, 7-10 um wide at the base, with 1-6

percurrent proliferations. CONIDIOGENOUS CELLS monoblastic, integrated,

terminal, percurrent, cylindrical, tapered to a truncate apex. CONIDIA

holoblastic, terminal, solitary, dry, smooth, obovoid to pyriform, thick-walled,

1-septate, the upper cell longer than the basal one, basal cell pale brown, apical

cell brown, 15-19 um long, 8-9 um wide, with a distinct basal frill of 0.5-1um

long, derived from the distal end of the conidiogenous cell.

SPECIMEN EXAMINED: CHINA. GUNGDONG PROVINCE: tropical forest of Nanling, on

dead branches of unidentified plant, 10 Dec 2010, Sh.C. Ren, HSAUP H8346 (duplicate

HMAS 146107).

Notes: Endophragmiella resinae is similar to a group of species with obpyriform

to subglobose and 1-septate conidia, such as E. angustispora S. Hughes,

E. bukkensis Révay, E. boewei (J.L. Crane) S. Hughes, E. bogoriensis Rifai,

E. cambrensis M.B. Ellis, E. globulosa (B. Sutton) S. Hughes, E. pinicola (M.B.

Ellis) S. Hughes, E. ramificata Hol.-Jech., E. tuberculata S.M. Leao & Gusmao,

and E. uniseptata (M.B. Ellis) S. Hughes (Ellis 1976, Holubova-Jechova 1986,

Hughes 1978, 1979; Ledo-Ferreira & Gusmao 2010, Révay 1987, Rifai 2008).

Endophragmiella resinae can be distinguished from most of these species by

its obovoid to pyriform conidia with an upper cell that is 1.5-3 times longer

than the lower. Endophragmiella globulosa, which has some morphological

similarities to E. resinae, differs in its subglobose to broadly obovoid conidia

with a brown to dark brown upper cell and pale brown to brown basal cell.

The morphological features of our specimen overlap with the type specimen as

described by Kirk (1981).

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

Castaneda RE 1988. Fungi Cubenses III. Instituto de Investigaciones Fundamentales en Agricultura

Tropical “Alejandro de Humboldt” ACC, La Habana.27pp.

Ellis MB. 1976. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey,

England. 507pp.

Holubova-Jechova V. 1986. Lignicolous hyphomycetes from Czechoslovakia. 8. Endophragmiella

and Phragmocephala. Folia Geobotanica et Phytotaxonomica 21: 173-198.

Hughes SJ. 1978. Endophragmiella globulosa. Fungi Canadenses 127: 1-2.

130 ... Ren, Ma & Zhang

Hughes SJ. 1979. Relocation of species of Endophragmia auct. with notes on relevant generic names.

New Zealand J. Bot. 17: 139-188.

Kirk PM. 1981. New or interesting microfungi I. Dematiaceous hyphomycetes from Devon. Trans.

Br. Mycol. Soc. 76: 71-87. http://dx.doi.org/10.1016/S0007-1536(81)80010-1

Kirk PM. 1982. New or interesting microfungi V. Microfungi colonizing Laurus nobilis leaf litter.

Trans. Br. Mycol. Soc. 78: 293-303. http://dx.doi.org/10.1016/S0007-1536(82)80013-2

Kirk PM. 1985. New or interesting microfungi XIV. Dematiaceous hyphomycetes from Mt. Kenya.

Mycotaxon 23: 305-352.

Leao-Ferreira SM, Gusmao LFP. 2010. Conidial fungi from the semi-arid Caatinga biome of Brazil.

New species of Endophragmiella, Spegazzina and new records for Brazil, South America and

Neotropica. Mycotaxon 111: 1-10. http://dx.doi.org/10.5248/111.1

Révay A. 1987. New or interesting hyphomycetes on forest litter from Hungary. Acta Botanica

Hungarica 33: 67-73.

Rifai MA. 2008. Endophragmiella bogoriensis Rifai, spec. nov. (hyphomycetes). Reinwardtia 12:

270276:

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

1-143.

Sutton BC, Hodges Jr CS. 1978. Eucalyptus microfungi. Chaetendophragmiopsis gen. nov. and other

hyphomycetes. Nova Hedwigia 29: 593-607.

Wu WP, Zhuang WY. 2005. Sporidesmium, Endophragmiella and related genera from China. Fungal

Divers. Res. Ser. 15: 1-351.

MY COTAXON

http://dx.doi.org/10.5248/117.131

Volume 117, pp. 131-135 July-September 2011

Two new Minimelanolocus species from southern China

JIAN Ma, Y1-DONG ZHANG, LI-Guo Ma & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

AsBsTRACT — Minimelanolocus linderae sp. nov. and M. mori sp. nov. were discovered from

dead branches in southern China. They are described, illustrated, and compared with closely

related taxa. The specimens are deposited in Herbarium of Shandong Agricultural University,

Plant Pathology (HSAUP) and Mycological Herbarium, Institute of Microbiology, Chinese

Academy of Sciences (HMAS).

KEY worpDs — anamorphic fungi, taxonomy

During ongoing surveys of tropical and subtropical microfungi from the forests

in southern China, two interesting fungi with euseptate conidia and polyblastic

conidiogenous cells were collected on dead branches of Lindera communis

and Morus alba. The morphologies of their conidia and conidiogenous cells

suggested they are new species of the genus Minimelanolocus.

Minimelanolocus linderae Jian Ma & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 561233

Fungus anamorphicus. CoLoNniAE in substrato naturali effusae, brunneae vel

atrobrunneae, pilosae. Mycelium partim superficiale, partim immersum in substrato,

ex hyphis ramosis, septatis, pallide brunneis vel brunneis, laevibus, 1-2 um crassis

compositum. CONIDIOPHORA macronematosa, mononematosa, singula, interdum

caespitosa, nonramosa, erecta, recta vel flexuosa, septata, laevia, brunnea, apice versus

pallidiora, 83-133 um longa, 2-4.5 um crassa. CELLULAE CONIDIOGENAE holoblasticae,

polyblasticae, in conidiophoris incorporatae, indeterminatae, sympodialiter extendentes,

terminales deinde intercalares, pallide brunneae. Loci conidiogeno inconspicuo vel

leviter prominentibus, refractivi. Conidiorum secessio schizolytica. Conrp1A solitaria,

acropleurogena, ellipsoidea vel cylindrica, basi truncata, pallide brunnea, cellulis extimis

subhyalinis, laevia, plerumque 4-euseptata, raro 3-euseptata, 26-35 um longa, 7-9 um

crassa, basi truncata 1-1.5 um lata.

Type: China, Fujian Province: Mount Wuyi, on dead branches of Lindera communis Hemsl.

(Lauraceae), 15 Aug. 2009, J. Ma, (Holotype HSAUP H5113; isotype HMAS146122).

ETyMo_oey: in reference to the host genus, Lindera.

132 ... Ma &al.

Fic. 1. Minimelanolocus linderae. A. Conidiophores with conidia. B, C. Conidia.

Anamorphic fungi. CoLonizs effused on natural substrate, brown to dark

brown, hairy. Mycelium partly superficial, partly immersed, composed

of branched, septate, pale brown to brown, smooth-walled hyphae, 1-2

uum thick. ConipiopHores differentiated, single, sometimes caespitose,

unbranched, erect, straight or flexuous, septate, smooth, brown, paler toward

the apex, 83-133 um long, 2-4.5 um thick. CONIDIOGENOUS CELLS holoblastic,

polyblastic, integrated, indeterminate, sympodially extending, terminal

becoming intercalary, pale brown. Conidiogenous loci inconspicuous or

slightly prominent, refractive. Conidial secession schizolytic. Conip1a solitary,

apical and lateral, simple, ellipsoidal or cylindrical, truncate at the base, pale

brown except for the end cells which are subhyaline, smooth-walled, mostly

4-euseptate, rarely 3-euseptate, 26-35 um long, 7-9 um thick in the broadest

part, 1-1.5 um wide at the truncate base.

Minimelanolocus spp. nov. (China) ... 133

CoMMENTS — Minimelanolocus linderae is similar in conidial shape to

M. magnoliae K. Zhang & X.G. Zhang (Zhang et al. 2009), M. bambusae (N.D.

Sharma) R.F. Castafieda & Heredia, and M. subulifer (Corda) R.F. Castaneda

& Heredia (Castaneda Ruiz et al. 2001). However, conidia of M. linderae are

narrower than those of M. magnoliae (conidia 10-11 um wide), and larger

than those of M. bambusae (conidia 15-19 x 6-7 um) and M. subulifer

(conidia 12-29 x 3.5-5.5 um). Moreover, in M. linderae mature conidia are

3-4-euseptate, while those of the three known species are mostly 3-septate. In

addition, the mature conidia of M. linderae are pale brown, but the cell at each

end is subhyaline, which is obviously different from those of M. magnoliae,

M. bambusae and M. subulifer.

Minimelanolocus mori Jian Ma & X.G. Zhang, sp. nov. FIG. 2

MycoBank MB 561234

Fungus anamorphicus. CoLoniaeE in substrato naturali effusae, brunneae vel atrobrunneae,

pilosae. Mycelium partim superficiale, partim immersum in substrato, ex hyphis

ramosis, septatis, pallide brunneis vel brunneis, laevibus, 1.5-3 um crassis compositum.

CONIDIOPHORA macronematosa, mononematosa, singula, nonramosa, erecta, recta vel

flexuosa, septata, laevia, atrobrunnea, apice versus pallidiora, 150-280 um longa, 3.5-

5.5 um crassa. CELLULAE CONIDIOGENAE holoblasticae, polyblasticae, in conidiophoris

incorporatae, indeterminatae, sympodialiter extendentes, terminales deinde intercalares,

brunneae. Loci conidiogeno inconspicuo vel leviter prominentibus, subobscurus.

Conidiorum secessio schizolytica. CoNipIA solitaria, acropleurogena, simplicia, ellipsoidea

vel cylindrica, apice rotundata, ad basim truncata, pallide brunnea, cellulis apicali

subhyalina vel pallide brunnea, laevia, plerumque 3-euseptata, raro 2-euseptata, 19-25

um longa, 7.5-10 um crassa, basi truncata 2.5-3.5 um lata, cellulis basi 8.5-11 um longa.

Type: China, Hainan Province: tropical forest of Bairentan, on dead branches of Morus

alba L. (Moraceae), 8 Dec 2010, J. Ma, (Holotype HSAUP H5499-1; isotype HMAS

146123).

ETyMOLoGey: in reference to the host genus, Morus.

Anamorphic fungi. CoLONtEs effused on natural substratum, brown to dark

brown, hairy. Mycelium partly superficial, partly immersed, composed of

branched, septate, pale brown to brown, smooth-walled hyphae, 1.5-3 um thick.

CONIDIOPHORES differentiated, single, solitary, unbranched, erect, straight or

flexuous, septate, smooth, dark brown, paler toward the apex, 150-280 um long,

3.5-5.5 um thick. CONIDIOGENOUS CELLS holoblastic, polyblastic, integrated,

indeterminate, sympodially extending, terminal becoming intercalary,

brown. Conidiogenous loci inconspicuous or slightly prominent, somewhat

obscure. Conidial secession schizolytic. Conip1A solitary, apical and lateral,

simple, ellipsoidal or cylindrical, apex rounded, base truncate, brown except

for the apical cell which is subhyaline or pale brown, smooth-walled, mostly

3-euseptate, rarely 2-euseptate, 19-25 um long, 7.5-10 um thick in the broadest

part, 2.5-3.5 um wide at the truncate base, basal cell 8.5-11 um long.

134 ... Ma &al.

A — B > C

20nm

Fic. 2. Minimelanolocus mori. A, B. Conidiophores and conidia. C. Conidia.

ComMENtTs - The conidia of Minimelanolocus mori resemble those of

M. magnoliae, M. endospermi Jian Ma & X.G. Zhang (Ma et al. 2008), and

M. hughesii (M.B. Ellis) R.F. Castafieda & Heredia (Castaneda Ruiz et al.

2001) in shape. However, M. mori differs from M. magnoliae (conidia 29-38 x

10-11 um, basal scar 1-2 um wide) in its smaller conidia with wider basal scar,

and from M. hughesii (conidia 12-18 x 4.5-6 um) in having larger conidia.

Moreover, M. mori can be separated from M. endospermi (conidia 20-30 x

9-12 um, mainly 2-euseptate) by its slightly smaller mainly 3-euseptate conidia.

In addition, the conidia of M. mori have a much longer basal cell than is seen

in the other three species.

Acknowledgments

The authors express gratitude to Dr W.B. Kendrick and Dr N.R. O’Neill 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. 2006FY 120100, 2006FY110500-5).

Minimelanolocus spp. nov. (China) ... 135

Literature cited

Castaneda Ruiz RE, Heredia G, Reyes M, Arias RM, Decock C. 2001. A revision of the genus

Pseudospiropes and some new taxa. Cryptog. Mycol. 22: 3-18.

http://dx.doi.org/10.1016/S0181-1584(01)01057-0

Ma J, Zhang K, Zhang XG. 2008. Two new species of the genus Minimelanolocus in China.

Mycotaxon 104: 147-151.

Zhang K, Fu HB, Zhang XG. 2009. Taxonomic studies of Minimelanolocus from Yunnan, China.

Mycotaxon 109: 95-101. http://dx.doi.org/10.5248/109.95

MY COTAXON

http://dx.doi.org/10.5248/117.137

Volume 117, pp. 137-143 July-September 2011

Two new species of Passalora and Pseudocercospora

from northeastern Uttar Pradesh, India

RAGHVENDRA SINGH , SHAMBHU KUMAR & KAMAL

Department of Botany, D.D.U. Gorakhpur University, Gorakhpur, U.P, India- 273 009

* CORRESPONDENCE TO: drsinghtaxon@gmail.com

ABSTRACT — During surveys of hyphomycetes from northeastern Uttar Pradesh, India, two

undescribed taxa were discovered, Passalora barringtoniigena sp. nov. on living leaves of

Barringtonia acutangula (Lecythidaceae), and Pseudocercospora miliusae sp. nov. on Miliusa

tomentosa (Annonaceae). These two species differ from similar species morphologically.

Illustrations and descriptions of the new species are provided.

Key worps — biodiversity, foliar diseases, phytopathogenic fungi, taxonomy

Introduction

We have encountered several plant species exhibiting leaf blights during our

survey of plant diseases. Critical examination and a thorough survey of the

literature revealed that two of these blights were caused by two undescribed

species of Passalora and Pseudocercospora. Passalora is characterised by septate

coloured conidia and conidiophores with thickened scars, and Pseudocercospora

is characterised by septate coloured conidia and conidiophores without

thickened scars (Crous & Braun 2003). After comparing the fungi with all other

species reported on allied hosts, we found them to be distinct and identified

them as new species of Passalora and Pseudocercospora.

Materials & methods

Infected leaf samples from different parts of northeastern Uttar Pradesh were placed

in separate polythene bags and taken to the laboratory. Surface scrapings and free-hand

cut sections were prepared from infected portions of the leaf samples and mounted

in lactophenol cotton blue on microscope slides. Fungal structures were drawn under

1000x magnification with the help of camera lucida to illustrate all possible details of

morphology and ontogeny of reproductive propagules. Measurements were taken with

the help of an ocular micrometer. Morphotaxonomic determinations were made with

the help of current literature and available resident expertise. Holotypes have been

138 ... Singh, Kumar & Kamal

deposited in HCIO (Herbarium Cryptogamiae Indiae Orientalis), Indian Agricultural

Research Institute, New Delhi, India; isotypes were retained in the departmental

herbarium for further reference.

Taxonomy

Passalora barringtoniigena R. Singh, Sham. Kumar & Kamal, sp. nov. FIG. 1

MycoBank MB 519071

Maculae hypogenae, atro-brunneae vel nigrae. Coloniae hypophyllae, effusae. Mycelium

internum. Stromata parva. Conidiophora macronematosa, ex hyphis oriunda singulata vel

2-6-fasciculata, simplicia, erecta vel procumbentia, recta vel flexuosa, geniculata, laevia,

crassitunicata, pallide brunnea vel atro-brunnea, 2-7-septata, 22-105 x 3-6 um. Cellulae

conidiogenae integratae, terminales vel intercalares, polyblasticae, cicatricatae, cicatrices

incrassatae. Conidia simplicia, sicca, acropleurogena, solitaria, cylindricata, 3-11-septata,

recta vel leniter curvata, olivacea vel olivaceo-brunnea, laevia, tenui-tunicata, apices

obtusae, ad basim obconicotruncata, hila incrassata, 35-82 x 2-6 um, hila 1.5-2.5 um

lata.

TyPE: On living leaves of Barringtonia acutangula (L.) Gaertn. (Lecythidaceae), Nichlaul

Forest, Mahrajganj (U.P.), India, January 2008, coll. Raghvendra Singh, HCIO No. 48783

(holotype), GPU Herb. No. KSR-257 (isotype).

EryMo_oey: the epithet is derived from the genus name of the host.

Infection spots hypogenous, minute, dark brown to black. Colonies

hyphophyllous, effuse. Mycelium internal. Stromata absent or poorly developed.

Conidiophores macronematous, arising singly from internal hyphae or in a

fascicle of 2-6 from poorly developed stromata, simple, erect to procumbent,

straight to flexuous, geniculate, smooth, thick-walled, light to dark brown,

2-7-septate, 22-105 x 3-6 um. Conidiogenous cells integrated, terminal,

becoming intercalary, polyblastic, cicatrized with thickened conidial scars.

Conidia simple, dry, acropleurogenous, solitary, cylindrical, 3-11-septate,

straight to slightly curved, olivaceous to olivaceous-brown, smooth, thin-

walled, apex obtuse, base obconico-truncate, hila thickened, 35-82 um long

and 2-6 um thick, germinating conidia present, hila 1.5-2.5 um wide.

Two Passalora species previously described on Barringtonia are P. barringtoni-

icola (Y.L. Guo) U. Braun & Crous (Crous & Braun 2003) on B. yunnanensis

from China and P. barringtoniae-acutangulae (Kamal et al.) Poonam Srivast.

(Srivastava 1994) on B. acutangula from India and Australia. Passalora

barringtoniicola differs from P. barringtoniigena in having obclavato-cylindrical,

catenate conidia with long curved tips and conidiophores that are densely

fasciculate and arise from well-developed subepidermal stromata (Crous &

Braun 2003). In P. barringtoniae-acutangulae the stromata are well developed,

the conidiophores are longer (< 240 um), and the conidia are shorter and wider

(17.5-38.6 x 4.2-11.7 um), 0-3-septate, and obclavate to broadly fusiform

(Srivastava 1994).

Passalora and Pseudocercospora spp. nov. (India) ... 139

Fic. 1. Passalora barringtoniigena.

a: symptoms; b: conidia, germinating conidia, and conidiophores.

(Scale bars: a = 20 mm, b = 20 um).

140 ... Singh, Kumar & Kamal

Pseudocercospora miliusae R. Singh, Sham. Kumar & Kamal, sp. nov. FIG. 2

MycoBank MB 519075

Maculae amphigenae, primo discretae, venulis definitae, demum coalescentes, irregulares

et plus minusve, nigrae vel brunneae, 1-21 mm in diam. Coloniae hypophyllae,

effusae, brunneae. Mycelium internum. Stromata subepidermalia vel erumpentia,

pseudoparenchymatica, 17-21 x 36-42 um. Conidiophora ex hyphis oriunda singulata

vel 2-50-fasciculata, macronematosa, simplicia, cylindricata, erecta vel procumbentia,

recta vel curvata, laevia, crassitunicata, ramosa, pallide brunnea, 1-2-septata, 10-70

x 2-5 um. Cellulae conidiogenae integratae, terminales, monoblasticae, cicatrices non

incrassatae. Conidia simplicia, tenui-tunicata, sicca, acropleurogena, recta vel curvata,

non ramosa, cylindricata vel obclavata, 0-4-septata, apices subacuta vel acuta, ad basim

obconicotruncata vel rotundata, pallide brunnea, 20-58 x 2-3 um, hila non incrassata,

1-1.5 um lata.

Type: On living leaves of Miliusa tomentosa (Roxb.) J. Sinclair (Annonaceae),

Vindhyavashini Park, Gorakhpur (U.P.), India, January 2008, coll. Raghvendra Singh,

HCIO No. 48786 (holotype), GPU Herb. No. KSR-320 (isotype).

EryMo_oey: the epithet is derived from the genus name of the host.

Infection spots amphigenous, discrete and vein-limited in the beginning but

coalescing to become irregular and more or less necrotic afterwards, black to

brown, 1-21 mm in diam. Colonies hypophyllous, effuse, brown. Mycelium

internal. Stromata subepidermal to erumpent, pseudoparenchymatous, 17-21

x 36-42 um. Conidiophores arising singly from internal hyphae or in a fascicle

of 2-50 from subepidermal or erumpent stromata, macronematous, simple,

erect to procumbent, cylindrical, smooth-walled, thick-walled, branched,

light brown, 1-2-septate, 10-70 x 2-5 um. Conidiogenous cells integrated,

terminal, monoblastic, scars unthickened. Conidia simple, thin-walled, dry,

acropleurogenous, straight to curved, unbranched, cylindrical to obclavate,

0-4-septate, apex subacute to acute, base obconicotruncate to rounded, light

brown, 20-58 x 2-3 um, hila unthickened, 1-1.5 um wide.

A thorough survey of the literature indicates that no other Pseudocercospora

species has been described on Miliusa tomentosa, and only one, Ps. annonacea

(Kamal et al.) U. Braun (Braun 1994), has been described on any Miliusa species.

In addition to the different host species, Ps. annonacea is separated by conidia

that are larger (57.5-115 x 3.45-4.6 uum), pluriseptate (up to 15-20-septate) and

hyaline. The only other cercosporoid species described on Miliusa tomentosa,

Passalora miliusae U. Braun & Crous (Crous & Braun 2003), has subhyaline

conidia and thickened conidiophore scars.

Pseudocercospora species known to occur the same host family (Annonaceae)

differ from Ps. miliusae as noted below.

In Ps. aethiopicae Deighton, conidiophores are shorter (10-40 um) and

always densely fasciculate, never solitary (Deighton 1976).

In Ps. annonae U. Braun & Crous conidia (50-150 um) and conidiophores

(20-110 um) are longer (Braun et al. 2002).

Passalora and Pseudocercospora spp. nov. (India) ... 141

Fic. 2. Pseudocercospora miliusae.

a: symptoms; b: conidia, stroma, and simple and branched conidiophores.

(Scale bars: a = 20 mm, b = 20 um).

142 ... Singh, Kumar & Kamal

In Ps. annonae-squamosae U. Braun & R.F. Castafieda, conidia are longer

(< 75(-85) um) and 3-7-septate, and the shorter (< 30(-45) um) conidiophores

arise singly from superficial hyphae on the lower leaf surface to form large

dense (sometimes almost sporodochial) fascicles with well-developed stromata

on the upper leaf surface (Braun & Crous 2008).

Pseudocercospora annonarum (Petr. & Cif.) U. Braun & Crous is readily

separated by the presence of synnemata (80-250 x 20-50 um) and conidia that

are ellipsoid to ovoid, variably (0-10)-septate, and broader [< 8(-9) um diam.]

(Braun & Crous 2008).

In Ps. annonifolii (Bat. & Peres) U. Braun & EO. Freire, the conidiophores

are shorter (< 20-(30) um) and conidia longer (< 220 um) (Braun & Freire

2003).

The shorter (6-20 um) conidiophores of Ps. asiminae (Ellis & Morgan)

U. Braun & Crous arise from both substomatal and superficial hyphae and the

conidia are variably shaped (ellipsoid-ovoid to fusiform), longer (20-80 um),

and with 1-9 transverse (plus occasionally 1-2 longitudinal or oblique) septa

and an obtuse apex (Braun & Crous 2008).

The conidia of Ps. asiminae-pygmaeae U. Braun are larger (25-140 x 3-6(-7)

um) (Braun & Crous 2008).

Pseudocercospora oblecta (Syd.) Crous & U. Braun possesses sporodochia,

shorter (30-50 um) verruculose conidiophores that taper to a rounded or

subtruncate apex of conidiogenous cells that proliferate 1-4 times percurrently,

and longer (40-120 um) verruculose conidia with 1-15 septa and guttules

(Braun & Crous 2008).

In Ps. polyalthiae J.M. Yen et al. the conidiophores are consistently solitary

and arise from superficial hyphae, and the conidia are 3-13-septate and longer

(40-156 um) (Yen et al. 1982).

Pseudocercospora scitula (Syd.) Deighton produces longer conidia (50-110 x

5-8 um) and conidiophores (60-220 x 4-6 um) (Deighton 1976).

In Ps. xenoannonicola Crous & Bench. both external as well as internal

mycelium is present, conidiophores are shorter (10-30 x 3-4 um), and conidia

are longer (30-100 x 2-3 um) and 5-7-septate (Crous et al. 2000).

Acknowledgments

The authors are grateful to Dr Eric H.C. McKenzie and Dr Roland Kirschner for

serving as pre-submission reviewers. We also express our deep thanks to Dr Shaun

Pennycook for nomenclatural review. Authors’ thanks are also due to the Curator,

HCIO, New Delhi, for accepting holotype specimens and providing accession numbers

thereof.

Passalora and Pseudocercospora spp. nov. (India) ... 143

Literature cited

Braun U. 1994. Studies on Ramularia and allied genera (VI). Nova Hedwigia 58(1-2): 191-222.

Braun U, Crous PW. 2008. Cercosporoid hyphomycetes on hosts of the Annonaceae: Cercospora

annonaceae and Isariopsis annonarum revisited. Mycotaxon 105: 207-224.

Braun U, Freire FO. 2003. Miscellaneous notes on some cercosporoid hyphomycetes. Bibliotheca

Lichenologica 86: 79-98.

Braun U, Crous PW, Pons N. 2002. Annotated list of Cercospora species (epithets a-b) described by

C. Chupp. Feddes Repertorium 113(1-2): 112-127.

http:..dx.doi.org10.1002/1522-239X(200205)113:1/2<112::AID-FEDR112>3.0.CO;2-H

Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs: 1. Names published in Cercospora

and Passalora. CBS, Utrecht, The Netherlands, Fungal Biodiversity Centre. 571 p.

Crous PW, Benchimol RL, Albuquerque FC, Alfenas AC. 2000. Foliicolous anamorphs of

Mycosphaerella from South America. Sydowia 52(2): 78-91.

Deighton FC. 1976. Studies on Cercospora and allied genera. VI. Pseudocercospora Speg., Pantospora

Cif., and Cercoseptoria Petr. Mycological Papers 140: 1-168.

Srivastava P. 1994. Recombinations in genus Passalora Fries. Journal of Living World 1(2):

112-119.

Yen JM, Kar AK, Das BK. 1982. Studies on hyphomycetes from West Bengal, India, I. Cercospora

and allied genera of West Bengal, 1. Mycotaxon 16: 35-57.

MY COTAXON

http://dx.doi.org/10.5248/117.145

Volume 117, pp. 145-148 July-September 2011

Lecanora wrightiana and Rhizocarpon inimicum,

rare lichens new to Turkey and the Middle East

KENAN YAZIcI'., ANDRE APTROOT? & ALI ASLAN?

‘Biology Department, Science Faculty, Karadeniz Technical University, 61080, Trabzon, Turkey

?ABL Herbarium G.v.d. Veenstraat 107, NL-3762 XK Soest, The Netherlands

*Biology Department, Kazim Karabekir Education Faculty, Atatiirk University, Erzurum, Turkey

*CORRESPONDENCE TO kcagri_1997@yahoo.com

ABSTRACT — During a recent excursion in Igdir region (Turkey), we sampled lichens from

two localities, among which Lecanora wrightiana and Rhizocarpon inimicum were determined

as new to Turkey and the Middle East. Geographic distribution, substrate, chemistry, and

comparisons with morphologically similar taxa are presented.

Key worps — Ascomycetes, biodiversity, parasitic

Introduction

Compared to other countries, not many lichen studies have been conducted

in Turkey, so the lichen flora of Turkey remains poorly known. Recently, many

lichen taxa have been recorded for Turkey (Yazici et al. 2010a,b,c, Candan &

Halici 2009, Candan et al. 2010, Kinalioglu & Aptroot 2010, Kinalioglu 2010a,b),

but more studies are needed to form a complete lichen flora. No lichenized

fungi have previously been reported for Igdir region.

To date, 27 Rhizocarpon species and 95 Lecanora taxa have been reported

from Turkey. At least 18 parasitic Rhizocarpon species are known from Europe

(Poelt & Hafellner 1983, Poelt & Vézda 1984, Timdal 1986), but only one

parasitic species (Rhizocarpon epispilum on Pertusaria rupicola) has been found

from Turkey (Breuss & John 2004).

Material & methods

Igdir, one of the poorest forested areas in Turkey, is dominated by steppe. Rich grassy

plants cover the surface of the province. Mountain Zordag (Igdir: Center) is a well-

lit, very high, windswept, open treeless area with gently sloped terrain with streams,

grass and rocks. Prunus, Pyrus, and Populus trees are dominant in Uskaya village (Igdir:

Tuzluca) with Salix and Elaeagnus occasionally present (Baytop & Denizci 1963). The

146 ... Yazici, Aptroot & Aslan

climate is characterized by hot and dry summers with moderate precipitation, and cold,

snowy winters with high precipitation. Mean annual temperature is 11.6°C. The mean

annual rainfall is about 257.6 mm. Mean annual humidity is 63%. (Akman 1999).

The lichen specimens were collected from Igdir region on 13-14 June 2010. Air-dried

samples were examined with Nikon SMZ1500 stereomicroscope and a Nikon Eclipse

80i light microscope. Identifications were determined by consulting keys (Guderley &

Lumbsch 1999, Miyawaki 1988, Poelt 1990, Poelt & Vézda 1984, Upreti 1997). Vouchers

are stored in the herbarium of the Biology Department, Karadeniz Technical University,

Trabzon, Turkey (KTUB).

Species recorded

Lecanora wrightiana Zahlbr., Catalogus Lichenum Univeralis 5: 600.1928 Fic 1

= Lecanora japonica Tuck., Sert. Lich. Trop. Labuan Singapore: 35. 1891,

nom. illegit., non Mill. Arg. 1879.

Thallus corticolous, to 1 cm diam., thin, continuous or rimose-areolate

to verruculose, slightly yellow-grey, tcream-white, to slightly greenish-grey,

epruinose. Soredia absent. Apothecia sessile to slightly constricted at the base,

circular to irregular, 0.2-0.7 mm diam., disc dark red-brown, bright red-brown

when wet, epruinose; margin smooth, concolorous with thallus, mostly cream-

white, entire. Cortex hyaline, gelatinous, with small crystals. Amphithecium

campestris-type with small crystals. Parathecium hyaline, without crystals,

15 um thick. Epihymenium glabrata-type, without crystals, red-brown,

egranulose. Paraphyses apically +reticulate and slightly thickened. Asci

cylindric-clavate, 12-spored. Ascospores ellipsoid, 13-15 x 7-8 um. Thallus

and margin of apothecium contains atranorin and chloroatranorin. K+ yellow,

C-, KC-, P + yellow.

SPECIMEN EXAMINED: TURKEY. Igdir: Tuztuca, Uckaya village, 39°58°13.11”N,

43°39°44.08”E, on Populus sp., 1456m, 14.06.2010, KTUB-2018.

REMARKS— By its campestris-type amphithecium, L. wrightiana is similar

to L. elapheia, which, however, contains eight spores per ascus and produces

triterpenoids.

Lecanora wrightiana occurs mainly on bark and more rarely on rocks in

mountain regions. Previously known from Japan and India, it seems to be

especially common in Japan (Miyawaki 1988, Guderley & Lumbsch 1999). New

to Turkey and the Middle East.

Rhizocarpon inimicum Poelt & Vézda, Herzogia 6(3-4): 471. 1984. Fig 2

Thallus to 3.5 mm in diam., develops its ascomata, disappears at maturity,

areolate and parasitic on Lecanora rupicola, black-grey, flat at first, soon with

massive curved areolae; areolae 0.5-1.5 mm diam., plane at first, later convex;

apothecia in small groups, 0.2-0.7(-1) mm; discs dark-brown; proper margin

Lecanora wrightiana and Rhizocarpon inimicum in Turkey ... 147

Fic. 1 (left). Lecanora wrightiana. Fic. 2 (right). Rhizocarpon inimicum. Scale: 1mm.

black, slightly + crenulate, curly-raised, +pruinose, areolate initially, superficial,

sessile, immarginate, later moderately marginate in rough, soon concave;

hypothecium irregularly, brown; hymenium 120-150 um high; epihymenium

black-brown; hypothecium brown; amphithecium discrete, black-brown,

cellular. Asci clavate, ascospores 19-26 x 10-14 um, ellipsoid, wavering in the

middle or constricted, grey-brown to black-brown, 1-septate at first, vertical to

crosswise septa later, 4-celled at maturity, photobiont trebouxioid, hypothallus

indistinct, pycnidia absent. Thallus K-, C-, KC-, P-. Medulla I + violet.

SPECIMENS EXAMINED: TURKEY. Igdir: CENTER, Zordag, 39°45°49.05”N, 43°53’20.13”E,

on Lecanora rupicola, 2400m, 13.06.2010, KTUB-2020.

REMARKS—Rhizocarpon species with dark brown or grey-green ascospores

placed in subg. Phaeothallus —R. advenulum, R. epispilum, R. inimicum,

R. santessonii and R. schedomyces— have 1-septate to submuriform spores

(Poelt & Hafellner 1983, Poelt & Vézda 1984), but R. santessonii has the

smallest ones (Poelt & Vézda 1981). Rhizocarpon epispilum does not have an

amyloid medulla or vegetative hyphae (Timdal 1986) and produces 2-septate

spores, while R. inimicum has 1-septate spores with vertical to crosswise septa.

Rhizocarpon advenulum, R. epispilum, and R. schedomyces grow on Pertusaria

spp.; R. santessonii grows on Tremolecia atrata; and R. inimicum grows on

Lecanora rupicola.

Rhizocarpon inimicum grows parasitically on Lecanora rupicola (Poelt and

Vézda 1984). Not many parasitic Rhizocarpon species have been reported in

Turkey, probably because little lichenological research has been performed. If

studies on the lichen flora of Turkey are continued, we hope that the number of

recorded Rhizocarpon species can be increased. Previously known from Spain

and Portugal. New to Turkey and Asia.

148 ... Yazici, Aptroot & Aslan

Acknowledgements

We are grateful to Dr. Paolo Giordani and Dr. Javier Etayo for revision of and helpful

comments on an earlier draft of this manuscript. This study was supported by TUBITAK

(108T566 coded project).

Literature cited

Akman Y. 1999. Climate and bioclimate (The methods of bioclimate and climate types of Turkey).

1*Edn., Kariyer Matbaacilik Ltd., Sti, Ankara.

Baytop A, Denizci R. 1963. Tirkiye’nin Flora ve Vejetasyonuna Genel Bir Bakis. Ege Univ. Fen Fak.

Monografiler Ser. 1, Ege Univ. Mat., izmir.

Breuss O, John V. 2004. New and interesting records of lichen from Turkey. Osterr. Z. Pilzk. 13:

281-294.

Candan M, Halici MG. 2009. Two new lichenicolous Arthonia species from Turkey. Mycotaxon

107: 209-213. http://dx.doi.org/10.5248/107.209

Candan M, Halici MG, Ozdemir-Tiirk A. 2010. New records of peltigericolous fungi from Turkey.

Mycotaxon 111: 149-153. http://dx.doi.org/10.5248/111.149

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MY COTAXON

http://dx.doi.org/10.5248/117.149

Volume 117, pp. 149-164 July-September 2011

A new volvate Macrolepiota (Agaricomycetes, Agaricales)

from Italy, with observations on the M. procera complex

ALFREDO VIZZINI’*, MARCO CONTU’, STEFANO GHIGNONE;,

& ELSE VELLINGA‘*

'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

° Istituto per la Protezione delle Piante, CNR Sezione di Torino,

Viale Mattioli 25, I-10125 Torino, Italy

* Department of Plant and Microbial Biology, 111 Koshland Hall #3102,

UC Berkeley, Berkeley CA 94720-3102, U.S.A.

* CORRESPONDENCE TO: alfredo. vizzini@unito.it

AxBstTRAcT — A new Macrolepiota taxon from Italy, M. rhodosperma var. velicopia, is

described and illustrated based on morphological and ITS rDNA data. It is characterized

by a well-developed volva and abundant, evident velar remnants on the pileus, a stipe with a

minutely squamulose covering, and very thick-walled elements in the pileipellis. A discussion

on its taxonomic position within Macrolepiota and notes on closely related taxa are provided.

DNA sequence analyses support the new taxon within the variability of M. fuliginosa sensu

Vellinga, a non-volvate taxon that differs from Barla’s original sense of M. fuliginosa. As Barla

did not indicate a holotype in his protologue of Lepiota procera var. fuliginosa and there are no

extant original herbarium specimens, Barla’s Fig. 5/PL. 9 (from Les champignons des Alpes

maritimes) is selected as a lectotype, and a recent collection from Liguria (Italy) is chosen as

the epitype. A recent collection from Sardinia is chosen as epitype for L. permixta. Finally,

M. fuliginosa and M. permixta are reduced in rank to forms of M. procera, based on their

morphology.

KEY worps — Agaricaceae, biodiversity, taxonomy, Volvolepiota

Introduction

Macrolepiota Singer is a genus in the Agaricaceae characterized by basidiomes

with a scaly pileus, a complex annulus, and a stipe with a smooth to granulose-

squamulose covering and by spores with a distinct germ pore covered by a

hyaline cap (Singer 1986). Species with a distinct volva were accommodated

in the genus Volvolepiota Singer, proposed in 1959 to replace the illegitimate

Lepiotella Rick (Rick 1938), a later homonym of Lepiotella (E.-J. Gilbert) Konrad.

150 ... Vizzini & al.

The type of Volvolepiota is the Argentinean species Lepiotella brunnea Rick.

Singer (1959) added a second species, V. albida Singer. In their monograph

on Volvolepiota Heinemann & de Meijer (1996) argued that the presence of

a well developed universal veil in this taxon was not sufficient to warrant a

genus separate from Macrolepiota but did not propose new combinations in

Macrolepiota.

Recent ITS-based molecular analyses by Vellinga (2003), Vellinga et al. (2003),

and Ge et al. (2010) place Volvolepiota species nested in the Macrolepiota clade

but in a basal position. Vellinga & Yang (2003), who considered Volvolepiota

a synonym of Macrolepiota, proposed new Macrolepiota combinations and

described M. velosa, a new volvate species from China. Ge et al. (2010) proposed

Macrolepiota sect. Volvatae Z.W. Ge et al. to accommodate the volvate species.

Four Macrolepiota taxa with a volva thus far known (from Australia, East Asia

and South America; Vellinga & Yang 2003) all have a tropical distribution and

relatively small spores.

Here we discuss our research on an Italian taxon with a copious universal

veil and well-developed volva to determine its position within the genus. In

the process we also studied M. fuliginosa, M. permixta, and the complex of M.

procera (Scop.) Singer.

Materials & methods

Morphology

Macromorphological features were described from fresh specimens. Colour notations

in the macroscopic descriptions are from Séguy (1936). Microscopical characters were

made from dried material rehydrated in 5% KOH and stained in Congo red, Cresyl

Blue, and Melzer’s reagent. Spore measurements are based on means of 30 spores

from two collections stained in Melzer’s reagent. Basidia were measured where widest

and from apex (sterigmata excluded) to basal septum. Author citations follow Index

Fungorum (http://www.indexfungorum.org/authorsoffungalnames.htm). Herbarium

abbreviations follow Thiers (2011). All examined material is housed at TO (Herbarium

generale del Dipartimento di Biologia Vegetale, Universita degli Studi di Torino, Italy).

The new Latin description is deposited in MycoBank (http://www.mycobank.org/).

The following abbreviations are used: Q = quotient of length and width of spores in

side view; Q_ = average quotient; L = number of entire lamellae; 1 = number of lamellulae

between each pair of entire lamellae; (Se) = Séguy (1936).

DNA extraction, PCR amplification, and DNA sequencing

Genomic DNA was isolated from 1 mg of herbarium specimens (TABLE 1) using

the DNeasy Plant Mini Kit (Qiagen, Milan Italy) according to the manufacturer's

instructions. Universal primers ITS1F/ITS4 (White et al. 1990; Gardes & Bruns 1993)

were used to amplify the ITS1+5.8S+ITS2 rDNA region between the small (18S gene)

and large (28S gene) subunits. Amplifications were performed in PE9700 thermal

cycler (Perkin-Elmer, Applied Biosystems) in a 25 ul reaction mixtures containing 5 ng

The Macrolepiota procera complex... 151

TABLE 1. Macrolepiota samples newly sequenced in this study.

! SPECIES ! CounTRY — ! COLLECTOR ! HABITAT ‘ITS

SAMPLE : i i : i

aia : ; LOCATION ; SAMPLING DATE; ; ACC. NO.

, i i i [herb. number] i

1 iM. fuliginosa Italy — i M. Contu : Coastal ? HM246501

: Sardinia, Olbia- : 15/X1/2009 _ pinewood

: Tempio, Rena Majore : [TOHG2006] _ :

2 M. fuliginosa Italy — : A. Vizzini : : Quercus ilex : : HM246502

: : Liguria, Savona, : 09/X/1998 litter :

: Borgio Verezzi : [TO AFM11]

3 M. permixta : Italy — M. Casula : Pinus sp. litter _ HM246503

i : Sardinia, M.S. : 07/XI1/2007 i i

EE Rel iis Solem as panera strait, suo AEM IZ] ne

4 M. rhodosperma i Italy — PA. Vizzini i Fagus sylvatica : : HM246505

: var. velicopia : Piedmont, Turin, : 07/X/2008 : litter :

: : Giaveno, Colletto : Te HG2003] :

: del Forno

5 M. rhodosperma Italy — : L. Latino ? Quercus robur _HM246506

: var. velicopia : Piedmont, Turin, : 15/X/2008 :&Q petraea :

: Venaria Reale, Parco :[TOHG2004]_ : litter

: Regionale La Mandria :

6 M. rhodosperma : Italy — i A. Vizzini i ? Quercus rubra /HIM246507

: Piedmont, Turin, : 20/X/2008 : & Corylus

Venaria Reale, Parco [TO HG2005] avellana litter

: Regionale La Mandria :

7 M. excoriata Italy — A. Vizzini Pasture HM246504

: (Schaeff.) Wasser — : Piedmont, Turin, : 16/VIII/2008 i i

i Colle del Frais [TO AFM13]

DNA, 1x PCR buffer (20 mM Tris/HCl pH 8.4, 50 mM KCl), 1 uM of each primer, 2.5

mM MgCl, 0.25 mM of each dNTP, and 0.5 unit of Taq polymerase (Promega). PCR

run times were 3 min at 95°C (1 cycle), 30 s at 94°C, 45 s at 50°C, 2 min at 72 °C (35

cycles), and 10 min at 72°C for 1 cycle. PCR products were resolved on a 1.0% agarose

gel and visualized by staining with ethidium bromide. PCR products were purified 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

(Gordon et al. 1998). The sequences have been deposited in GenBank (http://www.ncbi.

nlm.nih.gov/genbank/).

Sequence alignment and phylogenetic analysis

A representative set of the ITS1-5.8S-ITS2 regions of Eurasian and Australian

Macrolepiota taxa and two Leucoagaricus species was retrieved from Genbank. The

sequence alignment was performed by MAFFT version 6 (Katoh et al. 2002; Katoh &

Toh 2008) using default settings. After minimal visual fine-tuning of the alignment,

a final sequence set with a total 775 database length comprised 65 collections from

different studies.

A Bayesian Metropolis-coupled Markov chain Monte Carlo (BI) analysis was

conducted with MrBayes 3.1.2 (Huelsenbeck & Ronquist 2001). The model of nucleotide

substitution for the BI analysis was selected using ModelTest v. 3.7 (Posada & Crandall

1998). The transversional substitution model with gamma correction (TVM + G)

152 ... Vizzini & al.

was chosen as the best-fit model based on hierarchical likelihood tests, following the

Akaike Information Criterion (AIC). The general structure (lset) of the substitution

model was defined using nst=6 rates=gamma ngammacat=4;TVM + G model

was gained using the following parameters to define the priors: the four stationary

frequencies of the nucleotides (statefreqpr=fixed(0.2310, 0.2236, 0.2349, 0.3105)),

the six different nucleotide substitution rates (revmatpr=fixed(1.7916, 5.3304, 2.2627,

0.7366, 5.3304, 1.00)), the proportion of invariable sites (pinvarpr=fixed(0.00)), and the

shape (shapepr=fixed(0.3944)). Markov chains were run for 10° generations, with the

sampling frequency set every 100th generation; all other parameters were used at the

default settings. The outgroup Leucoagaricus nympharum was selected to root the tree.

A burnin value set to 25% of sampled trees was used to calculate the parameters and the

consensus tree.

The data set was also analyzed by a maximum likelihood (ML) method using

RAXML version 7.2.3 (Stamatakis et al. 2008). One hundred rapid ML bootstraps were

performed, using default settings. The phylogenetic trees were visualized and edited

with FigTree (http://tree.bio.ed.ac.uk/software/figtree). The alignment was deposited in

TreeBase (www.treebase.org) under study accession no. $10573.

Results & discussion

Phylogeny based on the ITS dataset

The ITS amplification products of the analysed samples ranged from 344

to 680 bp. A total of 65 sequences were used for the ITS phylogeny, including

the seven newly sequenced collections (Tas. 1) and all others retrieved from

GenBank (accession numbers are given in Fig. 3). The multiple sequence

alignment was 775 characters long in total.

The phylogenetic trees resulting from the two types of analysis do not

differ in topology. The Bayesian inference consensus tree (Fig. 3) indicates

that our volvate taxon clusters together with M. rhodosperma (P.D. Orton)

Migl. (AY243596) and with isolates formerly identified as M. fuliginosa, and

one accession under the name M. konradii (P.D. Orton) M.M. Moser. The two

sequences from our volvate Macrolepiota differ among themselves and from M.

fuliginosa and M. rhodosperma sequences in only1-2 bases in three different

combinations. This clade, supported by a 1.0 posterior probability value and

98% ML bootstrap value, is sister to all other species in sect. Macrolepiota,

except for M. clelandii Grgur.

ITS sequences of taxa identified as M. permixta and M. fuliginosa do not

differ from those of M. procera.

The group of M. mastoidea (Fr.) Singer, although homogeneous in sequences,

represents basidiomes with quite different morphologies, and these have been

accessed under a range of species names; an in-depth multigene analysis of this

complex is needed.

All three sections, sect. Macrolepiota, sect. Macrosporae (Singer) Bon, and

sect. Volvatae are recovered in the phylogenies, and well supported.

The Macrolepiota procera complex ... 153

d-f from TO HG2003). a. Mature basidiomes. b. Pileus surface with evident velar patches.

c. Flattened bulb and ornamented stipe. d. Young basidiomes covered with universal veil patches

and with well developed basal rhizomorphs. e-f. Submarginate bulb with free velar limbs (volva).

Scale bars: a-c = 5 cm; d-f = 2 cm.

Taxonomy

Macrolepiota rhodosperma var. velicopia Vizzini & Contu, var. nov. FIGs. 1-2

MycoBank MB518426

A typo differt velo albo membranoso ad instar Amanitarum in pileo et ad basim stipitis

manifesto, hyphis pilei cutis conspicue crassotunicatis, sporis minoribus, fibulisque haud

fraequentibus.

Ho.otypus — Italia, in regione Piemonte dicta, ad locum dictum Parco Regionale La

Mandria (Venaria Reale, Torino), 15.X.2008, leg. L. Latino (TO HG2004).

154 ... Vizzini & al.

EryMoLocy — the varietal epithet, derived from the Latin words velum, veli = veil,

velum; and copid, copiae = abundance, emphasizes the copious veil.

PILEUs up to 15 cm wide when expanded, at first parabolic then plano-convex,

with a low, wide, central umbo, dry, when young entirely covered with a

granulose velvety covering, later breaking into a central calotte, which may be

round to irregularly star-shaped, felted-subsquamulose, up to % of radius wide,

cream hazel (Se 134, 695) to light chestnut brown (Se 111, 131), surrounded

by 2-10 mm wide and easily removable lighter patches with curled edges,

on a radially coarsely floccose-fibrillose covering made up of chamois beige

to pale brown fibrillose strands on a white background; with white to dirty

white membranous velar remnants as patches on the surface; margin slightly

to distinctly fringed and exceeding lamellae. LAMELLAE, L = 80-140, | = 0-3,

free and remote from the stipe, with an evident collarium, moderately crowded,

up to 9 mm broad, at first whitish then with cream to pinkish tinges, with a

fimbriate-floccose concolorous edge. STIPE protruding into the pileus, (5-)

9-18(-22) x (0.5—)0.8-1.5(-1.8) cm, cylindrical to slightly tapering towards

the apex, hollow, bulbous at the base, with an oblong to flattened submarginate

bulb, (2-)2.5-3.2(-3.5) x 3-4 cm, often with long white rhizomorphs; with

adnate minute cream-hazel zigzagging bands over the whole length, on a white

background; bulb white tomentose with evident and membranous white velar

remnants on the margin. ANNULUS movable with age, complex, thickened with

a relatively broad edge, upperside creamy and fringed and underside hazel.

CONTEXT white, unchanging, both on handling and in ammonia vapour, up to

13 mm thick in the pileus; smell indistinct, and taste indistinct to mild. SpoRE-

PRINT whitish-cream.

SPORES (13—)13.5-15(-16.5) x 9-10.5(-11.2) um, on average 14.55 x 9.93 um,

Q=1.4-1.6,Q_ = 1.46, ellipsoid to oblong, ovoid, hyaline, smooth, thick-walled,

with walls easily swelling in 5% KOH, with an apical and central germ-pore

(1-1.5 um wide) and hyaline cap (callus) on it, with a small apiculus, dextrinoid,

cyanophilous, congophilous, with inner wall metachromatic in Cresyl Blue

(Fic. 2a); BASIDIA 25-42 x 9-16 um, 4-spored, some 1-2-spored, broadly clavate

(Fic. 2b); SUBHYMENIUM well-differentiated, cellular; HY MENOPHORAL TRAMA

subregular to slightly interwoven; PLEUROCYSTIDIA absent; LAMELLAR EDGE

sterile, consisting of tightly packed cheilocystidia; CHEILOCYSTIDIA 25-50(-60)

x 10-15 um, abundant, very variable in shape and size, narrowly clavate to

sub-fusiform, seldom also lageniform, utriform or cylindrical, sometimes

with a small capitulum, rarely with an apical excrescence, often catenulate

(pluricellular) septate in lower part, colourless and thin-walled, non-encrusted

(Fic. 2c); PILEUS COVERING (in calotte) a trichoderm of confusedly erect,

(4.5-)6-12 tum wide hyphae ending in a subfusiform (gradually tapering

toward the apex) terminal element with several secondary septa (pseudosepta),

The Macrolepiota procera complex... 155

FiGuRE 2. Macrolepiota rhodosperma var. velicopia. Microscopic features (from the holotype).

a. Spores. b. Basidia. c. Cheilocystidia. d. Pileus covering. d*. Pileus covering underlying elements.

e. Annulus elements. f. Velar elements. Scale bar: a—c = 20 um; d-f = 30 um.

156 ... Vizzini & al.

some very thick-walled (1.5-2 um thick), up to 250 um long, with a narrow

lumen, and then resembling the setiform terminal elements in the pileipellis

of Crinipellis species, other thin-walled and shorter (Fic. 2d); at the base of the

trichodermial elements there is a loose discontinuous layer made up of short

cylindrical to subcapitulate elements (Fic. 2d*). PIGMENTATION in the form

of brownish pigment, parietal and cytoplasmatic in the thick-walled hyphae,

parietal and minutely encrusting in the thin-walled hyphae, especially towards

their base. ANNULUS consisting of tightly packed, catenulate, articulate, and short

cylindroid hyphae, 4.5-9 um wide, with rare subglobular elements (Fic. 2e).

VELAR PATCHES made up of elongate, cylindrical hyphae, thin-walled, septate,

3-11 um wide, sometimes branched or nodulose, with scattered pseudoclamp

connections (proliferating clamps), intermixed with thick-walled hyphae (cell

wall up to 1.5 um thick), up to 10.5 um wide, with internal lumen, looking like

the crinipelloid elements present in the pileus covering, probably derived from

the underlying pileipellis (Fic. 2f). CLAMP CONNECTIONS rare, small, observed

at the base of the cheilocystidia; THROMBOPLEROUS HYPHAE absent.

HABITAT AND DISTRIBUTION Solitary, terrestrial, on litter of broad-leaved

trees, especially Fagaceae (Fagus and Quercus). So far known only from Italy.

ADDITIONAL MATERIAL STUDIED: Macrolepiota rhodosperma var. velicopia —ITALY,

PIEDMONT, TURIN, Giaveno, loc. Colletto del Forno, 7 Oct 2008, litter of Fagus sylvatica

L., 1000 m asl, leg. A. Vizzini (TO HG2003); M. rhodosperma var. rhodosperma — ITALY,

PIEDMONT, TuRIN, Venaria Reale, Parco Regionale La Mandria, 20 Oct 2008, litter of

Quercus rubra L. and Corylus avellana L., 230 m asl, leg. A. Vizzini (TO HG2005).

Macrolepiota rhodosperma var. velicopia was collected from two different

localities in northern Italy. A third collection turned out to be infected by

a hyphomycete species that covered the pileus and base of the stipe. Such

infections are very rare in the Agaricaceae.

Morphologically, M. rhodosperma var. velicopia is very different from the

other volvate Macrolepiota species because of such peculiar features as an

obviously decorated stipe, spores easily reaching 14-15 um in length, not strictly

cylindrical cheilocystidia, and long very thick-walled elements in the pileus

covering. This taxon is the first volvate Macrolepiota reported in Europe.

The South American M. pulchella de Meijer & Vellinga [= Volvolepiota

brunnea (Rick) Singer] and M. brunnescens Vellinga [= V. albida Singer]

differ in smaller spores that rarely reach 11 um in length, a stipe lacking any

decoration, and a less complex annulus; furthermore M. brunnescens has a

white context turning pink-brown after handling (Heinemann & de Meijer

1996). Macrolepiota eucharis Vellinga & Halling from northeast Australia has

a smaller pileus (< 6 cm broad when fully expanded), a pileus decorated with

tiny black scales, a simple ascending annulus that is white on both sides, a

uniformly fibrillose dark brown stipe, narrower spores, narrower cylindrical

cheilocystidia, and no clamp connections (Vellinga 2003).

AF482868 Leucoagaricus nympharum G

AF482865 Leucoagaricus leucothites NL

Country abbreviations:

AU - Australia

CHI - China

F - France

G - Germany

I - Italy

JAP - Japan

NL - the Netherlands

P - Portugal

SP - Spain

UK - United Kingdom 0.5

FiGurE 3. ITS-rDNA phylogeny inferred by Bayesian analysis. Bayesian posterior probability

values greater than 95%, derived from 10.000 Markov chain Monte Carlo sampled trees, are given

above branches; thick branches indicate >75% ML bootstrap values from the RAxML analysis. The

tree is rooted using Leucoagaricus nympharum as outgroup. Volvate Macrolepiota taxa are in bold.

* indicates samples sequenced in this work and reported in TABLE 1. Names appear as in GenBank

except for two recently described species from China (Ge et al. 2010). The bar indicates number of

substitutions per site.

The Macrolepiota procera complex ... 157

I DQI2TI11 dolichaula CHIN

/ i AF482839 dolichaula AU

AY 083193 dolichaula AU

1 AY 243586 detersa JAP

AF482851 detersa JAP

AY 243587 detersa JAP

HQ423285 procera SP

HQ423286 procera SP

HQ423287 procera SP

HQ412661 permixta I

AY 243590 procera F

0.9/7 HQ423289 procera SP

HQ412658 procera I

HM246501 fuliginosa 1*I

HM246502 fuliginosa 2*1

HM246503 permixta* I

HQ423291 procera NL

AF482848 procera NL

HQ423288 procera SP

HQ412657 procera |

AY243589 procera NL

HQ412663 rachodes 1

HQ423290 procera P

AY 243588 procera NL

AY 243591 aff. procera JAP

Sonne,

AY 243597 fuliginosa F

AY 243596 rhodosperma UK

HM246507 rhodosperma 6* 1

HQ423283 rhodosperma SP

HQ423284 rhodosperma SP

AF482841 fuliginosa NL

AY 243598 fuliginosa NL

AJ617494 konradii G

HQ423282 rhodosperma SP

AF482838 clelandii AU

4 AY083195 clelandii AU

1 AY 083203 clelandii AU

AY 083204 clelandii AU

foc)

sn LE SRS ND A IS ES MEN CTE IEEE ES LATO NIE SR

———

HQ412659 mastoidea 1

HQ412659 mastoidea I

U85314 gracilenta F

U85313 excoriata F

AY 243599 psammophila F

AY 243601 konradii NL

AY 243603 konradii NL

AY 243602 konradii NL

AY 243600 psammophila F

HQ412662 rickenii I

AY 243604 mastoidea NL

AY 243605 subsquarrosa 1

tp AY 243606 heimii UK

HM246504 excoriata* I

HQ412660 excoriata |

AF482840 excoriata NL

AY 243607 excoriata F

AF482850 orientiexcoriata CHI

A AF482847 phaeodisca F

atte RMS ANTES RPO IHL OR VET SAIS REISE IRT SEED

procera

HM246505 rhodosperma var. velicopia 4* 1

HM246506 rhodosperma var. velicopia 5* |

excoriata

incl. M. permixta

incl. M. fuliginosa

sect. Macrolepiota

pe OSS TR TT

rhodosperma

(fuliginosa

ss. Vellinga)

incl.

var. velicopia

satan

"ittetaae

=e ance RTS AD

“ey

sceiseimdhriieieieieeasstnarameteruecinimtan

sect. Vol vatae)

neon

mastoidea group \

TE TEE

sect. Macrosporae

158 ... Vizzini & al.

Macrolepiota velosa Vellinga & Zhu L. Yang, from south-western China and

northern Thailand, is characterized by a 7-9 cm wide pileus with dark brown to

purple scales, a finely fibrillose or squamulose stipe, a less complex annulus, very

small (8-10(-11) x 6-7 um) spores, longer and strictly cylindrical cheilocystidia

measuring 44-68 x 4.5-7.5 um, and the absence of clamp connections (Vellinga

& Yang 2003; Ge et al. 2010).

Among the poorly studied, insufficiently known taxa, “M. nordica var.

subvelata” Bon ad int. (Bon 1993) comes quite close to our taxon but differs in

a less decorated stipe and a different pileipellis structure comprising articulated

hyphae.

Our volvate taxon clusters with two Macrolepiota taxa that also exhibit thick-

walled elements in the pileus covering: M. fuliginosa sensu Vellinga (Vellinga

2001, Vellinga et al. 2003) and M. rhodosperma lacking such a volva and velar

remnants (Fic. 3); M. rhodosperma is considered a synonym of M. fuliginosa

sensu Vellinga (Vellinga 2001, Vellinga et al. 2003).

Our phylogenetic analyses (Fic. 3) clearly demonstrate that M. rhodosperma

var. velicopia is not closely related to the other known volvate taxa (M. eucharis,

M. velosa), which implies that velar structure acquisition or loss is a homoplastic

character that developed independently during the evolution of Macrolepiota,

which makes it unsuitable for a natural classification of these fungi and further

emphasizes the artificial nature of Volvolepiota.

Macrolepiota procera f. fuliginosa (Barla) Vizzini & Contu, comb. nov.

MycoBank MB518708

= Lepiota procera var. fuliginosa Barla, Champ. Alp. Marit.: 21, pl. 9 fig. 5, 1888.

= Leucocoprinus fuliginosus (Barla) Locq., Bull. Soc. Linn. Lyon 14: 92, 1945.

= Macrolepiota fuliginosa (Barla) Bon, Docum. Mycol. 7(27-28): 20, 1977.

= Macrolepiota procera var. fuliginosa (Barla) Bell & Lanzoni,

Beitr. Kenntn. Pilze Mitteleurop. 3: 190, 1987.

Type: FRANCE, ALpgEs-MariTiMEs, illustration in Les champignons des Alpes

maritimes (Barla 1888: pl. 9 fig. 5, iconotype, lectotype designated here). ITALY,

LicurIA, Savona, Borgio Verezzi, in Quercus ilex litter, 09/10/1998, leg. et det. A. Vizzini

(Fig. 5a; TO AFM11, epitype designated here).

Lepiota procera var. fuliginosa, described from Mediterranean France (Barla

1888), has been differently interpreted by subsequent authors due to the lack

of original material (Trimbach 1996). According to mycologists with a vast

Mediterranean experience (Bellu & Lanzoni 1987, Candusso & Lanzoni 1990,

Bon 1993, Migliozzi 1995), this taxon comes very close to Macrolepiota procera

from which it differs mainly in the darker tinged pileus and stipe, the hardly

disrupted stipe covering, and the browning context. These authors describe

the pileipellis of M. fuliginosa as a trichoderm made up of thin-walled to

only slightly thick-walled hyphae due to the presence of a parietal pigment.

The Macrolepiota procera complex ... 159

PL.9

wi ;

eur Wy NY Uf) Wl

KS sily D

UvsteiN ve

SNe grees,

i (3)

(

f .

7 ie Least AUTEN Scop 3 SF jraoreta, 14008 flip Nob

7 / Sf

Figure 4. Lepiota procera var. fuliginosa (lectotype, Barla 1888: pl. 9 fig. 5).

Other authors, including Vellinga (2001), Lange & Vellinga (2004), and Lange

(2008), described under the name “Macrolepiota fuliginosa” a different agaric,

characterised in part by a pileus with grey-brown, loose and easily removable

scales and a pileipellis comprising very thick-walled elements with brown

intracellular and encrusting pigments. This latter taxon is widespread and

known from Denmark southwards into Spain (see Fic. 3).

160 ... Vizzini & al.

FiGuRE 5. a. Macrolepiota procera f. fuliginosa (epitype, TO AFM11).

b. M. procera f. permixta (epitype, TO AFM12). Bars = 5 cm.

Our ITS sequence analysis, which includes two Mediterranean collections

(from Sardinia and Liguria, Italy; Tas. 1), shows that M. fuliginosa sensu

Vellinga differs from M. fuliginosa sensu auct. pl. (Fic. 3). We regard Lepiota

procera var. fuliginosa as a predominantly Mediterranean, xerophilous taxon

that is correctly delimited only when based upon Mediterranean records.

Consequently, we designate the original Figure 5/Pl. 9 included by Barla

(1888) in the protologue (Fic. 4) as its lectotype (iconotype) and select a recent

collection from Liguria - where the Mediterranean taxon is rather widespread

— as epitype. The basidiomata of the epitypical collection here selected are

identical, in gross characters, with those depicted by Barla (1888) and exhibit

microscopic features perfectly fitting the descriptions reported by many authors

(Bellu & Lanzoni 1987, Candusso & Lanzoni 1990, Bon 1993, Migliozzi 1995)

in the region. It is also clear from Fic. 3 that the ITS sequences do not separate

this taxon from M. procera. However, morphological comparisons do support

its recognition as a form of M. procera as noted by Bellu. & Lanzoni (1987),

Candusso & Lanzoni (1990), and Migliozzi (1995). Therefore we introduce the

necessary new combination.

The name Macrolepiota rhodosperma is available for M. fuliginosa sensu

Vellinga. Notes on the synonymy of the two are given by Vellinga (2001) and

Vellinga et al. (2003). A good picture is provided by Breitenbach & Kranzlin

(1995, fig. 250 as M. konradii). Macrolepiota rhodosperma var. rhodosperma,

which lacks the copious velar remnants of var. velicopia, also has longer spores

and a context that changes brown-red.

The Macrolepiota procera complex ... 161

Figure 6. Lepiota permixta (Barla 1888: pl. 11 figs 1-4).

Macrolepiota procera f. permixta (Barla) Vizzini & Contu, comb. nov.

MycoBank MB518709

= Lepiota permixta Barla, Bull. Soc. Myc. Fr. 2: 114, 1886.

= Leucocoprinus permixtus (Barla) Locq., Bull. Soc. Linn. Lyon 14: 92, 1945.

= Macrolepiota permixta (Barla) Pacioni, Micol. Ital. 8(3):13, 1979.

162 ... Vizzini & al.

= Macrolepiota procera var. permixta (Barla) Quadr., in Quadraccia

& Lunghini, Quad. Acc. Naz. Lincei 264: 110, 1990.

Type: FRANCE, ALpEs-MARITIMES, Bendejeun, 7/12/1893, Barla Myc.283 (NICE,

lectotype, designated in Trimbach 1996: 226). ITALY, Sardinia, M.S. Vittoria Esterzili,

in Pinus litter, 07/12/2007, leg. M. Casula, det. M. Contu (Fig. 5b; TO AFM12, epitype

designated here).

Barla (1886) described Lepiota permixta as a large long-stemmed species

characterised by a pileus that is “10-15” cm wide, “brun-cannelle-fauve au

centre, “brunatre” towards the margin, “a écailles plus ou moins apprimées,

fibrilleux, blanchatre et plus ou moins écorché vers la marge’, lamellae that are

yellowish-white or “charné-clair’, a very slender white stipe with “petites écailles

appliquées, irréguliéres, brunatres, écorché vers la marge’,a membranous

“fauve-brunatre” annulus, and a reddening context. He added that the described

basidiomes were collected in a submontane region and that the species was

rare. In the short notes accompanying the French description, Barla stated that

Lepiota permixta combined features from three species: the slender habit of

L. procera, the “écorché a la marge” pileus of L. excoriata, and the reddening

context of L. rachodes. The colour plate later published by Barla (1888) displays

basidiomes exhibiting the very same features as described in the protologue

and with an annulus that apparently differs from that typically described for

Macrolepiota procera complex (Fic. 6). Most mycologists, especially those in the

Mediterranean region, have treated Lepiota permixta as a species separate from

the M. procera complex, as supported by the clearly reddening context (e.g.,

Pacioni 1979 with a very good description of Sardinian material, Bon 1993,

Bellu & Lanzoni 1987, Pazmany, 1985). Others (Candusso & Lanzoni 1990,

Migliozzi 1995, 1997, Quadraccia & Lunghini 1990) consider M. permixta a

variety of M. procera, assuming that the presence or absence of reddening is not

sufficient to distinguish the two taxa at species level. Pazmany (1988) showed

the great macromorphological variability of M. procera by describing eight

infraspecific taxa. Since our selected L. permixta epitype and other collections

identified as M. permixta share nearly or completely identical ITS sequences

with M. procera and M. fuliginosa, we reduce M. permixta to a morphological

form of M. procera and propose a new taxonomic combination.

Acknowledgements

We would like to thank Prof. E. Grilli (Popoli, Italy) for improving the English text

and G. Eyssartier (Paris, France) and Prof. Fernando Esteve (Alcala de Henares, Spain)

for their pre-submission reviews. Our sincere thanks also to Dr. P.A. Moreau (Lille,

France) for providing helpful suggestions. SG benefited from a Compagnia di San Paolo

grant. Dr. Todd Osmundson and Lavinia Latino kindly provided us with pre-submission

Macrolepiota sequences and with M. rhodosperma var. velicopia pictures, respectively,

and ECV acknowledges funding from NSF grant DEB 0618293.

The Macrolepiota procera complex ... 163

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MY COTA XON

DOI: 10.5248/117.165

Volume 117, pp. 165-205 July-September 2011

Studies in Amanita (Amanitaceae) of Central America. 1.

Three new species from Costa Rica and Honduras

R.E. TuLLoss!a, R.E. HALLING> & G.M. MUELLERS

aP. O. Box 57, Roosevelt, New Jersey 08555-0057, USA

b The New York Botanical Garden, Bronx, New York 10458, USA

¢Chicago Botanic Garden, 1000 Lake Cook Road, Glencoe, Illinois 60022, USA

CORRESPONDENCE TO: ret@eticomm.net, rhalling@nybg.org,& gmueller@chicagobotanic. org

ABSTRACT—Amanita conara, A. costaricensis, and A. garabitoana are proposed as new

species. These taxa are added to twelve previously described species known from, or

reported here for the first time from, the region of study: A. advena, A. arocheae, A.

brunneolocularis, A. colombiana, A. eburnea, A. farinosa, A. flavoconia var. inquinata,

A. fuligineodisca, A. muscaria subsp. flavivolvata, A. polypyramis, A. sororcula, and A.

xylinivolva, Amanita flavoconia var. sinapicolor is proposed to be a taxonomic synonym

of A. flavoconia var. inquinata. An unusual species of Amanita subsection Vittadiniae is

given the code Amanita sp. HON] and treated only in a key to regional species of Ama-

nita section Lepidella. A gazetteer is provided for Costa Rican sites at which Amanita

species have been collected.

Key worps—Area de Conservacion Guanacaste, Cordillera Talamanca, Mexico,

Andean Colombia, taxonomy

Introduction and overview

This paper addresses the genus Amanita Pers. as a part of the extensive

study of macromycetes undertaken by Halling, Mueller, and numerous col-

leagues in Costa Rica.

The history of work investigating the diversity of Costa Rican macrofungi

[see discussions in (Halling & Franco-M. 1996; Halling & Mueller 1999,

2005; Baroni and Halling 2000; Ammiurati et al. 2007)] has been long, but

intermittent. Currently there is an emphasis on obtaining additional data on

the fungi of Costa Rica through the Costa Rican National Biodiversity Inven-

1. Research Associate (hons.), New York Botanical Garden, Bronx, NY.

166...Tulloss, Halling & Mueller

oe . 4 he

% A *

mY - a = =

— 4, - * i. 4

a o> js , ' i

—* i i aa

Fig. 1. Amanita garabitoana. a. Habit (Halling 8198) (<0.25*). b. Habit (holotype)

(<0.27). Photos by R.E. Halling (a) and R.E. Tulloss (b).

tory—a multinational project coordinated by the Costa Rican National Biodi-

versity Institute (INBio) (Mueller & Mata 2001). Work to date has

documented a highly diverse macrofungal community (Mueller & Halling

1995; Carranza & Mueller 1996; Halling & Mueller 1997, 2002; Mueller et al.

2006). To date our investigation of Costa Rican Amanitaceae indicates there

are at least 35 species of Amanita in the country; of these, a number will be

new to science.

Materials and Methods

Methods and terminology follow those of Tulloss et al. (1992) and expansions by

Tulloss (1993, 1994, 1998b, 2000a). A recent terminological summary appeared in

(Tulloss & Lindgren 2005) and is available on-line (Tulloss 201 1a).

Samples from selected exsiccata revised during research reported herein have been

supplied to three laboratories now carrying out phylogenetic studies related to Amanita

sections Caesareae and Lepidella.

Color codes of the form “6D6” are from (Kornerup & Wanscher 1978). Color

codes of the form “10YR 4/3” are from the Munsell soil color charts (Anonymous

1975). Color codes of the form created by the Inter-Society Color Council and the

U.S. National Bureau of Standards were translated to Munsell notation using (Kelly

1965; McKnight 1977). Color names with initial capital letters (e.g., Empire Yellow)

are from Ridgway (1912), and conversions from Ridgway’s names to Munsell color

codes is based on the work of Hamly (1949).

New Central American amanitas...167

The form of author citations follows Kirk & Ansell (1992, 2010).

“PN.” stands for “Parque Nacional.” The nonitalicized abbreviation “std.” stands

for “standard deviation.” With two exceptions (““HKAS”—Kunming Institute of Bot-

any, Academia Sinica, China; and “RET’—Tulloss’ personal herbarium), codes for

herbaria follow Index Herbariorum (Holmgren et al. 1990; Thiers 2010).

In the lists of collections examined, locality names in Costa Rica are provided in

abbreviated form. Known site data for these abbreviated names are provided in the

appendix to this paper.

Taxonomic part

Extensive collecting in Costa Rica in the last fifteen years has much

improved understanding of the genus Amanita in that country. We estimate

the number of species 1n the genus present in Costa Rica alone to be 35 to 40.

To date, 12 previously described taxa of Amanita have been collected and

determined or described as new from Costa Rica and Honduras. Previously

described taxa are provided in the following list—organized by section of the

genus. When range data in a list entry includes Colombia, the source is Tul-

loss et al. (1992).

Amanita sect. Amanita (three taxa):

¢ A. farinosa Schwein. — Costa Rica is the known southern limit of distri-

bution. It was described from North Carolina, USA; the range of this

mushroom extends northward into Prov. Québec, Canada (Pomerleau

1980).

¢ A. muscaria subsp. flavivolvata Singer — originally described from ca.

San Francisco, California, USA, this species now is known to have a

range extending north into the Alaskan panhandle and eastward across

Canada and the northern United States to Newfoundland in the north

and extending southward at least to Costa Rica. In the southeastern

USS. its distribution is unclear due to confusion with A. muscaria var.

persicina Dav. T. Jenkins (Jenkins 1977, 1986), which is endemic in

that region. In North America, a yellow-capped color-variant of subsp.

flavivolvata dominates in most of that part of the region of distribution

north and east of the Great Plains. The typical, red-capped color-vari-

ant dominates throughout the remainder of the range. In the neovolca-

nic region of Mexico, the subspecies is relatively common in mixed

montane forest; and is associated with Quercus in Costa Rica. If not

native in Colombia, it has been introduced there with Pinus or Quercus

from more northerly latitudes. The taxon is often misreported through-

out its range as the type subspecies or type variety of A. muscaria (L. :

Fr.) Lam.(Tulloss 2009c; Geml et al. 2008).

168...Tulloss, Halling & Mueller

° A. xylinivolva Tulloss, Ovrebo & Halling — This species was described

from Andean Colombia. Cochise Co., Arizona, is the known northern

distribution limit (Tulloss 201 1b).

Amanita sect. Caesareae Singer: no previously described taxa known.

Amanita sect. Vaginatae sensu Zhu L. Yang (Yang 1997) (three taxa):

¢ A. colombiana Tulloss, Ovrebo & Halling — Costa Rica is the known

northern limit of distribution. The southern limit is in Andean Colom-

bia.

° A. fuligineodisca Tulloss, Ovrebo & Halling — Honduras is the known

northern limit of distribution. The southern limit is in Andean Colom-

bia.

¢ A. sororcula Tulloss, Ovrebo & Halling — Costa Rica is the known

northern limit of distribution. The southern limit is in Andean Colom-

bia.

Amanita sect. Lepidella sensu Bas (1969) (two taxa):

¢ A. advena Tulloss, Ovrebo & Halling — Costa Rica is the known north-

ern limit of distribution. The southern limit is in Andean Colombia.

Previous to our work in Costa Rica, the only known collection of this

species was the Colombian holotype.

¢ A. polypyramis (Berk. & M. A. Curtis) Sacc. — Costa Rica 1s the south-

ern limit of known distribution. The known northern limit is in the

sandy Atlantic coastal plane of Cape Cod, Massachusetts (Tulloss

2009b). The species was described from North Carolina.

Amanita sect. Amidella (E.-J. Gilbert) Konrad & Maubl.: no previously

described taxa known.

Amanita sect. Phalloideae (Fr.) Quél. (two taxa):

¢ A. arocheae Tulloss, Ovrebo & Halling — No change in known range

(central Mexico to Andean Colombia).

¢ A. eburnea Tulloss (Tulloss 1989) — No change in known range (Belize

and Honduras).

Amanita sect. Validae (Fr.) Quél. (two taxa):

¢ A. brunneolocularis Tulloss, Ovrebo & Halling — This species was

originally described from Costa Rica and Colombia. It has now been

found under Pinus in North Carolina, USA, along a well-traveled high-

way (Tulloss 2009d).

¢ A. flavoconia var. inquinata Tulloss, Ovrebo & Halling — The known

northern limit of the range of this taxon is in the neovolcanic zone of

central Mexico. Field observation in Costa Rica has led us to conclude

that A. flavoconia var. sinapicolor Tulloss, Ovrebo & Halling com-

prises material of var. inquinata that has been exposed to direct sun-

New Central American amanitas...169

light at higher altitudes resulting in some desiccation and alteration of

pigment. As has been observed in other taxa of Amanita (e.g., Tulloss

& Borgen 1996; Tanghe & Hillhouse 1973), the desiccation also results

in production of smaller and more nearly globose spores. As previ-

ously observed, (Tulloss et al. 1992), there is a strong statistical corre-

lation between cap color and spore shape; but we now believe this was

wrongly interpreted as genetically determined by the authors of the

pale variety. Therefore, we propose reducing A. flavoconia var. sinapi-

color to synonymy with var. inquinata.

1. Section Caesareae

At present we believe that there are as many as 3 taxa of Amanita sect.

Caesareae in Costa Rica (Tulloss 2009e). These include no taxa known pre-

viously from Andean Colombia (Tulloss et al. 1992). In this paper we pro-

pose a new species easily distinguishable in the field, in Amanita stirps

Hemibapha (Tulloss 1998a, Tulloss 2009a)—A. garabitoana.

Amanita garabitoana Tulloss, Halling & G.M. Muell. sp. nov. Fics. 14

MyYcoBANK MB 518295

Fig. 2. Amanita garabitoana. Habit (Tulloss 6-16-95-D), (a. x0.38. b. x0.31.). Photos

by R.E. Tulloss.

170...Tulloss, Halling & Mueller

Pileus 60—212 mm latus, aurantiobrunneus vel olivaceoflavus vel flavobrunnea vel

aurantioflava, disco olivaceobrunneo vel aurantiobrunneo vel rubrobrunneo, saepe

zona brunnea circa ad extremum intimum striae marginalium, margine longistriato,

nonappendiculato. Lamellae subadnatae vel liberae, confertae, subroseocremeae vel

subaurantiocremeae vel pallidocremeae vel subalbae vel albae; lamellulae subtrun-

catae vel truncatae. Stipes 95—258 x 15—24 mm, subaurantioflavus vel pallidobuba-

linus vel subcremeus, bulbo nullo, annulatus, frustris coactis flavis vel sordidoflavis

vel olivaceoflavis vel brunneis vel aurantiis partis superae limbi interni veli univer-

salis ornatus; velum universali saccatum, robustum, membranosum, 48—96 x 30—45

mm, pagina externa alba, pagina interiori subaurantia vel pallidobrunnea. Fibulae

praevulgaris. Basidiosporae (7.5—) 8.0—-11.0 (—13.6) x (5.7—) 6.5—8.4 (-9.9) um,

(L = (8.3—) 8.7—10.0 (—10.3) um; L’ = 9.4 um; W = (6.6—) 6.9—7.7 (—7.8) um; W =

7.3 um; Q = (1.06—) 1.15—-1.43 (-1.73); Q = (1.22-) 1.24-1.37 (-1.40); Q’ = 1.29),

nonamyloideae, late ellipsoideae vel ellipsoideae. In quercetis tropicalibus montanis

costaricensibus et hondurensibus habitat. Amanita arkansana Amanitae stirpis

Hemibaphae simulanissima. Species nova ob distributionem geographicam, pileum

et stipem relative robustiores, ordinationem colores, colores variabiles ob aetatem

vel contusa, et magnitudinem sporarum.

HOLOTYPE — Costa Rica, Prov. San José, San Gerardo de Dota no. 1, Tulloss

6-21-95-G (USJ).

EtymMoLoGcy — In honor of Garabito, an indigenous military-political leader of the

Huetares, a people of the Central Valley of Costa Rica. Garabito resisted Spanish

occupation of Costa Rica until his capture in 1574. He fought to maintain the cultural

heritage of his people and to oppose both the mistreatment of indigenous peoples by

the Spanish and the establishment of Catholicism.

ILLUSTRATIONS: Rossman, et al. (1998: cover [Tulloss 6-16-95-D]); Tulloss (2000: 16

(fig. on right) [Tulloss 6-16-95-D]); Halling & Mueller (2005: 32 [Halling 8138],

173 [Halling 8438]).

PILEUS: 60-212 mm wide, orange-brown (e.g., 5C6-7) to olivaceous yellow or

yellowish brown (4B-C7) to olivaceous tan (more orange than 4B8) to orang-

ish yellow (4A6), darker over disc [e.g., olive brown (more olivaceous than

5D7-8) to orange-brown (6C8) to red-brown (6E6-7, 10F5)], often with dark

zone (brown to chestnut brown) at inner end of marginal striations, drying

dark brown, campanulate to strongly convex to convex to planoconvex with

slightly depressed disc, with large broadly subconic umbo, viscid to tacky to

dry (then subshiny), often dull, subglabrous to glabrous, silky fibrillose to

fibrillose to somewhat finely pruinose over disc; context white to pale sordid

white to pale yellowish white outside of disc, yellow (3A3) to pale yellow

under pileipellis and above stipe in disc or above lamellae, 4.5-12.5 mm thick

at stipe, thinning evenly for 75—85% of radius, then membranous to margin;

margin strongly striate to plicate-striate [(0.15R—) 0.5R-0.75R], incurved at

first, remaining at least somewhat decurved, rounded serrate to eroded, nonap-

pendiculate; universal veil absent.

New Central American amanitas...171

Rus a ss

David C. Tulloss del.

Fig. 3. Amanita garabitoana habit. a. Tulloss 6-16-95-D (x0.33). b. Tulloss 6-15-95-H

p.p. (<0.57). c. Tulloss 6-15-95-H p.p. (<0.39).

LAMELLAE: adnate to narrowly adnate to free, sometimes with short decurrent

line (rarely extending to partial veil, sometimes requiring 10x lens) on stipe,

172...Tulloss, Halling & Mueller

close to crowded, faintly pinkish cream to pale orangish cream to pale yellow-

ish cream to light cream to pale yellow to dull white to off-white in mass,

white to off-white to pale yellowish white in side view, 4.5-23 mm broad,

broadest at about 75% of radius from stipe, thin to moderately thick, with

entire and concolorous edge, some forked near margin, with occasional

reverse forking; /amellulae rounded truncate to subtruncate to truncate (less

frequently subattenuate to attenuate, but commonly so in one collection),

unevenly distributed, of diverse lengths, plentiful.

STIPE: 95-258 x 15-24 mm, with ground color light orangish yellow (ca. 4A5)

to pale yellow (3A3) to buff to yellowish white to off-white, yellow and pru-

inose at apex, unchanging or with ground taking on more orange tint or

becoming more sordid with age, cylindric or narrowing upward, barely or not

at all flaring at apex, below partial veil decorated with dry ochraceous to dull

brownish orange to light orange-brown (5C5) to yellow brown (ca. 4B7-8) to

yellow (2A3-4) floccose to fibrillose scales (becoming darker or more orange

from handling), below annulus minutely fibrillose or finely striatulate (espe-

cially with age); context white to pale yellow (2A2) to pale yellow-orange

(4A3), unchanging, with larval tunnels concolorous, stuffed with white glis-

tening fibrillose material moderately loosely packed, becoming hollow, with

central cylinder 3-10 mm wide; partial veil with upper surface light yellow to

slightly greenish yellow (3A4) to dull yellow (4A4-5) to sordid yellow (more

sordid than 4A8, more sordid than 4B4) to moderate yellow (not as green as

3A4), subsuperior to subapical to apical (e.g., attached for 5+ mm, with 3-15

mm free), membranous, thin, copious, skirt-like, persistent, eventually col-

lapsing on stipe and becoming sordid yellow, striate on upper surface, smooth

below; universal veil as saccate volva almost always connected at (or very

near to) base of stipe, often with considerable rather firm portion below stipe

base, 48-96 x 30-45 mm (often entirely below substrate surface), membra-

nous, soft, with outer surface white (sometimes with orange-brown discolor-

ation) and inner surface pale olivaceous tan to pale orangish white [may

become browner (paler than 1OYR 8/6) with age], with context white, 1.5—4+

mm thick, with limbus internus not always distinct or well preserved [when

present, largely white or largely concolorous with material decorating stipe, at

variable distance from point of attachment of volva and stipe, at first firmly

connected to sordid yellowish felted material (with latter eventually becoming

stipe-decorating squamules and patches)].

Odor mild, indistinct, or fungoid. Taste indistinct.

MACROCHEMICAL TESTS: Laccase test (syringaldazine): in material just matur-

ing, negative throughout basidiome. Tyrosinase test (paracresol): in material

New Central American amanitas...173

pe errr teres ene

‘

Fig. 4. Amanita garabitoana. a-d.Variation of stipe base attachment to universal veil and

variation of limbus internus (a. Tulloss 6-16-95-C. b. Tulloss 6-16-95-K. c. holotype. d.

Tulloss 6-15-95-H). e. Elements of hymenium and subhymenial tree (isotype). f. Ele-

ments of partial veil interior (Schmit 475). g. Elements of partial veil viewed from

underside (isotype). h. Basidium with clamp (Tulloss 6-16-95-C). Scale bars = 10 um.

just maturing, positive in cap context, pileipellis, lower half of stipe context,

universal veil except for very base of volva, partial veil, and on edges (includ-

ing breaks and cuts) of lamellae. Test voucher: Tulloss 6-21-95-G.

174...Tulloss, Halling & Mueller

PILEIPELLIS: 75-100 um thick; subpellis yellow-orange, 65-90 um thick;

suprapellis minimal, pallidly concolorous to colorless, extensively gelati-

nized, 10* um thick, intimately connected to collapsed hyphal detritus of uni-

versal veil (see below); filamentous undifferentiated hyphae 1.0-5.0 um wide,

branching, dominating, subradially oriented, densely packed vertically, some-

times with hyphal tips expanded slightly at apex; vascular hyphae 1.4-7.1 um

wide, common, sinuous, infrequently branching, without dominant orienta-

tion. PILEUS CONTEXT: filamentous undifferentiated hyphae 0.9-8.5 um wide,

plentiful, branching, frequently fasciculate, forming matrix loosely to densely

interwoven around acrophysalides; acrophysalides of two forms, away from

stipe apex plentiful narrowly clavate to elongate (e.g., 152 x 28 um, 95 x 42

um), above stipe apex dominating and densely packed with longitudinal ori-

entation like stipe context acrophysalides and ovoid to elongate ovoid to sub-

pyriform (up to 74 x 46 um or larger), in umbo greatly reduced in number;

vascular hyphae 2.5-17.8 um wide, branching (especially frequently in region

above stipe in disc), sinuous to looping and interweaving in loose “knots,”

common locally, especially common near pileipellis and in region above stipe

in disc; clamps plentiful. LAMELLA TRAMA: bilateral, divergent; we, = 25-45

um; subhymenial base dominated by curved intercalary inflated cells (up to

94 x 22 um) sometimes as concatenated pair (together up to 111 um long) and

then arising from filamentous undifferentiated hyphae arising in central stra-

tum, otherwise arising from short partially inflated clavate segment in manner

of chained pair, giving rise to cells of subhymenium; central stratum contain-

ing intercalary narrowly fusiform cells (e.g., 62 <x 14.0 um); filamentous

undifferentiated hyphae 3.24.9 um wide, with those in subhymenial base

sometimes giving rise to cells of subhymenium; vascular hyphae 2.0-11.3 um

wide, occasionally branching, often sinuous, uncommon to locally common;

clamps common in central stratum. SUBHYMENIUM: weg-near = 105-130 um

(crushed?); wgy-near = 95-105 (very good rehydration, estimated from mea-

surement and partially schematic drawing); wg-far = 150 um (crushed?);

We-far = 100-115 um (very good rehydration, estimated from measured data

and schematic drawing); cellular (pseudoparenchymatous) or dominated by

inflated cells, with cells in 2 to 3 layers (1 to 2 layers below longest basidia),

with some uninflated branched or unbranched elements arising from inflated

cells of subhymenium and giving rise in turn to basidia/-oles, otherwise with

basidia arising from inflated cells up to 12.0 x 10.0 um. BASIDIA: 29-56 (-59)

x (8.1—) 8.5-13.0 um, dominantly 4-sterigmate, rarely 5-sterigmate in imma-

ture material, with sterigmata up to 5.5 <x 2.0 um; clamps and proliferated

clamps plentiful, prominent. UNIVERSAL VEIL: On pileus: at least in moist con-

ditions persisting to maturity in thin layer (25— 45 um thick) and sometimes

New Central American amanitas...175

giving hoary appearance to pileus, at 1250 in cross-section giving appear-

ance of somewhat sparse “curly hair”; filamentous undifferentiated hyphae

dominant, collapsed, partially gelatinized, mostly under 2.0 um wide, curling

or coiling, branching, without dominant horizontal orientation; inflated cells

very infrequent, soon collapsed and gelatinized; vascular hyphae without

dominant orientation, of width similar to other hyphae, moderately frequent,

scattered. On stipe base, exterior surface: partially gelatinized, in a relatively

shallow layer, interior visible through occasional gaps; filamentous undiffer-

entiated hyphae 1.7-4.2 um wide, in fascicles up to 12 or more hyphae wide

or singly, densely criss-crossed and interwoven, with many larger fascicles

longitudinally oriented, with occasional openings giving (on whole) appear-

ance of openings in expanded net shopping bag. On stipe base, interior:

rather dense lattice-like structure of plentiful interwoven filamentous undiffer-

entiated hyphae, enclosing globose to ellipsoid to clavate cells (up to 66 x 38

um), with such cells plentiful to locally dominating in regions somewhat dis-

tant from exterior surface layer, enclosing smaller less frequent often clavate

cells near exterior surface layer. On stipe base, inner surface: \ike interior but

broken and gelatinized, indicating gelatinization takes place within universal

veil or at interface between universal veil and pileipellis. STIPE CONTEXT: lon-

gitudinally acrophysalidic; filamentous undifferentiated hyphae 2.0-7.0 um

wide, branching, rather common away from surface, becoming dominant and

strongly longitudinally oriented toward outer surface, at which forming dense

pellis-like layer; acrophysalides up to 182 <x 24 um, smaller near exterior sur-

face, with walls thin or up to 0.7 um thick, with some near surface having yel-

lowish walls; vascular hyphae 2.1-10.1 um wide, sometimes sinuous,

infrequent, unevenly distributed. PARTIAL VEIL: underside strongly gelati-

nized; upper side bearing plentiful remnants of inflated cells from former

interface to lamellae edges; filamentous undifferentiated hyphae 1.5-4.5 um

wide, branching, dominant in interior, dominantly subradially oriented, domi-

nantly fasciculate (with fascicles mostly 4 to 5 hyphae wide); inflated cells of

interior common, clavate, to elongate ellipsoid, thin-walled, up to 28 x 17.5

um, terminal, solitary, often with subradial orientation of longer axis,

unevenly distributed, occasionally in small clusters; vascular hyphae not

observed. LAMELLA EDGE TISSUE: Sterile.

BASIDIOSPORES: [558/27/16] (7.5—) 8.0-11.0 (-13.6) x (5.7-) 6.5-8.4 (-9.9)

um, (L = (8.3-) 8.7-10.0 (-10.3) um; L’ = 9.4 um; W = (6.6-) 6.9-7.7 (-7.8)

um; W’ = 7.3 um; Q = (1.06-) 1.15-1.43 (-1.73); Q = (1.22-) 1.24-1.37

(-1.40); Q’ = 1.29), hyaline, colorless, thin-walled, smooth, tnamyloid,

broadly ellipsoid to ellipsoid, rarely elongate, usually at least somewhat adax-

ially flattened, wand-like or very narrowly clavate in early development;

176...Tulloss, Halling & Mueller

apiculus sublateral (or rarely lateral in immature material), cylindric; contents

monoguttulate to multiguttulate, with or without small additional granules;

white in deposit.

EcoLocy: Costa Rica: Solitary to subgregarious, at 1000-2500 m elev. In

mixed forest with, and sometimes dominated by, Quercus (including Q. bren-

esii, O. copeyensis, QO. oocarpa, and Q. seemannii), with or without substan-

tial understory. Honduras: In undisturbed Quercus forest.

MATERIAL EXAMINED: COSTA RICA: ALAJUELA—Bosque del Nifio, 31.v.1996

R.E. Halling & J.L. Mata [Halling 7591] (NY; USJ). CARTAGo—Estrella,

28.vii1.1992 B.A. Strack & G.M. Mueller [Mueller 4433] (F 1102486), 4.vi.1996

R.E. Halling & J. Ammirati [Halling 7603] (NY; USJ); Tapanti, 20.vi.2001 R.E.

Halling & J. Carranza [Halling 8198] (NY; USJ). GUANACASTE—Cacao no. 2,

4.vi.1994 J.P. Schmit 475 (F; USJ). SAN Jost—La Chonta, 11.vii.1982 L.D.

Gomez 18172 (F 1100529), 7.vi.1994 R.E. Halling & T.J. Baroni [Halling 7237]

(NY; USJ), 16.vi.1995 K. Shanks, R.E. Halling, R.E. Tulloss & R.H. Petersen [Tul-

loss 6-16-95-C] (RET 333-6; USJ), R-.E. Tulloss 6-16-95-D (RET 332-7; USJ),

R.E. Halling s.n. [Tulloss 6-16-95-K] (RET 337-1; USJ), 11.vi.2001 R.E. Halling,

B. Buyck, R. Aldana-Gomez [Halling 8178] (NY;USJ); El Empalme, 15.vi.1995

R.E. Halling, R.H. Petersen, K. Shanks, R.E. Tulloss [Tulloss 6-15-95-H] (RET

333-2; USJ); Jardin de Dota, 13.vii.1993 G.M. Mueller 4501 (F 1110801); San

Gerardo de Dota no. 1, 8.vi.1994 R.E. Halling s.n. [G.M. Mueller 4659] (F

1112080; USJ), 21.vi.1995 R.E. Tulloss 6-21-95-G (holotype, USJ; isotype, NY),

24.v1.1997 R.E. Halling, G.M. Mueller, S. Huhndorf & D. Quist [Halling 7737]

(NY; USJ). HONDURAS: FRANCISCO MORAZAN—Tegucigalpa — P. N. La

Tigre, Sendero Bosque Nublado, 4.vii.1991 G.M. Mueller, B.A. Strack, R. & M.

Singer & R. Andino [Mueller 4119] (F 1098650).

COMMENTS — The (a) relatively shallow subhymenium; (b) subhymenial base

dominated by elongate, curved, intercalary cells; (c) plentiful clamps at bases

of basidia; (d) habit; (e) proportionately long marginal striations; (f) strongly

pigmented pileus and stipe decoration; and (g) broadly ellipsoid to ellipsoid

spores indicate that this entity has strong affinity to the group of taxa pheneti-

cally similar to A. hemibapha (Berk. & Broome) Sacc. (Saccardo 1887), A.

Jacksonii Pomerl. (Pomerleau 1984), A. arkansana H.R. Rosen (Rosen 1926),

A. hayalyuy Arora & Shepard (Shepard et al. 2008), etc —taxa of Amanita

stirps Hemibapha (Tulloss 1998a, 2009a).

The species of the above cited group that is most similar to A. garabitoana in

habit and in size and shape of spores is A. arkansana the known range of

which lies within the southeastern United States (Arkansas to the Gulf Coast

states—from Texas to Florida). The present taxon differs from A. arkansana

in at least the following:

¢ Pigmentation: The North American species lacks olivaceous tints in the

pileus at all stages of development and is not as deeply pigmented as 1s

New Central American amanitas...177

the red-brown disc of A. garabitoana, moreover, A. arkansana retains

its yellow-orange coloration as dried rather than becoming dark brown

to sordid or olivaceous tan. In addition, the patches on the stipe of A.

arkansana are concolorous with the stipe (yellow) at first and become

deeper yellow or more orange from handling; they are never olivaceous

or sordid yellow.

Fragile outer part of pileus: The basidiome of A. arkansana is notori-

ously fragile and difficult to collect without breaking, which is not the

case for A. garabitoana. The portion of pileus context with greater

thickness than a membrane extends half or less of the radius toward the

margin in examined material of A. arkansana, but 75% to 85% of the

radius in A. garabitoana.

Fragility of the stipe: The ratio of the diameter of the central cylinder to

the diameter of the stipe (both measured at midstipe) in specimens of

the two taxa are 0.5-0.56 for A. arkansana and 0.21-0.42 (-0.52) for A.

garabitoana. Hence, the context forms a supporting wall with thick-

ness of 22%-25% of the stipe diameter in the northern species and with

(24%—) 29%-40% of the stipe diameter in the mesoamerican species.

Tyrosinase spot test: In contrast to the strong positive reaction for

tyrosinase in A. garabitoana, that reaction is very limited in most of the

context of A. arkansana. In immature material of A. arkansana, using

paracresol, a positive reaction for tyrosinase was only seen in the limb

of the volva and in five, small and widely scattered spots on the stipe

and pileus context after 18 min. Another test with more mature mate-

rial, using L-tyrosine (only on pileipellis, stipe surface, and stipe con-

text), produced very faint positive reaction on the stipe surface and

pileipellis after 19 min. Voucher specimens: Tulloss 10-26-85-A and

7-16-87-C.

Spore size: Spores of A. arkansana are somewhat smaller than those of

A. garabitoana: [290/13/7] (7.0-) 7.7-10.5 (-15.0) x (5.6—) 6.0-8.0

(-10.2) um, (L = (8.0—) 8.3-9.9 um; L’ = 9.0 um; W = (6.3-) 6.5-7.3

um; W’ = 6.9 um; Q = (1.10-) 1.19-1.43 (-1.70); Q@= (1.22-) 1.24-

1.38; Q’= 1.30).

MATERIAL EXAMINED (Amanita arkansana): U. S. A.: ARKANSAS—Pulaski Co.

— Little Rock, Arkansas Dept. of Pollution Control & Ecology, 14.vi.1994 J. Justice

sn. (RET 136-6). Washington[?] Co. — E of Fayetteville, E of Mt. Sequoia,

13.x.1925 [packet marked “13.x.1926” (sic)] H.R. Rosen s.n. (holotype!, BPI; iso-

types TENN 21294 & 21299). FLORIDA—Alachua Co. — Gainesville, across

from Florida St. Mus., 11.viii.1985 A. Norarevian s.n. [Tulloss 8-11-85-AN1] (RET

137-5). MtIsstsstppt—Jackson Co. — Pascagoula R. Wildlife Mgt. Area,

16.vill.1987 D.C. & R.E. Tulloss 7-16-87-C (in herb. David T. Jenkins, Univ. Ala.,

Birmingham; RET 149-5). M1ssourt—Stoddard Co. — Mingo Nat. Wildlife

178...Tulloss, Halling & Mueller

Refuge, 19.1x.2008 J. Justice s.n. (RET 450-8). TExAs—Hardin or Tyler Co. —

Big Thicket Nat. Preserve, Turkey Crk. Unit, 26.x.1985 A. Norarevian & J. Justice

s.n. [Tulloss 10-26-85-A] (RET 139-10); Tyler Co. — 8 km E of Spurger, Forest

Lake Exp. For., off rd. FM1013, ca. plots 39 & 41, 26.vi.1994 D.P. Lewis 5302

(RET 283-5).

So far as is known (C. Bas pers. corresp.; Tulloss unpub. data), it is not

unusual to find vascular hyphae especially plentiful in the pileus context

above the stipe in Amanita. This is an item worthy of further study. Bas

(1969 and pers. corresp.) has noted that vascular hyphae are sometimes con-

centrated in damaged areas in Amanita. Since they arise from filamentous

undifferentiated hyphae (Tulloss unpub. data), 1t may be the case that they are

produced in response to damage or in areas of mechanical stress (such as the

stipe-pileus convergence region). One hypothesis might be that the (par-

tially?) insoluble material often seen when a vascular hypha is cut or broken

during sectioning may include an antibiotic or some other aid to maintaining

the integrity of the basidiome and, hence, reproduction of the species.

We also wish to make it clear that we make no claim for novelty with regard to

the arrangement of acrophysalides in the center of the pileus. However, it

does seem of interest to examine (in the future) the mechanical structure(s) by

which the joining of the stipe and cap takes place in Amanita.

Mueller 4501 was immature when dried; in measuring spores from a single

mount, three 5-sterigmate basidia were noted with the sterigmata bearing con-

tent-less, wand-like or very narrowly clavate spores. The single basidiome of

this collection is considered by us to be bearing abnormal spores.

Mueller 4119 was immature when dried.

2. Section Lepidella

2.1. Key to section Lepidella in Costa Rica and Honduras — This key treats only

those subsections of section Lepidella represented in the region of study. Spe-

cies marked by an asterisk (*) are not treated at length in this paper.

1. Pileus pinkish buff; universal veil on pileus in tightly appressed patches, white to

concolorous, comprising outer layer of interwoven filamentous undifferentiated

hyphae and interior dominated by chains of narrow, relatively large, inflated cells

with periclinal orientation, pileipellis poorly developed; spores (7.8—) 8.2—10.0

1. While there are three collections representing parts of the original collection, none is marked

as the holotype. The place of deposit of the holotype was not provided in the protolog. BPI was

indicated as the location of the “type” by Gilbert (1940-41: 98, caption for Tab. IX, fig. 2). This

is fortunate because the portion of the type in BPI has the most thorough and abundant representa-

tion of all parts of the included basidiomes of any of the three duplicates of the original collection.

New Central American amanitas...179

(—10.5) x (5.5—) 6.0-7.0 (—8.0) um, with Q = 1.34—1.39 (Amanita subsect. Vittadin-

LAE BIS) Ser eee tasks Pee ako boars onan borer Mee aay a hecasa taken denne haweendemaleccdece ead sek sient

A. species HON1*.

1. Universal veil as warts or irregular patches or as pyramidal warts; elements compris-

ing these warts disordered or having more or less anticlinal orientation, never as

chains of relatively large, narrow, inflated cells with strong periclinal orientation.

2. Outer layer of volva consisting mainly of hyphae; consequently, often with small,

incomplete volval limb on stipe’s basal bulb; lamellae ochraceous to yellow to pale

orange. (Amanita subsect. Gymnopodde Bas)..............c60cccccccctecc cee ceeeseeeeeeceeecneeees

3. Spores (7.8—) 8.1—10.0 (-11.5) x (5.3—) 6.0-7.5 (-8.7) um, with Q = 1.33-1.41

(—1.44); found at 1600-1720 m elev. in Prov. Cartago with Quercus oocarpa and

PISS CWT IN ada serscs det sa 7 ott cat Seth a se eas etd nea te Tost bee deatsh sul ade an SoltE mene st Adee ty

A. conara.

3. Spores (6.1—) 7.0-9.5 (-11.6) x (5.1—) 6.9-7.9 (-9.5) um, with Q = (1.11-)

1.15—1.23; found at 1350-1500 m elev. in Prov. Puntarenas with Q. corrugata

AMG) SCCM ITE. A NW NO EE ES OR Le ee Ne

A. species CR18*.

2. Volva never submembranous, never forming limb at base of stipe; lamellae never

ochraceous (Amanita subsect. Solitariae Bas).

4. Pileus rat gray to brown, micaceous gray to pallid in age or after rains wash

away pigment, with brownish gray pulverulent universal veil remains in pyrami-

dal warts or irregular patches, often darker than pileipellis; bulb of stipe napi-

form to radicating, often markedly radicating; spores (7.5—) 8.5—13.2 (—16.0) x

(5.1—) 6.0-8.5 (—12.6) um, with Q = 1.35—1.68 (-1.70) ooo. ceeeeees

A. costaricensis.

4. Pileus and universal veil white, at least at first.

5. Much of basidiome staining ochraceous buff to rust colored to chestnut brown

or yellow to cinnamon; universal veil on pileus as flattened to slightly erum-

pent large warts or small patches; all parts of basidiome liable to staining pale

yellowish and, eventually, cinnamon; bulb of stipe napiform, never abrupt or

subabrupt, decorated with narrow rings of volval tissue beginning on lower

stipe above bulb and continuing to approximately broadest part of bulb; spores

(8.5—) 9.0-12.5 (14.0) x (6.5—) 7.0-9.5 (-10.2) um, with Q = 1.29-1.32 .......

A. advena*.

5. Pileus not staining, often covered rather densely by minute pyramidal warts;

bulb slenderly subnapiform to napiform, subradicating, with either rings of

flocculence or minute pyramidal warts around top of bulb; spores (7.0—) 9.1—

13.0 (-17.5) x (5.2—) 5.9-7.8 (9.5) um, with Q = (1.35—) 1.50-1.81 oo...

A. polypyramis*.

180...Tulloss, Halling & Mueller

2.2. Descriptions of taxa —

Amanita conara Tulloss & Halling sp. nov. Fics. 5-8

MycoBAnk MB 518296

Pileus 60-150 mm latus, subochraceobubalinus vel subbubalinus vel aurantiocre-

meus vel griseoaurantius, labibus subaurantiis vel subbrunneis, margine longistri-

ato, nonappendiculato, reliquiis veli universalis verrucas vel assumenta formaniti.

Lamellae liberae, subconfertae vel confertae, subcremeae vel subaurantio-flavobub-

alinae; lamellulae subattenuatae vel rotundato-truncatae vel truncatae. Stipes 7\—

115+ x 11-—30+ mm, albus vel subroseus vel subbrunneoroseus, annulatus, bulbo

robusto 42—60 x 37-60 mm, interdum velo universali membranaceo pertenui super

bulbum. Basidiosporae (7.8—) 8.1—10.0 (—11.5) x (5.3—) 6.0-7.5 (—8.7) um, (L =

(8.6—) 8.8-9.4 um; L’ = 9.1 um; W = 6.5-6.8 (—7.0) um; W’ = 6.6 um; Q = (1.17-)

1.23—1.51 (—1.70); Q = 1.33-1.41 (—1.44); Q’ = 1.37), amyloideae, late ellipsoideae

vel ellipsoideae. Basidiae 30-58 x 9.1-13 um. Fibulae relative communis. In quer-

cetis tropicalibus montanis costaricensibus habitat. Species Amanitae subsectionis

Gymnopodae simulanissima. Species nova ob distributionem geographicam, magni-

tudinem et formam sporarum, consortionem Querco, colorem et formam et anato-

miam microscopicam veli universalis et limbi interni, et absentiam rhizoidei bulbo

Stipitis.

HoLotyPe — Costa Rica, Prov. Cartago, Estrella, 14.vi.1996 R.E. Halling & J.

Ammurati [Halling 7686] (USJ).

ETYMOLOGY — Kovapoc (Grk.), fat or well-fed, because of the large basal bulb of the

present species; proposed English name, “Fat Barefoot Lepidella.”

PILEUS: 60-150 mm wide, light ochraceous buff (ca. 5A2) with light orange

stains (ca. 6A4) or pale buff to faintly tannish or orangish cream (paler and

more tan than 7.5YR 8/4, paler than 1OYR 8/4) or grayish orange (6B4), occa-

sionally stained faintly brown here or there, convex at first, planoconvex and

somewhat undulate at maturity, not viscid when wet, dull at first, later shiny

when dry; context white to cream to slightly brownish cream, context

unchanging or slowly staining brown to reddish brown when cut or bruised,

7-15 mm thick at stipe, thinning evenly and slowly to margin, rather thick

even near margin; margin nonstriate, decurved, appendiculate (at first with

appendiculate material floccose and pale pinkish, at maturity only with scat-

tered crumb-like bits); universal veil as crumb-like warts or pyramidal warts

over disc, sometimes as eccentric patches, at first pale orangish white to pink-

ish (SA2, paler toward pileus margin) becoming sordid or brown or taking on

brownish tint, smooth surfaced, often easily taking fingerprint, pulverulent to

subfelted, detersile, with patches having membranous upper layer.

LAMELLAE: free, with decurrent lines on stipe apex connecting to striations on

upper surface of partial veil, close to subcrowded to crowded, pale cream

(immature) to slightly orangish yellowish tan in mass, more vividly colored in

New Central American amanitas...181

. aa

pes x 5) a Sa wes

Fig. 5. Amanita conara. Habit (Halling 7272) (<0.45). Photo by R E. Halling.

side view [light cream in immature material, then more yellow than 2.5Y 7/6

or more brown than 2.5Y 8/6 or more orange than 4B6 or more brown than

4A6 or pale yellow (4A4) or pale orange (5A3)], unchanging when cut or

bruised, 6.5—12 mm broad, occasionally forked, with edge minutely pulveru-

lent and white to approximately concolorous with partial veil; /amellulae

subattenuate to rounded attenuate to truncate, unevenly distributed, of diverse

lengths, plentiful.

STIPE: 71—115+ x 11-—30* mm, white, sometimes with ochraceous to light

orange stains, sometimes faintly brick colored or pale pink after handling or in

wounds, narrowing upward, flaring at apex, pulverulent above partial veil,

sometimes satiny below partial veil, longitudinally striatulate, with upward

pointing fibrillose scales or squamules [concolorous to light orange (ca. 6A4),

with tips sometimes faintly brick colored or concolorous with partial veil and

limbus internus of universal veil], with shaggy region beginning below annu-

lus and ending roughly 10 mm above bulb; bu/b 42-60 x 37-60 mm, robustly

clavate-subnapiform to robustly napiform, with white mycelial threads promi-

nent at very base; context solid or stuffed? (possibly appearing so due to larval

damage), concolorous with pileus context, with bruising/staining reaction as

in pileus context, with larval tunnels yellowish in center of stipe and reddish

brown elsewhere; partial veil apical, submembranous, shredding, sometimes

deciduous at least in part, pale orangish white (5A2), with underside bearing

182...Tulloss, Halling & Mueller

Fig. 6. Amanita conara. Habit (Tulloss 6-13-95-C) (<0.74). Photo by R.E. Tulloss.

warts and flocculence, with upper side striate, sometimes thickened at edge;

universal veil often present as short thin limb and/or in incomplete and some-

times slightly overlapping rows of triangular flaps (minute, weakly structured,

upward-pointing) 0.5 mm high on (or appressed to) upper part of bulb, with

edges of such structures sometimes appearing frayed and darkening with age

(viewed at 6—12%), often with nearly complete limbus internus partially encir-

cling (about half-circumference) base of stipe and in other small fragments on

lower stipe above bulb, 1-2 mm thick, nearly white to pale orangish white

(5A2).

Odor “earthy” in immature specimen; rich, strong, like “old ham” (but some-

times not unpleasantly) or like “unwashed athletic socks” in mature material.

Taste not recorded.

MACROCHEMICAL TESTS: Spot test for laccase (syringaldazine): negative

throughout basidiome. Spot test for tyrosinase (paracresol): positive in stipe

(excluding bulb) and pileipellis and often in large areas of lamellae and pileus

context (outside of disc) or strongly positive throughout basidiome except for

parts of lamellae. Test vouchers: Halling 7272, Mueller 4643, Tulloss

6-13-95-C.

New Central American amanitas...183

Fig. 7. Amanita conara (holotype). Elements of hymenium and subhymenial tree (gray lines

of subhymenial base are semischematic). Scale bars = 20 um.

PILEIPELLIS: narrow and with pronounced boundaries, bounded above by

densely placed anticlinally oriented hyphae binding upper surface to remains

of universal veil, bounded below by loosely interwoven and disordered tissue

of pileus context, with separation from universal veil at first via mechanical

rupture of hyphae; 30—55 um thick at approximately midradius in mature or

near mature material, lacking substantial suprapellis, with gelatinization

absent even at point of breaking of partial veil and in mature material limited

to surface, with elements densely packed vertically, yellow-orange; filamen-

tous undifferentiated hyphae 2.5—7.5 um wide, branching, disordered and

interwoven over disc, dominantly subradially arranged with some criss-cross-

ing hyphae away from disc, often yellow or yellowish, hyphal tips of broader

hyphae common; vascular hyphae 3.4—11.4 um wide, sinuous, sometimes in

tangles, yellow, scattered. PILEUS CONTEXT: filamentous undifferentiated

hyphae 2.5-8.9 um wide, occasionally branching, sometimes fasciculate,

common in disc, plentiful at mid-radius; acrophysalides broadly ellipsoid to

ellipsoid (up to 80 x 60 um in disc) or clavate (up to 131 <x 48 um), similarly

shaped and smaller at mid-radius, commonly with inflated subterminal cell, in

disc dominating and disordered, at mid-radius plentiful and with tendency to

184...Tulloss, Halling & Mueller

subradial orientation; vascular hyphae 2.8—17.8 um wide, sinuate to hypersin-

uate, rarely branching, common in region immediately below pileipellis,

rather common elsewhere, scattered in all mounts. LAMELLA TRAMA: bilateral,

divergent; We, = 50-65 um (good rehydration); central stratum comprising

interwoven filamentous undifferentiated hyphae 2.5—11.4 um wide, lacking

inflated intercalary segments; subhymenial base dominated by densely packed

divergent filamentous undifferentiated hyphae 3.8—11.4 um wide, with angle

of divergence shallow; terminal, inflated cells not observed; vascular hyphae

2.3-8.5 um, infrequently branching, sinuous, uncommon. SUBHYMENIUM:

Wg-near = 70-85 um (moderate to good rehydration); wg-far = (80-) 90-105

um (moderate to good rehydration); scant, often with only one (or no) cell(s)

between bases of longest basidia/-oles and subhymenial base, with basidia

arising from elongate cells of subhymenial base and subglobose to branched

to ellipsoid to clavate cells and uninflated hyphal segments, with such ele-

ments arranged approximately perpendicular to central stratum. BASIDIA:

(30—) 34-59 (—60) x 9.1—-13.0 (-14.4) um, 4-sterigmate, with sterigmata up to

7.5 x 2.0 um and sometimes subcylindric; clamps and proliferated clamps rel-

atively common. UNIVERSAL VEIL: On pileus, exterior surface: particularly

well-formed and in place on surface of patches, very thin, comprising lattice

of irregularly ordered fascicles of partially gelatinized orange-brown hyphae,

with fascicles up to 18 (or more) hyphal diameters wide; filamentous undiffer-

entiated hyphae 1.8—3.9 um wide, branching; with immediately underlying

thin membranous layer, comprising loosely interwoven filamentous undiffer-

entiated hyphae 2.2-8.9 um wide, disordered, commonly branching, anasto-

mosing, plentiful to dominating, sometimes in narrow fascicles, thin-walled,

colorless and hyaline to (infrequently) yellowish and subrefractive; inflated

cells uncommon, terminal, singly or in pairs, broadly clavate to clavate to

cylindric, thin-walled, hyaline, up to 57 x 13.0 um; vascular hyphae not

observed. On pileus, interior: all elements strongly vertically aligned, orang-

ish brown on wart surface; filamentous undifferentiated hyphae 1.0-12.1 um

wide, frequently branching, occasionally anastomosing, inside wart colorless

and hyaline or (occasionally) yellowish and with subrefractive walls; inflated

cells in terminal chains of up to at least five, thin-walled, plentiful, colorless

and hyaline inside wart, barrel-shaped to ovoid to broadly clavate to clavate to

narrowly fusiform and up to 48 x 25 um in lower third of wart above basal

part and subglobose to ellipsoid to clavate (up to at least 78 x 38 um) to nar-

rowly clavate or narrowly fusiform (up to 91 =x 19.0 um) to irregularly shaped

and sometimees connected with three or more hyphae via separate septa in

upper third of wart; vascular hyphae not observed except in upper part of wart

and there very infrequent and (e.g.) 2.24.9 um wide. On pileus, basal part:

New Central American amanitas...185

Fig. 8. Amanita conara. a. Cross-section of fragments of limbus internus on bulb and

lower stipe (Tulloss 6-13-95-C) (<0.82). b. Elements of universal veil (limbus internus)

viewed through—from above—remains of partial veil (not shown) (holotype). c. Ele-

ments of universal veil, just below gelatinized hyphal layer on top of pileal wart (Hal-

ling 7272). d. Elements of universal veil near base of pileal wart (Halling 7272). Scale

bars = 20 um.

densely packed, anticlinally aligned hyphae connecting intimately with

pileipellis. On underside of partial veil (1.e., limbus internus): filamentous

undifferentiated hyphae 2.2—10.8 um wide, occasionally branching, common;

inflated cells dominating, terminal singly or as concatenated pairs, globose to

subglobose to pyriform to broadly ellipsoid to ellipsoid to elongate to broadly

186...Tulloss, Halling & Mueller

clavate to clavate to narrowly clavate (up to 60 x 33 um), dominantly thin-

walled, occasionally with wall up to 0.8 um thick; vascular hyphae not

observed. On stipe base: with structure very like that of surface layers of uni-

versal veil on pileus; filamentous undifferentiated hyphae dominating, thin-

walled, slightly gelatinized at surface, hyaline and colorless or (occasionally)

with walls yellowish and subrefractive, 2.5—10.0 um wide, commonly branch-

ing, fasciculate, densely packed, with predominantly vertical orientation (1.e.,

not disordered as on pileus); inflated cells rather infrequent, scattered, termi-

nal, narrowly clavate to clavate (e.g., 30* x 12.5 um); vascular hyphae not

observed; clamps not observed. STIPE CONTEXT: longitudinally acrophy-

salidic; filamentous undifferentiated hyphae 3.2-17.8 um wide, branching

occasionally, plentiful in interior, dominant near surface, sometimes con-

stricted at septa; acrophysalides up to at least 395 x 32 um, plentiful in inte-

rior, common near surface; vascular hyphae 7.6—-11.4 um wide, sinuous,

infrequent, occasionally clustered; clamps not observed. PARTIAL VEIL: very

thin and sparse layer of criss-crossing, hyphae, in mounts dominated by

inflated cells of limbus internus attached to underside of hyphal framework;

filamentous undifferentiated hyphae 1.5—3.8 um wide, occasionally branch-

ing, in narrow fascicles, rapidly gelatinizing prior to maturity of basidiome;

inflated cells not observed; vascular hyphae not observed. LAMELLA EDGE

TISSUE: Sterile.

BASIDIOSPORES: [181/8/4] (7.8—) 8.1—10.0 (-11.5) x (5.3—) 6.0—7.5 (—8.7) um,

(L = (8.6—) 8.8-9.4 um; L’ = 9.1 um; W = 6.5-6.8 (—7.0) um; W’ = 6.6 um; Q

= (1.17—-) 1.23-1.51 (—1.70); Q = 1.33-1.41 (—1.44); Q’ = 1.37), hyaline, col-

orless, thin-walled, smooth, amyloid, broadly ellipsoid to ellipsoid, sometimes

swollen at one end, often at least somewhat adaxially flattened; apiculus sub-

lateral, cylindric; contents multi- to monoguttulate; color in deposit unknown.

Eco.Locy: Subgregarious to scattered, at 1600-1720 m elev. In thick, dark,

wet loam under shallow layer of leaf litter of mixed broadleaf forest including

Quercus oocarpa or in such a habitat with that tree and QO. seemannii.

MATERIAL EXAMINED: COSTA RICA: CARTAGO—Estrella, 14.vi.1996 R.E. Hal-

ling & J. Ammirati [Halling 7686] (holotype, USJ; isotype, NY); off Interamerican

Hwy., 6 km S of Cartago at rd. to Palo Verde, 28.vii.1992 B.A. Strack & G.M.

Mueller [Mueller 4434] (F 1102487); Palo Verde, 31.v.1994 G.M. Mueller 4643 (F

1112041; USJ), 11.vi.1994 R.E. Halling & T.J. Baroni [Halling 7272] (NY; USJ),

13.vi.1995 R.E. Tulloss 6-13-95-C (RET 153-4; USJ).

COMMENTS — Four taxa have been assigned to Amanita subsect. Gymnopodae

world wide, but all can be easily distinguished from the present species. In

addition to their general habit, three of these species have in common with A.

conara a number of distinctive characters among which are rubescent or dark-

New Central American amanitas...187

ening context, a distinctive odor, yellow or orange tints on the lamellae, and a

submembranous surface layer of the universal veil.

Amanita gymnopus Corner & Bas (Corner & Bas 1962), described from

Malaysian “jungle” and reported from southern China (Chen et al.

2001, Yang 2005) and Japan (Hongo 1982), has small (S—7 < 5-6.5 um

per Corner and Bas), globose to subglobose spores; a universal veil that

forms submembranous patches and not warts on the pileus; and marked

rhizoids attached to the stipe base (Bas 1969: fig. 378). Chen et al.

reported spores of 6.0—7.5 (—8.0) x (5.0—) 5.5-6.5 um from a single

specimen. Yang reported spore data as follows in specimens from at

least three localities: 6.0-8.5 x (5.0—) 5.5—7.5 um, (Q = (1.03—) 1.07—

1.24 (-1.42); Q= 1.14 + 0.08).1

Amanita ochraceobulbosa A. E. Wood (Wood 1997) has subglobose to

broadly ellipsoid spores 9.3—11.7 x (7.3—) 8.1—-9.6 (10.2) um with est.

Q= 1.2, acap that is cream to cream-buff to yellow-orange-cream with

universal veil remains as (often large) pyramidal warts, gills that are

white to cream, flesh that apparently is nonstaining when cut or

bruised, and pyramidal warts sometimes present on the stipe’s basal

bulb. Absence of a submembranous outer volval layer, presence of

warts on the bulb, and no mention of staining flesh or distinctive odor

suggest that this taxon is not appropriately placed in subsect. Gymnop-

odae.

Amanita ochrophylla (Cooke & Massee) Cleland (Cleland 1924),

described from edges of open forest in eastern and southeastern Austra-

lia (Bas 1969: 556), has larger, proportionately narrower spores (9-11 x

5.5—7 um) with Q = 1.3—2.0 and Q = 1.5—1.8; a limbus internus that is

apparently more coherent than that of A. conara and is frequently left

as a “second annulus” on the stipe just below the partial veil (Bas 1969:

fig. 374); flesh that becomes pink on exposure; etc. Wood (1997: 803)

gives spore measurements as follows: 9.3—10.8 (-12.6) x 5.4-7.4

(-8.4) um, with Q = 1.43—1.72 (—1.88). We estimate Q’ for Wood’s

material as 1.55—1.6.

Amanita ochrophylloides D. A. Reid (Reid 1978, 1980), described from

“under Eucalyptus sp. (of peppermint group)” in the state of Victoria,

Australia, has predominantly subglobose to broadly ellipsoid to ellip-

soid spores (7.0—10.0 x 5.0—8.0 um, with est. Q = 1.3); a pale brown

pileus with dark, “strobiliform” warts; a tendency to darken from

handling; pale golden yellow lamellae; a slight volval limb on the

Yang’s Q corresponds to the Q of our notation; and his Q, to our Q’.

188...Tulloss, Halling & Mueller

y 3

pie

Fig. 9. Amanita costaricensis. a. Habit (holotype) with weathered cap on right (<0.55).

b. Habit (Tulloss 6-23-95-M) (<0.37). Photos by R. E. Tulloss.

stipe’s basal bulb; an odor that is faint and mealy; abundant, highly

refractive hyphae in the universal veil on the pileus; etc.

Guzman (1975) reported A. ochrophylla from Quercus forest in Guerrero. His

macroscopic description fits A. conara rather well; and his measurements for

spores are 9-11 x (5.5—) 6.0-7.5 um. This is a relatively good match for our

spore data from A. conara. We estimate a Q’ of approximately 1.5 for spores

of Guzman’s material. We have not yet reviewed the material Guzman cited,

but it appears to us that 4. conara may occur in Mexico.

Mueller 4643 consists of a single, rather large, immature specimen of A.

conara. It comes from the same habit as the other material cited and is easily

recognizable because of excellent photographs and notes on fresh material

that accompany the exsiccatum; moreover, the thin membranous outer layer of

the universal veil can be seen on scalp sections of the universal veil remnants

on the pileus.

We have three collections of what appears to be a similar, but possibly distin-

guishable, taxon from Ctn. Coto Brus, Prov. Puntarenas (La Amistad sites nos.

1 and 2) in association with QO. corrugata, Q. seemannii, and other oaks and

belonging in subsection Gymnopodae (“species CR18” in the above key).

New Central American amanitas...189

These collections differ from A. conara in colors of the basidiome (not all col-

lections are fully annotated) and consistently have smaller subglobose to

broadly ellipsoid spores: [160/8/3] (6.1—) 7.0-9.5 (—-11.6) x (5.1—) 5.9-7.9

(—9.5) um, (L = 7.6—-8.2 (—8.3) um; L’ = 7.9 um; W = (6.3—) 6.7—7.0 um; W’ =

6.7 um; Q= (1.04) 1.08-1.29 (-1.41); Q@=(1.11-) 1.15—-1.23; Q’ = 1.17). All

three collections are deposited in NY (Halling 7803, 8148. 8360).

The present species was previously called “A. species CR10” in drafts and

keys circulated by Tulloss.

Amanita costaricensis Tulloss, Halling, G.M. Muell. & Singer sp. nov. Fics. 9-11

MycoBANK MB 518297

Pileus (43—) 50-120 (—145) mm latus, brunneus vel alutaceus vel griseus, aliquando

subroseotinctus, disco fuscior, hemisphaericus vel convexus demum applanatus disco

subumbonato; contexto albo, aliquando subochraceo ubi laeso; margine appendicu-

lato aliquando brevistriato; velo universali griseo vel brunneo vel atrobrunneo ver-

rucato. Lamellae liberae vel subadnatae, confertae, saepe subventricosae, 3—7.5 mm

latae, lamellulis truncatis vel attenuatis. Stipes 52-124 x 9-18 (—22) mm, subalbus

vel griseus, annulatus, pulverulentus vel flocculosus sub annulo; bulbo (30—) 57—

112+ x 17-40 mm, napiformi vel dauciformi, radicanti; velo universali pulverulento

vel verrucato, fuligineogriseo vel concoloro. Sporae amyloideae, (7.5—) 8.5—13.2

(—16.0) x (5.1—) 6.0-8.5 (—12.6) wm, (L = (9.1—) 9.4-12.3 um; L’ = 10.8 um; W =

6.4-7.7 (8.3) um; W = 7.2 um; Q = (1.10—) 1.27-1.83 (2.15); Q = 1.35-1.68

(—1.70); Q’ = 1.51), late ellipsoideae vel ellipsoideae vel elongatae. Fibulae vul-

garis. In quercetis tropicalibus montanis costaricensibus et hondurensibus habitat.

Amanita onusta et A. atkinsoniana Amanitae stirpis Microlepis simulanissimae. Spe-

cies nova ob magnitudinem sporarum, magnitudinem basidiomae, lamellarum non

flavescentem et non celeriter putrescentem, et ubi probata paracresolo non reagens.

HoLortyPe — Costa Rica, Prov. San José, Ctn. Dota, 21.v1.1995 R.E. Halling s.n.

[Tulloss 6-21-95-M] (USJ).

EtymMoLocy — In honor of the Republic of Costa Rica.

PILEus: (43—) 50-120 mm wide [up to 145 mm in Mueller 4368], brown (6C4-

5) to light brown (5-6D4-5) to light brown with slight pinkish cast (ca. 7D5) to

pale sordid tan to yellowish cream (then with sordid tan disc) to light brown-

ish gray (7.0YR 5.4/1.2) to brownish gray to mouse (or rat) gray, micaceous

gray in old material (GOmez & Alfaro 20641) or dull dark brown over disc

[Mueller 4368], with pigment at least sometimes washed out by rain, hemi-

spheric to convex at first, becoming applanate around subumbonate disc,

tacky when moist, sometimes viscid over disc, dull to satiny to subshiny,

shiny in age, sometimes becoming areolate and then with areolae centered on

warts; context white, usually with pale brownish gray to gray to brown region

under pileipellis, sometimes pale ochraceous when exposed, 3—10.5 mm thick

over stipe, thinning evenly to margin; margin nonstriate, sometimes short sul-

190...Tulloss, Halling & Mueller

D.C. Tulloss del.

Fig. 10. Amanita costaricensis. Habit (Tulloss 6-23-95-M) (0.63).

cate (0.05R) in age, incurved at first, then decurved, occasionally uplifted in

age, appendiculate with white and or gray floccose-felted shreds of partial

veil; universal veil as patches or flakes or warts (some rather broad and irreg-

ular, some small and pyramidal), often squamulose near margin, gray to

brownish gray to brown or dark grayish brown and darker than pileus

(6D-E4-6 to 7E6 to 7F7 to 5.6YR 3.4/0.9), occasionally as flat pinkish brown

(7B4) patches, pulverulent, darkening with age, verruculose (10x lens), deter-

Sile.

New Central American amanitas...191

LAMELLAE: free to narrowly adnate, close to subcrowded to crowded, lacking

decurrent lines on stipe apex, white to pale cream in mass, off-white to

slightly sordid white to very pale yellowish white in side view, reddish brown

where damaged, relatively thick, 3-7.5 mm broad, frequently subventricose,

entire, with thin gray edge; /amellulae truncate to rounded truncate to subat-

tenuate to attenuate to attenuate in steps, plentiful, unevenly distributed, of

diverse lengths or all very short [only reported for Mueller 4368].

STIPE: 52-124 x 9-18 (-22) mm [broadest in luxuriant mature specimen of

Mueller 4368], ground color white to off-white to pale grayish, pallid to gray

matte to whitish-gray and smooth to subglabrous above partial veil, below

partial veil (and extending downward to first row of warts of universal veil)

decorated with densely pulverulent to floccose or floccose-fibrillose universal

veil (q.v.), with such material mouse gray to light gray-brown (ca. 7C3), later

decorated with light gray-brown (5B3 to 9.7YR 6.4/2.5) granules and light

gray-brown to reddish brown fibrillose upward pointing scales, with such

material evenly distributed or in narrow concentric bands, finely striatulate

with some raised fibrils in undecorated areas below partial veil, with surface

decoration becoming reddish brown from handling, cylindric or narrowing

upward or broadest at mid-stipe or narrowest at mid-stipe, flaring at apex or

not; bulb (30-) 57-112+ x 17-40 mm [shortest in mature specimen of Mueller

4368], napiform to narrowly napiform or dauciform, radicating, varying from

poorly differentiated from stipe to subabrupt; context white to sordid white

[bruising light yellow only in Mueller 4368], solid (under some conditions

center of stipe becoming water soaked or gelatinized and pale sordid yellow-

ish tan), sometimes with lacunae in center (especially in large specimens),

infrequently becoming hollow [e.g., in Mueller 4368], concolorous to pale

pallid brown or pale pinkish brown or ochraceous or pale brick-colored in lar-

val tunnels; partial veil apical to superior, white becoming gray above, occa-

sionally with edge darker gray in age, below bearing gray pulverulence, striate

above, skirt-like, submembranous to floccose-felted, tearing, often left as floc-

cose patches over lamellae; universal veil absent or as pyramidal warts or

patches placed irregularly or subconcentrically arranged in few to many (up to

10 or more) rows near broadest point of bulb or for as much as upper three-

quarters of bulb length, often with underlying context forming white to pale

grayish recurved scales with universal veil remnants on tips of such scales,

rather dark brownish gray (5.6YR 3.4/0.9), with limbus internus sometimes as

thick, loosely attached, friable ring at stipe base (then concolorous with stipe).

Odor lacking or similar to nutmeg at first; in age, “old ham’-type, but more

penetrating and unpleasant. [“Musty” in Mueller 4368.| Taste not distinctive.

192...Tulloss, Halling & Mueller

Fig. 11. Amanita costaricensis. a. Elements of universal veil from pileus wart (Tulloss

6-23-95-M). b. Elements of universal veil from floccose material of stipe (isotype). c.

Elements of weakly structured partial veil (isotype). d. Elements of hymenium and sub-

hymenial tree. Scale bars 20 um.

MACROCHEMICAL TESTS: Spot test for laccase (syringaldazine): negative

throughout basidiome except for very few spots on margins of lamellae. Spot

New Central American amanitas...193

test for tyrosinase (paracresol): negative throughout basidiome. Test voucher:

Tulloss 6-20-95-A.

PILEIPELLIS: 315-460 um thick overall (175-235 um in old specimen); supra-

pellis 95-230 um thick (40-100 um in old specimen), partially gelatinized,

pale yellow to colorless except (occasionally) dark brownish orange at sur-

face; subpellis 160-260 um thick (130-135 um in old specimen), ungelati-

nized, dark brownish orange; filamentous undifferentiated hyphae 2.8-8.4 um

wide, branching, densely packed, dominantly subradially oriented, but with

some criss-crossing; vascular hyphae 7.4-21 um wide, scattered to locally

common, sinuous, with those of largest diameter from old (senescent?) speci-

men (Gomez & Alfaro 20641). PILEUS CONTEXT: filamentous undifferentiated

hyphae 2.0-20 um wide (many with width ca. 8.0 um), branching, dominat-

ing, singly and in fascicles interwoven in open lattice, occasionally with yel-

lowish subrefractive walls; inflated cells clavate to narrowly clavate to

subfusiform, up to 138 x 34 um, apparently mostly intercalary, one clavate

cell terminal [possibly fitting definition of “acrophysalide” per Bas (1975)]|

except for hypha branching from one side, with walls thin or slightly thick-

ened; vascular hyphae 8.4-31 um wide, occasionally branching, sinuous, scat-

tered to locally common especially just below pileipellis, locally in loose

knots, with those of largest diameter from old (senescent?) specimen (Gomez

& Alfaro 20641). LAMELLA TRAMA: bilateral, divergent; we, = 45-60+ um

(moderate rehydration); central stratum composed largely of rather broad

hyphae, apparently lacking intercalary inflated segments; subhymenial base

comprising filamentous undifferentiated hyphae and narrow intercalary

inflated cells (ellipsoid to obclavate to subfusiform to cylindric, up to 90 x 28

um, singly and in short chains, with width predominantly 17.0-28 um)

diverging in smooth curve and achieving marked angle to central stratum; fil-

amentous undifferentiated hyphae 2.8-10.2 um wide, branching, sometimes

constricted at septa; divergent, terminal inflated cells not observed; vascular

hyphae not observed. SUBHYMENIUM: Wg-near = 65-85 um (poor to moderate

rehydration) or 145-150 um (good rehydration); w.-far = 85—110* um (poor

to moderate rehydration) or 170-175 um (good rehydration); appearing

pseudoparenchymatous in some regions, comprising 2—4 layers of inflated

cells (sometimes dominating) and irregularly branched partially inflated to

inflated elements and partially inflated (proportionately small) hyphal seg-

ments, with basidia arising from cells of all types. BAsipIA: (21—) 39-63 x

(6.2—) 8.2-14.2 (-15.8) um, thin-walled, 4-sterigmate, with sterigmata up to

6.0 x 2.8 um or larger; clamps common. UNIVERSAL VEIL: On pileus: ele-

ments having strong vertical orientation except in base of wart; filamentous

undifferentiated hyphae 1.0-13.6 um wide, branching, common in upper part

194... Tulloss, Halling & Mueller

of wart, dominating in shallow interwoven layer at base of wart and there peri-

clinally oriented and more often in fascicles, sometimes constricted at septa,

colorless at first becoming orange-brown like inflated cells; inflated cells pale

brown to pale brownish gray to pale gray, becoming brownish orange to

orange-brown to brown, 28-128 x 11.5-50 um, dominating except in base of

wart, terminal in short chains, thin-walled, subglobose to broadly ellipsoid to

ellipsoid to broadly clavate (up to 83 x 50 um) or basidiiform to narrowly cla-

vate to subcylindric to cylindric to subfusiform (up to 128 x 37 um), all forms

occasionally rostrate when in terminal position in chain; vascular hyphae 7.2—

10.2 um wide sinuous, infrequent, scattered. On mid-stipe region: almost

entirely comprising scattered inflated cells, ungelatinized to partially gelati-

nized to gelatinized, subglobose (e.g., 32 <x 28 um), ellipsoid (e.g., 38 x 26

um), narrowly to broadly clavate (e.g., 56 x 33 um), bacilliform (e.g., 32 x 8.6

um). On bulb of stipe: comprising disordered elements, with greater propor-

tion of filamentous undifferentiated hyphae than in universal veil on pileus,

otherwise similar to remnants on pileus. STIPE CONTEXT: longitudinally acro-

physalidic; filamentous undifferentiated hyphae 1.9-9.9 um wide, plentiful,

occasionally branching; acrophysalides plentiful, up to 406 x 33 um; vascular

hyphae 7.0-16.5 um wide, infrequent, sinuous, yellowish, mostly near exte-

rior surface. PARTIAL VEIL: with hyphae, elongate cells, and rows of cells

sometimes subradially oriented; radially streaked on upper surface with par-

tially gelatinized collapsed remains of lamella edge tissue; filamentous undif-

ferentiated hyphae 1.5-7.5 um wide, moderately common, branching

occasionally, often single, sometimes in narrow fascicles, infrequently in fas-

cicles up to 17.5 um wide; inflated cells arising from nonvascular hyphae (see

below for inflated cells arising from vascular hyphae) thin-walled, terminal,

singly or in short chains (mostly of two), pyriform or ovoid or elongate or

ellipsoid or subfusiform or subcylindric or broadly clavate or clavate or nar-

rowly clavate, up to 113 <x 29 um, occasionally with yellowish walls, domi-

nantly colorless, hyaline; vascular hyphae 2.8-5.0 um wide, very infrequent,

infrequently giving rise to yellow-walled terminal cell (elongate to ellipsoid,

up to 26 x 22 um). LAMELLA EDGE TISSUE: Sterile.

BASIDIOSPORES: [321/16/10] (7.5—) 8.5-13.2 (-16.0) x (5.1—) 6.0-8.5 (-12.6)

um, (L = (9.1—) 9.4-12.3 um; L’ = 10.8 um; W = 6.4-7.7 (8.3) um; W’ = 7.2

um; Q =(1.10-) 1.27-1.83 (-2.15); Q= 1.35-1.68 (-1.70); Q’ = 1.51), hyaline,

colorless, smooth, thin-walled, amyloid, acyanophilous, dominantly ellipsoid

to elongate, infrequently broadly ellipsoid or cylindric, often at least some-

what adaxially flattened, occasionally expanded at one end; apiculus sublat-

eral, small, cylindric; contents mono- to multiguttulate to granular; white in

deposit.

New Central American amanitas...195

EcoLocy: Solitary to subgregarious. Costa Rica: At 1715-2500 m elev. In

Quercus-Magnolia forest or in mixed Quercus forest sometimes with Q.

oocarpa dominant. Honduras: At 1900+ m elev. In cloud forest with Quercus

trichodonta and other hardwoods.

MATERIAL EXAMINED: COSTA RICA: CARTAGO—San Cristobal, vii.1983 L.D.

Gomez & R. Alfaro 21222 (F 1100824). GUANACASTE—Cacao no. 1, sendero a

derrumbe, 3.vi.1996 C. Cano 197 (INBio; RET 325-4). SAN JosEt—La Chonta,

v.1982 L.D. Gomez 18147 (F 1051869); Jaboncillo de Dota, 20.vi.1995 R.E. Hal-

ling & K. Shanks s.n. [Tulloss 6-20-95-A] (RET 330-3; USJ); El Jardin de Dota,

vii.1983 L.D. Gomez & R. Alfaro 20641 (F 1073283); San Gerardo de Dota no. 1,

26.vil.1992 B. A. Strack, L.D. Gomez, G. Hewson & G.M. Mueller [Mueller 4398]

(F 1102451), 21.vi.1995 R_E. Halling s.n. [Tulloss 6-21-95-M] (holotype, USJ; iso-

type, RET 330-4), 23.vi.1995 Thomas O’Dell s.n. [Tulloss 6-23-95-M] (RET

330-5; USJ); San Gerardo de Dota no. 4, 24.vii.1992 B.A. Strack, J. Polishook,

L.D. Gomez, G. Hewson & G.M. Mueller [Mueller 4368] (F 1102421). HonpDv-

RAS: FRANCISCO MORAZAN—Mt. Uyuca, Valle Zamorano, 5.vii.1991 G.M.

Mueller, B.A. Strack, R. & M. Singer & R. Andino [Mueller 4138] (F 1098682).

OLANCHO— ca. Campamento, P.N. La Tigre, rd. to Finca Sapote, 9.vi1.1991 G.M.

Mueller, B.A. Strack, R. & M. Singer & R. Andino [Mueller 4182] (F 1098725).

COMMENTS — This species was described in the notebooks of the late Dr. Rolf

Singer under his herbarium name “costaricensis.” Since he originated the

name and had a clear concept of its application and since we utilize collec-

tions annotated by him, we have included Dr. Singer as an author of the spe-

cles.

According to the descriptions and keys of Bas (1969), A. costaricensis fits

well within his stirps Microlepis of Amanita [sect. Lepidella| subsect. Solitar-

iae Bas. Within this stirps, the present species is strikingly similar to A. atkin-

soniana Coker (Coker 1917) and A. onusta (Howe) Sacc. (Saccardo 1891).

Amanita atkinsoniana (Tulloss 2011d) is moderately common in a range

extending from southern Prov. Québec, Canada (Pomerleau 1980) to Michoa-

can edo., Mexico. Amanita atkinsoniana can be distinguished from the pres-

ent species by

¢ smaller spores—[230/10/10] (6.8—) 8.0-10.5 (-14.3) x (5.0—) 5.4-7.2

(—8.5) um, (L = (8.2-) 9.0-9.7 um; L’? = 9.2 um; W=5.7-6.6 (-6.9)

um; W’ = 6.2 um; Q = (1.17-) 1.27-1.71 (-1.95); Q = 1.36—1.57

(—1.67); Q’= 1.50)

* basidiome generally white or pallid except for universal veil, with uni-

versal veil in fine distinctly separated warts (grayish at first, then red-

dish brown) on pileus, with basal bulb usually napiform or turbinate,

with universal veil brownish gray becoming reddish brown and distrib-

uted in rather fine warts in at least six (6) concentric rings around the

stipe’s bulb from its top to (often) well below mid-length of the bulb,

196...Tulloss, Halling & Mueller

with fragile apical to subapical partial veil, which (when it persists)

may become slimy and yellow while remaining in a superior position

on the stipe

scattered, minimal, positive spot tests for tyrosinase—with L-tyrosine,

slowly positive on surfaces of lower stipe and upper bulb (possibly in

association with universal veil material in those areas) and in pileipel-

lis.

MATERIAL EXAMINED (Amanita atkinsoniana): MEXICO: MICHOACAN—Parq.

Nac. "Insurgente José Ma. Morelos," Mpio. Charo, 19.vii.1983 Reza Araujo s.n.

(FCME). U. S. A.: CONNECTICUT—Middlesex Co. — E. Haddam, Devil's Hop

Yard St. Pk., 25.1x.1999 Arnold s.n. [Tulloss 9-25-99-P] (RET 301-1); Meshomasic

St. For., 19.1x.1998 Peter Kukle s.n. [Tulloss 9-19-98-A] (RET 288-2). MAINE—

Cumberland Co. — S. Windham, 25.viii.1984 S.S. Ristich s.n. [Tulloss 8-25-84-

SSR-A] (RET 235-6). MASSACHUSETTS—Berkshire Co. — Pittsfield St. For.,

16.vill.1986 Wes Faust s.n. [Tulloss 8-16-86-G] (RET 137-7). NEw JERSEY

Morris Co. — Hacketstown Reservoir, 13.viii.1984 Robert Peabody & Roger Phil-

lips s.n. [Tulloss 8-13-84-PPA] (RET 113-8). NEw YorkK—Dutchess Co. —

Thompson Pond Preserve, Pine Plains, 4.viii.1996 Wm. Bakaitis s.n. (NYS

D1592). NORTH CAROLINA—Granville Co. — Butner, John Umstead Hospital

picnic area, 18.x1.1994 Owen L. McConnell s.n. (RET 139-6). McDowell Co. — ca.

Little Switzerland, Wildacres Resort, 27.1x.2008 J. Justice NC-AM12 (RET 445-

1). Orange Co. — Chapel Hill, Battle's Pk., 14.1x.1913 W.C. Coker 759 (holotype,

NCU, mixed collection segregated by C. Bas). WEST VIRGINIA—Greenbrier

Co. — Monongahela Nat. For., Lake Sherwood, 31.vii1.1982 R.E. Tulloss 8-31-82-D

(RET 226-2). Tucker Co. — Dolly Sods, 1.viii.1985 W. Sturgeon s.n. [Tulloss 8-1-

85-B] (RET 201-6).

Amanita onusta (Tulloss 201 1e) is a locally common species of eastern North

America with a range extending from Prov. Québec (Pomerleau 1980) and

Prov. Nova Scotia (Stewart & Grund 1974), Canada at least to Illinois and

Mississippi, USA. (Tulloss unpub. data). Amanita onusta can be distin-

guished from the present species by

¢ smaller spores—[215/12/12] (7.0—) 8.0-11.0 (-13.0) x (5.0—) 5.5-7.0

(—8.3) um, (L = 8.3—-10.5 (-11.0) um; L’=9.4 um; W=5.7-6.5 (-7.1)

um; W’'=6.1 um; Q = (1.14-) 1.28-1.85 (-2.21); Q =1.35-1.65

(—1.84); Q’=1.52)

* basidiomes paler and with little (if any) brown tint, frequently of

smaller size with pileus often less than 40 mm wide [13-76 mm wide in

Tulloss’ experience, although Bas (1969) reports at least one instance

of a specimen with pileus over 100 mm wide], with lamellae that rather

rapidly become yellowish and show signs of decay while the remainder

of the basidiome shows none, and often without a persistent partial veil

* positive spot tests for tyrosinase—with L-tyrosine, positive, throughout

the basidiome and, with paracresol, positive in scattered spots in very

New Central American amanitas...197

young material, positive in pileus context and context of stipe above

bulb in older material.

MATERIAL EXAMINED (Amanita onusta): U. S. A.: CONNECTICUT—Litchfield

Co. — Black Rock St. Pk., 10.viii.1983 D.C. & R.E. Tulloss 8-10-83-A (RET 104-

8). MAINE—Sagadahoc Co. — Woolwich, 7.x.1995 S.S. Ristich s.n. (RET 159-7).

New JERSEY—Mercer Co. — Hightstown, 22.vi1.1981 R.E. Tulloss 7-22-81-B

(RET 106-9). Monmouth Co. — Upper Freehold Twp., Assunpink Wildlife Met.

Area, Roosevelt Rd., 21.vii.1981 R.E. Tulloss 7-21-81-E (RET 163-3), 4.viii.1981

RE. Tulloss 8-4-81-A (RET 109-6), 18.ix.1981 M.A. King & RE. Tulloss

9-18-81-E (RET 164-7), 11.vii.1982 D.C., M.H., & R_E. Tulloss 7-11-82-B (RET

340-10). Morris Co. — Morristown, 27.vili.1981 Al Northrup s.n. [Tulloss

8-27-81-AN] (RET 166-10). VIRGINIA—Franklin Co. — ca. Ferrum, ca. Ferrum

College, 4.viii.1982 Gerald Bills, Pierre Dery, O.K. & H.H. Miller OKM19946

(VPI, as “A. peckiana’). WEST VIRGINIA—Marion Co. — Mill Fall Run,

21.ix.1992 R.P. Bhatt A2 (FWVA).

In two cases [Halling 7667 (NY; USJ) and Tulloss 6-21-95-E (RET 337-3;

USJ)], an amanita assignable to Amanita subgenus Lepidella (sect. Validae or

sect. Lepidella) was misdetermined in the field as A. costaricensis. The unde-

termined species has a gray stipe that is stuffed (becoming hollow), gray

underside and margin to the membranous annulus, gray warts on the gray-

brown to pale gray pileus, and short basidia [19-34 (44) um long] apparently

lacking clamps. The species is somewhat unusual for sect. Validae because of

its unpleasant smell (sometimes like the smell of disinfectant cleaning solu-

tion in hospitals). The misdetermined entity should be distinguishable by the

lack of a radicating bulb, the absence of basidial clamps, and by its smaller

and more nearly globose spores: [33/2/2] (7.3—) 7.5—9.0 (—10.0) x (6.5—) 7.0-

8.0 (-9.1) um, (L = 8.2 um; L’ = 8.2 um; W= 7.5 um; W’ = 7.5 um; Q=

(1.04—) 1.05-1.14 (-1.23); Q= 1.09-1.11; Q’ = 1.09). Especially, considering

the apparently parasitized collections of A. costaricensis discussed below, we

had some hesitation concerning whether or not Halling 7667 and Tulloss

6-21-95-E also comprised parasitized material. However, we are convinced

that they represent a distinct species because (1) the spores of these two col-

lections were both shorter (on average) and broader (on average) than those of

A. costaricensis [Compare values of L’ and W’. Reduction in spore volume

due to stress would have reduced both length and width. ]; (2) both exhibited a

membranous annulus; and (3) both apparently lack basidial clamps. Basidia

predominantly less than 30 um long, might support assignment to sect. Vali-

dae, but further study is required.

In one case {Estrella, 14.vi.1996 R.E. Halling & J. Ammirati [Halling 7684]

(NY; USJ)}, all the specimens of a rather large collection apparently assign-

able to A. costaricensis had an odor like pig manure. In addition, some had

rather distorted stipes; and the spores of all collections were uniformly smaller

198...Tulloss, Halling & Mueller

than normal ([100/5/1] (7.2—) 7.6-9.5 (—11.0) x (5.3—) 5.5—7.3 (—8.0) um, (L =

8.3-9.0 um; L’ = 8.7 um; W = 6.2-6.6 um; W’ = 6.4 um; Q = (1.18—) 1.23-

1.50 (—1.80); Q = 1.30—1.45; Q’ = 1.37). Observed at 1250x, many of the

spores were surrounded with small, hyaline, colorless, inamyloid spheres (1+—

3+ um diam.) connected to the spore walls by what appeared to be fine

tubules—the whole having the appearance of a pincushion. We hypothesize

that the noted alterations in morphology and odor are a result of “parasitiza-

tion.” The authors explicitly exclude Halling 7684 from the set of paratypes.

In another case of apparent parasitization (San Gerardo de Dota no. 4,

24 .vii.1992 B.A. Strack, J. Polishook, L.D. Gomez, G. Hewson & G.M. Muel-

ler [Mueller 4368] (F 1102421)), spores, basidia, and lamella trama were quite

normal for the species; however, the context of both the pileus and stipe

stained yellow when cut. Two other pigment-related characters were unusual

for A. costaricensis as we understand it: universal veil remnants on pileus,

stipe, and bulb of stipe were paler than usual and remained paler than the

pileus disc despite becoming brown with time, even in a mature specimen;

and the pileus disc was darker than usual. Further, lamellulae were reported to

all be very short; and the basidiomes were said to have had an unusual musty

odor. The yellow staining is very similar to that seen in apparently parasitized

specimens of A. subsolitaria (Murrill) Murrill from the Atlantic coastal plain

of the USA (Tulloss 1998b; 2000a: 57, fig. 5; Tulloss 2011c). In the latter

case, basidiomes are sometimes found that not only stain bright yellow when

cut, but release an orange-amber liquid from wounds. Spores of such speci-

mens are often, but not always, subnormal in size and distorted in shape—the

same yellowing phenomenon may be seen in specimens with spores nearly, or

quite, normal with regard to size and shape. The provisional name of Bas

(1969), A. “crassifolia,’’ was put forward for yellowing material that Bas

thought might be determinable as A. subsolitaria,; Tulloss is now convinced

this is the case.

It should be noted that two North American taxa assignable to sect. Lepidella

have been described by Bas (1969) because of their pronounced ability to

become bright yellow or yellow-orange when cut (A. cinereoconia vat. cro-

ceescens Bas and A. rhoadsii var. flavotingens Bas). Tulloss (1998b, 2000a)

questioned whether these entities might not be based on parasitized specimens

of the respective species. It would be interesting to see what information

about Amanita parasites might be determined from study of fresh material of

Costa Rican cases of apparent parasitization such as those presented above. If

the species with yellowing context reaction in sect. Lepidella may be grouped

together and the yellowing attributed to a single cause, or a set of common

causes, then A. costaricensis becomes the first representative of this group in

New Central American amanitas...199

Bas’ stirps Microlepis. We have included Mueller 4368 as a paratype because

microscopic examination produced observations completely consistent with

“normal” material of A. costaricensis. In the macroscopic description of A.

costaricensis, the unusual characters of Mueller 4368 are so labeled.

Mueller 4398 consists of a single immature specimen with the partial veil cov-

ering the lamellae.

Acknowledgments

We are grateful to Dr. Joseph Ammirati (University of Washington) and Dr. Zhu-

liang Yang (HKAS) for their prepublication reviews of the present paper. We particu-

larly wish to thank Dr. Yang for his comments on our observations of the anatomy of

the pileus context in A. garabitoana and for his indicating that he has made similar

observations in other taxa of the “Hemibapha group” within section Caesareae. We

are grateful to Dra. Julieta Carranza (USJ) for providing us with historical information

concerning Garabito and to Dr. J.L. Mata (Univ. S. Alabama, Mobile) for his sugges-

tion of honoring Garabito in the name of the first known species of “Slender Caesar’s

Mushroom” in the Americas south of Mexico. Mr. David C. Tulloss, Lambertville, NJ,

is thanked for preparation of habit illustrations of two newly described amanitas. Mrs.

Mary A. Tulloss, Roosevelt, NJ, is thanked for assisting in final preparation of the

paper for publication. The New York Botanical Garden and the Field Museum are

thanked for their support of field work in Costa Rica. The second and third authors are

grateful for financial support from the National Science Foundation in grants DEB-

9300798 (GMM, REH), DEB-9972018 (REH), and DEB-9972027 (GMM).

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New Central American amanitas...203

Appendix: Gazetteer of collecting sites in Costa Rica referenced in this

and planned future studies on Amanita

Localities are designated by uncomplicated names in the taxonomic part of

this paper. Full site descriptions with latitude, longitude, elevation, and the

species of Quercus present at the site (where data is available) are provided in

the following table. Full data was sometimes not available because of the dif-

ficulty of interpreting herbarium labels on collections made prior to the cur-

rent projects. Most of our data comes from the latter. Also, milestones along

the Interamerican Hwy. have been moved over time, and the mention of a

milestone in an old locality description may be less accurate than the appar-

ently less precise or more informal part of the description. The issue of

incompletely documented sites arises largely because of the number of good

quality “historic” collections in F. Latitude, longitude, and elevation for

recently explored sites were determined by use of a Garmin (GPS75) device.

Site Name

Prov. ALAJUELA

1. Bosque del Nifio

2. Palmira

PROV. CARTAGO

3. Cafion

4. Estrella

5. Palo Verde

6. [unnamed]

7. Prusia

8. San Cristobal

9. Sanatorio

Site Description

(including Oaks Present)

Ctn. Grecia, Grecia, Bosque del Nifio

- Q. seemannii (dominant)

- no data on oaks

ca. Cafion, E of Interam. Hwy., at

La Esperanza del Guarco

- QO. copeyensis (dominant)

5 km E of km 31 of Interam. Hwy.,

ca. Estrella

- Q. oocarpa, OQ. seemannii

4+ km E of km 31 of Interam. Hwy.,

ca. Palo Verde

- QO. oocarpa

off Interamerican Hwy., 6 km S of Cart-

ago at rd. to Palo Verde

- no data on oaks

Parque Prusia, W slope of Volcan Irazu

- Q. costaricensis

unknown site along Interam. Hwy.

- no data on oaks

Lat./Long.,

Elevation

10°9'4""N/84°14'42"W,

1900 m elev.

ca. 10°12'N/84°23'W,

no data on elev.

9°41'1"N/83°52'43"W,

2505 m elev.

9°46'4"N/83°S7'19"W,

1685-1717 m elev.

9°46'34"N/83°56'42"W,

1600 m elev.

159)

? m elev.

9°57'56"N/83°52'15"W,

2900 m elev.

2000 m elev.

? m elev.

204...Tulloss, Halling & Mueller

Site Name

10. Tapanti

PROV. GUANACASTE

11. Cacao no. 1

12. Cacao no. 2

Prov. HEREDIA

13. San José de la

Montafia

PROV. PUNTARENAS

14. Finca Las Alturas

15. La Amistad no. 1

16. La Amistad no. 2

17. San Vito

Prov. SAN JOSE

18. Alto de la Palma

19. C.A.T.LE. For.

Site Description

(including Oaks Present)

Guarco, Tapanti, P. N. Tapanti,

Macizo de la Muerte,

Area de Conservacion La Amistad

Pacifico

- O. copeyensis

Cerro Pedregal,

Estacion Biologia Cacao,

Area de Conservacion Guanacaste

- QO. brenesii

Cerro Cacao, Estacion Biologia Cacao,

Area de Conservacion Guanacaste

- QO. brenesii

Volcan Barva, Paso Llano, San José de la

Montafia

- no data on oaks

Ctn. Coto Brus, Finca Las Alturas,

Standford Biol. Field Stn.,

tr. to Cerro Echandi

- no data on oaks

Ctn. Coto Brus, La Amistad, Zona

Protectora Las Tablas, Finca La

Cafrosa, Camino EI Portones por El

Tajo

- Q. corrugata & QO. seemannii

Ctn. Coto Brus, Las Mellizas,

La Amistad Lodge,

ca. P. N. La Amistad

- Q. corrugata & QO. seemannii

Ctn. Coto Brus, San Vito de Coto Brus,

5 km W of Jardin Botanico Las Cruces

- no data on oaks

Ctn. Guadalupe, Alto de la Palma, ?

- no data on oaks

Ctn. Perez Zeledon, Villa Mills,

C.A.T.LE. Exp. For. of Villa Mills

- QO. costaricensis

Lat./Long.,

Elevation

9°4'6"N/83°52'30"W,

2600 m elev.

10°55'45"N/85°28'37"W,

1100 m elev.

10°56'8"N/85°27'14"W,

1000 m elev.

ca. 10°3'N/84°7'W,

2100 m elev.

1700 m elev.

8°55'34"N/82°46'0"W,

1350-1500 m elev.

8°54'52"N/82°46'50"W,

1380 m elev.

ca. 8°49'N/83°0'W,

1500 m elev.

eo)

? m elev.

9°33'3"N/83°40'55"W,

2880 m elev.

New Central American amanitas...205

Site Name Site Description Lat./Long.,

(including Oaks Present) Elevation

20. Copey de Dota Ctn. Dota, Copey de Dota 12 km S of 9°35'19"N/83°53'3"W,

Copey on rd. to Providencia 2800 m elev.

- QO. copeyensis

21. El Empalme Ctn. Dota, 200+ m W of Interam. Hwy. at 9°43'8"N/83°57'4"W,

El Empalme, La Guaria [Note: Site 2250 m elev.

has been completely logged. |

- QO. copeyensis

22. Finca Alejandrina = Ctn. Perez Zeledon, Villa Mills, Finca 9°33'43"N/83°44'22"W,

Alejandrina, Interam. Hwy. km 95, ca. 3000 m elev.

Hotel La Georgina

- QO. costaricensis

23. Jaboncillo de Dota | Ctn. Dota, Jaboncillo de Dota, 3.2 km 9°35'21"N/83°47'58"W,

from Interam. Hwy on rd. to San 2740 m elev.

Gerardo de Dota

- QO. copeyensis

24. Jardin de Dota Ctn. Dota, 3.5 km W of Interam. 9°42'52"N/83°58'28"W,

Hwy. at El Empalme 2220 m elev.

- QO. copeyensis, QO. seemannii (occas.)

25. La Chonta Ctn. Dota, S of Interam. 9°41'58"N/83°56'31"W,

Hwy. twd. Cerro Chonta 2400 m elev.

-Q. copeyensis, QO. seemannii, QO.

rapurahuensis

26. San Gerardo de Ctn. Dota, San Gerardo de Dota, 9°33'2"N/83°48'27"W,

Dota no. | 5+ km SW of Cerro de la Muerte, 2200 to ca. 2350 m elev.

Albergue de Montafia Savegre (Cabi-

nas Chacon)

- O. copeyensis, QO. seemannii, QO.

rapurahuensis

27. San Gerardo de Ctn. Dota, 500 m from Interam. Hwy. on | 9°36'13"N/83°47'26"W,

Dota no. 2 rd. to San Gerardo de Dota 3000 m elev.

- QO. costaricensis

28. San Gerardo Ctn. Dota, 1.5 km from Interam. Hwy. = 9°35'47"N/83°47'55"W,

de Dota no. 3 on rd. to San. Gerardo de Dota 2860 m elev.

- QO. copeyensis, Q. costaricensis

29. San Gerardo Ctn. Dota, off Interam. Hwy.,2-3kmS__ ?,

de Dota no. 4 of Cerro de la Muerte, rd. to San btwn. 2200 to 2860 m

Gerardo de Dota elev.

- no field data on oaks; Q. copeyensis and

Q. rapurahuensis likely, Q. seemannii

possible

MY COTAXON

http://dx.doi.org/10.5248/117.207

Volume 117, pp. 207-225 July-September 2011

Studies on Cephalotrichum from soils in China —

twelve new species and two new combinations

Yu-LAN JIANG??’, JUN-JiE Xu'?, YUE-MING Wu!, YUE-LI ZHANG',

Hu1-Me1 Liu!, Hao-QIn Pan! & TIAN- Yu ZHANG?"

'Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*Department of Plant Pathology, Guizhou University, Guiyang, 550025, China

College of Life Sciences, Linyi University, Linyi, 276005, China

* CORRESPONDENCE TO: tyzhang1937@yahoo.com.cn

ABSTRACT — A study of Cephalotrichum species diversity in soils of China resulted in twelve

new species: Cephalotrichum acutisporum, C. cylindrosporum, C. ellipsoideum, C. inflatum,

C. longicollum, C. macrosporum, C. oblongum, C. ovoideum, C. robustum, C. spirale,

C. terricola, C. verrucipes. These species are illustrated and described in this paper. The type

specimens (dried cultures) and living cultures are deposited in the Plant Pathology Herbarium

of Shandong Agricultural University (HSAUP). Two Doratomyces species are transferred to

Cephalotrichum.

KEY worps — anamorphic fungi, taxonomy

Introduction

The genus Cephalotrichum was established by Link (1809) with C. stemonitis

as type species. This genus produces its spores in a dry head at the apex of a

complex, erect conidiophore or synnema, which is often up to a millimeter

in height. The synnemata produce chains of powdery conidia with a ‘bottle

brush’ or ‘feather’ appearance. The ovoid conidia are produced from annellidic

conidiogenous cells covering the sporogenous area (Chlebicki 2008). About 50

taxa of Cephalotrichum have been reported (http://www.indexfungorum.org/

Names/Names.asp).

Doratomyces was proposed by Corda (1829), with D. neesii as type species.

However, the probable synonymy of Cephalotrichum and Doratomyces has been

a subject of debate for many years. Hughes (1958) and von Arx (1981) accepted

Doratomyces as a synonym of Cephalotrichum, whereas Morton & Smith (1963),

Y.L. Jiang and J.J. Xu contributed equally to this work.

208 ... Jiang & al.

Domsch et al. (2007), and Kirk et al. (2008) accepted Doratomyces as a separate

genus. In this paper, we treat Doratomyces as a synonym of Cephalotrichum.

During research on soil dematiaceous hyphomycetes in China, we obtained

twelve fungi that possessed the typical Cephalotrichum characters but did not

match other species in this genus. These fungi are described as new taxa and two

Doratomyces species are transferred to Cephalotrichum, as new combinations.

Materials & methods

Themethods ofsample collection, isolation and identification of fungi, and preparation

of dried cultures were as previously reported by Jiang & Zhang (2007). Conidia and

conidiophores were placed in a drop of lactic acid, examined, and photographed using

light microscopy. Mature conidia and conidiophores were measured by using a Nikon

90i microscope (Nikon Corporation, Japan) at 100x magnification. The fungi are

described from cultures grown on potato dextrose agar (PDA; 20 g white potato boiled

and filtered, 20 g dextrose, 20 g agar, 1 L distilled water).

Taxonomy

Cephalotrichum acutisporum J.J. Xu & T.Y. Zhang, sp. nov. FIG. 1

MycoBank MB 561106

Coloniae effusae, floccosae vel pulveraceae, fuscae vel atrae, in PDA crescentes lentissime

et attingentes diametrum 2-4 centimetri ad 25°C post duas hebdomadas, reverse atro-

brunneae vel fuscae. Mycelium plerumque superficiale, ex hyphis pallide brunneis vel

brunneis, laevibus, septatis, ramosis, 1.5-3.5 um crassis, in funiculos floccosos interdum

assurgens. Synnemata 120-820 wm alta, stipitata, atro-brunnea vel fusca, capitulo

cylindrica et ellipsoidea. Conidiophora pallide brunnea vel brunnea, ramosa, septata, laevia,

1-4 um lata, basin 1.5-3.5 um lata. Celluae conidiogenae percurrentes (annellophora),

ampuliformes, penicilliformiter dispositae, 4-10 x 2-3 um. Conidia catenata, ellipsoidea

vel cylindrica, basin truncata et apicem acuta, dilute brunnea vel brunnea, laevia, 5-6 x

2.5-3 um.

Ho.oryPeE: China. Fujian Province, Zhangping, from soil of a park, 22 October 2004, J.J.

Xu (HSAUPII, 2724, dried culture, holotype; HMAS196222, isotype).

EryMo oy: in reference to the conidia, which are pointed at the apex.

Colonies on PDA effuse, floccose or powdery, at first greenish grey, finally

blackish brown to black with the formation of abundantly sporing structures and

synnemata, reverse dark brown to blackish brown, growing slow, reaching 2-4

cm diameter at 25°C after two weeks. Mycelium mostly superficial, composed

of pale to mid brown, smooth, septate, branched, 1.5-3.5 um wide hyphae that

occasionally aggregate forming ropes. Synnemata 120-820 um high, stipes dark

brown to blackish brown, with cylindrical or ellipsoidal heads. Conidiophores

pale brown to brown, branched, septate, smooth, 1-4 um wide, 1.5-3.5 um

wide at the base. Conidiogenous cells (annellophores) ampuliform, penicillately

arranged, 4-10 x 2-3 um. Conidia catenate, ellipsoidal to cylindrical, truncate

at the base and pointed at the apex, pale to mid brown, smooth, 5-6 x 2.5-3

um, L/W = 2.

Cephalotrichum species new from China ... 209

Fic. 1. Cephalotrichum acutisporum: conidia, conidiogenous cells and synnema.

COMMENTS: C. acutisporum is most similar to C. stemonitis (Pers.) Link

(Link 1809) and C. microsporum (Sacc.) P.M. Kirk (Kirk & Spooner 1984).

However, conidia of C. acutisporum are ellipsoidal to cylindrical, but those of

C. microsporum and C. stemonitis are ovoid. Conidia are larger (6-8.5 x 4-4.5

um) in C. stemonitis and broader (3-4 um) in C. microsporum than those of

C. acutisporum, while synnemata are shorter (600 um) in C. microsporum and

longer (< 1200 um) in C. stemonitis (Ellis 1971, Morton & Smith 1963).

Cephalotrichum cylindrosporum Y.L. Zhang & T.Y. Zhang, sp. nov. FIG. 2

MycoBank MB 561114

Coloniae effusae, pulveraceae, glaucae vel nigro-virentes, reverse brunneae, in PDA

crescentes lentissime et attingentes diametrum 3-4 centimetri ad 25°C post duas

hebdomadas. Mycelium plerumque superficiale, ex hyphis dilute brunneis vel brunneis,

laevibus, septatis, ramosis, 2-2.5 um crassis. Synnemata 750-900 um alta, capitulo

210 ... Jiang & al.

Fic. 2. Cephalotrichum cylindrosporum: conidia, conidiogenous cells and synnemata.

elongato-fusiformi stipitata, atro-brunnea, 15-20 wm lata. Celluae conidiogenae

percurrentes (annellophora), ampuliformes, penicilliformiter dispositae, pallide brunneae

vel brunneae, laeves, 9-11.5 x 2.5-3.5 um. Conidia acrogena, solitaria vel catenata,

cylindrica vel subcylindrica, in extremitatibus rotundata, brunnea, laevia, 5.0-6.2 x

2.5-3.2 um.

Ho toryPe: China. Hainan Province, Tunchang, from a rice field soil, 1 November 2005,

Y.L. Zhang (HSAUPII,.2414, dried culture, holotype; HMAS196223, isotype).

EryMo_oey: in reference to cylindrical conidia.

Colonies on PDA effuse, powdery, glaucous to greenish black with the formation

of abundantly sporulating structures and synnemata, reverse brown, growing

slow, reaching 3-4 cm diameter at 25°C after two weeks. Mycelium mostly

superficial, composed of pale brown to brown, smooth, septate, branched,

2-2.5 um wide hyphae. Synnemata 750-900 um high, with long fusiform

heads, stipes dark brown, 15-20 um wide. Conidiogenous cells (annellophores)

ampuliform, penicillately arranged, pale brown to brown, smooth, 9-11.5 x

2.5-3.5 um. Conidia acrogenous, simple or occasionally catenate, cylindrical or

subcylindrical, rounded at the ends, brown, smooth, 5.0-6.2 x 2.5-3.2 um.

Cephalotrichum species new from China... 211

ComMENTs: Cephalotrichum cylindrosporum is similar to C. cuneiferum in

the shape of conidia, but in C. cuneiferum the conidia are longer (6-13 x 2.5-4

lum), synnemata are generally wider (10-40 um), and the conidiogenous cells

are longer (10-35 um) (Matsushima 1975).

Cephalotrichum ellipsoideum H.Q. Pan & T.Y. Zhang, sp. nov. FIG. 3

MycoBank MB 561108

Coloniae effusae, pulveraceae, griseo-brunnea, reverse brunnea, in PDA crescentes

lentissime et attingentes diametrum 2.5-4 centimetri ad 25°C post duas hebdomadas.

Mycelium plerumque superficiale, ex hyphis dilute brunneis vel brunneis, laevibus,

septatis, ramosis, 1.5-2.5 um crassis, in funiculos floccosos saepe assurgens. Synnemata

100-500 um alta, capitulo ellipsoidea et ovoidea, stipitata, atro-brunnea, 6-14 um lata.

Conidiophora brunnea, septata, ramosa. Celluae conidiogenae percurrentes (annellophora),

ampuliformes, penicilliformiter dispositae, 7.5-12.5 um longae, basi tumidulae 3-4 um

diam, quae in zonam annellatam, 1.5-3 um diam, dilute brunneae, laeves. Conidia

catenata, late ellipsoidea vel obovoidea, basin truncata et apicem rotundata, brunnea,

laevia, 6-8.5 x 3.5-6 um.

Ho.oryPe: China. Qinghai Province, Maduo, from a grassland soil, 8 June 2007, H.Q.

Pan (HSAUPII,.4053, dried culture, holotype; HMAS196224, isotype).

EryMo oy: in reference to the ellipsoidal conidia.

Fic. 3. Cephalotrichum ellipsoideum: conidia, conidiogenous cells and synnema.

212 ... Jiang & al.

Colonies on PDA effuse, powdery, greyish brown, reverse brown, growing

slow, reaching 2.5-4 cm diameter at 25°C after two weeks. Mycelium mostly

superficial, composed of pale brown to brown, smooth, septate, branched,

1.5-2.5 um wide hyphae which usually aggregate forming ropes. Synnemata

100-500 um high, with ellipsoidal to ovoid heads, stipes dark brown, 6-14

um wide. Conidiophores brown, septate, branched. Conidiogenous cells

(annellophores) ampuliform, penicillately arranged, 7.5-12.5 um long, swollen

base 3-4 um diam, tapering abruptly into the annellated zone 1.5-3 um diam,

pale brown, smooth. Conidia catenate, broadly ellipsoidal to obovoid, truncate

at the base and rounded at the apex, brown, smooth, 6-8.5 x 3.5-6 um.

ComMENtTs: The conidia of C. ellipsoideum resemble those of C. purpureo-

fuscum (Schwein.) S. Hughes (Hughes 1958), which, however, are slightly

smaller (5-7 x 3.5-4.5 um). In C. purpureofuscum the synnemata are also

differently shaped (globose or subglobose) and taller (< 900 um) (Ellis 1971).

Fic. 4. Cephalotrichum inflatum: conidia, conidiogenous cells and synnema.

Cephalotrichum species new from China... 213

Cephalotrichum inflatum Y.L. Jiang & T.Y. Zhang, sp. nov. FIG. 4

MycoBank MB 561110

Coloniae effusae, floccosae, griseo-brunneae, reverse brunneae. Mycelium partem

superficiale et partem immersum, ex hyphis hyalinis, ramosis, septatis, laevibus, 1.5-5.5

um crassis. Synnemata 160-700 ym alta, capitulo fusiformia et subcylindrica, stipitata,

atro-brunnea, 9-30 um lata. Conidiophora inflata. Celluae conidiogenae percurrentes

(annellophora), ampuliformes, penicilliformiter dispositae, 3-6.5 um longae, basi

tumidulae 2-3 um diam, quae in zonam annellatam, 1-2 um diam. Conidia solitaria vel

catenata, late obovoidea vel subellipsoidea, basin truncata, apicem rotundata vel leviter

acuta, laevia, pallide brunnea vel brunnea, 3-4.5 x 2-2.5 um.

Ho .oryPe: China. Sichuan Province, Mianyang, from a mountain soil, 8 August 2005,

Y.L. Jiang (HSAUPII,.0918, dried culture, holotype; HMAS196226, isotype).

EryMo oy: in reference to the inflated conidiophores.

Colonies on PDA effuse, at first powdery, finally floccose, greyish brown,

reverse brown. Mycelium partly superficial, partly immersed, composed of

hyaline, branched, septate, smooth, 1.5-5.5 um wide hyphae. Synnemata

160-700 um high, with fusiform or subcylindrical heads, stipes dark brown,

9-30 um wide. Conidiophores often inflated. Conidiogenous cells

(annellophores) ampuliform, penicillately arranged, 3-6.5 um long, swollen

base 2-3 um diam, tapering abruptly into the annellated zone 1-2 um diam.

Conidia solitary or occasionally catenate, broadly obovoid to subellipsoidal,

truncate at the base, rounded or slightly pointed at the apex, smooth, pale to

mid brown, 3-4.5 x 2-2.5 um.

ComMENTs: Cephalotrichum inflatum differs from all previously described

Cephalotrichum species by its distinctly inflated conidiophores.

Cephalotrichum longicollum Y.L. Jiang & T.Y. Zhang, sp. nov. FIG. 5

MycoBank MB 561111

Coloniae effusae, floccosae, fuscae, reverse brunneo-virens. Mycelium partem superficiale

et partem immersum, ex hyphis hyalinis, ramosis, septatis, laevibus, 1-3 um crassis.

Synnemata 340-750 um alta, capitulo elongato-cylindrica, stipitata, atro-brunnea, 6-25

um lata. Celluae conidiogenae percurrentes (annellophora) ampuliformes, longicollo,

penicilliformiter dispositae, 9-15.5 um longae, basi tumidulae 3-6 um diam, quae in

zonam annellatam, 1-2.5 um diam. Conidia solitaria vel catenata, ellipsoidea vel late

ellipsoidea, brunnea, basin truncata, apicem rotundata vel interdum leviter acuta, laevia,

4.5-5.5 x 2.5-4 um.

Ho oryPe: China. Sichuan Province, from a soil of Emei Mountain, 9 August 2005, Y.L.

Jiang (HSAUPII,.0802, dried culture, holotype; HMAS196228, isotype).

EryMo_oey: in reference to the annellophores with long necks.

Colonies on PDA effuse, at first powdery, finally floccose, blackish brown, reverse

greenish brown. Mycelium partly superficial, partly immersed, composed

of hyaline, branched, septate, smooth, 1-3 um wide hyphae. Synnemata

340-750 um high, with long cylindrical heads, stipes dark brown, 6-25 um wide.

214... Jiang & al.

25um

Fic. 5. Cephalotrichum longicollum: conidia, conidiogenous cells and synnema.

Conidiogenous cells (annellophores) ampuliform with long neck, penicillately

arranged, 9-15.5 um long, swollen base 3-6 um diam, tapering abruptly into

the annellated zone 1-2.5 um diam. Conidia solitary or catenate, ellipsoidal

to broadly ellipsoidal, smooth, mid brown, truncate at the base, rounded or

sometimes slightly pointed at the apex, 4.5-5.5 x 2.5-4 um.

ComMENTs: Cephalotrichum longicollum is distinguished from all previously

described Cephalotrichum species by annellophores with long neck and wider

(3-6 um) swollen base. Only C. cuneiferum (Berk. & Broome) Kuntze (Kuntze

1898) produces annellophores with a longer neck. However, shape of the

conidia distinguishes these two species (Matsushima 1975).

Cephalotrichum macrosporum Y.L. Jiang & T.Y. Zhang, sp. nov. FIG. 6

MycoBank MB 561112

Coloniae effusae, floccosae, griseo-brunneae, reverse fuscae. Mycelium partem superficiale

et partem immersum, ex hyphis hyalinis, ramosis, septatis, laevibus, 1-3 um crassis.

Synnemata 200-600 um alta, capitulo cylindrica vel elongato-cylindrica, stipitata, atro-

brunnea, 10-22 um lata. Celluae conidiogenae percurrentes (annellophora), ampuliformes,

penicilliformiter dispositae, 6.5-13 um longae, basi tumidulae 2.5-3.5 um diam, quae in

zonam annellatam, 1-2.5 um diam. Conidia solitaria vel catenata, cylindrica vel ovoidea,

Cephalotrichum species new from China... 215

longa ellipsoidea, brunnea, laevia, basin truncata, apicem rotundata, 5.5-12 x 2.5-4.5

um.

Ho .oryPe: China. Sichuan Province, from a forest soil of Jiuzhaigou, 18 August 2005,

Y.L. Jiang (HSAUPII,.0878, dried culture, holotype; HMAS196229, isotype).

EryMoLoey: in reference to the large conidia.

ab 0

25umM

Fic. 6. Cephalotrichum macrosporum: conidia, conidiogenous cells and synnema.

216... Jiang & al.

Colonies on PDA effuse, at first powdery, finally floccose, greyish brown, reverse

blackish brown. Mycelium partly superficial, partly immersed, composed of

hyaline, branched, septate, smooth, 1-3 um wide hyphae. Synnemata 200-600

um high, with cylindrical to long cylindrical heads, stipes dark brown, 10-22

um wide. Conidiogenous cells (annellophores) ampuliform, penicillately

arranged, 6.5-13 um long, swollen base 2.5-3.5 um diam, tapering abruptly into

the annellated zone 1-2.5 um diam. Conidia solitary or catenate, cylindrical to

ovoid or long ellipsoidal, mid brown, smooth, truncate at the base and rounded

at the apex, 5.5-12 x 2.5-4.5 um.

ComMENTs: Cephalotrichum cuneiferum also produces cylindrical conidia

but they are slightly larger than those of C. macrosporum. In addition, both

the annellophores and synnemata of C. macrosporum are shorter than

C. cuneiferum (Matsushima 1975).

Fic. 7. Cephalotrichum oblongum: conidia, conidiogenous cells and synnemata.

Cephalotrichum oblongum J.J. Xu & T.Y. Zhang, sp. nov. FiGs7

MycoBank MB 561113

Coloniae effusae, pulveraceae vel velutinae, griseae, brunneo-virens vel fuscae, reverse

fuscae, in PDA crescentes mediocris et attingentes diametrum 6.5 centimetri ad 25°C

post duas hebdomadas. Mycelium plerumque superficiale, ex hyphis dilute brunneis vel

brunneis, ramosis, septatis, laevibus, 2-2.5 um crassis, in funiculos floccosos interdum

assurgens. Synnemata 90-200 um alta, capitulo ellipsoidea, 10-21 um lata. Conidiophora

dilute brunnea vel atro-brunnea, laevia, septata, 2-3 um lata. Celluae conidiogenae

Cephalotrichum species new from China... 217

percurrentes (annellophora), ampuliformes, penicilliformiter dispositae, 6-10 x 2-3 um.

Conidia catenata, breve cylindrica vel oblonga, basi truncata, apicem rotundata, brunnea,

laevia, 3.5-7.5 x 2-2.5 um.

Ho toryPe: China. Yunnan Province, from a soil of Pingbian County, 11 October 2004,

J.J. Xu (HSAUPIL, 2723, dried culture, holotype; HMAS196230, isotype).

ETryMoLoey: in reference to the oblong conidia.

Colonies on PDA effuse, powdery or velvety, grey, greenish brown to blackish

brown with the formation of abundantly sporulating structures and synnemata,

reverse blackish brown, growing moderately fast, reaching 6.5 cm diameter

at 25°C after two weeks. Mycelium mostly superficial, composed of pale

brown to brown, branched, septate, smooth, 2-2.5 um wide hyphae which

may occasionally aggregate forming ropes. Synnemata 90-200 um high, with

ellipsoidal heads, 10-21 um wide. Conidiophores pale to dark brown, smooth,

septate, 2-3 tum wide. Conidiogenous cells (annellophores) ampuliform,

penicillately arranged, 6-10 x 2-3 um. Conidia catenate, short cylindrical or

oblong, truncate at the base and rounded at the apex, brown, smooth, 3.5-7.5

*2-2.5 um.

ComMENTs: Both C. purpureofuscum and C. microsporum also produce

oblong conidia, which, however, differ in size. In C. oblongum annellophores

are narrower than those of C. purpureofuscum and synnemata are shorter than

those of C. purpureofuscum and C. microsporum (Morton & Smith 1963).

Cephalotrichum ovoideum Y.L. Jiang & T.Y. Zhang, sp. nov. Fic. 8

MycoBank MB 561115

Coloniae effusae, floccosae, fuscae, reverse flavo-brunneae. Mycelium partem superficiale

et partem immersum, ex hyphis hyalinis, ramosis, septatis, laevibus, 1-2.5 um crassis.

Synnemata 250-800 um alta, ramaso, capitulo sphaerica vel subsphaerica, stipitata, atro-

brunnea, 9-25 um lata. Celluae conidiogenae percurrentes (annellophora), ampuliformes

vel obclavatae, penicilliformiter dispositae, 6-13 um longae, basi tumidulae 2.5-3.7 um

diam, quae in zonam annellatam, 1.5-2 um diam. Conidia solitaria vel catenata, dilute

flavo-brunnea vel flavo-brunnea, ovoidea, obpyriformia vel subsphaerica, apicem acuta

vel interdum rotundata, basin truncata, laevia, 4.5-7 x 3-4 um.

Ho .oryPe: China. Sichuan Province, Jiuzhaigou, from a forest soil, 18 August 2005, Y.L.

Jiang (HSAUPII,.0846, dried culture, holotype; HMAS196231, isotype).

ETYMOLOGy: in reference to the ovoid conidia.

Colonies on PDA effuse, at first powdery, finally floccose, blackish brown, reverse

yellowish brown. Mycelium partly superficial, partly immersed, composed

of hyaline, branched, septate, smooth, 1-2.5 um wide hyphae. Synnemata

250-800 um high, branched with spherical or subspherical heads, stipes dark

brown, 9-25 um wide. Conidiogenous cells (annellophores) ampuliform to

obclavate, penicillately arranged, 6-13 um long, swollen base 2.5-3.7 um diam,

tapering abruptly into the annellated zone 1.5-2 um diam. Conidia solitary

218 ... Jiang & al.

Fic. 8. Cephalotrichum ovoideum: conidia, conidiogenous cells and synnemata.

or catenate, pale yellowish brown to yellowish brown, ovoid, obpyriform or

subspherical, usually pointed at the apex or sometimes rounded, truncate at the

base, smooth, 4.5-7 x 3-4 um.

ComMENTSs: Cephalotrichum ovoideum resembles C. purpureofuscum and

C. byssoides in conidial morphology, but the conidia of C. purpureofuscum

are usually rounded at the apex and its synnemata are unbranched. Conidial

ornamentation and size clearly distinguish C. ovoideum and C. byssoides

(Morton & Smith 1963, Mason & Ellis 1953).

Cephalotrichum robustum Y.L. Jiang & T.Y. Zhang, sp. nov. FIG. 9

MycoBank MB 561116

Coloniae effusae, floccosae, brunneo-virens, reverse flavo-brunnea. Mycelium partem

superficiale et partem immersum, ex hyphis hyalinis, ramosis, septatis, laevibus, 1-3

um crassis. Synnemata 250-750 um alta, capitulo cylindrica et elongato-ellipsoidea,

stipitata, atro-brunnea, 6-25 um lata. Celluae conidiogenae percurrentes (annellophora),

subcylindricae, robustae, penicilliformiter dispositae, 4-8 um longae, basi tumidulae

2-4 um diam, zonam annellatam 1.5-2.5 um diam. Conidia solitaria vel catenata, late

ellipsoidea vel subsphaerica, basin truncata, apicem rotundata, flavo-brunnea, laevia, 5-7

x 3-5 um.

Cephalotrichum species new from China ... 219

25mm

Fic. 9. Cephalotrichum robustum: conidia, conidiogenous cells and synnema.

Ho.ortyPe: China. Sichuan Province, from a forest soil of Jiuzhaigou, 18 August 2005,

Y.L. Jiang (HSAUPII,.0875, dried culture, holotype; HMAS196232, isotype).

EryMo oy: in reference to the thickset annellophores.

Colonies on PDA effuse, at first powdery, finally floccose, greenish brown, reverse

yellowish brown. Mycelium partly superficial, partly immersed, composed of

hyaline, branched, septate, smooth, 1-3 um wide hyphae. Synnemata 250-750

um high, with cylindrical to long ellipsoidal heads, stipes dark brown, 6-25 um

wide. Conidiogenous cells (annellophores) subcylindrical, thickset, penicillately

arranged, 4-8 um long, swollen base 2-4 um diam, the annellated zone 1.5-2.5

um diam. Conidia solitary or catenate, broadly ellipsoidal to subspherical,

truncate at the base and rounded at the apex, yellowish brown, smooth, 5-7 x

3-5) lm,

220 ... Jiang & al.

ComMENTs: Cephalotrichum robustum morphologically resembles

C. ellipsoideum, C. longicollum, C. castaneum (Jiang & Zhang 2008), and

C. purpureofuscum. The annellophores of C. robustum are the shortest among

these five species. In addition, synnemata of C. robustum are shorter than in

C. purpureofuscum and C. castaneum, longer than in C. ellipsoideum (Ellis

1971), and about the same length as in C. longicollum. However, the conidia of

C. robustum are bigger than those of C. longicollum.

25um

Fic. 10. Cephalotrichum spirale: conidia, conidiogenous cells and synnema.

Cephalotrichum spirale H.M. Liu, H.Q. Pan & T.Y. Zhang, sp. nov. FIG. 10

MycoBank MB 561117

Coloniae effusae, floccosae, atrogriseae vel atrae, in PDA crescentes lentissime et attingentes

diametrum 4 centimetri ad 25°C post duas hebdomadas. Mycelium plerumque superficial,

ex hyphis hyalinis, ramosis, septatis, laevibus, 1-2.5 um crassis. Synnemata 700 um

alta, capitulo ellipsoidea et cylindrica, stipitata, atro-brunnea, 10-15 um lata. Celluae

Cephalotrichum species new from China... 221

conidiogenae percurrentes (annellophora), ampuliformes vel obclavatae, penicilliformiter

dispositae, 6-12 um longae, basi tumidulae 2-3.5 um diam, quae in zonam annellatam,

1-2 um diam. Conidia solitaria vel catenata, brunnea, late obovoidea vel late ellipsoidea,

basin truncata et apicem rotundata, spiraliter disposita verruculosa, 5-7 x 3-4.5 um, in

universum fusca.

Ho.otype: China. Qinghai Province, from a grassland soil of Dari County, 12 June

2007, H.Q. Pan (HSAUPII, 4033, dried culture, holotype; HMAS196233, isotype).

EryMo oy: in reference to the verruculose conidia with verrucae in a spiral.

Colonies on PDA effuse, floccose, central sporulating area at first pale grey, the

grey colour spreading and darkening to blackish, finally dark grey to black with

the formation of abundantly sporulating structures and synnemata, growing

slow, reaching 4 cm diameter at 25°C after two weeks. Mycelium mostly

superficial, composed of hyaline, branched, septate, smooth, 1-2.5 um wide

hyphae. Synnemata 700 um high, with ellipsoidal to cylindrical heads, stipes

dark brown, 10-15 um wide. Conidiogenous cells (annellophores) ampuliform

to obclavate, penicillately arranged, 6-12 um long, swollen base 2-3.5 um diam,

tapering abruptly into the annellated zone 1-2 um diam. Conidia solitary or

catenate, brown, broadly obovoid to broadly ellipsoidal, truncate at the base

and rounded at the apex, verruculose, verrucae arranged in a spiral, 5-7 x

3-4.5 um, blackish brown in mass.

ComMENTs: Based on the presence of ornamented conidia, C. spirale

is similar to C. stemonitis, C. byssoides, C. phillipsii, C. nanum (Ehrenb.)

S. Hughes (Hughes 1958), and C. verrucisporum (Jiang & Zhang 2008). However,

conidial shape distinguishes C. spirale from the other five species. In addition,

synnemata of C. spirale are longer than in C. phillipsii and shorter than in

C. stemonitis, C. nanum, C. byssoides, and C. verrucisporum (Mason & Ellis

1953, Morton & Smith 1963, Jiang & Zhang 2008).

Cephalotrichum terricola Y.L. Jiang & T.Y. Zhang, sp. nov. FIG. 11

MycoBank MB 561118

Coloniae effusae, floccosae, brunneo-virens, reverse fuscae. Mycelium partem superficiale

et partem immersum, ex hyphis hyalinis, ramosis, septatis, laevibus, 1-3 um crassis.

Synnemata 300-660 um alta, capitulo fusiformia vel elongato-cylindrica, stipitata, atro-

brunnea, 9-19 um lata. Celluae conidiogenae percurrentes (annellophora), ampuliformae,

penicilliformiter dispositae, 4.5-9.5 um longae, basi tumidulae 2-4.5 um diam, quae in

zonam annellatam, 1-2 um diam. Conidia solitaria vel catenata, obovoidea vel ellipsoidea,

flavo-brunnea, laevia, basin truncata, apicem rotundata vel interdum leviter acuta,

4.5-7x2.5-3.5 um.

Ho .oryPe: China. Sichuan Province, Panzhihua, from a mountain soil, China. 13 August

2005, Y.L. Jiang (HSAUPII,.0924, dried culture, holotype; HMAS196227, isotype).

ETYMOLOGY: in reference to the habitat.

Colonies on PDA effuse, at first powdery, finally floccose, greenish brown,

reverse blackish brown. Mycelium partly superficial, partly immersed,

222 ... Jiang & al.

Fic. 11. Cephalotrichum terricola: conidia, conidiogenous cells and synnema.

composed of hyaline, branched, septate, smooth, 1-3 um wide hyphae.

Synnemata 300-660 um high, with fusiform to long cylindrical heads, stipes

dark brown, 9-19 um wide. Conidiogenous cells (annellophores) ampuliform,

penicillately arranged, 4.5-9.5 um long, swollen base 2-4.5 um diam, tapering

abruptly into the annellated zone 1-2 um diam. Conidia solitary or catenate,

obovoid to ellipsoidal, yellowish brown, smooth, truncate at the base, rounded

or sometimes slightly pointed at the apex, 4.5-7 x 2.5-3.5 um.

Cephalotrichum species new from China... 223

ComMENts: The species most similar to C. terricola in conidial morphology

are C. ellipsoideum and C. purpureofuscum, except that those species produce

wider conidia. Additionally, synnemata of C. purpureofuscum are longer (< 900

um) with spherical or subspherical heads and those of C. ellipsoideum shorter

(100-500 um) with ellipsoidal to ovoid heads.

Cephalotrichum verrucipes Y.L. Jiang & T.Y. Zhang, sp. nov. FIG. 12

MycoBank MB 561119

Coloniae effusae, floccosae, brunneo-virens, reverse fuscae. Mycelium partem superficiale

et partem immersum, ex hyphis hyalinis, ramosis, septatis, laevibus, 2-4.5 um crassis.

Synnemata 2800 um alta, capitulo cylindrica vel ellipsoidea, stipitata, dilute brunnea,

verruculosa, 25-66 ym lata. Conidiophora verruculosa. Celluae conidiogenae percurrentes

(annellophora), longa ampuliformae vel obclavata, penicilliformiter disposita, 7-14 um

longa, basi leviter tumidula 1-2.5 um diam, quae in zonam annellatam, 1-2 um diam.

Conidia solitaria vel catenata, sphaerica, subsphaerica vel ellipsoidea, laevia, dilute flavo-

brunnea vel flavo-brunnea, 2-3.5 x 2-3 um, in universum atro-brunnea.

Ho.ortyPe: China. Sichuan Province, from a forest soil of Jiuzhaigou, 19 August 2005,

Y.L. Jiang (HSAUPII,.0849, dried culture, holotype; HMAS196234, isotype).

EryMo oy: in reference to the verruculose conidiophores.

eo \

0&9

QAQ9000

SHORVO

25um

25himM

Fic. 12. Cephalotrichum verrucipes: conidia, conidiogenous cells and synnema.

224 ... Jiang & al.

Colonies on PDA effuse, at first powdery, finally floccose, greenish brown,

reverse blackish brown. Mycelium partly superficial, partly immersed,

composed of hyaline, branched, septate, smooth, 2-4.5 um wide hyphae.

Synnemata 2800 um high, with cylindrical vel ellipsoidal heads, stipes pale

brown, verruculose, 25-66 um wide, stipes distinct, but density of branches

of stipes nonuniform, disperse branches often in the middle part or below the

stipes. Conidiophores verruculose. Conidiogenous cells (annellophores) long

ampuliform to obclavate, penicillately arranged, 7-14 um long, slightly swollen

base 1-2.5 um diam, tapering abruptly into the annellated zone 1-2 um diam.

Conidia solitary or catenate, spherical, subspherical to ellipsoidal, smooth, pale

yellowish brown to yellowish brown, 2-3.5 x 2-3 um, dark brown in mass.

ComMENTSs: Cephalotrichum verrucipes is the only species in the genus with

verruculose conidiophores.

New combinations

The following two species, previously placed in Doratomyces, both produce

spores in a dry head at the apex of a complex, erect condiophore synnema. For

that reason we transfer them to Cephalotrichum.

Cephalotrichum castaneum (YL. Jiang & T.Y. Zhang) Y.L. Jiang & T.Y. Zhang,

comb. nov.

MycoBank MB 561107

= Doratomyces castaneus Y.L. Jiang & T.Y. Zhang, Mycotaxon 104: 131, 2008.

Cephalotrichum verrucisporum (Y.L. Jiang & T.Y. Zhang) Y.L. Jiang & T.Y. Zhang,

comb. nov.

MycoBank MB 561120

= Doratomyces verrucisporus Y.L. Jiang & T.Y. Zhang, Mycotaxon 104: 133, 2008.

Acknowledgments

The authors are grateful for pre-submission comments and suggestions provided by

Dr. E.H.C. McKenzie and Prof. Y.R. Lin. This project was supported by the National

Science Foundation of China (no.30970011).

Literature cited

Arx JA von. 1981. The genera of fungi sporulating in pure culture. 3" edition. J. Cramer, Vaduz.

424 p.

Chlebicki A. 2008. Cephalotrichum stemonitis as a biofilm inhabitant in the gold mine in Poland.

Acta Mycologica 43(1): 67-70.

Corda ACJ. 1829. Deutschlands Flora, Abt. III. Die Pilze Deutschlands. 2(7): 37-70.

Domsch KH, Gams W, Anderson TH. 2007. Compendium of soil fungi. 2" edition. IHW- Verlag,

Eching. 672 p.

Ellis MB. 1971. Dematiaceous hyphomycetes. X. Mycological Papers 125. 30 p.

Hughes SJ. 1958. Revisiones Hyphomycetum aliquot cum appendice de nominibus rejiciendis.

Canadian Journal of Botany 36: 727-836. http://dx.doi.org/10.1139/b58-067

Cephalotrichum species new from China... 225

Jiang YL, Zhang TY. 2007. Notes on soil dematiaceous hyphomycetes from Shennongjia Natural

Conservation Area, Hubei Province I. Mycosystema 26: 17-21. http://dx.doi.org/CNKI:SUN:

JWXT. 0.2007-01-004

Jiang YL, Zhang TY. 2008. Two new species of Doratomyces from soil. Mycotaxon 104: 131-134.

Kirk PM, Spooner BM. 1984. An account of the fungi of Arran, Gigha and Kintyre. Kew Bulletin

38: 503-597.

Kirk PM, Cannon PF, David JC, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the

fungi, 10" edition. CABI Bioscience, Centre, Egham, UK. http://dx.doi.org/10.2307/4108573

Kuntze O. 1898. Revisio generum plantarum 3(3). 576 p.

Link HE. 1809. Observationes in ordines plantarum naturales. Dissertatio Ima. Magazin Gesellschaft

naturforschenden Freunde Berlin 3: 3-42.

Mason EW, Ellis MB. 1953. British species of Periconia. Mycological Papers 56. 127 p.

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Published by the author,

Kobe, Japan. 209 p.

Morton FJ, Smith G. 1963. The genera Scopulariopsis Bainier, Microascus Zukal, and Doratomyces

Corda. Mycological Papers 86. 96 p.

MY COTAXON

http://dx.doi.org/10.5248/117.227

Volume 117, pp. 227-237 July-September 2011

Morphology and phylogeny of Pseudocercospora kamalii sp. nov.,

a foliar pathogen on Terminalia from India

KUNHIRAMAN C. RAJESHKUMAR’, RAHUL SHARMA, RAHUL P. HEPAT,

SANTOSH V. SWAMI, PARAS NATH SINGH & SANJAY K. SINGH?

National Facility for Culture Collection of Fungi, MACS Agharkar Research Institute,

G.G. Agarkar Road, Pune, India

CORRESPONDENCE TO: ‘rajeshfungi@gmail.com & ’singhsksingh@rediffmail.com

ABSTRACT — Pseudocercospora kamalii (Mycosphaerellaceae, Capnodiales) is associated

with severe leaf spot disease of Terminalia chebula (Combretaceae) in natural forests of

Mahabaleshwar in the Western Ghats of India. Morphological characterization reveals that

the new species proposed has some affinity with the species of genus Prathigada. However,

based on the molecular sequence data (LSU and ITS 1-5.8S-ITS 2) generated in the present

study, we established its identity as a species of Pseudocercospora. The species is also compared

morphologically with other Pseudocercospora species reported on Terminalia from different

parts of the world.

Key worpDs — anamorphic fungi, Mycosphaerella, plant pathogen

Introduction

The microfungal diversity of northern Western Ghats has been well

explored in the recent past (Karandikar & Singh 2010, Rajeshkumar et al. 2010,

2011; Singh et al. 2009, 2010; Waingankar et al. 2008). During January 2011

a survey was conducted to explore the microfungal diversity in the natural

forests of Mahabaleshwar, situated in the northern part of the Western Ghats,

India, at 17°58'N 73°43'E. An unusual synnematous cercosporoid species was

discovered that caused a severe foliar disease in Terminalia chebula. The present

study aimed to identify the causal agent of the foliar infection on Terminalia in

northern Western Ghats, India using morphological and molecular methods.

Materials & methods

ISOLATES AND MORPHOLOGY— Synnemata of the fungus were directly isolated from

the surface of fallen fruits and observed under a Nikon Binocular stereo microscope

(Model SMZ-1500 with Digi-CAM, Japan). Single conidial cultures were established

228 ... Rajeshkumar & al.

on 2% potato dextrose agar plates (PDA; Crous et al. 2009a). For morphotaxonomic

studies and photomicrographs an Olympus (Model CX-41, Japan) microscope was

used. Conidia and conidiomata were mounted in lactic acid cotton blue and measured

using an ocular micrometer, with 30 observations per structure. Colony characteristics

in culture were studied on two different media: 2% malt extract agar (MEA) and PDA

(Crous et al. 2009a). Herbarium specimens were deposited in the Ajrekar Mycological

Herbarium (AMH); the culture was accessioned and preserved in the National Fungal

Culture Collection of India (NFCCI; WDCM-932), Agharkar Research Institute, Pune,

India under accession number NFCCI 2344.

POLYMERASE CHAIN REACTION (PCR) AND SEQUENCING— Total DNA was extracted

from cultures grown on PDA plates for two weeks at 25 °C, using a FastDNA°® SPIN kit

as per the manufacturer's instructions (MP Biomedicals GmbH, Germany). Fragments

containing the region encoding the 28S nrDNA (LSU) and ITS 1-5.8S nrDNA-ITS

2 (ITS) were amplified using primer pairs LROR (Rehner & Samuels 1994) and LR7

(Vilgalys & Hester 1990) for LSU; ITS4 and ITS5 (White et al. 1990) for ITS. DNA

amplification was performed in a 25 ul reaction using 2 ul of template DNA (10-25

ng), 0.5 U of Taq DNA polymerase (Genei, Bangalore, India), 2.5 ul of 10x Taq DNA

polymerase buffer, 0.5 ul of 200 uM of each dNTPs (Genei, Bangalore, India), 0.5 ul of

10 pmol primer, H,O (Sterile Ultra Pure Water, Sigma) to make up 25 ul. Amplification

in an Eppendorf Mastercycler AG used the following parameters: 5 min at 95 °C; 30

cycles of 1 min at 95 °C, 30s at 56 °C, and 1 min at 72 °C for the ITS region amplification;

and final 7-min extension step at 72 °C. DNA amplification of LSU followed the ITS

conditions except for a 52 °C annealing temperature. The PCR products were purified

with an Axygen PCR cleanup kit (Axygen Scientific Inc, CA, USA) and sequenced with

the same primers using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied

Biosystems, USA). The sequencing reactions were run on an ABI 3100 automated DNA

sequencer (Applied Biosystems, USA).

SEQUENCE ALIGNMENT AND PHYLOGENETIC ANALYSIS— LSU & ITS sequences from

P. kamaliiwere aligned manually using the text editor option of the Molecular Evolutionary

Genetics Analysis (MEGA) software v4.0. (Tamura et al. 2007). The manually edited

NFCCI 2344 sequences were deposited in the NCBI sequence nucleotide database (ITS:

JF 824126, LSU: JF 824127). They were also subjected to a BLAST search of the NCBI

Genbank nucleotide database. The ITS sequences were aligned using Clustal W together

with the homologous regions of ITS of closely related species of Pseudocercospora Speg.

and Mycosphaerella s.l. For ITS, the matrix was analyzed with the Maximum Parsimony

Fic. 1. Phylogenetic tree based on aligned LSU sequences of Pseudocercospora and its teleomorph,

Mycosphaerella s.1. The consistency index (0.566343), the retention index (0.821571), and the

composite index (0.479675) were calculated in MEGA for all sites and parsimony-informative sites

(in parentheses). The percentage of replicate trees in which the associated taxa clustered together

in the bootstrap test (1000 replicates) is shown next to the branches. The MP tree was obtained

using the Close-Neighbor-Interchange algorithm with search level 3 in which the initial trees were

obtained with the random addition of sequences (10 replicates). The tree is drawn to scale, with

branch lengths calculated using the average pathway method and are in the units of the number of

changes over the whole sequence. All positions containing gaps and missing data were eliminated

from the dataset (Complete Deletion option).

Pseudocercospora kamalii sp. nov. (India) ...

GU214673.1 Mycosphaerella cruenta

DQ204761.1 Pseudocercospora basiramifera

GU214478.1 Pseudocercospora macrospora

GU214672.1 Pseudocercospora chengtuensis

DQ204764.1 Pseudocercospora paraguayensis

GU214479.1 Pseudocercospora paraguayensis

GU214675.1 Pseudocercospora fuligena

72| GU214472.1 Pseudocercospora cordiana

AY875385.1 Mycosphaerella musicola

AY875387.1 Mycosphaerella musicola

82 AY875386.1 Mycosphaerella musicola

GU214680.1 Pseudocercospora pallida

AY875401.1 Mycosphaerella eumusae

? ! AY875400.1 Mycosphaerella eumusae

GU214671.1 Pseudocercospora atromarginalis

GQ852652.1 Pseudocercospora sphaerulinae

gg | 2Q204759.1 Pseudocercospora basitruncata

DQ204748.1 Mycosphaerella fori (Pseudocercospora fori)

GQ852631.1 Pseudocercospora crousil

GQ852630.1 Mycosphaerella bixae (Pseudocercospora bixae)

DQ204767.1 Pseudocercospora robusta

DQ204762.1 Pseudocercospora eucalyptorum

GQ852636.1 Pseudocercospora pseudoeucalyptorum

99 - GQ852596.1 Dothistroma pini

GQ852597.1 Mycosphaerella pini

EU710897.1 Ramularia pusilla

DQ885902.1 Ramularia miae

DQ470968.1 Mycosphaerella punctiformis

DQ267574.1 Mycosphaerella walkeri (Sonderhenia eucalyptorum)

99 © GQ852679.1 Mycosphaerella walkeri (Sonderhenia eucalyptorum)

GU214658.1 Cercosporella virgaureae

99 GQ852653.1 Ramulispora sorghi

GQ852654.1 Ramulispora sorghi

GU214443.1 Mycosphaerella latebrosa (Septoria aceris)

GU214490.1 Septoria apiicola

GQ852675.1 Septoria convolvuli

GQ852676.1 Septoria cucubali

DQ678091.1 Cercospora beticola

95! GQ852583.1 Cercospora apii

GU214663.1 Mycosphaerella dearnessii (Lecanosticta acicola)

ie F.J493206.1 Phaeophleospora eugeniae

99! FJ493207.1 Phaeophleospora eugeniae

EU041878.1 Zasmidium cellare

on CN Zasmigium cir

EU167610.1 Cymadothea trifolii(Polythrincium trifoli

99 'EU167611.1 Cymadothea trifoli (Polythrincium tnfolii)

DQ377868.1 Botryosphaeria visci

99 DQ377858.1 Botryosphaeria rhodina

JF824127 Pseudocercospora kamalii NFCCI 2344

Pseudocercospora

Dothistroma

Ramularia/Mycosphaerella s. str

Sonderhenia

‘| Cercosporella

Ramulispora

Septoria

Cercospora

‘] Lecanosticta

Phaeophleospora

Zasmidium

Polythrincium

229

aeaoeya1aeydsoo“, Ww

230 ... Rajeshkumar & al.

method using the Tamura model (Tamura et al. 2007) to calculate the sequence

divergence, and the bootstrap consensus tree inferred from 1000 replicates is taken to

represent the evolutionary history of the taxa analyzed. All positions containing gaps and

missing data were eliminated from the dataset (Complete Deletion option in MEGA4).

Results

DNA phylogeny

The alignment for the Pseudocercospora LSU phylogenetic analyses

comprised a total of 481 positions in the final dataset, of which 128 were

parsimony informative. The LSU sequence analysis presented here (Fic. 1)

reveals the systematic position of the new species within Pseudocercospora,

along with its 11 closely allied sister genera in Mycosphaerellaceae. The LSU

phylogenetic sequence analysis implied 12 main clades: Pseudocercospora,

Dothistroma Hulbary, Ramularia Unger/Mycosphaerella s. str., Sonderhenia

H.J. Swart & J. Walker, Cercosporella Sacc., Ramulispora Miura, Septoria Sacc.,

Cercospora Fresen., Lecanosticta Syd., Phaeophleospora Rangel, Zasmidium

Fr, and Polythrincium Kunze. All Pseudocercospora species clustered

together to form a major clade. The new species, P kamalii, clustered close to

Mycosphaerella musicola R. Leach ex J.L. Mulder in the major Pseudocercospora

clade. Botryosphaeria visci (Kalchbr.) Arx & E. Mull. (DQ 377868.1) and

B. rhodina (Berk. & M.A. Curtis) Arx (DQ 377858.1) in the Botryosphaeriaceae

served as outgroup taxa.

For the phylogenetic analysis of closely related Pseudocercospora species,

the ITS sequence data alignment of 487 bp included 452 positions in the final

data set. The analysis of ITS sequence shown in Fic. 2 reveals a significant

association of Pseudocercospora representatives, some with Mycosphaerella-

like teleomorphs. The ITS sequence analysis implied four major clades, with

Pseudocercospora eumusae Crous & Mour. (= Mycosphaerella eumusae Crous &

Mout.) forming a unique clade (94% bootstrap support). However, P tereticornis

Crous & Carnegie, P cruenta (Sacc.) Deighton, P. pallida (Ellis & Everh.)

H.D. Shin & U. Braun, P. casuarinae Crous & R.G. Shivas, P. elaeodendri (G.P.

Agarwal & Hasija) Deighton, P fuligena (Roldan) Deighton, P. chengtuensis

(EL. Tai) Deighton, and P atromarginalis (G.E Atk.) Deighton clustered

together to form a major group in the Mycosphaerellaceae. Pseudocercospora

musae (Zimm.) Deighton (= Mycosphaerella musicola) also formed a unique

clade (100% bootstrap support). Our new species, P kamalii, (66% bootstrap

support), separated as sister to the Pseudocercospora musae (AY 646475.1 & AY

646474.1) clade. Cladosporium herbarum (Pers.) Link (= Davidiella tassiana

(De Not.) Crous & U. Braun) (DQ 289799.2) was chosen as the outgroup, as

it belongs to the family Davidiellaceae that is allied to the Mycosphaerellaceae

(Crous et al. 2006).

Pseudocercospora kamalii sp. nov. (India) ... 231

GU168036.1 Mycosphaerella eumusae

GU168031.1 Mycosphaerella eumusae

GU168024.1 Mycosphaerella eumusae

AY923758.1 Mycosphaerella eumusae

GU168026.1 Mycosphaerella eumusae

AY923757.1 Mycosphaerella eumusae

GU168028.1 Mycosphaerella eumusae

AY923760.1 Mycosphaerella eumusae

AY923759.1 Mycosphaerella eumusae

GQ852769.1 Pseudocercospora tereticonis

GU214673.1 Mycosphaerella cruenta

GU214680.1 Pseudocercospora pallida

af 57 | HQ599603.1 Pseudocercospora casuarinae

30 GU980950.1 Pseudocercospora elaeodendri

GU060636.1 Pseudocercospora fuligena

GU214672.1 Pseudocercospora chengtuensis

56] GU214675.1 Pseudocercospora fuligena

GU214671.1 Pseudocercospora atromarginalis

JF824126 Pseudocercospora kamalii NFCCI 2344

66 AY646475.1 Mycosphaerella musicola

100! AY646474.1 Mycosphaerella musicola

DQ289799.2 Davidiella tassiana

aaa

0.02

Fic. 2. Phylogenetic tree based on aligned ITS sequences of Pseudocercospora and its teleomorph,

Mycosphaerella sl. and inferred using the Neighbor-Joining method in MEGA. The optimal

tree with the sum of branch length = 0.25106088 is shown. The percentage of replicate trees in

which the associated taxa clustered together in the bootstrap test (1000 replicates) is shown next

to the branches. The tree is drawn to scale, with branch lengths in the same units as those of

the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were

computed using the Maximum Composite Likelihood method and are in the units of the number

of base substitutions per site. All positions containing gaps and missing data were eliminated from

the dataset (Complete Deletion option).

Taxonomy

The new cercosporoid species on Terminalia chebula is easily distinguishable

from all Pseudocercospora species hitherto described on Terminalia by its

synnematous conidiomata, and hence it is described here as new species.

232 ... Rajeshkumar & al.

Pseudocercospora kamalii Rajeshkumar, Rahul Sharma & S.K. Singh, sp. nov.

MycoBank MB 561201 FIGS. 3-4

Synnemata 200-337.5 um longa, ex stromatibus oriunda. Stromata bene evoluta, globosa,

subglobosa vel irregularia, atrobrunnea. 50-72 um diam., ex cellulis oblongis, cylindraceis

composita. Conidiophora 130-263 x 5-6 um, modice brunnea vel atrobrunnea,

crassitunicata, pluriseptata. Conidia solitaria, sicca, in multitudine atro-brunnea, 25-57.5

x 5.5-8 um, laevia, raro verruculosa, crassitunicata, 2-7 septis crassis, basi truncate.

Type: India, Mahabaleshwar, Western Ghats, Maharashtra, on leaves of Terminalia

chebula Retz. (Combretaceae), January 2011, K.C. Rajeshkumar & Rahul Sharma (AMH

9425, holotype; ex-type culture NFCCI 2344.)

EryMo.ocy: kamalii, named in honour of Prof. Dr. Kamal, Emeritus Professor,

Department of Botany, D.D.U. Gorkhpur University for his major contribution to the

cercosporoid fungi of India.

LEAF SPOTS necrotic, amphigenous, circular, angular or irregular, forming

concentric dark and pale brown patterns on the spots, spreading eventually

covering most of the leaf, but not vein limited, 1-3 cm diam. CONIDIOMATA

synnematous and caespituli either exclusively hypogenous or amphigenous,

synnemata blackish brown 200-337.5 um long, arising from well developed

stroma, globose, subglobose or irregular, dark brown or blackish brown, 50-72

um diam, formed of elongated cylindrical cells. ContDIOPHORES medium to

dark brown, thick and dark-walled, formed as a continuation of stromatic cells,

130-263 um long and 5-6 um wide, 8-10 to multiseptate. CONIDIOGENOUS

CELLS integrated, terminal, cylindrical or sometime doliiform, geniculate,

slightly wider and paler towards truncate apex, which at times can be obtusely

rounded; conidial scars 1-2 in number, 2-3 um diam, slightly thicker and dark,

conidial loci not usually protuberant. Conrp1a solitary, holoblastic, dry, dark

brown to blackish brown in mass, pale brown when immature, later turning to

medium to dark brown, 25-57.5 x 5.5-8 um, straight or slightly curved, mostly

smooth, rarely minutely verruculose, thick-walled, thick septate, 2-7-septate,

conidial wall and septations are thicker and darker in the basal part, and pale

and thin towards apex, conidial base truncate, 2.0-2.2 um, and apex obtuse or

subacute 2.5-3.5 um

TELEOMORPH: not observed.

Cotonigs on MEA (Fic. 5) very slow growing, 0.5 to 0.8 mm diam after 7

days, grayish white or white initially, later turning dark grayish black to blackish

brown, velutinous, reverse blackish to dark blackish brown. Colonies become

2 to 2.3 mm after 30 days, forming a sulcate pattern, and sectored on MEA.

Fic. 3. Pseudocercospora kamalii (holotype). a. Habit. b. Symptoms on leaves and synnemata.

c-d. Synnemata side view. e. Synnemata top view. f-h. Conidioma with stroma. i. Well developed

stroma. j. Conidiophore tip and conidial scar. k-m. Conidia with thick wall and thick septations.

Bars: k, 1, m = 25 um.

293

Pseudocercospora kamalii sp. nov. (India) ...

234 ... Rajeshkumar & al.

Fig. 4. Pseudocercospora kamalii. (holotype). a. Conidiophore and conidial development.

b. Conidia. c. Synnemata

Culture on MEA not sporulating (lacking conidial development) but forming

distinct synnemata-like bunches of conidiophores and dark celled stroma-like

structures. The hyphae are medium to dark brown, highly verruculose and

interwoven, forming a thick mat.

Pseudocercospora kamalii sp. nov. (India) ... 235

Fic. 5. Pseudocercospora kamalii. (holotype) Ex-type culture on MEA. a. Top view b. Reverse view.

Discussion

The new species, Pseudocercospora kamalii, is morphologically quite

unusual and obscure within Pseudocercospora, as it exhibits many characters

that match the genus Prathigada. Among these, the thick conidial septations

and wall pattern are mostly identical with species of Prathigada. Only a few

cercosporoid species with similar morphological characters (thick-walled,

multi-septate, pigmented conidia containing slightly thickened and darkened

scars) have been transferred to Prathigada (Braun 1996; Furlanetto & Dianese

1999; Sutton 1994), most being retained in Pseudocercospora. ‘Thus far, 15

species are recorded under the genus Prathigada. Initial morphological studies

also made it evident that Pseudocercospora kamalii is morphologically close to

Prathigada terminaliae (Syd.) B. Sutton recorded on Terminalia spp. (Sutton

1994). However, P. kamalii has synnematous conidiomata, smaller conidia, and

fewer conidial septa. Furthermore, the LSU and ITS sequence analyses accurately

defined the species boundaries and placed P. kamalii into Pseudocercospora.

Crous et al. (2009b) recently stated that due to the unavailability of cultures,

no decision could yet be made on the phylogenetic placement of several less

well-known genera such as Prathigada Subram. within the Mycosphaerellaceae.

A recollection, epitypification and detailed phylogenetic analysis of the type

species, Prathigada cratevae (Syd.) Subram. on Crateva religiosa G. Forst.

from India, is needed to reveal the exact relation of the genus Prathigada with

Pseudocercospora and their placement in Mycosphaerellaceae.

Sutton (1994) also reported six Pseudocercospora taxa on Terminalia

spp. from India: P arjunae B. Sutton (on T! arjuna), P. brevis B. Sutton (on

T. bellerica), P catappae (Henn.) Y.L. Guo & XJ. Liu (on T. catappa,

T: tomentosa, T. arjuna), P. chebulae B. Sutton (on T. chebula), P. combretacearum

236 ... Rajeshkumar & al.

R.K. Verma & Kamal var. combretacearum (on T: bellerica, Terminalia sp.), and

P. combretacearum var. minima B. Sutton (on T. bellerica); he also reported two

other species: P. neodeightonii B. Sutton (on T! albida from Sierra Leone) and

P. zambiensis (Deighton) B. Sutton (on T. mollis from Zambia). Kamal (2010:

347) listed P. catappae, P. chebulae, and P. combretacearum var. combetacearum

from Indian T: chebula. These three species differ from P. kamalii in various

morphological characters; most conspicuously, they have mononematous,

fasciculate, or sporodochial conidiophores in contrast with the synnematous

conidiophores of P. kamalii, which is the only synnematous Pseudocercospora

species recorded on Terminalia.

In the present study, P kamalii is proposed as a new species based on the

LSU & ITS sequence analysis (Fic. 1, 2) and morphological characterization

(Fic. 3, 4). Phylogenetically, P kamalii is closely related to Pseudocercospora

musae (66% bootstrap support) on Musa spp. (Arzanlou et al. 2008, Braun et

al. 1999). With its distinct synnemata with a well-developed basal stroma and

larger conidiophores and conidia, P. kamalii is morphologically distinct from

Pseudocercospora musae and all other taxa with higher phylogenetic similarity.

Acknowledgements

We are indebted to Pedro W. Crous and Johannes Zacharias Groenewald

(Centraalbureau voor Schimmelcultures, Fungal Biodiversity Centre, Utrecht,

The Netherlands) and Uwe Braun (Martin-Luther-University, Institute of Biology,

Department of Geobotany and Botanical Garden, Herbarium, Halle (Saale), Germany)

for reviewing this manuscript. Thanks are also due to Department of Science and

Technology (DST), Government of India, New Delhi for providing financial support for

setting up the National Facility for Culture Collection of Fungi (No. SP/SO/PS-55/2005)

at MACS’Agharkar Research Institute, Pune, India and the Director, MACS'ARI for

providing facility.

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MY COTAXON

http://dx.doi.org/10.5248/117.239

Volume 117, pp. 239-246 July-September 2011

Mycena sect. Longisetae:

a new species, anew name, and an addition

DOLLYMOL M. ARAVINDAKSHAN & P. MANIMOHAN*

Department of Botany, University of Calicut, Kerala, 673 635, India

CORRESPONDENCE TO *: dollym.aravind@gmail.com & * pmanimohan@gmail.com

ABSTRACT — Mycena saloma sp. nov. is described and illustrated from Kerala State, India.

Mycena lomavritha nom. nov. is proposed for Mycena indica Manim. & Leelav., nom. illegit.

The Brazilian species M. rhaphidocephala, previously accepted in sect. Sacchariferae, is

accepted in sect. Longisetae.

Keyworps — Agaricales, Basidiomycota, nomenclature, taxonomy

Introduction

As currently accepted, Mycena sect. Longisetae A.H. Sm. ex Maas Geest.

(Basidiomycota, Agaricales, Mycenaceae) incorporates about eleven species that

are primarily characterised by several long, needle-like, thick-walled hairs on

the pileus (Desjardin et al. 2002). Additionally, these species have ascending,

narrowly adnate to free lamellae, a pruinose to hispid stipe with a typically

discoid base, ellipsoid basidiospores, 4-spored basidia, a hymenium devoid of

pleurocystidia, a pileus margin often beset with cystidia, dextrinoid pileal and

stipe trama, smooth caulocystidia that typically taper towards the apex, and a

pileipellis composed of either acanthocysts or spinulose hyphae. According to

Desjardin et al. (2002), southeast Asia could be the centre of diversity of this

group.

During our investigations on the agaric mycota of Kerala State, India, we

came across an as yet undescribed species belonging to this group that we

describe here. Mycena indica, a species originally placed by Manimohan &

Leelavathy (1988) in sect. Sacchariferae and subsequently transferred to the

amended sect. Longisetae by Desjardin et al. (2002), needs a replacement name

(nom. nov.) as the present one is an illegitimate later homonym (McNeill et

al. 2006: Art. 53.1). A nom. nov. for this species is proposed here, along with

some observations on the species. The Brazilian species M. rhaphidocephala,

240 ... Aravindakshan & Manimohan

originally placed by Maas Geesteranus & de Meijer (1998) in sect. Sacchariferae,

has all the characteristics of sect. Longisetae and is accepted in the latter section

here.

Materials & methods

Conventional morphology-based taxonomic methods were employed for this study.

Microscopic observations were made on material stained with 1% aqueous solution of

Congo red and mounted in 3% aqueous KOH. Melzer’s reagent was used to observe

whether the spores and tissues were amyloid. For evaluation of the range of spore-size,

twenty basidiospores each from one specimen of each collection cited were measured.

The herbarium acronyms follow Thiers (2011). The collections cited without herbarium

acronyms are in the personal herbarium of the second author. The concept of Desjardin

et al. (2002) is followed for Mycena sect. Longisetae.

Taxonomy

Mycena saloma Aravind. & Manim., sp. nov. PLaTE 1

MycoBank MB 519881

Basidiomata delicata, dispersa. Pileus 1.25-5 mm latus, hemisphericus vel convexus,

transluso-striatus, subsulcatus, albus, pilis erectis subtilis vestitus. Lamellae adnexae,

albae, subdistantae, lamellularum intermixtae. Stipes 3.5-22 x 0.25-0.5 mm, centralis,

cylindricus, transluso-albus, puberulus, basi discoideo instructus. Sporae 7.5-9.5(-11) x

3.75-5.5 um, inaequilateraliter ellipsoideae, amyloideae, leves. Basidia 12-19 x 9.5-13

um, subglobosa et pedicellata, 4-sporigera. Cheilocystidia et pleurocystidia nulla. Trama

pilei et lamellarum ex hyphis dextrinoidei composita. Hyphae pileipellidis 2-11.5 um

latae, echinulatae, spinulis minutis instructae. Pileosetae 60-780 x 15-30 um (ad basim)

x 10-11 um (ad apicem), hyalinae, crassitunicatae. Cystidia margine pilei 17-82 x 6.5-20

um, clavata vel fusoidea, tenuitunicata, spinulosa. Caulocystidia 92.5-125.5 x 15-33 um,

flexuoso-cylindrica vel fusoidia, hyalina, tenuitunicata, leves. Hyphae omnes fibulatae.

Type: INDIA, Kerala State, Idukki District, Munnar, Kanniamalai Estate: 14.VIII.2010,

holotype: Dollymol DM461 (L).

ErymMo oey: saloma (Sanskrit), hairy

Basidiomata very small, delicate. Pileus 1.25-5 mm diam., up to 3 mm high,

initially parabolic, becoming hemispherical to convex with age; surface initially

pure white, becoming off-white with age, with prominent white hairs up to

0.5 mm long that are denser at the centre, pellucid-striate, very faintly sulcate,

dry, finely pruinose; margin straight, finely fringed. Lamellae adnexed, pure

white, thin, 0.5-0.75 mm wide, subdistant, with lamellulae of 1-2 lengths; edge

entire, concolourous with the sides. Stipe 3.5-22 x 0.25-0.5 mm, central, terete,

tapering towards apex, hollow; surface translucent white, pubescent, more so

towards the base, almost glabrous at apex; base discoid, with finely radiating

basal mycelium. Context very thin, concolourous with the pileus surface.

Odour and taste not distinctive.

Additions to Mycena sect. Longisetae (India) ... 241

PLaTE 1: Mycena saloma. A, basidioma (holotype); B, basidiospores (DM486); C, basidium

(DM486); D, pileus marginal cystidium (DM486); E, caulocystidium (DM486); FE, pileipellis

(DM486); G, pileus hair (DM486). (Scale bars: A = 5 mm; B-G = 10 um.)

242 ... Aravindakshan & Manimohan

Basidiospores 7.5-9.5(-11) x 3.75-5.5 (8.57 + 0.68 x 4.45 + 0.44) um,

Q = 1.66-2.4, Qm = 1.9, pip-shaped to ovo-ellipsoid, thin-walled, hyaline,

smooth, with refractive guttules, strongly amyloid. Basidia 12-19 x 9.5-13

um, subglobose, pedicellate, hyaline, bearing 4 sterigmata up to 5 um long.

Lamella-edge fertile. Cheilocystidia and pleurocystidia not seen. Lamellar

trama regular to subregular; hyphae 2-15.5 um, often inflated up to 30 um,

thin-walled, hyaline, faintly dextrinoid in Melzer’s reagent; subhymenium

pseudoparenchymatous. Pileus trama subregular; hyphae 3-26 um wide, rarely

inflated up to 37.5 um, thin-walled, hyaline, distinctly dextrinoid in Melzer’s

reagent. Pileipellis a cutis of repent hyphae supporting scattered pileal hairs;

hyphae 2-11.5 um wide, thin- to slightly thick-walled (0.25 um), hyaline,

spinulose, with acanthophysoid terminal elements; excrescences 0.5-1.5 x

0.25-0.5 um; acanthophysoid terminal elements 12-37 x 7-19 um, repent or

ascending, clavate to narrowly clavate or cylindrical, with refractive guttules

and with excrescences 0.5-2 x 0.5-1 um; pileal hairs 60-780 um long, 15-30 um

broad at the base, 10-11 um broad at the apex, gradually tapering towards the

obtuse apex, smooth, hyaline, with a refractive wall 1-9 um thick. Pileus margin

beset with marginal cystidia; marginal cystidia 17-82 x 6.5-20 um, clavate or

fusiform, entirely spinulose or often with a smooth apical prolongation, thin-

walled, hyaline; excrescences 0.75-4 x 0.75-1 um. Stipitipellis a cutis; hyphae

2-8 um wide, thin-walled, hyaline, smooth. Caulocystidia 92-125 x 15-33 um,

flexuoso-cylindric, fusoid or lageniform with a subconical apex, thin-walled,

hyaline, smooth. Stipe trama strongly dextrinoid in Melzer’s reagent. Clamp

connections present on all hyphae.

HasitatT: On dicotyledonous leaves and twigs, scattered; altitude:

1800-2000 m.

ADDITIONAL SPECIMEN EXAMINED: INDIA, KERALA STATE, [DUKKI DISTRICT, Munnar,

Eravikulam National Park: 16. VIII.2010, Dollymol DM486 (L).

DiscussIon: This species has all the diagnostic features of sect. Longisetae

discussed earlier. The key by Desjardin et al. (2002) to species of Mycena

sect. Longisetae leads to a couplet differentiating M. breviseta Hohn. and

M. longiseta Hohn. Mycena breviseta is a poorly known species whose

holotype collection is reported to contain ‘no intact basidiomes and no pilei’

(Desjardin et al. 2002). The Desjardin et al. (2002) key describes this Javan

species as having an umbilicate pileus, pileal hairs that are up to 1000 um long,

and clavate marginal cystidia that are entirely spinulose. The closely related

M. longiseta, also from Java, has a grey pileus and pileal hairs that are up to 1000

um long. A perusal of Mycena literature revealed that the Brazilian species,

M. rhaphidocephala (whose infrageneric position is discussed below but hitherto

placed in sect. Sacchariferae), is strikingly similar to M. saloma in most macro-

and microscopic features. An examination of the M. rhaphidocephala holotype

Additions to Mycena sect. Longisetae (India) ... 243

PLATE 2: Mycena lomavritha. A, basidioma (AF70); B, pileus hair (DM423); C, cheilocystidia

(DM423); D, basidiospores (DM423); E, caulocystidium (DM423).

(Scale bars: A = 5 mm; B-E = 10 um.)

244 ... Aravindakshan & Manimohan

and protologue, however, revealed that it has pileal hairs that are much longer

(up to 3000 um), smooth pileipellis hyphae, well-developed acanthocysts,

consistently lageniform caulocystidia, and clampless hyphae.

Mycena lomavritha Manim., nom. nov. PLATE 2

MycoBank MB 519882

= Mycena indica Manim. & Leelav., Mycologia 80(6): 861 (1989,

“1988”), nom. illeg., non Sarwal & Rawla 1983.

EtyMo.Loecy: lomavritha (Sanskrit), covered with hair.

DESCRIPTION AND ILLUSTRATIONS: Manimohan & Leelavathy (1989).

SPECIMENS EXAMINED: INDIA, KERALA STATE, MALAPPURAM DistTRICT, Calicut

University campus: 6.VII.1987, Manimohan M383 (holotype TRTC 50991); 7.VI.2006,

Dollymol DM13; PataxKap District, Silent Valley National Park: 16.VI.2010,

Dollymol DM423.

Discussion: Thus far known only from southern India, M. lomavritha was

originally described based on a single collection. Several collections were

subsequently made by the present authors from Kerala State, India. All our

observations support the original description except that the pileal trama was

consistently dextrinoid. Also, while the original collection was found growing

on the bark of a jackfruit tree, we found the species also growing on the bark of

other trees. Mycena lomavritha is characterised by small off-white corticolous

basidiomata with a hairy pileus and discoid stipe base, amyloid basidiospores,

dextrinoid tramal tissues, ventricose-fusoid cheilocystidia with sparsely

spinulose lower middle region, a pileipellis composed of acanthocysts as well as

scattered thick-walled, aculeate hairs, and abundant obclavate caulocystidia.

Manimohan & Leelavathy (1988) placed this species in sect. Sacchariferae

despite the thick-walled pileus hairs because sect. Longisetae then was

characterized by inamyloid basidiospores, gelatinized pileipellis, and absence

of clamp connections. Subsequently both sect. Sacchariferae (Desjardin 1995)

and sect. Longisetae (Desjardin & Horak 2002) were redefined. As discussed

by Desjardin et al. (2002), the combination of characters exhibited by

M. lomavritha are clearly better accommodated in the redefined sect. Longisetae,

where it seems close to M. brunneisetosa Corner from Singapore.

Mycena rhaphidocephala Maas Geest. & de Meijer, Persoonia 17: 39 (1998). PLATE 3

DESCRIPTION AND ILLUSTRATION: Maas Geesteranus & de Meijer (1998: 39-40, Fig. 7).

SPECIMEN EXAMINED: BRAZIL, PARANA STATE, Morretes, Porto do Cima, Parque

Murumbi: 27.V1.1995, A.A.R. de Meijer MA-3098 (L, holotype).

Discussion: This Brazilian species has all the characteristic features of the

redefined sect. Longisetae such as the small off-white basidiomata, thick-

walled aculeate hairs on the pileus, discoid stipe base, ellipsoid basidiospores,

Additions to Mycena sect. Longisetae (India) ... 245

PLaTE 3: Mycena rhaphidocephala (holotype). A, primordium; B, mature basidioma; C, pileus hair;

D, basidiospores; E, pileus marginal cystidia.

(Scale bars: A, B = 1 mm; C = 100 um; D, E = 10 um.)

246 ... Aravindakshan & Manimohan

acanthocysts on the pileipellis, spinulose marginal cystidia with a smooth apical

prolongation, and smooth caulocystidia that taper towards the apex. We have

verified these features on the holotype of this species. What Maas Geesteranus &

de Meijer (1998) had described as cherocytes are actually the spinulose marginal

cystidia typical of the sect. Longisetae (see Desjardin 1995: 5 for the history

and definition of the term cherocyte). Our examination of these entities in the

holotype revealed that they are thin-walled —not “somewhat thick-walled” as

noted by Maas Geesteranus & de Meijer (1998)— and hence cannot be called

cherocytes. Maas Geesteranus & de Meijer (1998), who originally placed this

species in sect. Sacchariferae, noted that its combination of characters was not

known in any other species of that section. We accept M. rhaphidocephala as a

member of sect. Longisetae.

Acknowledgment

We are indebted to Prof. D.E. Desjardin and Dr D.J. Lodge for presubmission reviews

of our manuscript. We are also thankful to Dr M.E. Noordeloos and the staff of the

Nationaal Herbarium, Leiden Branch, for arranging a loan of the holotype of Mycena

rhaphidocephala. Mr V. Adnaan Farook is thanked for letting us use his field photograph

of M. lomavritha in this paper.

Literature cited

Desjardin DE. 1995. A preliminary accounting of the worldwide members of Mycena sect.

Sacchariferae. Bibliotheca Mycologica 159: 1-89.

Desjardin DE, Horak E. 2002. Agaricales of Indonesia. 4. Mycena sect. Longisetae with comments

on allied species. Sydowia 54: 142-156.

Desjardin DE, Boonpratuang T, Hywel-Jones NL. 2002. An accounting of the worldwide members

of Mycena sect. Longisetae. Fungal Diversity 11: 69-85.

McNeill J, Barrie FR, Burdet HM, Demoulin V, Hawksworth DL, Marhold K, Nicolson DH, Prado

J, Silva PC, Skog JE, Wiersema JH, Turland NJ (eds). 2006. International Code of Botanical

Nomenclature (Vienna Code): adopted by the Seventeenth International Botanical Congress,

Vienna, Austria, July 2005. A.R.G. Gantner Verlag, Ruggell.

Manimohan P, Leelavathy KM. 1989 [“1988”]. Mycena indica, a new species from southern India.

Mycologia 80(6): 861-862. http://dx.doi.org/10.2307/3807569

Maas Geesteranus RA, de Meijer AAR. 1998. Further mycenas from the State of Parana, Brazil.

Persoonia 17: 29-46.

Thiers B. 2011 [continuously updated]. Index Herbariorum: A global directory of public herbaria

and associated staff. New York Botanical Garden's Virtual Herbarium.

http://sweetgum.nybg.org/ih/

MY COTAXON

http://dx.doi.org/10.5248/117.247

Volume 117, pp. 247-253 July-September 2011

Three new hyphomycetes from southern China

JIAN Ma, Li-Guo Ma, Y1I-DONG ZHANG & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Three new hyphomycetes were collected from dead branches from tropical and

subtropical forests in southern China: Ellisembia schimae from Schima superba, Linkosia

hibisci from Hibiscus mutabilis, and Stanjehughesia micheliae from Michelia skinneriana. The

new species are described, illustrated, and compared with similar species.

KEY worDs — anamorphic fungi, taxonomy

Introduction

The mycota of tropical and subtropical forests of southern China is very

rich in wood-inhabiting fungi (Ma et al. 2008, Dai et al. 2009, Dai & Li 2010,

Zhang et al. 2009a). During ongoing mycological surveys in these forests, three

hyphomycetes with morphological features typical of Ellisembia Subram.,

Stanjehughesia Subram. (Subramanian 1992), and Linkosia A. Hern. Gut. &

B. Sutton (Hernandez-Gutiérrez & Sutton 1997) were collected on dead

branches. However, these collections differ significantly from the currently

accepted species of these genera, and are therefore proposed as new taxa.

The specimens are deposited in the Herbarium of Shandong Agricultural

University, Plant Pathology (HSAUP) and the Mycological Herbarium, Institute

of Microbiology, Chinese Academy of Sciences (HMAS).

Taxonomy

Ellisembia schimae Jian Ma & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 561476

Fungus anamorphicus. COLoNIAE in substrato naturali effusae, brunneae vel atrobrunneae,

pilosae. Mycelium partim superficiale, partim immersum in substrato, ex hyphis ramosis,

septatis, pallide brunneis, laevibus, 1.5-3 um crassis compositum. CONIDIOPHORA

macronemata, mononematica, singula vel fasciculata, erecta, nonramosa, recta vel

flexuosa, cylindrica, atrobrunnea vel nigra, laevia, septata, 16-39 x 6.5-9 um. CELLULAE

CONIDIOGENAE monoblasticae, integratae, terminales, determinatae, lageniformes

248 ... Ma & al.

) J

20m | ees od

Fic. 1. Ellisembia schimae.

A. Conidiophores with conidia. B. Conidiophores. C-D. Conidia.

vel cylindricae, brunneae vel atrobrunneae, laeves. Conidiorum secessio schizolytica.

Conrp1A holoblastica, solitaria, acrogena, recta vel curvata, obclavata, laevia, brunnea vel

atrobrunnea, 14-21-distoseptata, 122-155 um longa, 4.5-6.5 um crassa, apicem acutum

gradatim versus attenuata, basi truncata 3.5-4.5 um lata.

Type: China, Guangdong Province: Liuxihe National Forest Park, on dead branches of

Schima superba Gardn. & Champ. (Theaceae), 16 Oct 2010, J. Ma, (holotype HSAUP

H5380—2; isotype HMAS146141).

ETryMoOLoey: in reference to the host genus, Schima.

Anamorphic fungi. COLONIES on natural substrate effuse, brown to dark

brown, hairy. Mycelium partly superficial, partly immersed, composed of

branched, septate, pale brown, smooth-walled hyphae, 1.5-3 um thick.

CONIDIOPHORES differentiated, single or in groups, erect, unbranched, straight

or flexuous, cylindrical, dark brown to black, smooth, septate, 16-39 x 6.5-9

um. CONIDIOGENOUS CELLS monoblastic, integrated, terminal, determinate,

Ellisembia, Linkosia & Stanjehughesia spp. nov. (China) ... 249

lageniform or cylindrical, brown to dark brown, smooth. Conidial secession

schizolytic. Conrp14 holoblastic, solitary, acrogenous, straight or curved,

obclavate, smooth-walled, brown to dark brown, 14—21-distoseptate, 122-155

um long, 4.5-6.5 um thick in the broadest part, tapered gradually toward the

acute apex, 3.5-4.5 um wide at the truncate base.

ComMMENTS - Subramanian established the genus Ellisembia, proposing

twelve new combinations for twelve species of Sporidesmium Link and

designating E. coronata (Fuckel) Subram. as the type species (Subramanian

1992). Subsequently, Wu & Zhuang (2005) merged Imicles Shoemaker &

Hambl. (Shoemaker & Hambleton 2001) into Ellisembia and expanded the

generic concept. Ellisembia was mainly characterized by differentiated, single,

unbranched, septate conidiophores, and monoblastic, integrated, terminal,

lageniform, doliiform or cylindrical, determinate or percurrently extending

conidiogenous cells that produce solitary, holoblastic, distoseptate conidia.

Assigning species within Ellisembia is primarily based on conidial features such

as shape, size and septation (Subramanian 1992, Wu & Zhuang 2005, McKenzie

2010).

Our specimen is clearly a species of Ellisembia from its conidiophore and

conidial morphology. The new species bears some resemblance to E. britannica

(B. Sutton) W.P. Wu and E. carrii (Morgan-Jones) W.P. Wu (Wu & Zhuang

2005) in conidial shape. However, E. schimae differs from E. britannica (conidia

50-130 x 3-5 um, 10-13-distoseptate) in its larger conidia with more numerous

septa, and from E. carrii (conidia 90-130 x 8-10 um) in having longer and

narrower conidia. In addition, conidia of E. schimae have an obvious pointed

apex, while those of E. britannica and E. carrii have rounded apices.

Linkosia hibisci Jian Ma & X.G. Zhang, sp. nov. FIG. 2

MycoBank MB 561477

Fungus anamorphicus. CoLONIAE in substrato naturali effusae, brunneae. Mycelium

superficiale, ex hyphis ramosis, septatis, brunneis vel atrobrunneis, laevibus, 1.5-3 um

crassis compositum. CONIDIOPHORA nulla. CELLULA CONIDIOGENA determinatae,

monoblastica, solitaria, simplicia, lageniformia vel ampulliformia, brunnea vel

atrobrunnea, laevia, 9-22 x 4-5 um, ad apicem 4-4.5 um crassa et truncatae. Conidiorum

secessio schizolytica. Conrp1A holoblastica, solitaria, acrogena, recta vel curvata, obclavata

vel obclavata-rostrata, atrobrunnea vel brunnea, laevia, 16-21-distoseptata, 160-210 um

longa, 7.5-9.5 um crassa, apicem versus ad 1.5-2.5 um attenuata; cellula apicalis pallide

brunnea, rotundata; cellula basalis cylindrica vel conico-truncata, ad basim 2-4.5 um

crassa.

Type: China, Hainan Province: tropical forest of Bawangling, on dead branches of

Hibiscus mutabilis L. (Malvaceae), 8 Dec 2010, J. Ma (holotype HSAUP H5199-1;

isotype HMAS 146142).

EryMo_oey: in reference to the host genus, Hibiscus.

250 ... Ma & al.

A C D E F

a |

;

: a

a ' 20m

Fic. 2. Linkosia hibisci.

A. Conidiogenous cells with conidia. B. Conidiogenous cells. C-G. Conidia.

—

wiz

Anamorphic fungi. COLONIES on natural substrate effuse, brown. Mycelium

superficial, composed of branched, septate, brown to dark brown, smooth-

walled hyphae, 1.5-3 um thick. ConrpiopHoREs absent. CONIDIOGENOUS

CELLS determinate, monoblastic, solitary, simple, lageniform or ampulliform,

brown to dark brown, smooth, 9-22 x 4-5 um, 4-4.5 um wide at the truncate

apex. Conidial secession schizolytic. Conrp1< holoblastic, solitary, acrogenous,

straight or curved, obclavate to obclavate-rostrate, dark brown to brown,

smooth, 16-21-distoseptate, 160-210 um long, 7.5-9.5 um thick in the broadest

part, tapering to 1.5-2.5 um near the apex; apical cells pale brown, rounded;

basal cell cylindrical, truncate, 2-4.5 um wide.

ComMENTS—The genus Linkosia, with L. coccothrinacis (A. Hern. Gut. &

J. Mena) A. Hern. Gut. & B. Sutton as the type species, was characterized by

absence of conidiophores, and solitary, simple, short, monoblastic, determinate,

lageniform or ampulliform conidiogenous cells producing distoseptate conidia

(Hernandez-Gutiérrez & Sutton 1997, Wu & Zhuang 2005). These characters

separate Linkosia from other similar genera including Stanjehughesia and

Janetia M.B. Ellis (Ellis 1976). Six species are currently included in this genus, of

Ellisembia, Linkosia & Stanjehughesia spp. nov. (China) ... 251

which four have previously been described from China (Hernandez-Gutiérrez

& Sutton 1997, Wu & Zhuang 2005, Zhang et al. 2009b). All six species are

reported to survive saprobically on dead leaves or branches of bamboo.

Of the known species, Linkosia hibisci is similar to L. obclavata W.P. Wu

(Wu & Zhuang 2005) and L. mori K. Zhang & X.G. Zhang (Zhang et al. 2009)

in conidial shape, but differs in conidial dimensions (L. obclavata 122-177 x

12-16 um, L. mori 110-125 x 9-12 um). Moreover, L. hibisci conidia have more

septa than those of L. obclavata (12-14 distosepta). In addition, the mature

conidia of L. hibisci are more darkly pigmented than those of the other two

species.

Stanjehughesia micheliae Jian Ma & X.G. Zhang, sp. nov. FIG. 3

MycoBank MB 561478

Fungus anamorphicus. CoLONIAE in substrato naturali effusae, nigrae. Mycelium

superficiale, ex hyphis ramosis, septatis, pallide brunneis vel brunneis, laevibus, 1.5-3

um crassis compositum. CONIDIOPHORA nulla vel brevis, 1-3-septata, brunnea vel

atrobrunnea, 11-28 x 4.5-5 um. CELLULA CONIDIOGENA monoblastica, determinatae,

solitaria, simplicia, lageniformia vel ampulliformia, brunnea vel atrobrunnea, laevia,

4.5-6.5 x 3.5-5 um, ad apicem 3-4.5 um crassa et truncatae. Conidiorum secessio

schizolytica. Conrp1A holoblastica, solitaria, acrogena, recta vel curvata, obclavata vel

obclavata-rostrata, atrobrunnea vel brunnea, laevia, 13-19-distoseptata, 130-190 um

longa, 7-9 um crassa, apicem versus ad 2-3 um attenuata; cellula apicalis rotundata;

cellula basalis cylindrica vel conico-truncata, ad basim 3.5-4.5 um crassa; Appendicibus

lateralibus 0-2, brunneae, septata, cylindricae, surgentibus ex cellulla e apicem 2nd vel

3rd.

Type: China, Guangdong Province: Chebaling National Nature Reserve, on dead

branches of Michelia skinneriana Dunn (Magnoliaceae), 16 Oct 2010, J. Ma (holotype

HSAUP H5414; isotype HMAS146143).

EryMo_oey: in reference to the host genus, Michelia.

Anamorphic fungi. COLONIES on natural substrate effuse, black. Mycelium

superficial, composed of branched, septate, pale brown to brown, smooth-

walled hyphae, 1.5-3 um thick. Conip1opHoREs absent or short, 1-3-septate,

brown to dark brown, 11-28 x 4.5-5 um. CONIDIOGENOUS CELLS monoblastic,

determinate, solitary, simple, lageniform or ampulliform, brown to dark brown,

smooth, 4.5-6.5 x 3.5-5 um, 3-4.5 um wide at the truncate apex. Conidial

secession schizolytic. Conip14 holoblastic, solitary, acrogenous, straight or

curved, obclavate to obclavate-rostrate, dark brown to brown, smooth, 13-19-

distoseptate, 130-190 um long, 7-9 um thick in the broadest part, tapering

to 2-3 um near the apex; apical cells rounded; basal cell cylindrical, truncate,

3.5-4.5 um wide; lateral appendages 0-2, brown, septate, cylindrical, arising

from the 2nd or 3rd cells from the apex.

ComMENTS - ‘The genus Stanjehughesia was established by Subramanian

(1992) to accommodate five new combinations from Sporidesmium. The

252 ...Ma &al.

A G

20m \

wiz

witgz

woz

wiz

Fic. 3. Stanjehughesia micheliae.

A-B. Reduced conidiophores or conidiogenous cells with conidia. C-F. Conidia.

genus is characterized by very reduced or absent conidiophores; determinate,

monoblastic, lageniform or ampulliform, solitary, short, simple conidiogenous

cells that produce obclavate to obclavate-rostrate, euseptate conidia. Additional

species have been described or transferred to Stanjehughesia (McKenzie 1995,

Mena-Portales et al. 2001, Wu & Zhuang 2005, Delgado 2008, Marincowitz

2008). At present, the genus comprises 11 species, most of which grow as

saprobes on rotten wood, dead branches, stems or decaying leaves.

Stanjehughesia micheliae is unique among known species of Stanjehughesia

by its obclavate to obclavate-rostrate conidia with 0-2 cylindrical appendages

arising from the 2nd or 3rd cell from the apex.

Acknowledgments

The authors express gratitude to Dr WB. Kendrick and Dr N.R. O’Neill 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. 2006FY 120100, 2006FY110500-5).

Ellisembia, Linkosia & Stanjehughesia spp. nov. (China) ... 253

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Divers. Res. Ser. 15: 1-351.

Zhang K, Fu HB, Zhang XG. 2009a. Taxonomic studies of Corynespora from Hainan, China.

Mycotaxon 109: 85-93. http://dx.doi.org/10.5248/109.85

Zhang K, Ma LG, Zhang XG. 2009b. A new hyphomycete species from Guangxi, China. Mycotaxon

108: 123-125. http://dx.doi.org/10.5248/108.123

MY COTAXON

http://dx.doi.org/10.5248/117.255

Volume 117, pp. 255-260 July-September 2011

A new species of Junghuhnia

(Basidiomycota, Meruliaceae) from tropical China

HalI-SHENG YUAN

State Key Laboratory of Forest and Soil Ecology, Institute of Applied Ecology,

Chinese Academy of Sciences, Shenyang 110164, P. R. China

CORRESPONDENCE TO: yuanhs911@yahoo.com.cn

ABSTRACT — A new polypore, Junghuhnia minor sp. nov., is described and illustrated from

the tropical forest of Yunnan Province, southwestern China. The new species is characterized

by very thin annual resupinate basidiocarps, a cream to pale buff pore surface with small

and slightly lacerated pores, small (2.7-3 x 1.9-2.2 wm) ellipsoid basidiospores, and

skeletocystidia.

Key worps — lignicolous fungi, Meruliaceae, taxonomy

Introduction

Lignicolous fungi are important components of forest ecosystems, where

they decompose forest litter and maintain energy flow and material circulation

(Lonsdale et al. 2008). Many, including some traditionally used for medicinal

purposes in Asia, are edible while others are plant pathogens (Dai et al.

2007, 2009b). New species have been described after recent investigations on

lignicolous fungi in subtropical and tropical forests in China (Cui & Dai 2008,

Cui et al. 2008, 2011, Dai et al. 2009a, 2010, Dai & Li 2010, Du & Cui 2009, Li

& Cui 2010, Li et al. 2008, Wei & Dai 2008, Xiong & Dai 2008, Zhou & Dai

2008).

Characters noted for two polypores collected on fallen angiosperm twig

during a survey of the lignicolous fungi in southern China (dimitic hyphal

system, encrusted skeletocystidia, small ellipsoid basidiospores) suggest an

affinity with Junghuhnia Corda. ‘They differ morphologically from existing

Junghuhnia species, and so I describe and illustrate them as a new species.

Materials & methods

The specimens are deposited at the biological herbarium of Institute of Applied

Ecology, Chinese Academy of Sciences (IFP). The microscopic procedure follows Dai

256 ... Yuan

(2010). The microscopic studies were made from sections mounted in Cotton Blue

(CB): 0.1 mg aniline blue dissolved in 60 g pure lactic acid; CB+ = cyanophily, CB- =

acyanophily. Amyloid and dextrinoid reactions were tested in Melzer’s reagent (IKI):

1.5 g KI (potassium iodide), 0.5 g I (crystalline iodine), 22 g chloral hydrate, aq.

dest. 20 ml; IKI- = neither amyloid nor dextrinoid reaction. 5% KOH was used as

reagent. Sections were studied at magnifications up to x1000 using a Nikon Eclipse

E600 microscope and phase contrast illumination, and dimensions were estimated

subjectively with an accuracy of 0.1 um. In the spore measurements, the apiculus was

excluded. In presenting the variation of spore size, 5% of the measurements out of each

end of the range are given in parentheses. The following abbreviations are used in the

text: L = mean spore length (arithmetical mean of all spores), W = mean spore width

(arithmetical mean of all spores), Q = extreme values of the length/width ratios among

the studied specimens, and n = the number of spores measured from a given number of

specimens. Special color terms are from Anonymous (1969) and Petersen (1996).

Taxonomy

Junghuhnia minor HS. Yuan, sp. nov. Fic. 1

MycoBank MB 561532

Carpophorum annuum, resupinatum. Facies pororum cremeum vel bubalinum; pori

angulati, 5-6 per mm. Systema hypharum dimiticum, hyphae generatoriae fibulatae,

hyphae skeletales subiculi 2-4 um diam. Sporae hyalinae, ellipsoideae, 2.7-3 x 1.9-2.2

ym.

Type. — China. Yunnan Province, Mengla County, Wangtianshu Forest Park, on fallen

angiosperm twig, 16.[X.2007 Yuan 3268 (holotype in IFP).

ETYMOLOGY — minor (Lat.): refers to small basidiospores.

FruitBopy — Basidiocarps annual, resupinate, coriaceous, without special

odor or taste when fresh, corky when dry, up to 15 cm long, 1 cm wide and

1 mm thick; sterile margin thinning out, membranous, cream, up to 3 mm

wide. Pore surface cream to pale buff-yellow upon drying; pores angular, 5-6

per mm, dissepiments thin, slightly lacerate. Subiculum very thin to almost

lacking, cream to pale buff, ca. 0.1 mm thick. Tubes concolorous with pore

surface, corky, ca. 0.4 mm long.

HyYPHAL STRUCTURE — Hyphal system dimitic; generative hyphae bearing

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

SUBICULUM — Dominated by skeletal hyphae; generative hyphae hyaline,

thin-walled, occasionally branched, 1.8-3.5 um diam; skeletal hyphae hyaline,

thick-walled with a distinct lumen to subsolid, straight to flexuous, occasionally

branched, interwoven, 2-4 um diam.

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

branched, 1.5-2.5 um diam; skeletal hyphae dominant, hyaline, thick-walled

with a distinct lumen to subsolid, occasionally branched, interwoven, 1.8-2.8

Junghuhnia minor sp. nov. (China) ... 257

Set ter ORO OKaIe-Or®

10 pm

Fic. 1. Microscopic structures of Junghuhnia minor (drawn from the holotype).

a: Basidiospores. b: Basidia and basidioles. c: Skeletocystidia.

d: Hyphae from trama. e: Hyphae from subiculum.

258 ... Yuan

um diam. Skeletocystidia numerous, clavate, thick-walled, originated from

trama, embedded or projecting, heavily encrusted, 15-50 x 6-11 um (with

encrustation); basidia clavate, bearing four sterigmata and a clamp connection

at the base, 9-12 x 4-5 um; basidioles in shape similar to basidia, but slightly

smaller.

SporES — Basidiospores small, ellipsoid, hyaline, thin-walled, smooth,

IKI-, CB-, (2.5-)2.7-3 x (1.8-)1.9-2.2(-2.3) um, L = 2.85 um, W = 2.04 um,

Q = 1.38-1.41 (n=60/2).

ADDITIONAL SPECIMEN EXAMINED — CHINA. Yunnan Province, Mengla County,

Wangtianshu Forest Park, on fallen angiosperm twig, 16.1X.2007 Yuan 3215 (IFP).

TYPE OF ROT — White rot.

REMARKS — Junghuhnia minor is characterized by annual, resupinate and very

thin basidiocarps, pale buff-yellow pore surface with white to cream margin,

small ellipsoid basidiospores (2.7-3 x 1.9-2.2 um), and skeletocystidia.

Junghuhnia luteoalba (P. Karst.) Ryvarden also is characterized by resupinate,

pale-buff colored basidiocarps and slightly lacerate dissepiments, but differs in

its cylindrical basidiospores and larger pores (4-8 per mm) (Nufiez & Ryvarden

2001). In addition, J. luteoalba is distributed in boreal and temperate zones.

Junghuhnia crustacea (Jungh.) Ryvarden resembles the new species in its

resupinate, thin basidiocarps and cream to pale-buff pore surface but has more

shallow pores, distinctly lacerate pores and larger basidiospores (Nunez &

Ryvarden 2001, Ryvarden & Johansen 1980).

Junghuhnia minuta I. Lindblad & Ryvarden, which has similar basidiospores

(2.5-3 x 2-2.5 um), differs from J. minor by having distinctly pileate basidiocarps

and tiny pores (10-12 per mm, Lindblad & Ryvarden 1999).

Junghuhnia microspora Rajchenb. also has small basidiospores but is

distinguished from J. minor by its orange to chestnut colored pore surface and

narrower basidiospores (2.6-3.6 x 1.0-1.6 um, Rajchenberg 1983).

OTHER SPECIMENS EXAMINED: Junghuhnia crustacea — CuinaA. Yunnan Province,

Mengla County, Wangtianshu Forest Park, on fallen angiosperm branch, 17.[X.2007

Yuan 3638 (IFP); KeNyA. Western Province, Kakamega district, Kakamega Forest,

substrate unknown, 20.1.1970 Ryvarden 5431 (O); THAILAND. Cangwat Chiang Mai,

Amphoe Mae Rim, Kong Hae, on fallen angiosperm trunk, 15.11.1979 Ryvarden 17568

(O).

J. microspora — ARGENTINA. Missiones, Iguazu Falls, Apepu Reservation, substrate

unknown, 4.1I.1982 Rajchenberg 3504 (O).

J. minuta — PuERTO Rico. Luquillo, Beasley water shed, on deciduous tree, 6.V1.1997

Ryvarden 40279 (O).

J. luteoalba — Cuina. Heilongjiang Province, Ning’an County, Volcano forest, on fallen

trunk of Populus, 14.1X.2004 Yuan 618 (IFP); Liaoning Province, Kuandian County,

Baishilazi Nat. Res., on rotten wood of Quercus, 1.VIII.2004 Cui 1090 (IFP).

Junghuhnia minor sp. nov. (China) ... 259

Acknowledgements

I appreciate Drs. Zheng Wang and Bao-Kai Cui who reviewed the manuscript. The

author also thanks Dr. Yu-Cheng Dai (IFP, China) and Dr. Cony Decock (MUCL,

Belgium) for the company in the field trips. The research was financed by the National

Natural Science Foundation of China (Project Nos. 31070023 & 30700004).

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MY COTAXON

http://dx.doi.org/10.5248/117.261

Volume 117, pp. 261-267 July-September 2011

Caloplaca allochroa (lichenized Ascomycetes),

a new saxicolous lichen species from South Korea

YOGESH JOSHI”, JAN VONDRAK?®, OLGA VONDRAKOVA‘,

Tui THUuy NGUYEN? & JAE-SEOUN Hur”

‘Department of Botany, S.S.J. Campus, Almora 263601, Uttarakhand, India

*Korean Lichen Research Institute, Sunchon National University

Sunchon 540-742, South Korea

*Institut of Botany of the ASCR, Zamek 1, Prithonice 252 43, Czech Republic

‘Institute of Steppe (Urals Branch of Russian Academy of Sciences),

Pionerskaya st. 11, Orenburg 460000, Russia

CORRESPONDENCE TO *: ***dryogeshcalo@gmail.com, *j.vondrak@seznam.cz,

“mer.os@mail.ru, *pthuydhtn@yahoo.com, *jshur1@sunchon.ac.kr

ABSTRACT — ‘The saxicolous lichen species, Caloplaca allochroa, is described as new from

subtropical inland localities of Gangwon and Jeollanam Province (South Korea). It is

characterized by occurring as two colour variants, yellowish and gray, based on presence vs.

absence of anthraquinone pigments. Its colour contrast between brownish-red to brownish-

black apothecial disc and yellowish-orange apothecial margin is very characteristic. Further

diagnostic characters are a hymenium with numerous oil-drops and specific ecology. The

species is related to Caloplaca flavorubescens and C. flavovirescens.

Key worps — anthraquinones, ITS nrDNA, phylogeny, taxonomy, Teloschistaceae

Introduction

A study of specimens of the genus Caloplaca Th. Fr. collected during field

trips in the mountains of South Korea by L. Lékés and Y. Joshi, revealed an

interesting and unusual saxicolous species, which is described here as new

to science. The new species has been found only at inland localities in three

mountains, Mt Jobong, Mt Taebaek, and Mt Gyeokja. It grows in open woodlands

on siliceous and non-calcareous rocks together with abundant members of

Aspicilia, Cladonia, Heterodermia, Lecanora, Leptogium, Myelochroa, Pertusaria,

Phaeophyscia, Porpidia, and Ramalina.

Although we obtained only three specimens of this new taxon, we describe

it without any hesitation because 1) the species is morphologically well defined,

262 ... Joshi & al.

2) one may wait for too a long time to obtain more material of this rare taxon

when conservationists and scientists could benefit from the data, 3) DNA

evidence supports our hypothesis, and 4) the material was collected in ample

amount and duplicates sent to several herbaria.

Materials & methods

Studied specimens are deposited in CBFS, KH, KoLRI, UCRand the private herbarium

of J. Halda (Czech Republic). Description and photographs of external morphology

are based on air-dried material observed under a dissecting stereomicroscope (Nikon

SMZ645). Sections were made with a razor blade under the stereomicroscope and

mounted in lactophenol cotton blue. Anatomical descriptions are based on these

preparations under a compound microscope (Nikon Eclipse E200). The mycological

terminology follows Kirk et al. (2008). Measurements of ascospores were made at

x400 magnification mounted in water; only free ascospores lying outside the asci were

measured. The results are recorded as 95% confidence interval of the measured values.

Number of measurements (n) is within square brackets. Chemical constituents were

identified by HPLC (Sochting 1997).

DNA EXTRACTION AND AMPLIFICATION. Direct PCR was used for PCR-amplification

of the ITS regions including the 5.8S gene of the nuclear rDNA following Arup (2006).

Primers for amplification were ITSIF (Gardes & Bruns 1993) and ITS4 (White et al.

1990). PCR cycling parameters follow Ekman (2001).

PHYLOGENETIC ANALYSIS. Five newly obtained ITS sequences were included in the

phylogenetic analysis along with six sequences from GenBank (Tas. 1) to illustrate

the phylogenetic position of Caloplaca allochroa within the Caloplaca flavorubescens/

C. flavovirescens clade. Caloplaca crenularia and C. ferruginea were selected as outgroup.

Sequences were aligned using BioEdit and manually cut to eliminate the unaligned

TABLE 1. Sequences used in phylogenetic analysis.*

SPECIES : VOUCHER DATA i GENBANK ACCESSION NUMBER

Caloplaca allochroa South Korea; CBFS JV7987 HQ415800

Y. Joshi, Vondrak & Hur

C. crenularia (With.) J.R. Laundon Arup et al. 2007 : EF643512

C. egeana Cl. Roux & Nav.-Ros. : Gibraltar; CBFS JV6262 ? HQ644198

C. flavorubescens

(Huds.) J.R. Laundon

C. flavovirescens

(Wulfen) Dalla Torre & Sarnth.

C. juniperina Tomin

C. stantonii W.A. Weber ex Arup California; UCR 55976 HQ644197

* Newly obtained sequences shown in bold.

Caloplaca allochroa sp. nov. (Korea) ... 263

ends and ambiguously aligned regions of ITS1 and ITS2; 539 positions were retained.

Bayesian phylogenetic analysis was carried out using the program MrBayes 3.1.1

(Ronquist & Huelsenbeck 2003). The optimal nucleotide substitution model (GTR+G)

was found using the program MrModeltest v2.3 (Nylander 2004) with the Akaike

Information Criterion and the hierarchical likelihood ratio test (Posada & Crandall

1998). The MCMC analysis was run for one million generations, performed in two runs,

each with four chains starting from a random tree and using the default temperature of

0.2. Every 100th tree was sampled, and the first 215,000 generations were discarded as

burn-in, using standard deviation of splits between runs less than 0.01 as a convergence

criterion.

New species

Caloplaca allochroa Y. Joshi, Vondrak & Hur, sp. nov. Fic. 1

MycoBank MB 561554

Thallus crustaceus, continuus ad rimosus ad disperso-areolatus, laevigatus, tenuis,

luteus vel cinereus, UV-. Prothallus praesentia, nigrescens. Apothecia biatorina, sessilia,

rotundata ad angulatus, 0.3-0.8 mm. Discus planiusculus ad +convexus, porphyreus ad

fuscus, +pruinosus. Proper margine flavoaurantiacus, persistens. Hymenium inspersum.

Hypothecium indistinctus cellularis. Proper margine tenuis, prosoplectenchymatous.

Thalline margine absentia. Paraphyses simplices, +furcatus ad apicis, septatus. Asci

8-spori, ascosporae polarilocularis, ellipsoideus ad late ellipsoideus, 15-20 um longae et

7.5-10 ym latae.

Type: South Korea. Gangwon Prov.: Yangyang-gun, Seo-myeon, Hwang-ri, Mt Jobong,

37°56'10"N, 128°33'74"E, alt. 980 m, on siliceous rock, 14 May 2009, Y. Joshi, X.Y. Wang,

J.A. Ryu, J.Y. Hur 090296 (HOLOTYPE KoLRI; isotypes, CBFS JV7987, KH).

EryMoLocy — The species epithet derives from the Latin word allochrous, which means

changing colour.

THALLUS saxicolous, crustose, continuous to rimose to dispersed areolate,

smooth, indeterminate, effuse, 6-8 cm in diam., 70-90 um thick [n = 20],

whitish-gray (Fic. 1A) or yellow (Fic. 1B). Correx thin to moderately thick,

8-23 um thick [n = 20], paraplectenchymatous, necral layer absent. Algal layer

continuous, 25-50 um high [n = 20]. Medulla hyaline, of densely interwoven

hyphae, without crystals, 12.5-40 um high [n = 20]. ProrHattus blackish-

gray.

APOTHECIA biatorine, numerous, scattered to +aggregated, 0.3-1.0 mm

in diam. [n = 20], sessile to +constricted at the base, slightly concave when

young, later plane to slightly convex; disc rounded to irregular, brownish-red

to brownish-black, + greenish-yellow pruinose (Fic. 1B); margin egg yolk

coloured to yellowish-orange (distinctly paler than disc) to orange-brown,

variously thick, flush, smooth to +wavy, entire, persistent. EPIHYMENIUM

dark golden brown, with numerous yellowish brown granules, 12.5-20 um

high [n = 20]. HyMenrum hyaline, with numerous oil droplets not dissolving

in K, 87.5-100 um high [n = 20]. HyporHecium hyaline, of indistinct

264 ... Joshi & al.

Fic. 1 Caloplaca allochroa (isotype). A. White-gray thallus with apothecia (Scale = 1 mm); B. Yellow

thallus with apothecia (Scale = 1 mm).

cells, 75-110 um [n = 20]. PROPER MARGIN thin to thick, 25-75 um wide

[n=20], prosoplectenchymatous, with outer region yellowish-brown pigmented.

Algal layer +continuous below the hypothecium. THALLINE MARGIN absent.

PaRAPHYSES hyaline, simple to +furcated at the apices, septate, 1.5-2 um thick,

tips 2.5-3 um broad [n = 20]. Ascosporgs 8 per ascus, hyaline, polarilocular,

ellipsoid to broadly ellipsoid, 12.5-21.5 x 6-11.5 um, isthmus 2.5-7.5 um

[n = 20]. Pycnrp1a rarely present, ostioles reddish-orange; conidia bacilliform,

2.1-2.5 x 0.7-0.9 um [n = 20].

CHEMISTRY — Spot test reactions: thallus K- (grayish regions) or K+

purple (yellowish regions), C-, KC-, P-, UV-. Epihymenium K+ purple, C-.

Apothecia contain parietin and fragilin (major anthraquinones), emodin and

7-chloroemodin (traces); fragilin was observed as the major pigment in yellow

thalli together with traces of parietin, emodin and 7-chloroemodin. Only traces

of parietin and fragilin were observed in white thalli. No acetone insoluble

pigments were identified in darkened apothecial discs.

PHYLOGENY — Caloplaca allochroa is placed in the clade containing well-

known species C. flavorubescens (Huds.) J.R. Laundon and C. flavovirescens

(Wulfen) Dalla Torre & Sarnth. (PP=1.00). The clade is not internally well-

resolved with C. egeana, C. flavovirescens and C. stantonii in the basal polytomy

(FIG! 2):

ECOLOGY AND DISTRIBUTION — Caloplaca allochroa is so far known only

from three mountains (Mt Jobong, Mt Taebaek, and Mt Gyeokja) in South

Korea. The species occurs on non-calcareous rocks in subtropical inland areas

at elevations from 350-980 m. It grows both on subvertical and horizontal faces

of the rocks, preferring situations with sufficiency of diffuse sunlight, under

open canopied stands. Both yellow and gray-white morphotypes are reported

from Mt Jobong and Mt Gyeokja, while in Mt Taebaek only the gray-white

morphotype has been collected.

Caloplaca allochroa sp. nov. (Korea) ... 265

Caloplaca ferruginea FJ866808

C. crenularia EF643512

C. stantonii HQ644197

——C. egeana HQ644198

C. flavovirescens AF 353966

C. allochroa HQ415800

C. juniperina HQ644199

1.00

C. juniperina HQ644200

ORS

VOD

C. flavorubescens EU266111

ij C. flavorubescens AY 143394

1.00

C. flavorubescens AF279887

0.1

Fic. 2 Bayesian phylogenetic tree based on ITS nrDNA sequences showing the alliance of

Caloplaca allochroa with C. flavorubescens and C. flavovirescens. Posterior probabilities are

present at nodes.

Its three known communities have rather different lichen compositions. At

the type locality Caloplaca allochroa was growing in a community with species

of Cladonia, Heterodermia, Leptogium, Myelochroa, Pertusaria, Porpidia and

Ramalina. The new species was accompanied only by Aspicilia sp. on Mt Taebaek

but by Leptogium, Phaeophyscia, Myelochroa, Pertusaria, and Lecanora on Mt

Gyeokja. Even though phorophytes, such as Acer, Fraxinus, Rhododendron,

Pinus, and Quercus occur in all three C. allochroa localities, the new taxon has

not yet been found growing on bark.

ADDITIONAL SPECIMENS EXAMINED: SOUTH KOREA. GANGWON PRov.: Taebaek-si,

Mungoksodo-dong, Mt Taebaek, 37°06'08.3"N, 128°57'05.4"E, alt. 970 m, on rock, 14

October 2005, L. Lok6és 050734 (KoLRI). JEOLLANAM PRov.: Wando Co., Bogil-myeon,

Bogil Island, Buyong-ri, Mt Gyeokja, en route from Keungiljae to Suribong, 34°08'50"N,

126°32'90"E, alt. 368 m, on rock, 05 February 2010, Y. Joshi, H. S. Jeon & M . H. Jeong

100152 (KoLRI).

REMARKS — This is a distinctive species characterized by the colour variation

shown by its thallus (yellow to gray), dark brown-red biatorine apothecia with

yellow-orange margin, and inspersed hymenium. The colour of the areoles is

266 ... Joshi & al.

yellow and the discs are orange brown without traces of black colour in less sun-

exposed sites but on the strongly exposed rock faces, the thallus is distinctly

gray and the apothecia are brownish-black.

The yellow morphotype of the new species is most likely to be confused

with the saxicolous Caloplaca flavovirescens or the corticolous C. alnetorum

Giralt et al., C. flavorubescens, and C. gordejevii (Tomin) Oxner ex Khodos., but

none of the latter possesses such a strong colour contrast between apothecial

disc and margin. Gray-white thallus morphotypes are similar to saxicolous

Caloplaca egeana Cl. Roux & Nav.-Ros. and C. subochracea (Wedd.) Werner, and

corticolous C. aegatica Giralt et al., which also differ in apothecial coloration.

Caloplaca lypera Poelt & Hinter., a similar species from temperate or sub-

alpine Himalayan regions, cannot be confused with this taxon, as it differs in

having yellowish medulla and ochraceous-gray thallus (Poelt & Hinteregger

1993; Joshi & Upreti 2011). The combination of parietin and fragilin as main

anthraquinones in the new species is rare within Caloplaca; it is known (as

in C. flavovirescens and C. flavorubescens; Santesson 1970) and described as

chemosyndrome B1 by S@chting (2001).

Existence of two colour thallus variants based on presence vs. absence of

anthraquinones is probably not exceptional in Teloschistaceae; it has been

recently described in Caloplaca phlogina (Ach.) Flagey (Vondrak et al. 2010)

and also occurs in the North American C. stanfordensis H. Magn. (Vondrak,

unpublished data).

Acknowledgments

This work was supported by a grant from the Korea National Research Resource

Center Program through National Research Foundation of Korea (NRF) and Korean

Forest Service Program (KNA 2011) through Korea National Arboretum. Sequencing

was financed by the Visegrad Fund (Grant 51000067). The authors are thankful to Drs

A. Khodosovtsev and U. Sechting for reviewing the manuscript and providing valuable

comments. Josef Halda and Kerry Knudsen kindly provided their lichen samples and

Pavel Hrouzek performed HPLC.

Literature cited

Arup U. 2006. A new taxonomy of the Caloplaca citrina group in the Nordic countries, except

Iceland. Lichenologist 38: 1-20. http://dx.doi.org/10.1017/S0024282905005402

Arup U, Akelius, E. 2009. A taxonomic revision of Caloplaca herbidella and C. furfuracea.

Lichenologist 41(5): 465-480. http://dx.doi.org/10.1017/S0024282909008780

Arup U, Grube M. 1999. Where does Lecanora demissa (Ascomycota, Lecanorales) belong?

Lichenologist 31(5): 419-430.

Arup U, Arneng E, Sochting U. 2007. Caloplaca fuscorufa - a misunderstood species in northern

Europe. Lichenologist 39(5): 409-414. http://dx.doi.org/10.1017/S0024282907007098

Ekman S. 2001. Molecular phylogeny of the Bacidiaceae (Lecanorales, lichenized Ascomycota).

Mycological Research 105: 783-797.

Caloplaca allochroa sp. nov. (Korea) ... 267

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes. Application

for the identification of mycorrhizae and rust. Molecular Ecology 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Joshi Y, Upreti DK. 2011. Four new records of Caloplaca (lichenized Ascomycetes) from India.

Mycotaxon 116: 53-60. http://dx.doi.org/10.5248/116.53

Kasalicky T, Déring H, Rambold G, Wedin M. 2000. A comparison of ITS and LSU nrDNA

phylogenies of Fulgensia (Teloschistaceae, Lecanorales), a genus of lichenized ascomycetes.

Canadian Journal of Botany 78(12): 1580-1589. http://dx.doi.org/10.1139/cjb-78- 12-1580

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

10" ed. Wallingford, Oxon, CAB International.

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology

Centre, Uppsala University.

Poelt J, Hinteregger E. 1993. Beitrage zur Kenntnis der Flechtenflora des Himalaya. VII. Die

Gattungen Caloplaca, Fulgensia und Ioplaca (mit englischem Bestimmungsschliissel).

Bibliotheca Lichenologica 50: 1-247.

Posada D, Crandall KA. 1998. Modeltest—testing the model of DNA substitution. Bioinformatics

14: 817-818. http://dx.doi.org/10.1093/bioinformatics/14.9.817

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

Santesson R. 1970. Anthraquinones in Caloplaca. Phytochemistry 9: 2149-2166.

http://dx.doi.org/10.1016/S0031-9422(00)85380-7

Sochting U. 1997. Two major anthraquinone chemosyndromes in Teloschistaceae. Bibliotheca

Lichenologica 68: 135-144.

Sechting U. 2001. Chemosyndromes with chlororinated anthraquinones in the lichen genus

Caloplaca. Bibliotheca Lichenologica 78: 395-404.

Vondrak J, Soun J, Sogaard M, Sochting U, Arup U. 2010. Caloplaca phlogina, a lichen with two

faces; an example of infraspecific variability resulting in the description of a redundant species.

Lichenologist 42: 685-692. http://dx.doi.org/10.1017/S0024282910000435

Wedin M, Baloch E, Grube M. 2002. Parsimony analyses of mtSSU and nITS rDNA sequences

reveal the natural relationships of the lichen families Physciaceae and Caliciaceae. Taxon 51(4):

655-660. http://dx.doi.org/10.2307/1555020

Wei X, Jeon H-S, Han KS, Koh YJ, Hur J-S. 2008. Antifungal activity of lichen-forming fungi

isolated from Korean and Chinese lichen species against Colletotrichum acutatum causing

anthranose on hot pepper. Plant Pathology Journal 24(2): 202-206.

White TJ, Bruns TD, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

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http://dx.doi.org/10.2307/1555020

MY COTAXON

http://dx.doi.org/10.5248/117.269

Volume 117, pp. 269-278 July-September 2011

Four anamorphic fungi (with two new species)

from forests of Western Ghats, India

J. PRATIBHA", D.J. BHAT * & S. RAGHUKUMAR™

'Myko Tech Pvt. Ltd., Plot no. 12, Mapusa Industrial Estate, Mapusa Goa - 403507, India

*Department of Botany, Goa University, Goa - 403 206, India.

CORRESPONDENCE TO: “jalmipratibha@rediffmail.com, *bhatdj@rediffmail.com,

*s_raghukumar@mykotech.com,

ABSTRACT — Anaselenosporella indica and Arachnophora goanensis, two new species of

anamorphic fungi isolated from decaying plant litter collected from the forest of Goa, India,

are described and illustrated. Anaselenosporella indica, found growing on dead twig of an

unidentified plant, is characterized by polyblastic, sympodial, discrete, conidiogenous cells

and cylindrical, rarely curved, aseptate, hyaline conidia. Arachnophora goanensis, collected

from dry decaying bark of an unidentified tree, is characterized by blastic pigmented

stauroconidia and blastic hyaline fusiform synanamorphic conidia. Two other species from

the monotypic genera Catenosynnema and Cheiromyceopsis are also reported for the first time

from India.

Key worps — biodiversity, taxonomy

During research on microfungi from forests of Western Ghats in Goa, four

interesting fungi belonging to the anamorphic genera Anaselenosporella

Heredia et al., Arachnophora Hennebert, Catenosynnema Kodsueb et al. and

Cheiromyceopsis Mercado & J. Mena were isolated from fallen and decaying

plant litter. Two are described and illustrated as new species, Anaselenosporella

indica and Arachnophora goanensis. Collections of new species are conserved

in the herbarium of Indian Agricultural Research Institute, New Delhi (HCIO)

and new records at Department of Botany, Goa University, Goa.

Anaselenosporella indica Pratibha, Bhat & Raghuk. sp. nov. Fics 1, 2

MycoBank MB561104

Ad fungus anamorphicus. Conidiophora macronemata, mononemata, singula,

erecta, laevia, recta vel leviter flexuosa, ramosa ad apicum, atrobrunnea ad basim,

pallid brunnea ad apicum, multiseptata, 85-140 x 4.5-8 ym. Cellulae conidiogenae

polyblasticae, lageniformes, leviter geniculatus et elongatus versus apicum, 15-30 x 2-4

270 ... Pratibha, Bhat & Raghukumar

Fic. 1. Anaselenosporella indica.

Conidiophores, conidiogenous cells and conidia.

um, indeterminatus, sympodialis, discretae ex ramis metuloideis, cuneiformibus, 10-14x

3-6 um. Conidia solitaria, cylindrica, raro curvus, aseptata, utrinque rotandata, hyalina,

laevia, 8-12 x 1.5 um.

TyPeE: India, Goa, Ponda, Bondla, on dead twig of unidentified tree, 20/08/2010, Pratibha

J., HOLOTYPE: HCIO 50176.

EryMo_oey: indicus (Latin), referring to the country where the type was collected.

Anamorphic fungus. Colonies on natural substrate effuse, brown; mycelium

partly superficial, partly immersed, composed of smooth, light brown,

branched, septate, 2-3.5 um wide hyphae. Colonies on PDA dark green,

Anaselenosporella indica & Arachnophora goanensis spp. nov. (India) ... 271

Fic. 2. Anaselenosporella indica: a—e. conidiophores, conidiogenous cells, and conidia;

f. conidiogenous cells and conidia; g. conidia.

272 ... Pratibha, Bhat & Raghukumar

wooly, reverse black, margin serrated, attaining a diam. of 1.8 cm in 10 days.

Conidiophores macronematous, mononematous, single, erect, smooth,

straight to slightly flexuous, branched at the apex, dark brown at the base, pale

brown towards the tip, multiseptate, 85-140 x 4.5-8 um. Conidiogenous cells

polyblastic, lageniform, slightly geniculate and elongated towards the apex, 15-

30 x 2-4 um, indeterminate, sympodial, discrete, formed in a compact cluster

on cuneiform, dark brown, smooth, thick-walled, 10-14 x 3-6 um metuloid

branches. Conidia solitary, cylindrical, rarely curved, aseptate, rounded at both

ends, hyaline, smooth, 8-12 x 1-1.5 um.

Notes: Heredia et al. (Castafieda et al. 2010) established the monotypic genus

Anaselenosporella with A. sylvatica as type species to accommodate a fungus

with polyblastic, discrete, lageniform, sympodial conidiogenous cells formed

on metuloid cells with acicular, filiform, fusiform to semi-circular, unicellular,

hyaline conidia. Anaselenosporella indica differs from the type species in

conidiophore and conidiogenous cell dimensions and conidiophore branching.

In A. sylvatica the conidiophores are much larger (700-1200 x 12-28 um) and

dichotomously branched and the conidiogenous cells smaller (5-10 x 2-2.5

um) than for A. indica. The type species is further differentiated by conidia

that are acicular, curved to semicircular, truncate at the base, and not solitary,

cylindrical, rarely curved, rounded at both ends as in the new species.

Arachnophora goanensis Pratibha, Bhat & Raghuk. sp. nov. Fics 3, 4

MycoBank MB561105

Ad fungos anamorphicus. Conidiophora macronemata, mononemata, apicem versus

attenuata, singula, erecta, laevia, recta vel leviter flexuosa, non-ramosa, atrobrunnea ad

basim, pallide brunnea ad apicem, multiseptata, 80-200 x 4.5-7 um. Cellulae conidiogenae

monoblasticae, cylindricae, terminales, integratae, determinatae vel interdum percurrentes,

pallide brunneae, 5-8 x 2.5-4 um. Conidia solitaria, blastica, acrogena, staurosporia,

septata, sicca, laevia, 25-40 x 20-25 um, cum unicus, atrobrunnea, 6-9 x 5.5-8.5 um

cellulae basali et 1-3, atrobrunneae, 3-7.5 x 5.5-8.5 um cellulae centrales, edens ortus ad

pallide brunnae cellulae laterales, ortus ad 1-4, hyalina ad pallide brunnea, 4-8 x 1-3

um brachiis. Synanamorpha conidia hyalina, fusiformia, aseptata, 5-8 x 1 um, in cellulae

peripherales, oriunda.

Tye: India, Goa, Sanguem, Netravali, on dry and decaying bark of unidentified tree,

28/09/10, Pratibha J., HOLOTYPE: HCIO 50177.

ETYMOLOGY: goanensis (Latin), referring to the state where the type was collected.

Anamorphic fungus. Colonies on natural substrate effuse, brown; mycelium

partly superficial, partly immersed, composed of smooth, light brown,

branched, septate, 2-3 um wide hyphae. Colonies on PDA dark green,

flat, reverse black, margin serrated, attaining a diam. of 1.4 cm in 10 days.

Conidiophores differentiated, mononematous, tapering towards apex, single,

erect, smooth, straight to slightly flexuous, unbranched, dark brown at the base,

Anaselenosporella indica & Arachnophora goanensis spp. nov. (India) ... 273

Fic. 3. Arachnophora goanensis.

Conidiophores, conidiogenous cells, stauroconidia, and Selenosporella-like synanamorph.

pale brown towards the apex, multiseptate, 80-200 x 4.5-7 um. Conidiogenous

cells monoblastic, cylindrical, terminal, integrated, determinate or sometimes

percurrently proliferating, light brown, 5-8 x 2.5-4 um. Conidia solitary,

274 ... Pratibha, Bhat & Raghukumar

Fic. 4. Arachnophora goanensis: a-e. conidiophores, conidiogenous cells and conidia;

f-i. stauroconidia, with Selenosporella-like synanamorph.

Anaselenosporella indica & Arachnophora goanensis spp. nov. (India) ... 275

blastic, acrogenous, staurosporous, septate, dry, smooth, 25-40 x 20-25 um,

with one, dark brown, 6-9 x 5.5-8.5 um basal cell and 1-3, dark brown, 3-7.5

x 5.5-8.5 um central cells; each central cell either gives rise directly to 2 or

more fertile, hyaline to pale brown arm-cells, or to a light brown lateral cell,

which in turn gives rise to 1-4 fertile arms; fertile arms hyaline to pale brown,

4-8 x 1-3 um, producing synanamorphic conidia at the tip. Selenosporella-like

synanamorphic conidia hyaline, fusiform, aseptate, 5-8 x 1 um, formed on tip

of the peripheral cells of each conidial arm.

Notes: Hennebert (1963) established Arachnophora to accommodate a

fungus having monoblastic, terminal, integrated, percurrently proliferating

conidiogenous cells and staurosporous, solitary, pigmented, acrogenous conidia

and with usually two central cells in the axis and several radial, recurved, conical

arms that arise from the central cells. Of the eight species previously included

in the genus, only six are currently accepted (TABLE 1): Arachnophora crassa,

A. excentrica (B. Sutton) S. Hughes [= Digitoramispora excentrica (B. Sutton)

R.F. Castafeda & W.B. Kendr.], A. fagicola (type species), A. hughesii, A.

polyradiata, A. pulneyensis, A. simplex Ichinoe [= Uberispora simplex (Ichinoe)

Piroz. & Hodges], and A. uberisporoides (Hennebert 1963, Pirozynski & Hodges

1973, Révay & Goncz6l 1989, Castafeda & Kendrick 1990, Castaneda et al.

1996, 1997; Castafieda & Guarro 1998).

TABLE 1: Synopsis of accepted species of Arachnophora.

SPECIES

A. crassa

Révay & Génczél

: 2-celled central body, 16-20 x 10-13 wm, each

A. fagicola

Hennebert

2-4-celled central body, 25-40 x 20-25

2-3-celled central body, 27-30(-37) x

A. hughesii

R.F. Castafieda

& Guarro

: 3-celled central body, 28-40 x 25-38 um,

: each cell bearing 2—3-septate, radiating arms :

A. polyradiata (Mercado

& R.E Castafieda) R.E

Castafieda & W. Gams

2-3-celled central body, 16-24 x 6.4-8.1 um,

A. pulneyensis (Subram.

& Bhat) R.F. Castafieda

2-celled central body, 17-20 x 9-13

A. uberisporoides

R.E Castafieda

: CONIDIA

2-celled central body, 16-22 x 8-9.6 um, each

cell bearing 1-2-celled lateral branches

cell bearing several lateral protuberances,

each bearing 1 or more inwardly

curved, claw- or spine-like processes

uum; each central cell bearing aseptate,

hyaline to pale brown arms, 4-8 x

1-3 um, borne directly on either central

cell or light brown lateral cells

17-24(-30) um, with 2-5 tapered, 0-1-

septate horn-like or conical arms

with up to 4 arms

um, with 4-6 dome-shaped arms

: Selenosporella-L1KE

SYNANAMORPH

: Borne directly on the

apex of lateral branches

: Conidia blastic, hyaline,

fusiform, aseptate, 5-8

x lum

: Conidia blastic, fusiform,

aseptate, colourless,

3-4 x 0.5-1 um

: Conidia hyaline, aseptate,

4.5-7 x 0.5-1 um

276 ... Pratibha, Bhat & Raghukumar

Arachnophora goanensis resembles A. polyradiata in conidial shape and the

Selenosporella-like synanamorph at the tip of the arms with fusiform, hyaline,

1-celled conidia. However, it differs in conidiophore dimension and number of

conidial cells and arms. ‘The A. polyradiata conidiophores are shorter (< 50 um

long) and the conidia are composed of two dark brown central cells, with each

cell giving rise to one 2—3-septate, radiating arm.

Catenosynnema micheliae Kodsueb et al. Cryptog. Mycol. 28: 239. 2007. Fras 5a, b

Anamorphic fungus. Conidiomata sporodochial, superficial, light brown.

Conidiophores branched, light brown, 40-70 x 2-4 um. Conidiogenous cells

blastic, integrated, determinate, sub-hyaline. Conidia in simple to branched

acropetal chains, aseptate, light brown, smooth, ellipsoidal to fusiform, 8-12

x 3-5 um.

SPECIMEN EXAMINED: INDIA. MAHARASHTRA: Amboli, on dead and decaying twigs of

unidentified tree, 18/08/09, Pratibha J..GUBH MTO1.

Cheiromyceopsis echinulata Mercado & J. Mena, Acta Bot. Cubana 53: 2. 1988.

Fics. 5c, d

Anamorphic fungus. Conidiomata sporodochial, superficial, light brown.

Conidiophores macronematous, brown, 20-40 x 5-7 um. Conidiogenous cells

monoblastic, integrated, terminal, determinate. Conidia cheiroid, dark brown,

22-30 um long, branched; branches 3-7, verrucose, septate, 15-20 x 5-7.5

um.

SPECIMEN EXAMINED: INDIA. KaRNaTAKA: UTTAR KANNADA, Kathlekan, on dry and

decaying bark of unidentified tree, 17/11/09, Ashish P., GUBH MT02.

Discussion

This paper is the first report of the genera Anaselenosporella, Catenosynnema,

and Cheiromyceopsis from India, and of the genus Arachnophora from the

Western Ghats. The genus Anaselenosporella was monotypic, reported only

from Mexico. Catenosynnema and Cheiromyceopsis are monotypic, previously

reported from Cuba and Thailand, respectively. The genus Arachnophora is

widely distributed. The collections from Western Ghats form an interesting

observation.

Acknowledgments

DJB thanks the CSIR, MoEF, and UGC, New Delhi, for financial support in the form of

research grants. We are indebted to Dr R.F. Castafieda-Ruiz, Instituto de Investigaciones

Fundamentales en Agricultura Tropical Alejandro de Humboldt (INIFAT), Cuba for

providing literature, valuable suggestions and reviewing the manuscript and to Dr Eric

McKenzie, Landcare Research, New Zealand, for kindly reviewing the manuscript for

Mycotaxon.

Anaselenosporella indica & Arachnophora goanensis spp. nov. (India) ... 277

Fic. 5. a-b: Sporodochia and conidial chain of Catenosynnema micheliae.

c-d: Cheiroid conidia of Cheiromyceopsis echinulata.

Literature cited

Castafieda-Ruiz RF, Guarro J. 1998. Two new hyphomycetes from rainforests of Cuba. Canadian

Journal of Botany 76(9): 1584-1588. http://dx.doi.org/10.1139/cjb-76-9-1584

Castafieda-Ruiz RF, Kendrick B. 1990. Conidial fungi from Cuba: II. University of Waterloo,

Biology Series 33: 1-62.

278 ... Pratibha, Bhat & Raghukumar

Castafieda-Ruiz RF, Guarro J, Cano J. 1996. Notes on conidial fungi. X. A new species of

Ceratosporella and some new combinations Mycotaxon 60: 275-281.

Castafieda-Ruiz RF, Gams W, Saikawa M. 1997. Three new conidial fungi (hyphomycetes) from

Cuba. Nova Hedwigia 64: 473-483.

Castafieda-Ruiz RF, Heredia AG, Arias RM, Stadler M, Saikawa M, Minter DW. 2010.

Anaselenosporella sylvatica gen. & sp. nov. and Pseudoacrodictys aquatica sp. nov., two new

anamorphic fungi from Mexico. Mycotaxon 112: 65-74. http://dx.doi.org/ 10.5248/112.65

Hennebert GL. 1963. Un hyphomycéte nouveau, Arachnophora fagicola gen. nov. spec. nov.

Canadian Journal of Botany 41(8): 1165-1169. http://dx.doi.org/10.1139/b63-097

Pirozynski KA, Hodges CS. 1973. New hyphomycetes from South Carolina. Canadian Journal of

Botany 51(1): 151-173. http://dx.doi.org/10.1139/b73-024

Révay A, G6ncz6l J. 1989. Some dematiaceous hyphomycetes from woody litter in Hungary. Nova

Hedwigia, 48: 237-245.

MY COTAXON

http://dx.doi.org/10.5248/117.279

Volume 117, pp. 279-285 July-September 2011

Taxonomic studies of Endophragmiella

from southern China

L1-Guo Ma, JIAN Ma, YI-DONG ZHANG & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Two new species, Endophragmiella clausenae from dead branches of Clausena

lansium and E. pentaphylacis from dead branches of Pentaphylax euryoides, are described

and illustrated. Endophragmiella corticola is a new record for China. These three species were

collected from tropical and subtropical forests in southern China.

Key worps — hyphomycetes, taxonomy

Introduction

Sutton (1973) established Endophragmiella B. Sutton to accommodate two

species, E. pallescens B. Sutton and E. canadensis (Ellis & Everh.) B. Sutton.

The genus is characterized by macronematous, branched or unbranched

conidiophores with terminal, integrated, percurrently proliferating and

monoblastic conidiogenous cells producing solitary and euseptate or

distoseptate conidia with rhexolytic secession (Sutton 1973, Ellis 1976, Hughes

1979, Wu & Zhuang 2005). These characters separate Endophragmiella from

similar genera, Phragmocephala E.W. Mason & S. Hughes, Chaetendophragmia

Matsush., and Melanocephala S. Hughes. Of the 80 species now assigned to

Endophragmiella, most are saprobes, occurring on rotten wood, dead branches,

and decaying leaves (Wu & Zhuang 2005).

Fungal diversity in southern China is high, and many wood-inhabiting

fungi have been discovered (Dai et al. 2009, Dai and Li 2010, Ma et al. 2011).

In our study on conidial fungi on dead wood from the tropical and subtropical

forests in southeast China, two hitherto undescribed species and a new record

of Endophragmiella were collected. The 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).

280 ... Ma & al.

20um

@ @ @®. ®D DD wD

Fic. 1. Endophragmiella clausenae. a. Conidiophores and conidia. b. Conidia.

Taxonomy

PiG#4.

Endophragmiella clausenae L.G. Ma & X.G. Zhang, sp. nov.

MycoBank MB 561597

COLONI4E effusae, brunneae. Mycelium partim immersum, partim superficiale ex hyphis

septatis, ramosis, laevibus, pallide brunneis, 2-4 um latis, compositum. CONIDIOPHORA

Endophragmiella spp. nov. (China) ... 281

solitaria, macronemata, mononemata, recta vel flexuosa, erecta, simplicia, multiseptata,

laevia, brunnea, 135-410 x 4.5—8.5 um. CELLULAE CONIDIOGENAE monoblasticae,

terminals, in conidiophoris incorporatae, cum proliferationibus percurrentes, cylindricae,

laeviae, pallide brunneae. Secessio conidiorum rhexolytica. ConrpIA solitaria, acrogena,

3-euseptata, ellipsoidea, clavata vel pyriformia, simplicia, sicca, laevia, brunnea vel

atrobrunnea, cellula basalis 4.5—6 um crassa, pallide brunneis, 31.5—42.5 x 14—16.5um.

Type: China, Yunnan Province: the Forbidden Forest of Banna, on dead branches of

Clausena lansium (Lour.) Skeels (Rutaceae), 17 Oct. 2008, L.G. Ma (holotype HSAUP

H0074; isotype HMAS 146108).

EryMoLoey: in reference to the host genus, Clausena.

CoLoniEs on the natural substrate effuse, brown. Mycelium partly immersed,

partly superficial, composed of septate, branched, smooth-walled, pale

brown hyphae, 2-4 um thick. ConipIiopHorREs solitary, macronematous,

mononematous, straight or flexuous, erect, simple, multiseptate, smooth,

brown, 135-410 x 4.5-8.5 um. CONIDIOGENOUS CELLS monoblastic, terminal,

integrated, with conspicuous percurrent proliferations, cylindrical, smooth,

pale brown. Conidial secession rhexolytic. Conip1A solitary, acrogenous, 3-

euseptate, ellipsoid, clavate to pyriform, simple, dry, smooth, brown to dark

brown, basal cells 4.5-6 um wide, pale brown, 31.5-42.5 x 14-16.5 um.

Endophragmiella clausenae is similar to E. bisbyi (Hughes 1978a) and

E. mexicana (Mercado et al. 1995) in conidial shape. Endophragmiella bisbyi

differs from the new species by its smaller (9-13 x 5-6.5 um) conidia with

the thick distal septum and brown distal cell, while E. mexicana is easily

distinguished by its much smaller (11-16 x 5.8-7.8 um) predominantly

4-septate conidia with paler distal and basal cells.

Endophragmiella pentaphylacis L.G. Ma & X.G. Zhang, sp. nov. FIG. 2

MycoBank MB 561604

COLONI4E effusae, brunneae. Mycelium partim immersum, partim superficiale ex hyphis

septatis, ramosis, laevibus, subhyalinis vel pallide brunneis, compositum. CONIDIOPHORA

solitaria, macronemata, mononemata, recta, erecta, simplicia, septata, laevia, pallide

brunnea vel brunnea, 56-92 x 4.5—7.5 um. CELLULAE CONIDIOGENAE monoblasticae,

terminals, in conidiophoris incorporatae, cum proliferationibus percurrentes, cylindricae,

laeviae, pallide brunneae. Secessio conidiorum rhexolytica. ConipIA solitaria, acrogena,

3-septata, late ellipsoidea vel pyriformia, simplicia, sicca, laevia, brunnea vel atrobrunnea,

cellula basalis 2.5—5 um crassa, pallide brunneis, 25—32.5 x 13-16.5um.

Type: China, Yunnan Province: the Forbidden Forest of Banna, on dead branches of

Pentaphylax euryoides Gardner & Champ. (Pentaphylaceae), 18 Oct. 2008, L.G. Ma

(holotype HSAUP H0042; isotype HMAS 146109).

EryMo_oey: in reference to the host genus, Pentaphylax.

CoLonies on the natural substrate effuse, brown. Mycelium partly

immersed, partly superficial, composed of septate, branched, smooth-walled,

subhyaline to pale brown hyphae. Conrp1oPHoRESs solitary, macronematous,

282 ... Ma &al.

Oum

Fic. 2. Endophragmiella pentaphylacis. a. Conidiophores and conidia. b. Conidia.

mononematous, straight, erect, simple, septate, smooth, pale brown to brown,

56-92 x 4.5—7.5 um. CONIDIOGENOUS CELLS monoblastic, terminal, integrated,

with conspicuous percurrent proliferations, cylindrical, smooth, pale brown.

Conidial secession rhexolytic. Conrp1a solitary, acrogenous, 3-septate, broadly

ellipsoid to pyriform, simple, dry, smooth, brown to dark brown, basal cells

2.5-5 um wide, pale brown, 25-32.5 x 13-16.5 um.

Endophragmiella pentaphylacis resembles E. bisbyi (Hughes 1978a),

E. ontariensis (Hughes 1978b), E. ellisii (Hughes 1979), and E. suttonii (Kirk

1981) in conidial shape, but it can be differentiated from E. suttonii, E.

ontariensis and E. ellisii by its larger, predominantly 3-septate and dark brown

to black conidia. E. bisbyi can be separated from E. pentaphylacis, which has

smaller (9-13 x 5-6.5 um), versicolored conidia.

Endophragmiella corticola P.M. Kirk, Trans. Br. Mycol. Soc. 78(1): 60, 1982. Fic. 3

CoLonigs on the natural substrate pale brown to brown, effuse. Mycelium

mostly superficial composed of branched, septate, subhyaline to pale brown,

smooth-walled hyphae, 1.5-3 um thick. CONIDIOPHORES macronematous,

Endophragmiella spp. nov. (China) ... 283

Fic. 3. Endophragmiella corticola. a-b. Branched conidiophores and conidia. c. Conidia.

mononematous, arising terminally and laterally from the mycelium, straight or

flexuous, erect, branched, septate, smooth, pale brown, up to 350 um high, 2.5-4

um wide. CONIDIOGENOUS CELLS monoblastic, terminal, integrated, percurrent,

cylindrical, tapered to a truncate apex, smooth, subhyaline. Conidial secession

rhexolytic. Conip1a solitary, acrogenous, 2-3-septate, narrowly obclavate to

obclavate, simple, smooth, basal cells 1.5-3 um wide, pale brown to brown,

apical cells subhyaline, 16.5-25 x 5.5-7.5 um.

284 ... Ma &al.

SPECIMENS EXAMINED: China, Fujian Province: the National Forest Park of Wuyishan,

on dead branches of Ficus gibbosa Blume (Moraceae), 14 Aug. 2009, L.G. Ma, HSAUP

H1026 (duplicate HMAS 146110).

Endophragmiella corticola is similar to E. eboracensis (Sutton 1975), E. acuta

(Wu & Zhuang 2005), E. verticillata (Hughes 1978c), E. curvata (Hughes

1979), and E. cesatii (Hughes 1979). Ellipsoidal, shorter, 3-septate conidia

and verticillately branched conidiophores distinguish E. verticillata, while

E. eboracensis can be separated by its much shorter and mainly 3-septate conidia.

Endophragmiella acuta has wider, 3-septate conidia with a tapered and rostrate

apex and occasionally branched conidiophores; E. curvata has predominantly

2-septate conidia and unbranched conidiophores; and E. cesatii has wider,

predominantly 3-septate conidia with brown central cells and unbranched

conidiophores. The type material differs from the Chinese specimen in generally

longer (14-42 um) conidia and shorter (< 60 um) conidiophores. ‘This is the

first record from China.

Acknowledgments

The authors are grateful 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

Dai YC, Li HJ. 2010. Notes on Hydnochaete (Hymenochaetales) with a seta-less new species

discovered in China. Mycotaxon 111: 481-487. http://dx.doi-org/10.5248/111.481

Dai YC, Cui BK, Yuan HS. 2009. Trichaptum (Basidiomycota, Polyporaceae) from China with a

description of three new species. Mycol. Prog. 8: 281-287.

http://dx.doi.org/10.1007/s11557-009-0598-0

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew,

Surrey, England.

Hughes SJ. 1978a. Endophragmiella bisbyi. Fungi Canadenses 124: 1-2.

Hughes SJ. 1978b. Endophragmiella ontariensis. Fungi Canadenses 128: 1-2.

Hughes SJ. 1978c. Endophragmiella verticillata. Fungi Canadenses 130: 1-2.

Hughes SJ. 1979. Relocation of species of Endophragmia auct. with notes on relevant generic names.

New Zealand J. Bot. 17: 139-188.

Kirk PM. 1981. New or interesting microfungi II. Dematiaceous hyphomycetes from Esher

Common, Surrey. Trans Br. Mycol. Soc. 77(2): 279-297.

http://dx.doi.org/10.1016/S0007-1536(81)80031-9

Ma J, Wang Y, O’Neill NR, Zhang XG. 2011. A revision of the genus Lomaantha, with the description

of a new species. Mycologia 103(2): 407-410. http://dx.doi.org/10.3852/10-176

Mercado Sierra A, Heredia G, Mena Portales J. 1995. New species of dematiaceous hyphomycetes

from Veracruz, Mexico. Mycotaxon 55: 491-499.

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

1-143.

Endophragmiella spp. nov. (China) ... 285

Sutton BC. 1975. Two undescribed dematiaceous hyphomycetes. Naturalist 933: 69-72.

Wu WP, Zhuang WY. 2005. Sporidesmium, Endophragmiella and related genera from China. Fungal

Divers? Res. Ser 152 122351.

MY COTAXON

http://dx.doi.org/10.5248/117.287

Volume 117, pp. 287-290 July-September 2011

Rhexodenticula zhengii sp. nov. from fallen leaves from China

DE-WEI LI*", JINGYUAN CHEN? & YIXUN WANG?

"The Connecticut Agricultural Experiment Station, Valley Laboratory

153 Cook Hill Road, Windsor, CT 06095

?Institute of Forest Disease and Insect Control, Hubei Academy of Forestry

I Lion Peak, Jiufeng, Wuhan, Hubei, 430075, China

*CORRESPONDENCE TO—dewei.li@ct.gov

ABSTRACT— A new species, Rhexodenticula zhengii from leaf litter of Sycopsis sinensis

collected from Houhe National Nature Reserve, Wufeng, Hubei, China is described and

illustrated with cymbiform conidia 18.5-22.0 x 4.5-5.0 um, constricted at proximal and

distal septa. A key to the species of Rhexodenticula is provided.

KEY worps— anamorphic fungi, rhexolytic secession, taxonomy

Introduction

Sycopsis sinensis is a native broadleaved evergreen woody plant species

in China. It grows in mixed evergreen broadleaved forests or bushes and is

sporadically distributed at elevations of 1000-2000 m in southern China (Li et

al. 2003). However, the largest population of S. sinensis as the dominant species

was reported in the Houhe National Nature Reserve by Liu et al. (1999).

A field trip was made to the Houhe National Nature Reserve (29°59'-

30°10'N 110°22'-110°52'E) in Wufeng Tujia Autonomous County, Hubei,

China to collect hyphomycetes in August 2008. A species of Rhexodenticula

new to science was discovered on decaying leaves of Sycopsis sinensis.

Materials & methods

Conidiophores and conidia of the fungus were mounted in 85% lactic acid. Tape

lifts were prepared for wet mounting using Mainstays Crystal Clear Tape (Imported by

Walmart Canada, Mississauga, Ontario). Microscopic observations were made using

bright field and Nomarski differential interference contrast optics. Photomicrographs

were taken with an Olympus Microfire digital camera (Goleta, CA). Herbarium

acronyms follow the Index Herbariorum (Holmgren & Holmgren 1998). Measurements

of the fungal structures were statistically analyzed with Microsoft Office Excel 2007

288 ... Li, Chen & Wang

with 95% confidence interval of means. The results were presented as ranges and mean

+ standard deviation. Q is length/width ratio and n, the number of fungal structures

measured.

Results

Rhexodenticula zhengii D.W. Li & Jing Y. Chen, anam. sp. nov. Fics 1-6

MycoBank MB 561153.

Conidiophora distincta, determinata vel percurrenter et sympodialiter proliferentia, erecta,

solitaria vel fasciculata, non-ramosa, laevia, brunnea, 28-132(-205) x (3.5-)3.5-4.5(-4.5)

um, 1-9-septata. Cellulae conidiogenae polyblasticae, terminales, integratae, denticulatae.

Conidia solitaria, obclavata, 3-septata, brunnea, verruculosa, (13.0-)18.5-22.0 x

(4.0-)4.5-5.0(-5.5) um ad basim margine, protrudente praedita. Teleomorphosis ignota.

Type: China. Hubei: Wufeng, Houhe National Nature Reserve, superficie in folio

emortuo Sycopsis sinensis Oliv. (Hamamelidaceae) Coll. xxviii-viii-2008, De-Wei Li,

[BPI 881495, holotype].

Erymotoey: in honor of Prof. Ruyong Zheng, a fellow of The Chinese Academy of

Sciences, for her contributions to the systematics of powdery mildews and Zygomycota.

CONIDIOPHORES differentiated, single or in groups, determinate or with

percurrent and sympodial extension, erect, unbranched, straight or flexuous,

sometimes geniculate, , dark brown, smooth, 1-9-septate, varying in length,

30-135(-205) x (3.4-)3.5-4.5(-5) um (mean = 79 + 53, x 4+ 0.5, n = 21),

more or less uniform in width with enlarged subglobose basal cell and 0-3

nodes, thick-walled, wall 0.5-0.8 um thick. CONIDIOGENOUS CELLS integrated,

terminal, polyblastic, denticulate, (12.5-)16.5-30.0(-33.5) x (3.0-)3.5-4.0

(-4.5) (mean = 23.5 + 6.5 x 3.7 + 0.3, n = 20) um; denticles (0.6-)0.8-1.2(-1.5)

x 0.7-1.1(-1.5) (mean = 1 + 0.2 x 0.9 + 0.2, n = 17) um. CONIDIA 3-septate,

apical and lateral, single, obclavate, brown, verruculose, (13.0—)18.5-22.0

(mean = 20.1 + 1.7, n = 30) x (4.0-)4.5-5.0(-5.5) (mean = 4.6 + 0.3, n = 30)

um, Q = (4.0)4-5.0(5.5) (mean = 4.4 + 0.4, n = 30), the middle cells darker

and longer than the apical and basal cells, constricted at proximal septum and

slightly constricted at distal septum, basal cell rounded with a narrow vestigial,

marginal frill at the base derived from the rhexolytically split separating cell at

the middle, the frill (0.5—)0.7-—1.1(-1.2) x (0.8-)0.9-1.1(-1.3) (mean = 0.9 + 0.2

x 1+0.1,n= 30) um.

TELEOMORPH: unknown.

DISTRIBUTION AND HABITAT: Known from Hubei, China on fallen leaves of

Sycopsis sinensis.

Discussion

Rhexodenticula was established by Baker & Morgan-Jones (2001) and typified

by Rhexodenticula cylindrospora (R.F. Castaneda et al.) W.A. Baker & Morgan-

Jones. Segregation of Rhexodenticula from Nakataea was based on its key

Rhexodenticula zhengii sp. nov. (China) ... 289

r > . a

Fics 1-6. Rhexodenticula zhengii. 1-2. Conidiophores and conidiogenous cells; arrows in 2

indicate separating cell walls. 3. Basal portions of conidiophores. 4-6. Conidia; the arrow in 5

indicates a separating cell wall. Scale bars: 1, 4-6 = 10 um, 2-3 = 5 um.

290 ... Li, Chen & Wang

character of releasing conidia by rhexolytically splitting a separating cell. The

genus previously had two species, Rhexodenticula elegiae Melnik et al. (Mel'nik

et al. 2004) being the second. Rhexodenticula zhengii adds a third species.

All three species have four-celled conidia. Rhexodenticula cylindrospora has

cylindrical conidia with four cells of equal length (Castafeda et al. 1996, Baker

et al. 2001), while conidia of R. elegiae and R. zhengii are much shorter with

paler end cells. The conidia of R. zhengii are obclavate, larger, and constricted at

basal and distal septa: these characters can differentiate it from R. elegiae, which

has fusiform conidia with unconstricted septa.

Key to species of Rhexodenticula

1; Conidia-eylindtical 1 3=21-x ARBs ike sa bo sed shi eie tthe tal R. cylindrospora

bar Comidia-obclavate-or FUSifOrtite sci dpb sckoz dpi scboa dyke scten dy gama dy gon dye g-dlacayhea Eeube deg ae 2

2. Conidia constricted at proximal and distal septa, obclavate,

NSS ee ero aa Era Sheek aden he Ped aN th A le R. zhengii

2a. Conidia not constricted at septa, fusiform, 16-19 x 5um .............. R. elegiae

Acknowledgments

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

Castafieda Ruiz for their critical review of the manuscript and to Dr. James A. LaMondia

for his pre-submission review. The authors are grateful to the Connecticut Agricultural

Experiment Station for supporting the collaboration. Financial support by the Resources

Sharing Platform of Natural Sciences and Technology, China (2005DKA2120715) to

JYC is acknowledged. The authors also thank Dengkui Tang, Zhizhang Zheng, Yeqing

Wang, Sanshan Cai, and Jihong Yang for their assistance. Dr. Lorelei L. Norvell’s editorial

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

Literature cited

Baker WA, Partridge EC, Morgan-Jones G. 2001. Notes on hyphomycetes. LXXXIV.

Pseudotrichoconis and Rhexodenticula, two new monotypic genera with rhexolytically

disarticulating conidial separating cells. Mycotaxon 79: 361-373.

Castaneda Ruiz RE, Saikawa M, Hennebert GL. 1996. Some new conidial fungi from Cuba.

Mycotaxon 59: 453-450.

Holmgren PK, Holmgren NH. 1998. [continuously updated]. Index Herbariorum: A global directory

of public herbaria and associated staff. New York Botanical Garden’s Virtual Herbarium.

http://sweetgum.nybg.org/ih/

Li X., Hu LL, Huang HD, Jiang MX. 2003. Structures and spatial distribution patterns of dominant

populations in Sycopsis sinensis community in Houhe Nature Reserve. J. Appl. Ecol. 14(6):

849-852.

Liu SX, Xu KY, Yang F-S. 1999. The biggest community Sycopsis sinensis was found in Houhe Nature

Reserve, Hubei Province, China. J Huazhong Nor Univ (Nat Sci) 33(4): 588-589.

Melnik V, Lee S, Groenewald JZ, Crous PW. 2004. New hyphomycetes from Restionaceae in fynbos:

Parasarcopodium ceratocaryi gen. et sp. nov., and Rhexodenticula elegiae sp. nov. Mycological

Progress 3(1): 19-28. http://dx.doi.org/10.1007/s11557-006-0072-1

MY COTAXON

http://dx.doi.org/10.5248/117.291

Volume 117, pp. 291-296 July-September 2011

Septobasidium atalantiae sp. nov. (Septobasidiaceae)

and S. henningsii new to China

SUZHEN CHEN?” & LIN GUO?*

‘Ocean University of China, Qingdao 266003, China

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

Chinese Academy of Sciences, Beijing 100101, China

CORRESPONDENCE TO *: Chensz2009@gmail.com & *guol@im.ac.cn

ABSTRACT —A new species, Septobasidium atalantiae on Atalantia buxifolia associated with

nymphal stage of a scale insect, and a new Chinese record of Septobasidium henningsii on

Memecylon floribundum are reported. They were collected from Hainan Province, China.

Key worps —Pucciniomycetes, Septobasidiales, taxonomy

In 2010 some Septobasidium specimens were collected by the second author

and her colleagues from Hainan Province. Among them a new species of

Septobasidium on Atalantia buxifolia, which is associated with the nymphal

stage of a scale insect, is described as follows:

Septobasidium atalantiae S.Z. Chen & L. Guo, sp. nov. Fras. 1-7

MycoBank MB 561544

Basidiomata resupinata, 0.2-7 cm longa, 0.15-1.4 cm lata, alba vel cinnamomeo-brunnea,

margine indeterminata, superficie laevia vel velutina, saepe protuberantia, interdum

maturitate fissurata, in sectione 480-1200 um crassa. Subiculum brunneum, 25-100

yum crassum. Columnae brunneae, 20-50 um altae, 20-70 um latae. Strata hypharum

420-830 um alta. Ab strato hymenii hyphae saepe repullulantes tum stratum hypharum

et hymenium secundum formantes. Hymenium hyalinum, 60-70 wm crassum. Basidia

cylindrica, curvata, 4-cellularia, 43-53 x 7.5-8.5 um, hyalina. Sterigmata cylindrica vel

conica, 6-7 x 3-5 um. Sine probasidio. Basidiosporae non visae. Haustoria ex hyphis

irregulariter spiralibus vel parallelis constantia.

Type: On Atalantia buxifolia (Poir.) Oliv. (Rutaceae): China, Hainan Province, Haikou,

alt. 420 m, 1.XII.2010, L. Guo 11536, HMAS 251151 (holotype), associated with

nymphal stage of a scale insect.

Basidiomata on branches, spreading to leaves, resupinate, 0.2-7 cm long,

0.15-1.4 cm wide, forming small isolated or confluent patches, white or

292 ... Chen & Guo

——

| —-s

10 pm

Fic. 1. Basidia of Septobasidium atalantiae (HMAS 251151, holotype).

cinnamon-brown; margin indeterminate, surface smooth or velutinous,

frequently with round protuberance, sometimes cracked at maturity. In section

480-1200 um thick. Subiculum brown, 25-100 um thick. Pillars brown, 20-50

um high, 20-70 um wide. Towards the top the pillars branch out to form a

hyphal layer, 420-830 um high, sometimes with a bunch of hyphae in holes

in the old stage. Hymenium hyaline, 60-70 um thick. In the old stage from

hymenial layer the fungal hyphae often renew growth to form the second

hyphal layer and hymenium. Basidia arising directly from the hyphae without

a probasidial cell, cylindrical, curved, 4-celled, 43-53 x 7.5-8.5 um, hyaline.

Sterigmata cylindrical or conical, 6-7 x 3-5 um. Basidiospores not seen.

Haustoria consisting of irregularly coiled hyphae, occasionally with closely

packed parallel hyphae.

Fics. 2-7. Septobasidium atalantiae (HMAS 251151, holotype). 2. Basidiomata on branches and

leaves. 3-4. Sections of basidiomata. 5-6. Basidia (arrows). 7. Haustoria.

Septobasidium atalantiae sp. nov. (China) ... 293

10 um

294 ... Chen & Guo

Fic. 8. Basidia of Septobasidium henningsii (HMAS 251152).

REMARKS: Morphologically, Septobasidium atalantiae is similar to S. natalense

Couch ex L.D. Gomez & Henk, but the latter has thinner section (250-500 mm

high), taller pillars (100-200 um high), and a non-stratose hymenial layer in

the old stage (Couch 1938).

Septobasidium henningsii Pat., Monsunia 1: 138, 1899. Figs. 8-13

Basidiomata on branches, resupinate, (0.5-)5-12 cm long, 0.3-3 cm

wide, brown or greyish brown; margin determinate, fibrillose, surface

smooth, frequently with round protuberance, cracked at maturity. In section

1460-2200(-2500) um thick. Subiculum brown, 25-50 um thick. Pillars brown,

530-1100um high, 30-40 um wide, entangled, arisingina slanting direction, often

with a horizontal layer at the base of pillar. Hymenia brown, 650-850 um thick,

1-4 strata, with closely packed parallel upright hyphae. Basidia at first pyriform

or subglobose, cylindrical at maturity, straight or slightly curved, 4-celled,

23-38 x 5-6 um, hyaline or yellowish, without a probasidial cell. Basidiospores

not seen. Haustoria consisting of subfusiform cells or hyphae.

SPECIMEN EXAMINED: On Memecylon floribundum Blume (Melastomataceae): CHINA,

HAINAN PROVINCE, Bawangling Natural Reserve, alt. 370 m, 26.XI.2010, Y.K Zhu & F.

He 522, HMAS 251152.

REMARKS: The Chinese specimen coincides morphologically with the original

description of Septobasidium henningsii (Couch 1938), except that the original

has larger (36-54 x 7-9.8 um) basidia.

Fics. 9-13. Septobasidium henningsii (HMAS 251152). 9. Basidiomata on branches. 10. Hymenia.

11. Basidium (arrow). 12. Haustoria.13. Section of basidioma.

Septobasidium atalantiae sp. nov. (China) ... 295

9 10

I] 12,

296 ... Chen & Guo

To date, 33 species of Septobasidium have 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 2011), including the two

species reported in this paper.

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 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. 2011. Septobasidium sichuanense sp. nov. (Septobasidiaceae) from China.

Mycotaxon 115: 481-484. http://dx.doi.org/10.5248/115.481.

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/10.5248/117.297

Volume 117, pp. 297-316 July-September 2011

Revision of Glomeromycetes with entrophosporoid and

glomoid spore formation with three new genera

FRITZ OEHL’, GLADSTONE ALVES DA SILVA’, IVAN SANCHEZ-CASTRO},

BRUNO TOMIO GOTO‘, LEONOR CosTA MalIA?’,

HELDER ELfsIo EVANGELISTA VIEIRA’, JOSE-MIGUEL BAREA3,

EWALD SIEVERDING & JAVIER PALENZUELA?

‘Federal Research Institute Agroscope Reckenholz-Tanikon ART, Organic Farming Systems,

Reckenholzstrasse 191, CH-8046 Ziirich, Switzerland

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

Av. Prof. Nelson Chaves s/n, Cidade Universitaria, 50670-420, Recife, PE, Brazil

*Departamento de Microbiologia del Suelo y Sistemas Simbidticos, Estacién Experimental

del Zaidin, CSIC, Profesor Albareda 1, 18008 Granada, Spain

‘Departamento de Botanica, Ecologia e Zoologia, CB, Universidade Federal

do Rio Grande do Norte, Campus Universitario, 59072-970, Natal, RN, Brazil

SInstitute for Plant Production and Agroecology in the Tropics and Subtropics,

University of Hohenheim, Garbenstrasse 13, D-70599 Stuttgart, Germany

“CORRESPONDENCE TO: fritz.oehl@art.admin.ch

ABSTRACT — New ribosomal gene analyses reveal that Entrophospora is non-monophyletic

and its type species E. infrequens closely related to Claroideoglomus species, which supports

transfer of the Entrophosporaceae from Diversisporales to Glomerales as well as the ‘ancestral’

Claroideoglomus spp. to Albahypha gen. nov. Entrophospora baltica, supported as a separate

clade within Diversisporales, is designated as type species for the new monospecific

Sacculosporaceae. Entrophospora nevadensis, phylogenetically close to Diversipora spp. and

Otospora bareae, is transferred to Tricispora gen. nov. (Diversiporaceae). Entrophospora,

Sacculospora, and Tricispora are morphologically distinguished by spore wall structure,

pattern of the two spore pore closures proximal and distal to the sporiferous saccule, and

relative spore and sporiferous saccule sizes. The shape of the white hyphae subtending the

spore base separates Albahypha spp. from Claroideoglomus spp.

Key worpbs — otosporoid, tricisporoid, evolution, molecular phylogeny, rDNA

Introduction

The genus Entrophospora was initially characterized according to intrahyphal

spore formation within the neck ofa sporiferous saccule (e.g. Ames & Schneider

1979, Blaszkowski et al. 1998, Palenzuela et al. 2010). Sieverding & Oehl (2006)

298 ... Oehl & al.

have transferred three species originally placed in Entrophospora (Schenck et al.

1984, Sieverding & Toro 1985, 1987, Wu et al. 1995) to new genera as Intraspora

schenckii (Sieverd. & S. Toro) Oehl & Sieverd., Kuklospora colombiana (Spain &

N.C. Schenck) Oehl & Sieverd., and K. kentinensis (C.G. Wu & Y.S. Liu) Oehl

& Sieverd. Support for the re-classification included i) clear morphological

differences from the type species, Entrophospora infrequens (Hall 1977; Ames &

Schneider 1979; Sieverding & Oehl 2006); ii) morphological aspects placing the

three species unequivocally in the Acaulosporaceae and Archaeosporaceae; and

iii) neither spore morphology nor genetic analysis of E. infrequens support that

species in Acaulosporaceae, where Kuklospora species phylogenetically belong

(Sieverding & Oehl 2006).

Phylogenetic analyses showed that the recently described E. nevadensis

belonged in Diversisporaceae (Palenzuela et al. 2010). To date Diversisporaceae

includes 13 Diversispora and six Redeckera species that form diversisporoid

spores on subtending hyphae (Oehl et al. 2011b) and one species with otosporoid

spores formed laterally on the neck of sporiferous saccules (Palenzuela et al.

2008). Another newly described species, Acaulospora colliculosa Kaonongbua

et al., was also believed to form its spores within the neck of sporiferous

saccules (Kaonongbua et al. 2010) but the illustrations rather indicate a mixture

of specimens of different species, isolated directly from field soil samples. Since

no saccule was presented in the latter study, further observations are needed

to confirm the existence of this species, which is assumed to form pacisporoid

spores instead of kuklosporoid spores as indicated by the images.

Recent morphological and molecular analyses suggest that the genus

Entrophospora, currently comprising three species, is not monophyletic, since

E. infrequens and E. nevadensis phylogenetically belong to different orders,

according to the recently revised Glomeromycetes (Oehl et al. 2011d). This

means that of the original species in the genus, only E. infrequens and E. baltica

(Blaszkowski et al. 1998) might remain in Entrophospora. Thus, one objective of

the current study was to analyze and re-classify Entrophospora species based on

combined morphological and phylogenetic analyses. In particular, we hoped

to resolve the confusion regarding the sequence variabilities for E. infrequens

noted by Millner et al. (2001) and Rodriguez et al. (2001). Another objective

was to confirm the phylogenetic position of Otospora bareae (Palenzuela et al.

2008) within the Diversisporaceae.

Material & methods

Specimens analyzed by morphological means

Type and non-type material representing 30 arbuscular mycorrhizal (AM)

fungal species currently placed in Entrophosporaceae, Claroideoglomeraceae and

Diversisporaceae (Sieverding & Oehl 2006, Schiifsler & Walker 2010, Oehl et al. 2011b)

were analyzed (TABLE 1). Source material of Entrophospora infrequens was originated

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 299

from field samples and from trap and pure cultures obtained from several countries,

with the pure cultures established in three different laboratories —EEZ, Granada, Spain;

CIAT, Cali, Colombia; and Swiss Collection of Arbuscular mycorrhizal fungi (SAF)

at Agroscope ART, Zurich, Switzerland (see Sieverding & Toro 1985, Palenzuela et

al. 2010, Oehl et al. 2010a, 2011a). Entrophospora baltica was obtained from the type

location in Poland and from several montane and alpine locations in Switzerland and

Spain. Entrophospora nevadensis and O. bareae have so far been reported only from their

type locations in the Sierra Nevada National Park and Sierra de Baza Natural Park in

Andalucia (Spain), respectively (Palenzuela et al. 2008, 2010).

TABLE 1. Collections analysed for reorganizing Entrophosporaceae,

Claroideoglomeraceae, and Diversisporaceae.

BAasIONYM

(SPECIES)

Entrophosporaceae

Glomus infrequens

Entrophospora baltica

E. nevadensis

Claroideoglomeraceae

Glomus candidum

G. claroideum

G. drummondii

G. etunicatum

G. lamellosum

G. luteum

G. viscosum

G. walkeri

Diversisporaceae

Diversispora celata

Glomus arenarium

G. aurantium

G. eburneum

G. epigaeum

G. gibbosum

TYPE MATERIAL

(EXAMINER)

Type OSC (Oehl)

Type (Oehl)

Type (Oehl & Palenzuela;

Palenzuela et al. 2010)

No access

Type OSC #40252 (Oehl)

Type (Oehl), ex type (Goto)

Holotype OSC (Oehl)

Isotype OSC #50183 (Oehl)

Type OSC, ex type

INVAM (Oehl)

Ex type (Oehl)

Type (Oehl), ex type (Goto)

Ex type, inclusive pure

cultures (Oehl; April

2009; deposited at SAF)

Type at OSC (Oehl),

ex type (Goto)

Type (Oehl)

Ex type at INVAM (Oehl)

Holotype OSC #39475 (Oehl)

Type (Blaszkowski

online pages)

NON-TYPE MATERIAL

(EXAMINER; COLL. DATE OR PUBLICATION)

Specimen from USA, Brazil, Chile

(Castillo et al. 2006), Colombia, Spain

(Palenzuela; July 2004; pure culture

established in 2006), Bolivia, Switzerland

(Oehl et al. 2004, 2011a; August 2005)

Specimen from Chile (Castillo et al. 2006), Spain

(Palenzuela & Oehl; July 2007), Switzerland

(Sieverding & Oehl 2006; July 2004)

Specimen from Brazil, Europe (Oehl

et al. 2010b); cultures from Benin

(Oehl; Tchabi et al. 2010)

Specimen from Benin, Bolivia, Paraguay,

Europe, Mexico (Oehl; Bashan et al. 2007)

Specimen from Poland, Germany (Oehl

& Sieverding; Oehl et al. 2003, 2005)

Specimen from Switzerland

(Oehl; April 2009)

Specimen from Chile and UAE

(Sieverding; Castillo et al. 2006)

Specimen from Germany (Oehl)

Specimen from Bolivia & Oman

(Oehl; Al-Yahyaei et al. 2011)

Specimen from UAE

(Sieverding, unpublished)

300 ... Oehl & al.

TABLE 1, CONCLUDED.

BASIONYM TYPE MATERIAL NON-TYPE MATERIAL

(SPECIES) (EXAMINER) (EXAMINER; COLL. DATE OR PUBLICATION)

G. insculptum Type (Oehl)

G. przelewicense No access Specimen and pure pot cultures from

Switzerland (Oehl & Sieverding; Oehl et

al. 2009, published as Glomus sp. BR12)

G. pustulatum Holotype OSC #46721 (Oehl) Btaszkowski collection (Oehl)

G. spurcum Ex type at INVAM (Oehl) Specimen from Bolivia (Oehl; Nov. 2000)

G. avelingiae No access

G. canadense Thaxter collection, Trappe collection (Oehl)

G. fragile Trappe collection (Oehl)

G. fulvum Trappe collection (Oehl), specimen

from Brazil (Goto)

G. megalocarpum Ex type (Oehl)

G. pulvinatum Trappe collection (Oehl)

G. tenerum Specimen from Australia (Oehl;

McGee & Trappe 2002)

G. trimurales Holotype OSC #49584 (Oehl)

Endogone versiformis Specimen at INVAM; specimen from Central

Europe (Oehl & Sieverding; Oehl et al. 2003)

Otospora bareae Type (Oehl & Palenzuela;

Palenzuela et al. 2008)

Morphological analyses

Morphological observations of spores (including sporiferous saccules and subcellular

structures), were based on freshly prepared specimens mounted in polyvinyl! alcohol-

lactic acid-glycerol (PVLG), in a mixture of PVLG and Melzer’s reagent (Brundrett

et al. 1994), in a 1:1 (v/v) mixture of lactic acid and water, in Melzer’s reagent, or in

water. Spore structure terminology for species with acaulosporoid, entrophosporoid, or

glomoid spore formation follows Goto & Maia (2006), Sieverding & Oehl (2006), Oehl

et al. (2006, 2011b), Spain et al. (2006), and Palenzuela et al. (2008, 2011). Photographs

were taken using an Olympus digital camera (model DP70-CU) mounted on a Zeiss

Axioplan compound microscope.

Molecular and phylogenetic analyses

Materials selected for molecular analyses included: E. infrequens— spores from

trap and pure culture material maintained at EEZ (Granada, Spain) and SAF (Zurich,

Switzerland); E. baltica— trap cultures and field samples from Mulhacen in Sierra Nevada

(Southern Spain, 3200 m asl) and Piz Corvo (Passo del Lugmagno, Swiss Central Alps,

2700 m asl); E. nevadensis— EEZ pure culture (Palenzuela et al. 2010); O. bareae— EEZ

maintained trap cultures (Palenzuela et al. 2008).

DNA extraction, amplification and sequencing from Spanish isolates were performed

at EEZ (Palenzuela et al. 2011) and from non-Spanish isolates at UFPE (Recife; see Goto

et al. 2011, Oehl et al. 2011c).

For sequence alignment we first verified the National Center for Biotechnology

Information (NCBI) databases using the BLASTn program to ensure that the E. infrequens

sequences were affiliated to Claroideoglomeraceae (Glomerales), and that the sequences

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 30.1

from E. baltica, E. nevadensis and O. bareae correctly aligned with Diversisporales. AM

fungal rRNA sequences (SSU = ~1770 bp; LSU = 733 bp) obtained in our laboratories

were then aligned with GenBank glomeromycotean sequences using ClustalX (Larkin et

al. 2007) and edited with BioEdit (Hall 1999) to obtain a final alignment. The sequences

were deposited at GenBank under the accession numbers FR865452—FR865456 and

JN113035-JN113037 for E. infrequens, FR865457-FR865462 and FR865449-FR865451

for E. baltica, FR865465-FR865467 and FR865446-FR865448 for E. nevadensis, and

FR8655463-FR865464 and FR865444-FR865445 for O. barea.

Maximum parsimony (MP) and neighbor joining (NJ) analyses with 1000 bootstrap

replications were performed using the Phylogenetic Analysis Using Parsimony (PAUP)

vers. 4 (Swofford 2003). Bayesian (two runs over 1 x 10° generations with a burnin

value of 2500) and maximum likelihood (1000 bootstrap) analyses were performed,

respectively, in MrBayes 3.1.2 (Ronquist & Huelsenbeck 2003) and PhyML (Guindon

& Gascuel 2003), launched from Topali 2.5. The nucleotide substitution model was

estimated using Topali 2.5 (Milne et al. 2004). Sequences from Neurospora crassa Shear

& B.O. Dodge and Boletus edulis Bull. were used as outgroups for Glomeromycota.

Results

Molecular phylogenetic analyses

The phylogenetic analyses on the nearly complete SSU of the ribosomal gene

and on partial sequences of the LSU show that E. infrequens, type species of

the Entrophosporaceae (Sieverding & Oehl 2006), is related to Claroideoglomus

species in the Glomerales (Fics 1-2). However, E. infrequens is phylogenetically

unrelated to the two other Entrophospora species that form two monophyletic

clades in the Diversisporales (Fics 1-2). Entrophospora nevadensis groups next

to O. bareae and several Diversispora species, while E. baltica groups in its own

clade, well distant from the other major clades representing families within the

Diversisporales. The phylogenetic analyses also render the genus Diversispora

paraphyletic (Fics 1-2).

Morphological comparisons

For the E. infrequens, E. baltica, E. nevadensis, and Otospora bareae clades

and the two Claroideoglomus clades, some morphological characters are

congruent with the phylogenetic findings. These characters are principally the

sporiferous saccule characteristics, pore closure patterns at the spore bases for

species forming spores within/on the saccule necks (Fics 3-19), and spore base

and subtending hyphae characteristics for species forming spores terminally on

hyphae (Fires 20-31).

Entrophospora infrequens forms saccules that are regularly larger than the

spores formed beneath, and the pore formed by the subtending hypha distal

to the saccules is closed by hyaline evanescent outer spore wall layers; the

next-inner structural pigmented persistent layer does not continue into the

distal part of the hyphal neck and never forms a pore distally to the saccule

302 ... Oehl & al.

Boletus edulis DQ534675

a Neurospora crassa AY046271

100 Paraglomus occultum DQ322629

700 P. brasilianum AJ301862

1.00 P. laccatum Am295493

00 Archaeospora trappei AM114274

ee A. trappei Y17634

: 100

sa e 50001" Geosiphon pyriforme AJ276074

4.0 100 G. pyriforme X86686

0% 78 00

G3] 1001.00 A mbispora fennica AM268195

A. callosa AB047308

Paraglomeraceae

Paraglomus

Archaeosporaceae

Archaeospora

Geosiphonaceae

Geosiphon

Ambisporaceae

Ambispora

100 1 A. appendicula AB047302

100) : ,

oes A. appendicula ABOt S082 Viscospora viscosa Y17652

Claroideoglomus etunicatum Y17639

100 #94 C. etunicatum AJ852598

1.00 EM; C. lamellosum AJ276083

oog?— C. /uteum AJ276089

‘ Entrophospora ety ig FR865453

“al E. infrequens FR865456

a E. infrequens FR865455

69 0.88 E. infrequens FR865452

= . E. infrequens FR865454

0.77 4 100 Simigiomus sp. AJ301857

: S. hoi AF485889

Septoglomus constrictum AJ534309

A se 7 S. africanum HM153416

r > a2 Funneliformis coronatum AJ276086

F. mosseae Z14007

57| ¢ F. caledonium Y17653

F. fragilistratum AJ276085

100) F. geosporum AJ245637

109 F. verruculosum AJ301858

ons) Glomus indicum GU059541

58 G iranicum HM153420

99 G proliferum AF213462

Enppphosporacese

Claroideog!

Claroi lomus

Entrophospora

Glomeraceae

Simiglomus

Septoglomus

Funneliformis

Glomus

100 G coremioides AJ249715

4 700) G sinuosum AJ133706

1.00 1.00 G._ clarum AJ852597

, 87 G manihotis Y17648

3 G. vesiculiferum L20824

0.97 G fasciculatum Y17640

98 G irregulare FJO09618

96 G intraradices AJ852526

96 Scutellospora calospora AJ306445 Scutellosporaceae

100 61.00 S. aurigloba AJ276093 Scutellospora

100 G gigantea AJ852602 Gigasporaceae

100 84 G rosea AJ852606 Gigaspora

1.00 86 G. albida AJ852599 Racocetraceae

99 1° Racocetra castanea AF038590 Racoc

97 99 Dentiscutata cerradensis AB041345 2 |

97 99 67 Fuscutata heterogama AJ852609 Dentiscutata

97 1.00 g F. heterogama AJ306434 Fuscutata

1.00 97 100 Pacispora scintillans AJ619944 Pacteponcest

0.99 100 P scintillans AJ619948 Pacispora

100 a0 Entrophospora baltica FR865457

=. 1,00 toot E. baltica FR865459

= too} &. baltica FR865458

51 1.00 E. baltica FR865462

0.92) E. baltica FR865460

eal E. baltica FR865461

Sacculosporaceae fam. nov.

Sacculospora gen. nov.

— 100 Kuklospora colombiana Z14006 Acaulosporaceae

A 100 91-— Acaulospora spinosa 214004 Kuklospora

1.00 A. laevis Y17633 Acaulospora

™ rae A. scrobiculata AJ306442

ane : A. rugosa 214005

3 A. longula AJ306439

0.68) 4.00 06 Redeckera fulva Am418543 | Diversisporaceae

sal Diversispora versiformis X86687 Redeckera

700 D. versiformis AJ276088 Diversispora

1.00 00 D. spurca AJ276077

700 D. eburnea AM713429

93 1.00 D. eburnea AM713431

: i D. celata AM713421

95 D. celata AM713422 Otospora

3) Otospora bareae FR865463

73 64} "|+ O. bareae FR865464

0.98 499 99 E. nevadensis FR865465 Tricispora gen. nov.

99 96 E. nevadensis FR865466

99 99 E. nevadensis FR865467

0.01 1.00 700

1.00

Fic. 1. A phylogeny of the Glomeromycota based on partial SSU rDNA sequences (~1800 bp).

NJ (neighbor joining), ML (maximum likelihood), and Bayesian analyses were performed with

GTR+G+I substitution model. Sequence labels correspond to their database accession numbers.

Support values are from NJ, MP (maximum parsimony), ML, and Bayesian analyses. New

sequences obtained in this study are indicated in bold. Only topologies with bootstrap values

= 50% are shown. Consistency Index = 0.47; Retention Index = 0.84.

(Fics 3-7). Thus, on the spores (particularly once the outer wall layers have

become degraded) just one single opening/pore forming a ‘cicatrix’ is visible

proximal to the saccules; the pore is closed by a plug-like wall material derived

apparently from the structural wall layer (ow13; Fics 3-4). The structural

wall layer extends into the saccule over 10-30 um, with the spore wall layer

100

65 1.06

85) =

93 00

1.00 98

1.00

0.1

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 303

Neurospora crassa AF286411

Boletus edulis AF336240

Paraglomus laccatum AM295494

2 occultum DQ273827

Geosiphon pyriforme AM183920

Ambispora fennica AM268202

=.A. gerdemannii AJ271712

: Claroideoglomus drummondii AJ972466

64 C. walkeri AJ972468

0.96 95 98, Entrophospora infrequens FJ461806

20 95|“ E. infrequens FJ461807

100 9 E. infrequens JN113037

1.00) 0.51-—} E. infrequens JN113035

96 E. infrequens JN113036

90 87|/- C. luteum FM876809

8174 C. etunicatum AF 145749

e C. etunicatum AJ623310

+ C. claroideum AJ271929

C. lamellosum AY541863

C. claroideum AF235007

78 Glomus diaphanum AJ972461

: G. diaphanum AJ972460

92 G sinuosum F 5461846

1.00 G_ intraradices AF396797

G intraradices AY842577

G. proliferum FM992402

95 G proliferum FM992398

100 G manihotis AM158947

00 G. clarum AJ510243

1.00) G. clarum AJ510242

Septoglomus deserticola AJ746249

7001 S. constrictum FJ461827

1.00 S. xanthium AJ849467

Funneliformis coronatum AF 145739

F. mosseae AY639273

100 F. mosseae AF 145735

100), F. fragilistratum AF 145747

1,00 F. caledonium AJ628059

63 F. caledonium AJ510239

55 F_coronatum AF145740

ane F. geosporum AF 145742

4 F. geosporum AF 145743

87 Cetraspora pellucida AF396784

92 84 [5p C._ pellucida AY639313

= Racocetra gregaria AJ510232

0.854n. | 100 R. castanea Y12076

mY 100 NI 4.00° R. verrucosa AY900508

Dentiscutata nigra AY900494

Bt D. nigra AY900498

88] 6 | Y Fuscutata heterogama AY900500

3 F, heterogama AY900503

ae Re Gigaspora rosea Y12075

58 G_marganita AF396783

84 G margarita AF396782

00 G. gigantea AY900504

1.00 100 gigantea AY900506

100 Pacispora scintillans FM876832

100 P. scintillans FM876831

Kuklospora colombiana FJ461804

5 Acaulospora lacunosa AJ510230

po A, laevis AJ510229

0° 60 A. longula AJ510228

0.85 A. paulinae AY639263

400 A. paulinae AY639328

4007 Entrophospora baltica FR865451

qooy- _E. baltica FR865449

1.00 | E. baltica FR865450

120 Diversispora trimurales FJ461851

D. aurantia EF581861

0.91 100 ©6100 n

00 00 D. aurantia EF581864

100 Otospora bareae FR865445

98 1.00 a O. bareae FR865444

100, D. versiformis AM947664

98 100 og 0. versiformis AY842573

0.98 © 100/(| 97° _E. nevadensis FR865448

opr sa} E. nevadensis FR865446

: p.98' E. nevadensis FR865447

D. eburnea EF067888

$2 D. eburnea EF067886

s7 D. celata AY639233

‘s x0 D. celata DQ350453

Paraglomeraceae

Paraglomus

Goceiemonacene

Ampisparaceae

Ambispora

Entrophosporaceae

Albahypha gen. nov.

Entrophospora

Claroideoglomus

Glomeraceae

Glomus

Septoglomus

Funneliformis

Racocetraceae

Cetraspora

Racocetra

Dentiscutataceae

Dentiscutata

Fuscutata

Gigasporaceae

Gigaspora

Pacisporaceae

Pacispora

Acaulosporaceae

Kuklospora

Acaulospora

Sacculosporaceae fam. nov.

Sacculospora gen. nov.

Diversisporaceae

Diversispora

Otospora

Tricispora gen. nov.

Fic. 2. A phylogeny of the Glomeromycota based on partial LSU rDNA sequences (~600 bp).

NJ (neighbor joining), ML (maximum likelihood), and Bayesian analyses were performed with

GTR + G substitution model. Sequence labels correspond to their database accession numbers.

Support values are from NJ, MP (maximum parsimony), ML, and Bayesian analyses. New

sequences obtained in this study are indicated in bold. Only topologies with bootstrap values

= 50% are shown. Consistency Index = 0.41; Retention Index = 0.79.

ornamentation apparently continuing to the pore closure area, although not

within the saccule.

Entrophospora baltica and E. nevadensis (Fics 8-19) have smooth proximal

cicatrices; here the ornamentation does not continue at the area of the generally

wide pore proximal to the sporiferous saccule. Both the distal and proximal

304 ... Oehl & al.

spore pores are closed by a smooth, rather thin septum of the structural,

laminate wall layer, and the structural wall layer continues for a small distance

into the distal hypha of the saccule neck (Fics 8, 12, 16). In both species the

saccules are often slightly (E. baltica) or substantially (E. nevadensis) smaller

than spores formed beneath since they sometimes equal half or a quarter of

the spore diameter, respectively. Remarkably, detailed spore wall structure

analyses revealed that E. baltica has three walls (Fics 17-19), while spores of

E. nevadensis (and Otospora bareae) are clearly bi-layered (Fic. 13, and

Palenzuela et al. 2010).

Typical Claroideoglomus species (e.g., C. etunicatum, C. claroideum,

C. lamellosum (Fics 20-25), C. luteum) form hyaline subtending hyphae, which

are ‘funnel-shaped’ — their width at the spore base is regularly > 2.5 times

wider than 10-50 um distant from the spore. However, two species that are

basal within the Claroideoglomeraceae — C. drummondii, C. walkeri (Fic. 2,

Oehl el al. 2011b)— have only slightly funnel-shaped to sometimes cylindrical

subtending hyphae that are regularly < 2.0 times broader at spore base than the

supporting hypha (Fics 26-31).

Based on our phylogenetic analyses, we propose to include the

Entrophosporaceae and its type species E. infrequens in the Glomerales. The

LSU rRNA sequence analyses indicate that Claroideoglomus should be further

divided into two genera, which is supported by the morphological differences

between the two recognized clades.

Since E. nevadensis forms a clade within the Diversisporaceae, a new genus

is indicated with it serving as type species. Entrophospora baltica, which forms

a monophyletic major clade within the Diversisporales, should serve as the type

species of a new family in the order.

Sequence analyses further imply that Diversispora is paraphyletic. However,

here the molecular and morphological databases do not yet provide sufficient

information to reclassify accurately the Diversispora species.

Taxonomic revision

Glomerales J.B. Morton & Benny, emend. Oehl, Palenz., G.A. Silva & Sieverd.

EMENDED DESCRIPTION: Spores form terminally on or intercalary in hyphae

or within the necks of sporiferous saccules in soil (or sometimes roots) singly

or (when glomoid) also in spore clusters or multi-spored loose to compact

sporocarps; when in compact sporocarps (with or without peridium), spores

randomly distributed or organized around a central hyphal plexus. Glomoid

spores with one single or multiple-layered wall. Entrophosporoid spores with

two walls: outer structural wall and inner (germinal) wall. In glomoid spores,

wall of the subtending hyphae (sH) conspicuously continuous with the spore

wall, sH funnel-shaped, cylindrical, or constricted and concolorous with

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 305

Fics 3-16. Examples of intra-hyphal spore formation within the neck of a sporiferous saccule.

Fics 3-7. Entrophospora infrequens: Spores have two walls (ow & Iw); saccule generally larger than

attached spore. ow is triple-layered (OwL1-3) and has a characteristic ornamentation (‘orn’) on

structural layer OwL3. At spore bases two cicatrices are formed: proximal and distal to the saccule.

The persistent proximal cicatrix (‘proci’) formed by the persistent owL3 (Fics 5-6) wall material

resembles a plug. The evanescent distal cicatrix (‘disci’) is formed by evanescent OWL2 (Fic. 7)

material. Figs 8-12. Sacculospora baltica: spores with two cicatrices; saccule (sac) substantially

smaller than attached spore. The proximal cicatrix (proci) is persistent and formed by the structural

ow layer (Fics 10-11); the corresponding spore pore is closed by a septum that does not resemble

a plug. The distal cicatrix (‘dici’) is also persistent and formed by the same layer (Fic. 12). Fics

13-16. Tricispora nevadensis: spores have two walls (ow & Iw) and two cicatrices; saccule (sac)

substantially smaller than the attached spore. The proximal cicatrix (proci) is persistent and formed

by the structural ow layer (Fics 14-15); the corresponding spore pore is closed by a septum that

does not resemble a plug. The distal cicatrix (dici) is also persistent and formed by the same layer

(Fic. 16).

306 ... Oehl & al.

BSc EM8

. a §

Ca ~< .

’ oe”. Dee

Py aos Sy , "4

wiv ; rv —>, .

SAG ¢

~ 30 um

Fics 17-19. Sacculospora baltica spore wall structure showing peridial hyphal mantle (peri),

conspicuous proximal cicatrix (proxy), and warts on the outer wall (ow) surface; middle wall (mw)

is 1(-2)-layered and inner wall is triple-layered (1twL1-3). The three rw layers generally tightly

adhere, making Iw] and 1w13 difficult to observe (Fic. 18); the separation of the three layers in

Fic. 19 is an artifact of cover-slip pressure.

spore, slightly paler, or (sub-)hyaline. In entrophosporoid spores, structural

pigmented outer wall layer discontinuous with the hyphal wall distal to the

saccule; forming typical vesicular-arbuscular mycorrhiza with mycorrhizal

structures that stain blue to dark blue in trypan blue.

TYPE FAMILY: Glomeraceae Piroz. & Dalpé

OTHER FAMILY: Entrophosporaceae Oehl & Sieverd.

Entrophosporaceae Oehl & Sieverd., emend. Oehl, Sieverd., Palenz. & G.A. Silva

= Claroideoglomeraceae C. Walker & A. Schiissler, The

Glomeromycota — a species list: 21. 2010.

EMENDED DESCRIPTION: Spore formation is of glomoid or entrophosporoid

type. Glomoid spores form in soil or (rarely) in roots singly, in clusters with

few spores, or (extremely rarely) in sporocarps; sH hyaline to white, rarely

subhyaline, often conspicuously funnel- or bill-shaped. Spores with one wall

of 1-4 layers; spore base pore closure often with a septum that arises from the

structural layer, an adherent thin inner layer, or both layers. Entrophosporoid

spores form singly in soils or rarely in roots, subterminally or intercalary within

the neck of a tightly attached sporiferous saccule that generally is larger in size

than the underlying spore; they have two walls: outer and inner. Outer, hyaline,

semi-persistent to evanescent layers of the outer spore wall are the hyphal neck

and sporiferous saccule wall layers. The pigmented structural layer does not

continue within the hyphal wall but only within the saccule terminus for some

distance. Pore closed by a plug towards the saccule. The inner wall is generally

thick, finely laminated and forms de novo. No inner wall layer has a beaded

appearance or stains in Melzer’s reagent. Fungal structures in roots stain blue

with trypan blue; forming vesicular-arbuscular mycorrhizae.

TYPE GENUS: Entrophospora R.N. Ames & R.W. Schneid.

OTHER GENERA: Claroideoglomus C. Walker & A. Schiissler, Albahypha Oehl et al.,

Viscospora Sieverd. et al.

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 307

FiGs 20-31. Examples of terminal spore formation on subtending, hyaline to white hyphae (sH);

spores with 2-4 layers (swL1-4); spore pores regularly closed by a septum (sp) at the spore base.

Fics 20-25. Claroideoglomus (Fics 20-22: C. etunicatum, FiGs 23-24: C. claroideum, Fic. 25:

C. lamellosum) — Claroideoglomus spores have a significantly funnel-shaped hypha with structural

wall layer (generally sw12 or sw13) that is >2.5 times thicker at spore base than 10-25 um distant

at transition between SH and mycelia hypha. Fics 26-31. Albahypha (Fics 26-28: A. drummondii,

Figs 29-31: A. walkeri) — Albahypha spores have a slightly funnel-shaped to cylindrical hypha

with structural wall layer (generally sw12) that is <2.0 times thicker at spore base than 10-25 um

distant.

Entrophospora R.N. Ames & R.W. Schneid., emend. Oehl, Sieverd., Palenz. & G.A.

Silva Figs 1-5

EMENDED DESCRIPTION: Sporocarps unknown. Entrophosporoid spores form

within the hyphal neck of tightly attached terminal or intercalary sporiferous

saccules, singly in soils, or (rarely) in roots. Sporiferous saccules generally are

308 ... Oehl & al.

larger in size than the underlying spores. Entrophosporoid spores are globose

to subglobose and have two walls: an outer and an inner. Outer, semi-persistent

to evanescent layers of the outer spore wall are the wall layers of the hyphal

stalk and the sporiferous saccule. The structural, pigmented layer beneath

does not continue within the hyphal wall but only for a short distance within

the saccule terminus. Thus, spores have only one persistent cicatrix, which is

proximal to the globose saccule terminus. A plug closes the pore towards the

saccule. The inner wall is thick, finely laminated wall and forms de novo. No

inner wall layers have a beaded appearance. Fungal structures in roots stain

blue with trypan blue; forming vesicular-arbuscular mycorrhizae.

TYPE SPECIES: Entrophospora infrequens (I.R. Hall) R.N. Ames & R.W. Schneid.

Claroideoglomus C. Walker & A. Schiissler, emend. Oehl, Sieverd., B.T. Goto

& G.A. Silva FIGS 20-25

EMENDED DESCRIPTION: Spores formed on subtending hyphae (su), generally

singly in soil or rarely in roots; sH are hyaline to white, rarely subhyaline, and

funnel- or bill-shaped with widths > 2.5 times greater at the spore base than

at 10-20 um from the spore. Spores with one wall of 1-4 layers; pore closure

at spore base often with a septum that arises species-specifically from the

structural layer, an adherent thin innermost layer, or both innermost layers.

TYPE SPECIES: Claroideoglomus claroideum (N.C. Schenck & G.S. Sm.) C. Walker

& A. Schiissler

Albahypha Oehl, G.A. Silva, B.T. Goto & Sieverd., gen. nov. Figs 26-31

MycoBank MB 561639

Sporae singulariter efformatae; tunica sporarum cum tunica hypharum coniuncta; hyphae

albae, tunica hyphae 1.2-2.0 maior ad basem sporarum quam in 10-20 um distantia

basae sporarum; porum sporarum occlusum septa tunicae sporarum, rarum apertum.

Mycorrhizas vesiculares-arbusculares formans caeruleas colorantes cum ‘trypan blue’.

TyPE SPECIES: Albahypha drummondii (Blaszk. & Renker) Sieverd. et al.

ErymMo.oey: derived from the Latin: alba = white; hypha = hypha; referring to the

white, slightly funnel-shaped subtending hypha which is characteristic for species of

this genus.

KEY CHARACTERS: Spores formed generally singly in soil or rarely in roots;

SH white, rarely subhyaline, 1.2-2.0 times wider at spore base than their width

10-20 tm distance from the spore, giving a slightly funnel-shaped or cylindrical

appearance. Spores with one wall of 1-4 layers; spore base pore closure often

with a septum that may arise from the structural layer, an adherent innermost,

(semi-)flexible layer, or both innermost layers.

Albahypha drummondii (Btaszk. & Renker) Sieverd., Oehl, B.T. Goto & G.A. Silva,

comb. nov.

MycoBank MB 561640

= Glomus drummondii Blaszk. & Renker, Mycol. Res. 110: 559. 2006.

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 309

= Claroideoglomus drummondii (Blaszk. & Renker) C. Walker & A.

Schiissler, The Glomeromycota — a species list: 22. 2010.

Albahypha walkeri (Btaszk. & Renker) Sieverd., Oehl, B.T. Goto & G.A. Silva,

comb. nov.

MycoBank MB 561641

= Glomus walkeri Blaszk. & Renker. Mycol. Res. 110: 563. 2006.

= Claroideoglomus walkeri (Blaszk. & Renker) C. Walker & A.

Schiissler, The Glomeromycota — a species list: 22. 2010.

Viscospora Sieverd., Oehl & G.A. Silva, Mycotaxon 116: 108. 2011.

TYPE SPECIES: Viscospora viscosa (‘T.H. Nicolson) Sieverd. et al.

Diversisporaceae C. Walker & A. Schiissler, emend. Oehl, Palenz., I.C. Sanchez, G.A.

Silva, B.T. Goto & Sieverd.

EMENDED DESCRIPTION: Spore formation diversisporoid, otosporoid, or

entrophosporoid sensu lato (‘tricisporoid’ sensu stricto). Diversisporoid spores

formed singly, in clusters, or in large disorganized sporocarps with high spore

numbers. In pigmented spores, subtending hyphae (sH) conspicuously change

color, becoming hyaline to white behind the septum, (immediately or at a very

short distance from this septum); sH generally straight, cylindrical, in some

species constricted or inflated. Spores with 1-3 wall layers; pore often closed

with a septum that may arise from innermost wall lamina, an overlaying

laminate layer, or from both; su pore rarely open. Otosporoid and tricisporoid

spores with two multiple-layered walls; otosporoid spores formed laterally on

the persistent neck of a terminal or intercalary sporiferous saccule at some

distance from the saccule terminus; spore pore generally closed by a septum at

spore base. Tricisporoid spores formed within the evanescent neck of a tightly

attached terminal or intercalary sporiferous saccule, closely attached to the

saccule terminus which is often smaller in size than the mature spores attached,

rarely equal in size; tricisporoid spores with two cicatrices formed by the outer

wall pigmented structural layer.

TYPE GENUS: Diversispora C. Walker & A. Schiissler

OTHER GENERA: Redeckera C. Walker & A. Schiissler, Otospora Oehl et al., Tricispora

Oehl et al.

Diversispora C. Walker & A. Schiissler, Mycol. Res. 108: 982. 2004.

TYPE SPECIES: Diversispora spurca (C.M. Peiff. et al.) C. Walker & A. Schiissler

Otospora Oehl, Palenz. & N. Ferrol, Mycologia 100: 297. 2008.

TYPE SPECIES: Otospora bareae Palenz. et al.

Redeckera C. Walker & A. Schiissler, The Glomeromycota — a species list: 44. 2010.

TYPE SPECIES: Redeckera fulva (Berk. & Broome) C. Walker & A. Schiissler

310 ... Oehl & al.

Tricispora Oehl, Sieverd., G.A. Silva & Palenz., gen. nov. Fics 13-16

MycoBank MB 561642

Sporocarpia ignota. Sporae singulatim efformatae subterminaliter vel intercalariter in

hypha inflata anguste adiacetum ad sacculum sporiferum terminalem vel intercalarem.

Sacculus sporiferus frequenter minor quam sporae globosae vel subglobosae; sporae duabus

tunicis stratis pluribus. Stratum exterior tunicae exterioris coniunctum tunica hyphae et

sacculi. Stratum interiorem tunicae exterioris laminatum, duas poras sporae occludens.

Stratum exterior tunicae interioris non granulatum. Formans mycorrhizas vesicular-

arbusculares. Structurae fungorum colorantes caeruleae cum ‘trypan blue’.

TYPE SPECIES: Tricispora nevadensis (Palenz. et al.) Oehl et al.

ErymMo_oey: derived from the Latin: (cica-)trix = cicatrix: and spora = spore; referring

to the two conspicuous cicatrices left on the structural wall layer of the spores, even

when the sporiferous saccules and the hyphal neck distal to the saccule have detached

completely from the spores.

KEY CHARACTERS: Sporocarps unknown. Spores formed within the hyphal

neck of closely adherent terminal or intercalary sporiferous saccules. The

globose saccule terminus generally is substantially smaller than the attached

mature spore. Spores have an outer and an inner wall. At least two layers

(including the outer wall structural layer) are continuous with the sporiferous

saccule wall. The outer layer of the outer wall is evanescent, the inner layers are

permanent. After the hyphal neck connections break off, spores show two, often

opposite, cicatrices that are closed by the permanent sublayers of the outer wall

structural layer. The inner wall forms de novo, consists of several layers without

granular (‘beaded’) appearance and does not stain with Melzer’s reagent. The

fungal structures in the roots stain blue to dark blue with trypan blue; forming

vesicular-arbuscular mycorrhiza.

Tricispora nevadensis (Palenz., N. Ferrol, Azcén-Aguilar & Oehl) Oehl, Palenz.,

G.A. Silva & Sieverd., comb. nov.

MycoBank MB 561644

= Entrophospora nevadensis Palenz., N. Ferrol, Azcon-

Aguilar & Oehl, Mycologia 102(3): 627. 2010.

Sacculosporaceae Oehl, Sieverd., G.A. Silva, B.T. Goto, I.C. Sanchez & Palenz., fam.

nov.

MycoBank MB 561645

Sporae singulatim efformatae in hypha inflata anguste adiacetum ad sacculum sporiferum

terminalem vel intercalarem. Sporae tribus tunicis stratis pluribus. Stratum interiorem

tunicae exterioris laminatum, duas poras sporae occludens. Stratum exterior tunicae

interioris non granulatum.

KEY CHARACTERS: Sporocarps unknown. Spores formed within the hyphal neck

of closely adherent terminal or intercalary sporiferous saccules. Spores have

three walls: outer, middle and inner. At least two layers (including the outer

wall structural layer) are continuous with the sporiferous saccule wall. After the

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 311

hyphal neck connections break off, spores show two, often opposite, cicatrices

that are closed by the permanent sublayers of the outer wall structural layer.

Middle and inner wall form de novo. Middle wall is 1-2-layered. Inner wall

consists of several layers, none of which has a granular (‘beaded’) appearance,

and does not stain in Melzer’s reagent.

TYPE GENUS: Sacculospora Oehl et al.

Sacculospora Oehl, Sieverd., G.A. Silva, B.T. Goto, I.C. Sanchez & Palenz., gen. nov.

MycoBank MB 561646 Fics 8-12, 17-19

Sporocarpia ignota. Sporae singulatim efformatae subterminaliter vel intercalariter in hypha

inflata anguste adiacetum ad sacculum sporiferum terminalem vel intercalarem. Sporae

globosae vel subglobosae, tribus tunicis stratis pluribus. Stratum exterior tunicae exterioris

coniunctum tunica hyphae et sacculi. Stratum interiorem tunicae exterioris laminatum,

duas poras sporae occludens. Stratum exterior tunicae interioris non granulatum.

TYPE SPECIES: Sacculospora baltica (Biaszk. et al.) Oehl et al.

ErymMo oey: derived from the Latin: sacculus = saccule: and spora = spore; referring to

the spore formation within the neck of sporiferous saccules.

KEY CHARACTERS: Sporocarps unknown. Spores formed within the hyphal neck

of closely adherent, terminal or intercalary sporiferous saccules. Spores have

three walls: outer, middle and inner. At least two layers (including the outer

wall structural layer) are continuous with the sporiferous saccule wall. Inner

layers of the outer spore wall are permanent. After the hyphal neck connections

break off, spores show two, often opposite, cicatrices that are closed by the

permanent sublayers of the outer wall structural layer. Middle and inner wall

form de novo. Middle wall is 1-2-layered. Inner wall consists of several layers,

none of which have a granular (‘beaded’) appearance (Fic. 18-19), and does

not stain in Melzer’s reagent. ‘The inner wall may be germinal in function, but a

germination structure has not yet been found.

Sacculospora baltica (Btaszk., Madej & Tadych) Oehl, Palenz., I.C. Sanchez,

B.T. Goto, G.A. Silva & Sieverd., comb. nov.

MycoBank MB 561647

= Entrophospora baltica Blaszk., Madej & Tadych, Mycotaxon 68: 167. 1998.

Discussion

New molecular and morphological analyses elucidated the molecular

phylogenetic and morphological congruencies within a group of species that

have long been considered heterogeneous (e.g. Morton & Benny 1990, Sieverding

& Oehl 2006). Species with entrophosporoid (sensu lato) spore formation

are now found within Entrophosporaceae (E. infrequens), Acaulosporaceae

(K. colombiana, K. kentinensis), Diversisporaceae (T: nevadensis), Sacculo-

sporaceae (S. baltica), and Archaeosporaceae (Intraspora schenckii).

Sequence analyses of E. infrequens, with a unique entrophosporoid spore

formation, place this species so close to Claroideoglomus that we regard

312... Oehl & al.

the recently described Claroideoglomeraceae as a heterotypic synonym of

Entrophosporaceae. Morphological congruencies were found to support the

phylogeny and helped identify a fourth genus, Albahypha, to include with the

type genus Entrophospora, Claroideoglomus, and Viscospora within the revised

Entrophosporaceae.

The Diversisporaceae now comprise four genera: two with diversisporoid

spore formation (Diversispora and Redeckera; Oehl et al. 2011b) and one each

with otosporoid (Otospora) and tricisporoid (Tricispora) spore formation.

There are now several phylogenetic clades that form spores terminally on

hyphae as well as on or within sporiferous saccules (spore formation type

glomoid s.l., acaulosporoid s.l., entrophosporoid s.l.). These are represented

in the Entrophosporaceae (Glomerales), Diversisporaceae (Diversisporales),

and Archaeosporaceae and Ambisporaceae (Archaeosporales) clades but only

Diversisporaceae, Archaeosporaceae and Ambisporaceae are represented by all

three major spore formation types.

Further studies are still needed to determine whether the entrophosporoid

(sensu lato) genus Kuklospora is monophyletic. Most phylogenetic analyses split

the type species, K. colombiana, off at the base of Acaulosporaceae (Schifsler et

al. 2001, Palenzuela et al. 2008, 2010, Oehl et al. 2011d). Thus, we conclude that

the synonymization of Kuklospora with Acaulospora (Kaonongbua et al. 2010)

is not justified and needs to be substantiated (Oehl et al. 2011e). Moreover,

a revision of the phylogenetically and morphologically heterogeneous

Acaulosporaceae is urgently needed (Spain 1992, Oehl et al. 201 1e).

Our analyses supports Tricispora nevadensis (= E. nevadensis) and

Otospora bareae as sister taxa within the Diversisporaceae (Fics 1-2). Both

Entrophospora and Otospora can readily be distinguished by spore formation

type, distance between the saccule terminus and the differentiated spore within

(Entrophospora) or laterally (Otospora) on the saccule neck, and the relative size

differences between saccule and spore. Morphologically, these genera have little

in common with the type genus of their family, Diversispora, whose species

form spores simply on subtending hyphae. We emphasize that there have

never been genera in Glomeromycetes (sensu Oehl et al. 2011d) that encompass

species with both glomoid and entrophosporoid or acaulosporoid spore

formation. This strongly supports the proposition of Otospora and Tricispora.

However, phylogenetically, both T: nevadensis and O. bareae render the genus

Diversispora polyphyletic.

We regard the Diversispora molecular database as still rudimentary, as the

genus comprises several species that yet to be molecularly analyzed (Oehl et al.

2011b). Additionally, the morphological differences among Diversispora clades

(Fics 1-2) are not yet clear. Before the genus Diversispora can confidently be

reclassified, additional molecular and morphological studies are needed to

Albahypha, Sacculospora & Tricispora gen. nov. (Glomeromycetes) ... 313

determine relationships accurately within this heterogeneous family.

Glomus hyderabadense Swarupa et al. also deserves mention, for it regularly

forms a small terminal spore and an adjacent, subterminal large spore on

subtending hyphae. Since the terminal spore is generally much smaller than

the subterminal spore (Swarupa et al. 2004), its spore formation superficially

resembles that of T. nevadensis. Although the G. hyderabadense sequence

(AY211274) aligns adjacent to G. clarum (Swarupa et al. 2004), the deposited

sequence does not represent a glomeromycotan fungus. Glomus hyderabadense

possibly forms its own clade within the Glomeromycetes and thus may represent

an undescribed fungal genus with a unique, diagnostic spore formation and

morphology.

Acknowledgments

We acknowledge the help of Joey Spatafora and Richard Halse for giving the

possibility to visit the Oregon State University herbarium (OSC) in Corvallis (Oregon)

in 2002 and 2005, and for providing us with type and non-type material of many

specimens. We especially thank Joseph B. Morton and William Wheeler (West Virginia

State University; Morgantown, USA) for giving F. Oehl the opportunity to investigate,

during his visit in 2002, several Glomeromycetes species maintained at INVAM. Also

James M. Trappe (USDA-Corvallis, Oregon, USA) and Janusz Blaszkowski (Department

of Plant Pathology, Academy of Agriculture, Szczecin, Poland), provided type or non

type specimen of several species. We acknowledge the valuable comments and revisions

of Dr. Renata Gomes de Souza (UFPE, Recife, Brazil), Dr. Fabien C.C. Hountondji

(University of Parakou, Benin), Danny Coyne (IITA-Nigeria), and PD Dr. Reinhard

Berndt (ETH Zurich, Switzerland) on the manuscript and appreciate the corrections

by Shaun Pennycook, Nomenclatural Editor, and suggestions by Lorelei L. Norvell,

Editor-in-Chief. This study was supported by the Swiss National Science Foundation

(SNSF, Project 315230_130764/1), Conselho Nacional de Desenvolvimento Cientifico

e Tecnolédgico (CNPq, who provided a fellowship to L.C. Maia and a grant through

the INCT-Herbario Virtual da Flora e dos Fungos), and by the Fundacao de Amparo a

Ciéncia e Tecnologia do Estado de Pernambuco (FACEPE) and the Universidade Federal

de Pernambuco (UFPE) which provided grants to F. Oehl as ‘visiting professor’.

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MY COTAXON

http://dx.doi.org/10.5248/117.317

Volume 117, pp. 317-320 July-September 2011

Puccinia cortusae (Basidiomycota; Uredinales) on

Cortusa brotheri (Primulaceae), new to southern Asia

(Fairy Meadows, Pakistan)

A.N. KHALID & M. SABA *

Department of Botany, University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

* CORRESPONDENCE TO: rustflora@gmail.com, 'drankhalid@gmail.com

ABSTRACT — Puccinia cortusae has been found parasitizing Cortusa brotheri from northern

Pakistan. This first report from Pakistan and southern Asia also establishes C. brotheri as

a new host for the rust fungus. We believe this is the first report of P cortusae since it was

first described in 1904. New observations of one-celled teliospores and variable germ pore

position supplement the original description.

Key worps — Bial Camp, Cortusa matthioli

Introduction

Worldwide, the Primulaceae comprises ca. 24 genera and 1000 species, (most

of which represent herbs confined mainly to north temperate regions), among

which only around 28 species have been recorded as alternate or (sometimes)

primary hosts for 18 rust species and five genera (Arthur & Cummins 1962,

Wilson and Henderson 1966, Hiratsuka et al. 1992, Farr & Rossman 2011).

Although 60 species (eight genera) representatives of the Primulaceae have

been reported from Pakistan, not a single species has been from the country as

a host for rust fungi.

During the present study, Cortusa brotheri was found parasitized by Puccinia

cortusae. With this addition, the number of rust fungi has been raised to 73

from Fairy Meadows, Pakistan (Afshan et al. 2009, 2011).

Materials & methods

During the survey of rust fungi from Pakistan, Cortusa brotheri was collected from

Northern Pakistan, Fairy Meadows, 3036 ma.s.l., occupying rock crevices along a running

stream. A healthy flowering plant was also collected for accurate host identification.

Free hand sections of infected material and spores were mounted in lactophenol. Semi-

permanent slides were prepared by cementing cover slips with nail lacquer (Dade &

Gunnell 1969). Preparations were observed under a Nikon YS100 microscope and

318 ... Khalid & Saba

photographed with a digipro-Labomed (USA). Spore drawings were made using a Leitz

Camera Lucida (Wetzlar, Germany). Twenty-five spores were measured using a Zeiss

ocular micrometer (St Albans, England). Measurements include range and arithmetic

means with extremes shown in parenthesis.

Taxonomy

Puccinia cortusae Tranzschel, in Sydow & Sydow, Monogr. Uredin.

(Lipsiae) 1(5): 886 (1904).

Fic. 1. Puccinia cortusae. A: Infected host plant, Cortusa brotheri (arrows showing telia along

and between veins); B-C: Teliospores (arrows showing sub-apical position of germ pore in distal

cell and next to septum in proximal cell with hyaline papillae); D-E: Aberrant teliospores with

additional oblique and perpendicular septa. Scale bars: A = 2 cm, B-E = 10 um.

Puccinia cortusae, new to South Asia (Pakistan) ... 319

SPERMOGONIA, AECIA and UREDINIA not seen. TELIA hypophyllous, rarely

epiphyllous, circinate, pulverulent, covered by ruptured epidermis, naked, along

or between veins, 70-140 x 280-400 um. TELIOsPorREs pale brown to chestnut

brown, ellipsoid, broadly ellipsoid or obovoid, rounded or attenuated at base,

slightly or not constricted at septum, chestnut brown at septum, 16-25(-31) x

24-37 um (mean 22.2 x 29 um); wall chestnut brown, smooth, 1-1.6 um; germ

pore one per cell, with variable position, mostly apical, sub apical or sometimes

adjacent to septum in distal cell; in proximal cell mostly adjacent to septum,

sometimes shifted % to % towards hilum, germ pores with hyaline papillae;

apex not thickened, rounded, sometimes conical; pedicel hyaline, deciduous,

short, sometimes obliquely attached, 5-11 x 3-20 um. One-celled teliospores

and aberrant teliospores with additional oblique or perpendicular septa present

but rare.

Fic. 2. Lucida drawings of Puccinia cortusae showing one- and two-celled teliospores.

Scale bar = 10 um.

MATERIAL EXAMINED: PAKISTAN, Fairy Meadows, Bial Camp, on Cortusa brotheri Pax

ex Lipsky (Primulaceae), 24 July 2010, Malka Saba 07, with III stage (LAH 1147).

COMMENTS: Among new observations made for P cortusae not mentioned

in the original description are the presence of both two-celled and one-celled

teliospores that are broader (15-21 um in the original). Teliospore germ pores

were observed in different positions, e.g., apical, subapical, or sometimes

adjacent to the upper cell septum although they were mostly observed in the

lower cell adjacent to the septum or sometimes shifted half or three-quarters of

the distance toward the hilum.

Puccinia cortusae is the only rust fungus reported on Cortusa L., and

has previously been known only from the type collection on C. matthioli L.

from the Alai Range in southern Kyrgyzstan (Sydow & Sydow 1904: “in jugo

Alaico Turkestaniae”; Farr & Rossman 2011). Our collection is a new record of

P. cortusae for southern Asia and C. brotheri is a new host for this rust fungus.

The two hosts are closely related, with C. brotheri sometimes treated as a form

of C. matthioli, C. matthioli f. brotheri (Pax ex Lipsky) R. Knuth.

320 ... Khalid & Saba

Gaumann (1959) described Puccinia cortusae as a microform (telial stage

only) of P primulae Grev. Teliospores resemble those of P. primulae in shape

although they differ in size, those of P cortusae being longer and broader

(16-25(-31) x 24-37 um vs. 15-18 x 22-30 um) with chestnut brown wall,

variable position of germ pores and apically unthickened.

Acknowledgments

We are thankful to Dr. Jan Alam (Department of Botany, Hazara University,

Mansehra, Pakistan) for his help in the identification of host plant. We are sincerely

thankful to Dr. Reinhard Berndt (Curator of Fungus Collections, Herbaria Z+ZT, ETH

Zurich, Switzerland) for his help in the identification of rust fungus. We also thank Dr.

Berndt and Dr. Najam-ul-Sehar Afshan (Centre for Undergraduate studies, University of

the Punjab, Lahore, Pakistan) for acting as presubmission reviewers. We are grateful to

Dr. Shaun R. Pennycook and Dr. Lorelei L. Norvell for their critical reviews to improve

the manuscript. We are highly indebted to the Pakistan Science Foundation for financial

assistance under project No. 405.

Literature cited

Afshan NS, Khalid AN, Iqbal SH, Niazi AR, Sultan A. 2009. Puccinia subepidermalis sp. nov. and

new records of rust fungi from Fairy Meadows, Northern Pakistan. Mycotaxon 110: 173-182.

http://dx.doi.org/10.5248/110.173

Afshan NS, Khalid AN, 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

Arthur JC, Cummins GB. 1962. Manual of the rusts in United States and Canada. Hafner Publishing

Company, New York.

Dade HA, Gunnell J. 1969. Class work with fungi. Commonwealth Mycological Institute Kew,

Surrey, England.

Farr DF, & Rossman AY. 2011. Fungal Databases, Systematic Mycology and Microbiology

Laboratory, ARS, USDA. http://nt.ars-grin.gov/fungaldatabases/

Gaumann EA. 1959. Die Rostpilze Mitteleuropas. Beitrage zur Kryptogamenflora der Schweiz 12.

1407 p.

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y,

Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba Shuppankai, Ibaraki.

Nasir YJ. 1984. Primulaceae. 1-77, in: E Nasir & SI Ali. (eds). Flora of Pakistan, vol. 157.

Sydow P, Sydow H. 1904. Monographia uredinearum, vol. 1. Genus Puccinia. Fratres Borntraeger,

Leipzig.

Wilson M, Henderson DM. 1966. British rust fungi, Cambridge University Press, Cambridge.

MY COTAXON

http://dx.doi.org/10.5248/117.321

Volume 117, pp. 321-330 July-September 2011

Gymnopus fuscotramus (Agaricales), anew species

from southern China

ARMIN MESIC!, ZDENKO TKALCEC!, CHUN- YING DENG”,

Tar-Hu1 LY’, BRUNA PLESE’? & HELENA CETKOVIC?

"Ruder Boskovié Institute, Bijenicka 54, HR-10000 Zagreb, Croatia

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

Guangdong Institute of Microbiology, Guangzhou 510070, China

*School of Bioscience and Biotechnology, South China University of Technology,

Guangzhou, 510641, China

CORRESPONDENCE TO *: amesic@irb.hr, * ztkalcec@irb.hr, mycolab@263.net,

bplese@irb.hr & cetkovic@irb.hr

ABSTRACT — A new species, Gymnopus fuscotramus, is described from China. It is

characterized by brown-incarnate colors in pileus and lamellae, sulcate pileus, free and

distant lamellae, floccose-squamulose, mostly black stipe, well-developed black rhizomorphs,

repent and diverticulate pileipellis hyphae, abundant clamp connections, diverticulate to

coralloid cheilocystidia, moderately thick-walled caulocystidia with obtuse apex, dextrinoid

hyphae in cortex of stipe, and gray-brown pileal and hymenophoral trama. Color images of

basidiomata and microscopic elements accompany the description. Gymnopus fuscotramus

is compared with similar species and its systematic position is also inferred using the ITS

rDNA sequence data.

Key worps — Basidiomycota, biodiversity, Omphalotaceae, taxonomy

Introduction

During field research on marasmioid fungi in southern China (Guangxi

Zhuang Autonomous Region), the third author collected an interesting

fungus which is described here as a new species of the genus Gymnopus

(Pers.) Roussel. According to morphological characters, it belongs to section

Androsacei (Kiihner) Antonin & Noordel., which is characterized by small

and marasmioid basidiomata, thin, insititious and mostly dark colored stipe,

usually well-developed black rhizomorphs, dextrinoid trama of stipe and

non-hymeniform pileipellis (Antonin & Noordeloos 2010). Members of this

section were traditionally placed in the genus Marasmius (Fries 1838). Kithner

(1933) described section Androsacei under the same genus. On the basis of

322 ... Medic & al.

its non-hymeniform pileipellis, Antonin (1987) excluded members of the

section Androsacei from Marasmius, and described a new genus Setulipes.

Molecular phylogenetic studies published by Moncalvo et al. (2002), Mata et

al. (2004) and Wilson & Desjardin (2005) showed that the type species of the

genus Setulipes, S. androsaceus (L.) Antonin, belongs to the gymnopoid clade.

Therefore, Noordeloos & Antonin (2008) transferred section Androsacei to the

genus Gymnopus.

Materials & methods

The description of Gymnopus fuscotramus is based on one collection consisting of

seven basidiomata, which were photographed in the field. Color codes in the macroscopic

description (given in brackets) are cited according to Kornerup & Wanscher (1981).

Microscopic features were observed with a light microscope (brightfield and phase

contrast - PhC) under magnification up to 1500x and photographed with a digital

camera. Description and images of microscopic characters were made from rehydrated

specimens mounted in 2.5% potassium hydroxide (KOH) solution. Amyloidity and

dextrinoidity were tested in Melzer’s reagent (Erb & Matheis 1983). Basidiospore

measurements were calculated from mounts of lamellae and based on calibrated

digital images. A total number of 50 randomly selected basidiospores from two mature

basidiomata were measured. Spore measurements (length, width) are given as: (min.) stat.

min. — av. — stat. max. (max), where “min.” = minimum (lowest measured value), “stat.

min.” = statistical minimum (arithmetic average minus two times standard deviation),

“av. = arithmetic average, “stat. max.’ = statistical maximum (arithmetic average plus

two times standard deviation), “max.” = maximum (highest measured value). Standard

deviation (SD) of spore length and width is also given. The length/width ratio of spores

is given as the “Q” value (min. - av. - max.). The holotype is deposited in the Herbarium

of Guangdong Institute of Microbiology (GDGM), while an isotype is deposited in

the Croatian National Fungarium (CNF). Comparison of Gymnopus fuscotramus with

similar taxa is based on revision of type specimens of Marasmius nigroimplicatus Corner

(E 206719) and M. subrigidichorda Corner (E 206861), as well as descriptions in the

following literature: Petch 1948, Singer 1976, 1989, Pegler 1986, Desjardin 1987a,b,

Corner 1996, and Antonin 2007.

Genomic DNA was isolated from dried material with E.Z.N.A. forensic kit (Omega

bio-tek) according to manufacturer's protocol for isolation of DNA from hair, nails and

feathers. The primers ITS1F and ITS4 (White et al. 1990) were used for amplification

and sequencing of ITS region, containing the ITS1, 5.8S and ITS2 regions of rDNA.

PCR amplifications were performed in a total volume of 25 ul. The initial denaturation

step at 95 °C for 85 s, was followed with 35 cycles of 94 °C for 35 s, 55 °C for 45 s, and

72 °C for 60 s. PCR products were subcloned into pGEM-T vector (Promega) according

to the manufacturer's instructions. Three positive clones were sequenced using the

pUC or T7 vector primers with the ABI BigDye Ready Reaction Kit on an ABI 3100

automated sequencer. Sequencing reads were assembled using Lasergene processing

software (DNASTAR Inc., Madison, USA) and checked manually for sequencing errors.

Sequence, submitted to GenBank with accession number JF303730, was compared

Gymnopus fuscotramus sp. nov. (China) ... 323

Fics 1-2. Gymnopus fuscotramus (holotype). 1. Basidiomata in situ. 2. Pileipellis (PhC). Bars:

1= 10mm; 2= 10 um.

324 ... Medié & al.

by homology searches with known sequences using BLAST (Benson et al. 2003). ITS

sequences from 25 species taken from NCBI were used for further analysis. An alignment

of the sequences was performed using Clustal X (Thompson et al. 1994). Ambiguously

aligned regions were determined and excluded from further analyses using the online

version of the program Gblocks 0.91b, under less stringent parameters (Castresana

2000). Final alignment was 620 bp long (available upon request). Jmodeltest (Posada

2008, Guindon & Gascuel 2003) was used to select the best-fit model of nucleotide

substitution. Phylogenetic analyses were performed using Bayesian MCMC, maximum

parsimony (MP) and Neighbor-joining (NJ) methods. Four species of Marasmius were

selected as outgroup taxa for rooting purposes. Bayesian inference of the phylogeny

using Metropolis coupled Markov chain Monte Carlo analyses (Geyer 1991) was

performed using MrBayes, version v. 3.1.2. (Ronquist & Huelsenbeck 2003), under the

Hasegawa-Kishino- Yano + gamma (HKY+G) model, which incorporates different rates

for transitions and transversions and rate variation across sites. MCMC sampling was

performed as implemented in MrBayes with the default settings (two runs of four chains

each) for 10,000 generations, with the first 10% discarded as burn-in. MrBayes was used

to compute a 50% majority rule consensus of the remaining trees to obtain estimates for

the posterior probabilities (PPs) of the groups. Branch lengths were computed as the

mean values over the trees sampled after burn-in. MP and NJ analyses were performed

in Mega version 4 (Tamura et al. 2007). Relative robustness of individual branches was

estimated by bootstrapping (BS), using 1000 replicates.

Taxonomy

Gymnopus fuscotramus MeSi¢, Tkaléec & Chun Y. Deng, sp. nov. FIGS 1-7

MycoBank MB 519324

Pileus 12-21 mm latus, campanulatus, sulcatus, brunneolo-incarnatus usque incarnato-

brunneus. Lamellae liberae, distantes, brunneolo-incarnatae. Stipes 15-30 x 1-1.5 mm,

floccoso-squamulosus, apice aurantio- usque rubro-brunneus, humilius nigro-brunneus

usque niger, insiticius. Sporae (6.7-)6.8-8.2-9.6(-9.8) x (3.0-)3.0-3.7-4.4(-4.8) um,

oblongae, subcylindricae, amygdaliformes vel lacrimiformes, hyalinae. Cheilocystidia

20-45(-60) x 5-15(-25) um, subcylindrica usque irregulariter clavata et diverticulata

vel coralliformia. Pleurocystidia absentia. Caulocystidia 5-100 x 4-12 ym, plerumque

cylindrica, subcylindrica vel anguste clavata, apice obtusa, crasse tunicata (0.5-2 um).

Pileipellis cutis, hyphis diverticulatis. Trama pilealis et hymenophoralis fusca. Fibulae

abundantes. Hyphae in corticali stipite dextrinoideae.

Erymotoecy: Named for the gray-brown color of its pileal and hymenophoral trama.

Ho.otype: CHINA, GUANGXI: Maoershan Nature Reserve, 72 km N of Guilin,

25°54’32”N, 110°27°30"E, alt. 1500 m, 29 May 2009, leg. C.-Y. Deng, GDGM 26313.

ISOTYPE: CNF 1/6044.

PiLEus 12-21 mm broad, campanulate, sometimes with applanate or slightly

depressed center, sulcate almost to the center, weakly translucently striate

when moist, hygrophanous, brownish incarnate to incarnate-brown when

moist (from 10B4 to 11D5), pale incarnate on drying, with darker, mostly dark

red-brown to blackish-brown center, surface dull, dry. LAMELLAE free (without

Gymnopus fuscotramus sp. nov. (China) ... 325

Fics 3-7. Gymnopus fuscotramus (holotype). 3. Spores (PhC). 4. Cheilocystidia (PhC). 5.

Hymenophoral trama. 6. Pileal trama. 7. Caulocystidia. Bars: 3 = 5 um; 4 = 20 um; 5-7 = 30 um.

326 ... Medsi¢ & al.

a collarium or pseudocollarium), distant (L = ca. 12, 1 = 1), <2 mm broad,

ventricose, reaching the margin of the pileus or almost so, sometimes slightly

intervenose, brownish incarnate, with entire, concolorous edge. STIPE 15-30 x

1-1.5 mm, subcylindrical, orange- to red-brown in the upper part, downward

brown-black to black, entirely pale brown floccose-squamulose, dry, hollow,

insititious, arising directly from substrate (not from rhizomorphs). CONTEXT

grayish-brown and very thin in pileus, whitish in stipe medulla, concolorous

with surface in stipe cortex. SMELL and TASTE not recorded. RHIZOMORPHS

abundant, filiform, unbranched, <135 mm long and <0.5 mm thick, glabrous,

black, with black to black brown inner context, hollow, apex with white globule

or tapering and concolorous.

SPORES [50/2/1] (6.7—)6.8-8.2-9.6(-9.8) x (3.0-)3.0-3.7-4.4(-4.8) um, SD

= 0.71 x 0.34, Q = 1.73-2.23-2.70, oblong to subcylindrical, in side view often

amygdaliform or even lacrymoid, smooth, hyaline, thin-walled, non-amyloid,

non-dextrinoid. BAsip1a 23-36 x 4.5-6 um, narrowly clavate, (2)4-spored,

thin-walled, hyaline, clamped. BAstpDIOLEs narrowly clavate, cylindrical or

fusoid. LAMELLAR EDGE Sterile, composed of repent, diverticulate, hyaline

to gray-brown, 1.5-8 um broad hyphae, with clusters of cheilocystidia.

CHEILOCYSTIDIA 20-60 x 5-15(-25) um, subcylindrical to irregularly clavate

with irregular finger- to knob-like projections or coralloid, thin- to moderately

thick-walled (<0.8 tm thick), hyaline to pale gray-brown. PLEUROCYSTIDIA

absent. HYMENOPHORAL TRAMA irregular, composed of 1-6(-8) um broad,

thin- to moderately thick-walled (<1 um thick), subhyaline to pale gray-brown

(gray-brown in mass) hyphae, pigment intracellular. PILEIPELLIs a subregular

cutis composed of 1.5-10 um broad, mostly thin-walled, less frequently

moderately thick-walled (<0.8 um thick), subhyaline to brown, mostly

diverticulate hyphae, with occasional coralloid elements, dark brown pigment

often coarsely encrusted. PILEAL TRAMA composed of pale gray-brown (gray-

brown in mass), non-gelatinized, thin- to moderately thick-walled (<0.8 um

thick), 1-7 um broad hyphae, pigment intracellular. STIPITIPELLIs a cutis of

parallel, 1.8-7 um broad, brown hyphae with intracellular, sometimes also

encrusted pigment. CAULOCYSTIDIA very abundant, 5-100 x 4-12 um, mostly

cylindrical, subcylindrical or narrowly clavate, with obtuse apex, sometimes

diverticulate, moderately thick-walled to thick-walled [walls 0.5-2(-2.5) um

thick], sometimes with one septa, often in groups, subhyaline to brown, with

intracellular, sometimes also encrusted pigment. STIPE TRAMA composed of

parallel, thin- to thick-walled (<1.2 um thick), 2-10 um broad hyphae, hyaline

in stipe medulla. CLlamMPp CONNECTIONS present and abundant in all tissues.

CHEMICAL REACTIONS: all parts of basidioma non-amyloid and non-dextrinoid

except hyphae in cortical layer of stipe and caulocystidia, which are dextrinoid

(red-brown in Melzer’s reagent).

Gymnopus fuscotramus sp. nov. (China) ... 327

Habitat — In small clusters on a twig, in humid subtropical forest of Fagus

longipetiolata and Castanopsis lamonitii.

DISTRIBUTION — Known only from the type locality in China.

Discussion

Gymnopus fuscotramus is characterized by a brown-incarnate colors

in pileus and lamellae, sulcate pileus, free and distant lamellae, floccose-

squamulose, mostly black stipe, well-developed black rhizomorphs, growing

only on woody substrate (not arising from rhizomorphs), non-hymeniform

pileipellis (cutis with diverticulate hyphae), abundant clamp connections,

diverticulate to coralloid cheilocystidia, moderately thick-walled caulocystidia

with obtuse apex, dextrinoid hyphae in cortex of stipe, and gray-brown pileal

and hymenophoral trama.

Among other species in the section Androsacei with clamp connections,

black rhizomorphs, non-glabrous stipe, and colored pileus and lamellae (not

white or cream), Marasmius bactrosporus Singer, M. campinaranae Singer,

and Setulipes brevistipitatus Antonin have (among others) much smaller

basidiomata and more elongated spores. M. nigroimplicatus has thinner

(<0.2 mm) and minutely pubescent rhizomorphs, adnate lamellae sometimes

attached to a pseudocollarium, more elongated spores, and caulocystidia with

thicker walls (1.8-5 um). M. rigidichorda Petch has basidiomata arising also

from the rhizomorphs, adnate lamellae, white context, hyaline hymenophoral

trama, and caulocystidia with pointed apex. M. subrigidichorda has basidiomata

arising also from the rhizomorphs, lamellae attached to a pseudocollarium,

longer and wider rhizomorphs (<410 x 0.9 mm), hyaline hymenophoral

trama, and caulocystidia with thicker walls (2-5 um). M. thiersii Desjardin has

crowded, adnate to adnexed lamellae, rare or absent rhizomorphs, and lacks

cheilocystidia. Setulipes rhizomorphicola Antonin has much shorter (<3 mm)

and eccentric stipe arising from rhizomorphs, adnate lamellae, and longer

spores.

According to morphological characters, Gymnopus fuscotramus belongs to

section Androsacei. However, our phylogenetic analysis using only ITS rDNA

sequences do not place our taxon on the same clade with the two sequenced

species from that section, G. androsaceus (L.) J.L. Mata & R.H. Petersen and

G. quercophilus (Pouzar) Antonin & Noordel. However, G. fuscotramus is

placed close to G. peronatus (Bolton) Gray (sect. Vestipedes). To further clarify

phylogenetic relationships between G. fuscotramus and other Gymnopus

species (especially from sect. Androsacei), more species (reliably identified)

and more DNA sequences should be included in analyses. Our gymnopoid

clade also includes Marasmiellus and Rhodocollybia species, which is in

accordance with results of Moncalvo et al. (2002), Mata et al. (2004) and Wilson

328 ... Mesié & al.

Rhodocollybia maculata AF505756

71*-0.95*

Rhodocollybia butyracea EU486454

Marasmiellus juniperinus AY256708

--0.97*

Gymnopus subpruinosus DQ450026

--0.89"

Gymnopus brunneigracilis AY 263434

74*99*1.00*

Gymnopus gibbosus AY263437

-86*1.00*

--0.99" | Gymnopus luxurians AY256709

Gymnopus trogioides AY263428

Gymnopus dichrous AF505766

77*71*1.00*

Marasmiellus ramealis DQ450030

Gymnopus confluens AF505773

98*99*1.00*

Gymnopus menehune AY 263426

Gymnopus fuscotramus JF303730

73*96*1.00*

Gymnopus peronatus AF505760

—1.00*

Gymnopus fusipes AY256710

71*-1.00*

Gymnopus quercophilus AF335435

100*91*1.00*

Gymnopus androsaceus GU234007

Gymnopus polyphyllus FJ596895

-88°0.99*

Gymnopus impudicus AF505779

100*-1.00* cel

Marasmiellus foetidus AF505780

Gymnopus aquosus AY256691

81*98*1.00*

Gymnopus indoctoides AY263424

Marasmius haematocephalus EU935534

100*100*-

Marasmius purpureostriatus FJ904978

81*75*1.00*

Marasmius pellucidus EU935508

89*100*1.00*

Marasmius bambusiniformis EU935521

0.1

Fic. 8. Phylogenetic tree based on the complete ITS rDNA region (ITS1, 5.8S rDNA, and ITS2),

showing mean branch lengths of a 50 % majority-rule consensus tree from a Bayesian MCMC

analysis. NJ, MP bootstrap values (>70%), and Bayesian PP values are given above nodes.

Gymnopus fuscotramus sp. nov. (China) ... 329

& Desjardin (2005). All these results indicate that Gymnopus and Marasmiellus

(as conceived recently) are polyphyletic and that new taxonomic concepts

inferred from DNA sequences should be proposed. However, since past

phylogenetic analysis included only a small number of species from these

two genera, further taxonomic solutions should be based on more extensive

phylogenetic research with more species and DNA sequences included.

Acknowledgments

We would like to thank Dr. David Harris, curator of Herbarium of Royal Botanic

Garden Edinburgh (E) for loan of Corner’s holotype collections. We are very grateful

to Dr. Vladimir Antonin (Moravian Museum, Brno, Czech Republic) and Dr. Roy E.

Halling (New York Botanical Garden, New York, USA) for their critical review of the

manuscript. Field research was supported by the National Natural Science Foundation

of China (No. 30770004, 30870019, 30970023).

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MY COTAXON

http://dx.doi.org/10.5248/117.331

Volume 117, pp. 331-342 July-September 2011

Biogeographical patterns in pyrenomycetous fungi and

their taxonomy. 2. Additions to the Grayan disjunction

LARISSA N. VASILYEVA’* & STEVEN L. STEPHENSON?

‘Institute of Biology & Soil Science, Far East Branch of the Russian Academy of Sciences,

Vladivostok 690022, Russia

*Department of Biological Sciences, University of Arkansas, Fayetteville, AR 72701, USA

*CORRESPONDENCE TO: vasilyeva@biosoil.ru

ABSTRACT — We report additional examples of pyrenomycetous fungi that display the

biogeographical pattern known as the Grayan disjunction. Daldinia grayana and Diatrypella

informis are described as new species, and Nemania pseudoillita is proposed as a new

combination.

KEY worps — Ascomycota, biogeography, new species

Introduction

Additional collecting trips and visits to several different herbaria have

revealed a number of specimens that warranted a re-identification. The data

available on their distribution indicate that they display the biogeographical

pattern known as the Grayan disjunction, which was discussed in a previous

paper (Vasilyeva & Stephenson 2010).

Two types of this distribution pattern have been observed in pyrenomycetous

fungi—vicariance and non-vicariance. In addition, there are two other groups

of fungi restricted to either eastern Asia or eastern North America that do not

have close relatives in the other region. Although these two groups of species are

seemingly pertinent to the two regions under consideration, the biogeographical

patterns they exhibit have different explanations, which will be considered in a

forthcoming paper. The purpose of the present paper is to provide additional

information relating to the Grayan disjunction in pyrenomycetes and to discuss

certain aspects of this pattern.

Materials & methods

The specimens included in this study are deposited in FH, MICH, and VLA. The maps

used as the basis for illustrations were taken from the web sites commons.wikimedia.org

332 ... Vasilyeva & Stephenson

(Fics 1-3) and www.paleoportal.org (Fic. 4); all maps were modified to show our data.

Photographs of ascomata were obtained using a Nikon D40x digital camera.

Examples of the non-vicariance pattern

Daldinia grayana Lat.N. Vassiljeva & S.L. Stephenson, sp. nov. Fig. 1

MycoBank MB 561654

Stromata solitaria vel aggregata et confluentia, sessilia vel breve stipitata, 0.5-2.5 cm

diam. x 0.8-2.5 cm alta, atro-lateritia, demum denigrata et laccata, rugosa, epapillata

et sine tumulis peritheciorum, pigmenti griseo- vel veriduli-olivacei, interdum atro-gresei

vel plumbei in KOH dissoluti; textura sub peritheciis zonis alternantibus composita, zonis

fuscis 0.3-0.6 mm crassis et zonis cremeo-fuscis 0.1-0.3 mm crassis praedita. Perithecia

tubularia, 0.1-0.2 mm diam. x 0.4-0.8 mm alta. Asci partibus sporiferis 80-100 x 7-8 um,

stipitibus 80-100 um longitudine, annulo apicali in liquore iodato Melzeri cyanescente,

discoideo, 3-4 x 0.8-1 um. Ascosporae brunneae, unicellulares, brevi- et late-ellipsoideae

vel anguste-ellipsoideae, apicibus anguste-rotundatis, (10-)12-14(-16) x 6-7.5 um, rima

germinativa recta longa praeditae; perisporium in KOH dehiscens, leve.

TyPE: Russia: Primorsky Territory, Lazo Nature Reserve, on Alnus hirsuta (Spach) Turcz.

ex Rupr. (Betulaceae), 4 Aug 1986, L. Vasilyeva (Holotype VLA P-2547).

Stromata solitary to aggregated and confluent, sessile or short stipitate,

0.5-2.5 cm diam., 0.8-2.5 cm high, dark reddish brown, blackened and

varnished in age, wrinkled, lacking papillae and perithecial mounds, KOH-

extractable pigments gray-olivaceous or greenish-olivaceous, sometimes dark-

gray or plumbeous; tissue below the perithecial layer composed of alternating

zones, the darker zones dark brown, 0.3-0.6 mm thick, the lighter zones

creamy-brown, 0.1-0.3 mm thick. Perithecia tubular, 0.1-0.2 mm diam. x

0.4-0.8 mm high. Asci in the spore-bearing part 80-100 x 7-8 um, the stipe

80-100 um long, with apical ring bluing in Melzer’s iodine reagent, discoid, 3-4

x 0.8-1 um. Ascospores brown, unicellular, variable in shape, broadly ellipsoid

to more narrow-ellipsoid, (10-)12-14(-16) x 6-7.5 um, with a straight germ

slit spore-length; perispore dehiscent in 10% KOH, smooth.

ADDITIONAL SPECIMENS EXAMINED: CHINA: HEILONGJIANG PROVINCE, Raohe,

on Alnus hirsuta, 7 Aug 2004, L. Vasilyeva (VLA P-2551). CANADA: Nova Scotia,

Salmon River, 14 Jul 1931, L.E. Wehmeyer (MICH 71984, as Daldinia concentrica). USA:

NEw York, Ithaca, Aug 1902, C.H. Kauffman (MICH 71986, as Daldinia concentrica;

substrate not indicated); VERMONT, Pawlett, on Betula papyrifera Marshall (Betulaceae),

C.W. Dodge & D.H. Linder (FH, as Daldinia concentrica); MICHIGAN, Vermilion Point,

30 Jul 1914, A.H. Povah (MICH 71987 & 71988, as Daldinia concentrica); Rock River,

on Alnus sp., 24 Aug 1927, A.H. Povah (MICH 71989, as Daldinia concentrica); SE of

Stockbridge, on dead wood, 20 Aug 1928, E. Whitney & B.B. Kanouse (MICH 71992,

as Daldinia concentrica); Rock Harbor, on Betula papyrifera, 20 Apr 1930, A.H. Povah

(MICH 71990, as Daldinia concentrica); Todd Harbor, on Alnus sp., 24 Aug 1930, A.H.

Povah (MICH 71985, as Daldinia concentrica); McCargoe Cove, on Alnus sp., 1930 (date

not indicated), A.H. Povah (MICH 71982, as Daldinia concentrica); Conway Lake Road,

on Betula alleghaniensis Britton, 12 Aug 1970, K.A. Harrison (MICH 71983, as Daldinia

concentrica); Pine Lake, on Betula sp., 23 Jul 1971, K.A. Harrison (MICH 71991, as

Grayan disjunction of pyrenomycetes ... 333

Fic. 1. Approximate biogeographical distribution of Daldinia grayana. Scale bars: A = 10 mm,

B=5mm.

Daldinia concentrica); TENNESSEE, Smoky Mountains National Park, Newfound Gap,

on Betula sp., 13 Aug 1968, D.A. Reid (K, as Daldinia vernicosa).

ComMENTS-The appearance of Daldinia grayana is similar to that of

D. gelatinosa Y.M. Ju et al. as the latter is illustrated (Ju et al. 1997, Fic. 44) and

described (Ju et al., 1997, p. 270: “Stromata... sessile or short stipitate, solitary or

aggregated, wrinkled, ...surface dark brick, blackened and varnished in age...”).

It is also noteworthy that D. gelatinosa appears to have a similar distribution

in eastern North America and is reported from Ontario (Canada), Idaho, and

New Hampshire. The latter record is associated with Betula alleghaniensis, as is

one record of D. grayana.

The resemblance of Daldinia gelatinosa to D. grayana is reinforced by the

fact that one piece of the exsiccate of ‘Daldinia concentrica’ in Ellis & Everhart’s

334 ... Vasilyeva & Stephenson

“Fungi Columbiani 213” was identified as D. gelatinosa (Ju et al. 1997: WSP

1781), whereas we identify another piece (MICH 71993) as Daldinia grayana.

Except for this questionable material, D. gelatinosa differs from D. grayana in

having dark-livid KOH-extractable pigments rather than gray- or greenish-

olivaceous pigments, as well as in the nature of the inner tissue of the stromata.

This tissue in D. gelatinosa is similar to that of D. fissa Lloyd in having the

alternating zones of dark brown and white, which are initially gelatinous but

disintegrate and become loculate when dry. In contrast, D. grayana has stromata

that are solid inside, and their alternating zones are only darker and lighter

tinges of brown, although sometimes the lighter zones are cream-colored.

Based on the abundant material from Michigan, Daldinia grayana might be

more of a northern or high elevation species (cf. the record from Newfound

Gap) than D. gelatinosa; in this respect, D. grayana is similar to D. loculata

(Lév.) Sacc., which occurs on birch (Betula spp.) in the more northern parts

of the Holarctic; when D. loculata is found in more southern regions, it occurs

primarily in the mountains.

In terms of the substrate preferences (Alnus spp., Betula spp.), Daldinia

grayana should be also compared with D. decipiens Wollw. & M. Stadler,

which is known to occur on Betula spp. in Europe (Stadler et al. 2001b), and

D. petriniae Y.M. Ju et al. described on Alnus also from Europe (Ju et al. 1997)

but later found to be circumpolar (Stadler et al. 2001b, p. 175). Only one locality

for D. petriniae in the United States is from the Pacific coast region (Stadler et

al. 2001a, c). This species differs from D. grayana in having larger, more fragile

stromata, livid purple or dark livid pigments, and a finely papillate stromatal

surface (similar to the condition illustrated by D. childiae J.D. Rogers & Y.M. Ju

as seen on the website http://mycology.sinica.edu.tw/Xylariaceae). We have a

specimen of D. petriniae on Alnus hirsuta from China (Heilongjiang Province)

collected in 2004. The pigments in its stromata display pale-vinaceous-grey

pigment in 10% KOH instead of livid purple or dark livid, but it is the same

portion of the color spectrum in contrast to greenish-olivaceous tints observed

in D. grayana.

Daldinia decipiens is similar to D. petriniae in having purple, dark livid

or vinaceous purple KOH-extractable pigments (Stadler et al. 2001b).

Morphologically, these two species are sometimes difficult to distinguish,

although D. decipiens appears to have larger ascospores; it also differs from D.

petriniae in the smaller, often stipitate, stromata with a smooth surface and

more solid inner tissue with brown zones of very close tinges. In addition, the

stromata of D. decipiens are reddish brown, especially the stalks, whereas those

in D. petriniae are mostly brown vinaceous. ‘The color is shared by D. decipiens

with D. grayana, but the stromatal pigments and consistency easily distinguish

them.

Grayan disjunction of pyrenomycetes ... 335

Diatrypella informis Ellis & Everh. ex Lar.N. Vassiljeva & S.L. Stephenson, sp. nov.

MycoBank MB 561655

Stromata solitaria vel aggregata, e cortice erumpentia, linea nigra substrato circumdata,

verrucaeformia, plano-convexa, suborbicularia vel ellipsoideae, 4-10 x 1.5-4 mm,

laeviuscula vel leviter rugosa, nigra, intus albida. Perithecia subglobosa, 0.1-0.2 mm diam.,

profunde immersa, cum ostioli integri, umbilicati vel leviter papillati, Asci longe ellipsoidei

vel clavati, polyspori, aparaphysati, partibus sporiferis 35-60 x 5-7 um, stipitibus ad 75

um longitudinus, tunica apice incrassata, annulo apicali nullis. Ascosporae allantoideae,

unicellulares,brunneolae vel flavidae, 4-6 x 1-1.2 um.

Type: Ellis & Everhart's North American Fungi N 2530: “Diatrypella informis E. & E. n.

sp., on dead Carpinus, London, Canada, Apr. 1890, J. Dearness”. [Holotype BPI].

Stromata usually densely scattered over the branches of the host tree, surrounded

by a black zone, erumpent from the bark with a black, robust, rounded or

ellipsoid, convex, smooth or somewhat wrinkled ectostromatic disc 4-10 x

1.5-4 mm in diam. Perithecia subglobose, 0.1-0.2 mm diam., deeply immersed,

the tissue above perithecial layer white, ostioles umbilicate or slightly papillate,

non-sulcate. Asci long ellipsoid or slightly clavate, polysporous, in the spore-

bearing portion 35-60 x 5-7 um, stalks up to 75 um, wall thickened at the

apex, without an apical ring and paraphyses. Ascospores unicellular, allantoid,

brownish yellow when crowded in asci, pale yellow when scattered, 4-6 x 1-1.2

um.

ADDITIONAL SPECIMENS EXAMINED: RUSSIA: PRIMORSKY TERRITORY, Vladivostok

vicinity, on Carpinus cordata Blume (Corylaceae), 6 Jun 1958, I. Bunkina (VLA P-1975);

11 Oct 1991, L. Vasilyeva (VLA P-1978 & P-2129); 5 Oct 2006, L. Vasilyeva (VLA P-

1108); Sikhote-Alin Nature Reserve, on Carpinus cordata, 24 Oct 1985, L. Vasilyeva

(VLA P-2126); Kedrovaya Pad Biosphere Reserve, on Carpinus cordata, 4 Oct 1987 & 25

Oct 1987, L. Vasilyeva (VLA P-2127 & P-2128); Ussuriysk Nature Reserve, on Carpinus

cordata, 19 Aug 1989, L. Vasilyeva (VLA P-2265); Anisimovka, on Carpinus cordata, 20

Sep 1996, L. Vasilyeva (VLA P-2130).

ComMENtTs- Diatrypella informis was illustrated in the previously published

paper on biogeographic patterns in pyrenomycetous fungi (Vasilyeva &

Stephenson 2010), but its name was never validly published. Therefore, a Latin

description is provided herein to validate the name.

The rather large robust stromata and non-sulcate ostioles of D. informis

are similar to those found in D. verruciformis (Ehrh.) Nitschke, which differs

by possessing larger asci (120-200 x 8-12 um: Nitschke 1867) and longer

ascospores (6-8 um long). The 4-6 um range in ascospore length is used

to segregate many species in the Diatrypaceae; examples include Diatrype

hypoxyloides De Not., Diatrypella decorata Nitschke, Eutypa mela (Schwein.)

Cooke, Eutypella kochiana Rehm. The regularly repetitive size ranges in different

genera of the family are always very helpful in delimiting certain species within

these families.

336 ... Vasilyeva & Stephenson

Examples of the vicariance pattern

Hypoxylon dearnessii and H. massulatum

Collections carried out in the vicinity of the University of Michigan

Biological Station (UMBS, located near Douglas Lake, Michigan) in 2010

revealed a Hypoxylon species with rather variably shaped, often undulate

stromata. This species was associated with Acer rubrum. Some of its superficial

features suggested Hypoxylon dearnessii (Ju & Rogers 1996: Fics. 11A-B),

recorded from maples only in eastern temperate North America and originally

described from the vicinity of the city of London (Ontario, Canada) in the same

general Great Lakes region as the UMBS area.

The KOH-extractable pigment in the Michigan specimen was the color of

sienna (as in H. dearnessii), and the ascospores exhibited a similar size range

(9-11.5 x 4.5-6 um), although with a slightly larger range in length (9-12.5 x

4.5-5.5 um) and with many more obovoid with one end narrowly rounded and

the other broadly rounded, as in H. fuscopurpureum (Schwein.) M.A. Curtis.

Hypoxylon fuscopurpureum differs in having greenish-olivaceous pigments

and larger ascospores, but it and the Michigan specimen both have perispores

that are indehiscent in 10% KOH. The perispore is important in delimiting

Hypoxylon species and, in fact, prevents our assigning the Michigan specimen

to H. dearnessii, which has a dehiscent perispore. Nonetheless, the specimen

is provisionally identified as H. dearnessii (VLA P-2519) and is accessible for

future comparative studies.

Hypoxylon dearnessii is known from Canada (Ontario, Quebec) and the

United States (Colorado, Maine, New York) (Ju & Rogers 1996; Stadler et

al. 2008). Specimens deposited in the Farlow Herbarium (all as “Hypoxylon

rubiginosum’), were collected in Connecticut, Massachusetts, New Hampshire,

Rhode Island, and Vermont, mostly on Acer rubrum. The eastern North

American temperate zone is home to H. dearnessii, but there are some collections

from eastern Asia that resemble this species. Especially suggestive is a specimen

collected in 2000 on Acer ukurunduense from the Big Khekhtsir Nature Reserve

(Khabarovsk Territory, Russia), because its stromata have the same shape as

H. dearnessii, and the substrate was maple. However, specimens with the same

type of stromata were collected later on Ulmus spp. in the Primorsky Territory

(Russia) and in Heilongjiang Province (China).

A close examination of the stromata of specimens on Acer ukurunduense

and Ulmus spp. showed their KOH-extractable pigment to be bright red instead

of sienna, and they were also found to have smaller ascospores (mostly 7.5-8.5

x 3.8-5 um) with an indehiscent perispore. Further comparisons showed that

these stromata are very similar to those in Hypoxylon massulatum described

from Alnus hirsuta (Vasilyeva 1998).

Grayan disjunction of pyrenomycetes ... 337

aan.

Fic. 2. Approximate biogeographical distribution of Hypoxylon massulatum (left) and H. dearnessii

(right). A-B. H. massulatum on Alnus (A) and Ulmus (B). C-D. H. dearnessii on Acer rubrum

from University of Michigan Biological Station area. Scale bars: A-B = 15 mm, C-D = 10 mm.

Hypoxylon massulatum has been considered as a synonym of H. howeanum

Peck (Ju et al. 2004), probably because of the similar ascospore size and

338 ... Vasilyeva & Stephenson

stromatal pigments, but its resemblance to H. dearnessii (also acknowledged

as showing affinities to H. howeanum: Stadler et al. 2008) is more evident.

Hypoxylon howeanum has mostly globose stromata, which are sometimes

rather large (up to 2 cm), brightly orange and very smooth at the surface and

silky brown inside.

Hypoxylon dearnessii Y.M. Ju & J.D. Rogers, Mycol. Mem. 20: 106 (1996).

SPECIMENS EXAMINED: USA: MICHIGAN, CHEBOYGAN Co., Douglas Lake, University of

Michigan Biological Station, on Acer rubrum L. (Aceraceae), 12 Aug 2010, L. Vasilyeva

(VLA P-2519); CONNECTICUT, New Haven, on Acer sp., about 1888, R. Thaxter (FH, as

Hypoxylon rubiginosum); MASSACHUSETTS, Waverly, on Acer rubrum, 23 Apr 1892, R.

Thaxter (FH); Medford, Middlesex Fells, 13 Oct 1935, G. Darker 5479 (FH, as Hypoxylon

rubiginosum); VERMONT, Middlebury, Great Swamp, 26 Nov 1896, E. Burt (FH, as

Hypoxylon rubiginosum); NEw HAMPSHIRE, near Sawyer's River, on Acer sp., Aug 1901,

R. Thaxter 7112 (FH, as Hypoxylon rubiginosum); Chocorua, on Acer rubrum, 9 Aug

1907, W. Farlow (FH, as Hypoxylon rubiginosum); RHODE ISLAND, Glochester, substrate

not indicated, 6 Jun 1922, D. Linder & S. Cook (FH, as Hypoxylon rubiginosum).

Hypoxylon massulatum Lar.N. Vassiljeva, Nizshie Rasteniya, Griby i

Mokhoobraznye Dalnego Vostoka Rossii, Griby 4: 182 (1998)

Stromata pulvinate, sometimes convex on top, rounded, ellipsoid or irregular,

often with crenate margins, 0.4-1 cm diam., surface dark brick, bright orange or

ochre; bright red granules immediately beneath surface, with KOH-extractable

pigments orange-red. Perithecia ovoid, 200-300 um diam., ostioles umbilicate.

Asci cylindrical, the spore-bearing portion 45-50 x 5-7 um, the stipes 30-40

uum long, with discoid apical ring bluing in Melzer’s iodine reagent. Ascospores

one-celled, ellipsoid, pale brown, 7.5-9(-10) x (3.5-)3.8-5 um, with straight

germ slit spore length; perispore indehiscent in 10 % KOH, smooth.

SPECIMENS EXAMINED: RUSSIA: KHABAROVSK TERRITORY, Big Khekhtsir Reserve,

on Acer ukurunduense Trautv. & C.A. Mey., 20 Jun 2000, L. Vasilyeva (VLA P-937);

PRIMORSKY TERRITORY, Nadezhdino District, Sirenevka, on Alnus hirsuta, 29 Aug

1991, L. Vasilyeva (VLA P-2511); Khanka Nature Reserve, on Ulmus sp., 21 Jun 2003,

L. Vasilyeva, P-936. CHINA: HEILONGJIANG PROVINCE, Xingkaihu Nature Reserve, on

Ulmus sp., 1 Sept 2003, L. Vasilyeva (VLA P-1623).

Nemania illita and N. pseudoillita

The genus Nemania Gray has received attention from a number of

mycologists who work with pyrenomycetous fungi (Granmo et al. 1999, Ju &

Rogers 2002) but remains rather difficult taxonomically. Seven species in this

genus were reported previously from the Russian Far East (Vasilyeva 1998) but

as members of the genus Hypoxylon Bull.

Hypoxylon pseudoillitum was described as differing from H. illitum

(Schwein.) M.A. Curtis in having larger ascospores (12-16 against 9-12 um).

The same difference exists between many Hypoxylon species (Vasilyeva 1983,

1985). Their stromata also differ (Fic. 3), although it is difficult to differentiate

Grayan disjunction of pyrenomycetes ... 339

these species in keys and descriptions. Most probably it is possible to count

the number of papilla per unit of surface area, but this feature was usually

not taken into account. A new combination for and English description of H.

pseudoillitum are provided below.

Nemania pseudoillita (Lar.N. Vassiljeva) Lar.N. Vassiljeva & S.L. Stephenson,

comb. nov. FIG. 3

MycoBank MB 561656

= Hypoxylon pseudoillitum Lar.N. Vassiljeva, Nizshie Rasteniya, Griby i

Mokhoobraznye Dalnego Vostoka Rossii, Griby 4: 190 (1998)

Stromata effuse-pulvinate, plane, with a densely and finely papillate surface,

brownish, becoming darker with age; perithecia spherical, 600-800 um diam.

Fic. 3. Approximate biogeographical distribution of Nemania pseudoillita (left) and N. illita

(right). North American localities listed by Miller (1961) include Alabama, Georgia, Kentucky,

Louisiana, Maine, Maryland, Missouri, Nebraska, New Hampshire, New York, North Carolina,

Pennsylvania, and Virginia in United States. Scale bar = 5 mm.

340 ... Vasilyeva & Stephenson

Asci cylindrical, the spore-bearing portion 80-90 x 5-7 um, the stipes 40-50

um long, with apical ring bluing in Melzer’s iodine reagent, cubiform, 2 x 2

um. Ascospores one-celled, diamond- or wedge-shaped, very light brown,

almost hyaline, brownish in mass, (11.5-)13-15(-16.5) x (2.5—)3-4.5 um, with

a ventral germ slit.

SPECIMENS EXAMINED: RUSSIA: KHABAROVSK TERRITORY, Big Khekhtsir Nature

Reserve, on wood, 16 Sept 1981, L. Vasilyeva (VLA P-1842); PRIMORSKY TERRITORY,

Lazo Nature Reserve, on wood, 26 Jul 1986, L. Vasilyeva (VLA P-1378); Kedrovaya Pad

Reserve, on wood, 28 Oct 1987, L. Vasilyeva (VLA P-1380); Ussuriysk Nature Reserve,

on wood, 28 Aug. 1989, L. Vasilyeva (VLA P-194); Vladivostok vicinity, on wood, 25

May 2003, L. Vasilyeva (VLA P-1379).

Discussion

We did not discuss the Grayan disjunction in pyrenomycetous fungi as part

of an overview of biogeographical problems in our previous paper (Vasilyeva

& Stephenson 2010). Especially interesting in this context is the vicarious

pattern. The occurrence of the same species in two widely separate regions of

eastern Asia and eastern North America could be attributed to the persistence

of Tertiary relics following continental fragmentation, but the occurrence

of intercontinental vicarious pairs of species (or varieties) is not so easily

understood.

Vicarious taxa usually have very minor morphological differences (e.g.,

ascospore size), and it is dificult to explain why Biscogniauxia maritima

Lar.N. Vassiljeva (ascospores 13-16 um long) occurs only in the southern

portion of the Russian Far East, north-east China, Japan, and Korea, whereas

B. atropunctata (Schwein.) Pouzar (ascospores 24-33 um long) is restricted to

eastern North America. Both species occur on Quercus spp., and the ascospore

size difference seems unlikely to have adaptive significance. We cannot identify

any association between the environmental conditions in the two regions

and the observed differences. Moreover, in some vicarious pairs, for example

Nemania illita and N. pseudoillita (see above) or Hypoxylon notatum Berk. &

M.A. Curtis and H. ulmophilum Lar.N. Vassilyeva (Vasilyeva & Stephenson

2010), the species with larger ascospores were found in East Asia. In the latter

case, the substrate restriction of H. notatum and H. ulmophilum to Quercus spp.

and Ulmus spp., respectively, also appears very strange, since representatives

of both host genera are abundant in the two disjunctive areas. Although both

species have plentiful host resources, they do not seem to jump from one host

genus to another in eastern Asia or eastern North America.

Despite their close similarity, vicarious taxa cannot be ancestors and

descendants to each other, since, if such is the case, there should be some

common area of speciation. And even if such an area existed and became

fragmented later, it is inexplicable how such fragmentation has so strictly

Grayan disjunction of pyrenomycetes ... 341

ARS ATLANTIGZS ~

OCEANS SKS

% 64; SRD

vj Africa Y

—.

Wks \-47~ TETHYS OCEAN | eh 4.

2 (fs |7,

Me SOUTH Bs {7 INDIAN

. ot v OCEAN

Fic. 4. The position of continents during the Cretaceous (A) and Tertiary (B) periods. White circles

indicate the present landmasses of eastern Asia and eastern North America; it is evident that

the original isolation of these regions has increased over time.

divided the populations in such a way that all ‘ancestors’ (with the same

ascospore size) appear to be confined to either Asia or North America, whereas

all ‘descendants’ occur on the other region. An additional hypothesis to account

for their origin might suggest that only one species existed before continent

fragmentation, and the second species evolved as a deviation of the first after

long period of isolation of the two regions. However, once again it is not clear

why the ancestral species disappeared completely from one fragment of the

disjunctive region.

The most probable explanation of the vicariance pattern might be attributed

to the position of continents during the Cretaceous and Tertiary periods (Fic.

4). Even during these earlier times, the present land masses of East Asia and

eastern North America were widely separated. Therefore, the isolation of

vicarious taxa was an original occurrence that only increased over time. The

vicarious taxa may have been peripheral populations of some species otherwise

widely distributed over Laurasia (the northern part of Pangaea).

It is particularly common for isolated populations at the margins of a range

for a given species to exhibit certain distinct features, and clearly distinct

peripheral populations are often observed on islands (Ridley 2003). During the

Tertiary, the last phase of the breakup of Pangaea, widely separated areas with

their own distinctive populations would have became biogeographical ‘islands,

and the vicarious taxa that exist today may simply represent fragments of the

same ancestral species.

Acknowledgments

This biogeographical project is being supported in part by the Russian Foundation

for Fundamental Studies (grant # 09-05-00245), with additional funding from several

other sources. We thank the curators of the mycological herbaria of Michigan University

342 ... Vasilyeva & Stephenson

and Harvard University (Farlow Herbarium), Dr Timothy James and Dr Donald Pfister,

as well as collection manager Patricia Rogers and curatorial assistant Genevieve Lewis-

Gentry for their help during the study of specimens in those herbaria. We are also

grateful to Dr. H.H. Burdsall (Fungal and Decay Diagnostics, LLC) and Dr. W.-Y. Zhuang

(Institute of Microbiology, Chinese Academy of Sciences) for serving as presubmission

reviewers and for providing helpful comments and suggestions. An early draft of this

manuscript was kindly looked over by Dr. R.H. Petersen (University of Tennessee), and

his comments are greatly appreciated.

Literature cited

Granmo A, Lessge T, Schumacher T. 1999. The genus Nemania s.l. (Xylariaceae) in Norden.

Sommerfeltia 27: 1-96.

Ju YM, Rogers JD. 1996. A revision of the genus Hypoxylon. Mycologia Memoir 20: 1-365.

Ju YM, Rogers JD. 2002. The genus Nemania (Xylariaceae). Nova Hedwigia 74: 75-120.

http://dx.doi.org/10.1127/0029-5035/2002/0074-0075

Ju YM, Rogers JD, San Martin F. 1997. A revision of the genus Daldinia. Mycotaxon 61: 243-293.

Ju YM, Rogers JD, Hsieh HM. 2004. New Hypoxylon species and notes on some names associated

with or related to Hypoxylon. Mycologia 96: 154-161. http://dx.doi.org/10.2307/3761997

Miller JH. 1961. A monograph of the world species of Hypoxylon. University of Georgia Press,

Athens. 158 p.

Nitschke T. 1867. Pyrenomycetes Germanici. Verlag von Eduard Trewendt, Breslau. 320 p.

Ridley M. 2003. Evolution. 3rd ed. Wiley-Blackwell, Hoboken. 792 p.

Stadler M, Baumgartner M, Grothe T, Mihlbauer A, Seip S, Wollweber H. 2001a. Concentricol,

a taxonomically significant triterpenoid from Daldinia concentrica. Phytochemistry 56:

787-793.

Stadler M, Baumgartner M, Wollweber H, Ju YM, Rogers JD. 2001b. Daldinia decipiens sp. nov. and

notes on some other European Daldinia spp. inhabiting Betulaceae. Mycotaxon 80: 167-177.

Stadler M, Wollweber H, Mihlbauer A, Henkel T, Asakawa Y, Hashimoto T, Ju YM, Rogers JD,

Wetzstein HG, Tichy HV. 2001c. Secondary metabolite profiles, genetic fingerprints and

taxonomy of Daldinia and allies. Mycotaxon 77: 379-429.

Stadler M, Fournier J, Granmo A, Beltran-Tejera E. 2008. The “red hypoxylons” of the temperate

and subtropical Northern hemisphere. North American Fungi 3(7): 73-125.

http://dx.doi.org/ 10.2509/naf2008.003.0075

Vasilyeva LN. 1983. On systematics of the genus Hypoxylon Fr. I. Mikologiya i fitopatologiya 17:

21-27. (in Russian)

Vasilyeva LN. 1985. The combinatorial principle in the pyrenomycetous systematics. Komarov's

Readings 32: 14-56. (in Russian)

Vasilyeva LN. 1998. Pyrenomycetes and loculoascomycetes. Lower plants, fungi, and bryophytes of

the Russian Far East. Vol. IV. Nauka, Saint-Petersburg. 419 p. (in Russian).

Vasilyeva LN, Stephenson SL. 2010. Biogeographical patterns in pyrenomycetous fungi and their

taxonomy. 1. The Grayan disjunction. Mycotaxon 114: 281-303.

http://dx.doi.org/10.5248/114.381

MY COTAXON

http://dx.doi.org/10.5248/117.343

Volume 117, pp. 343-349 July-September 2011

Lignincola conchicola from palms with a key to the species

of Lignincola

JIAN-Kut1 Liu’, E.B. GARETH JONES’, EKACHAI CHUKEATIROTE’,

A.H. BAHKALI? & KEVIN. D. HYDE”

"School of Science, Mae Fah Luang University, Chiang Rai, 57100, Thailand

? BIOTEC Central Research Unit, National Center for Genetic Engineering and Biotechnology,

113 Thailand Science Park, Bangkok, 12120, Thailand

* College of Science, Botany and Microbiology Department, King Saud University,

PO. Box: 2455, Riyadh 1145, Saudi Arabia

*CORRESPONDENCE TO: Ijiankui@gmail.com

ABSTRACT — During studies of palm fungi, a new Lignincola species was found in Thailand.

It differs from other members of the genus in ascospore dimensions and the occurrence of its

ascomata on the adhesive pad of a marine invertebrate. Lignincola conchicola is described and

illustrated, and a key to Lignincola species is provided. The palm Phoenix is a new substratum

for marine fungi.

Key worps — Halosphaeriaceae, mangrove, taxonomy, tropic

Introduction

During research on palm fungi in Thailand (Pinnoi et al. 2004, 2006,

2009, 2010, Pilantanapak et al. 2005, Pinruan et al. 2004a,b, 2007, 2010a,b,

Rungjindamai et al. 2008, Liu et al. 2010) a new taxon collected on Phoenix

paludosa in Ranong mangrove, was discovered. The taxon is characteristic of

Lignincola (Halosphaeriaceae).

Hohnk (1955) introduced Lignincola for a single species L. laevis Héhnk,

which was characterized by black perithecioid ascomata, persistent clavate

to fusiform, thin-walled asci lacking an apical apparatus, and 1-septate

hyaline ascospores lacking appendages. The genus differs from Aniptodera,

Halosarpheia, and Phaeonectriella in lacking appendaged ascospores (Pang et al.

2003a,b, Jones et al. 2009). Three other Lignincola species have been described.

One, initially described as Gnomonia longirostris Cribb & J.W. Cribb (Cribb

& Cribb 1956), was later transferred to Lignincola by Kohlmeyer (1984). Pang

et al. (2003a) subsequently transferred the species to a new genus Neptunella

344 ... Liu & al.

based on ultrastructural and molecular evidence. Asci in N. longirostris have

an apical pore, retraction of the plasmalemma, and ascospores with a thin

mucilaginous sheath (Pang et al. 2003a). A second species, Lignincola tropica

Kohlm., was described from mangrove wood (Kohlmeyer 1984) and has larger

ascospores than L. laevis. The identity of this species is confused as initially the

asci were described as possessing an apical pore, but subsequently Kohlmeyer

& Volkmann-Kohlmeyer (1988) revised the description by stating that there

was no pore to the ascus. In addition, sequence data show that collections with

asci with an apical pore do not belong in Lignincola (Pang unpublished data).

A third species, L. nypae K.D. Hyde & Alias (Hyde et al. 1999), was described

from the brackish water palm Nypa fruticans and has clavate asci with a

refractive apical thickening with a pore and cylindrical ascospores, features not

found in Lignincola. A molecular study is required to resolve the placement of

both L. tropica and L. nypae.

The new species Lignincola conchicola is described and illustrated and a key

to Lignincola species is provided.

Materials & methods

Palm fronds were collected from Ranong mangrove, Ranong Province, Thailand.

Samples were processed and examined following the methods outlined by Hyde

et al. (2000) and Taylor & Hyde (2003). Several unsuccessful attempts were made to

isolate and grow the fungus from single ascospores. Morphological measurements and

photomicrographs were made according to Liu et al. (2010). The holotype is deposited

in the herbarium of Mae Fah Luang University (MFLU), Chiang Rai, Thailand.

Taxonomy

Lignincola conchicola J.K. Liu, E.B.G. Jones & K.D. Hyde, sp. nov. PLATE 1

MycoBank: MB 561207

Ascomata superficialia, 120-160 um alta, 50-75 um diametro globosa, subglobosa vel

ellipsoidea, uniloculata, atro-brunnaea vel nigra. Asci 50—75(—90) x 13-16 um, clavati

vel cylindricae-clavati, unitunicati, 8-spori. Ascosporae 10.5-16 x 5—7 um, hyalinae,

ellipsoideae vel fusiformae, bicellulares, non-appendiculatae.

Type: Thailand, Ranong Province, Ranong mangrove, on adhesive disc of an invertebrate

on submerged fronds of Phoenix paludosa Roxb. (Arecaceae), 26 January 2010, E.B.G.

Jones and J.K. Liu, JKA0027 (MFLU11 0032 holotype).

Erymo ocy: Referring to its habitat on the adhesive disc of an invertebrate.

Ascomata 120-160 x 50—75 um, superficial, globose, subglobose or ellipsoidal,

uniloculate, dark brown to black, rounded at the base and developing on

the adhesive pad of a marine invertebrate (Plate 1 A-B). Peridium 12-22

um thick, membranous, comprising several layers, the outer stratum of 1-3

layers comprising dark brown, thick-walled cells, the inner 3—5 layers, textura

prismatica comprising hyaline, thick-walled cells. Paraphyses and catenophyses

Lignincola conchicola sp. nov. ... 345

PiaTE 1. Lignincola conchicola (MFLU11 0032 holotype). A-B. Ascomata on host surface.

C. Section of ascoma. D-F. Asci, G-K. Ascospores. C-D, G-H, in water, E-F, I-K, in cotton

blue. Scale bars. A = 1000 um, B = 500 um, C = 50 um, D-F = 30 um, G-K = 10 um.

absent. Asci 50—75 (—90) x 13-16 um (Mean = 62 x 14 um, n = 25), 8-spored,

unitunicate, thin-walled, clavate to cylindrical-clavate, short stipitate, lacking

346 ... Liu & al.

an apical apparatus and apical pore, persistent, arising from the base of ascoma.

Ascospores 10.5—16 x 5—7 um (Mean = 13 x 5.5 um, n= 50), 1-septate, hyaline,

ellipsoidal, frequently fusiform, with a slightly pale rose hue in transmitted

light, appendages absent.

HasiraT: Saprobic on adhesive disc of an invertebrate on submerged

Phoenix paludosa fronds.

DISTRIBUTION: Thailand.

Key to species of Lignincola:

1. Asci with an apical apparatus, ascospores cylindrical .................... L. nypae

1. Asci without an apical pore, ascospores ellipsoidal ............. 0... sce e eee ee eee 2,

2 ASCOSPOres, China walled (Ae Me mae Me st Med pls reece Winer eae nae edie eas 2

2. Ascospores: thick-walled, 22-36 x 12—16'tin .65.03 eee eee eka L. tropica

3; Ascospotes IS 24 x ST SHMIM a oo. a4:die 3d cn ed doer Quarter BA Oar BIO SiON L. laevis

3. Ascospores 10.5—16 x 5—7 um; on the adhesive pad of a marine invertebrate

ey ae eee re CO ee eee mee Veen reer een ree | L. conchicola

Discussion

Most taxa referred to Halosphaeriaceae are marine, usually with well-

developed perithecioid ascomata that are globose, light or dark, and usually

with an ostiole lined with periphyses (Miller & Arx 1962, Kohlmeyer 1972).

Paraphyses are absent, catenophyses generally deliquesce if present, asci are

clavate to ellipsoidal, thin-walled, unitunicate and deliquesce early, generally

lacking an apical apparatus or pore, and ascospores are hyaline to light brown,

unicellular or one to several septate, often with appendages or surrounded by a

mucilaginous sheath (Shearer & Miller 1977, Jones 1995, Sakayaroj et al. 2011).

A few transitional species of this family have been found in freshwater and

brackish water habitats, including some species of Aniptodera, Ascosacculus,

Halosarpheia, Lignincola, Luttrellia and Nais (Shearer & Miller 1977, Shearer

1989, Jones 1995, Pang et al. 2003a,b, Jones et al. 2009, Sakayaroj et al. 2011).

The morphological characteristics of Lignincola conchicola fit well within this

family.

Jones et al. (2009) suggested that the genus Lignincola has only one

unifying character — the hyaline 1-septate ascospores without appendages. In

L. conchicola, asci are persistent and thin-walled and lack an apical apparatus

(Plate 1 D-F) and ascospores are thin-walled and lack appendages (Plate 1

G-K). Lignincola conchicola differs from other species in the genus in ascospore

dimensions and the persistent occurrence of ascomata on the adhesive pad of

a marine invertebrate.

Lignincola conchicola is most similar to L. laevis, the type species of the

genus. The two taxa have similarly shaped asci and ascospores. They differ in

that L. laevis has larger ascospores (13-24 x 5-8 um) and the ascomata are

Lignincola conchicola sp. nov. ... 347

necked. Lignincola tropica differs from L. conchicola in having larger ascospores

(22-36 x 12-16 um) and in being thick-walled (Jones et al. 2009), Lignincola

nypae can be distinguished from the new species by the presence of an apical

apparatus in the ascus and cylindrical ascospores (Hyde et al. 1999, Pang et al.

2003a, Jones et al. 2009).

Three genera —Aniptodera, Gesasha, Nais— are also morphologically similar

to Lignincola with some species with 1-septate, unappendaged ascospores

(Shearer & Miller 1977, Kohlmeyer 1984, Crane & Shearer 1986, Abdel-Wahab

& Nagahama 2011). In the type species of Aniptodera, the ascus is clavate with an

apical pore and apical retraction of the plasmalemma, while the ascospores are

thick-walled. Three species were described when Abdel-Wahab & Nagahama

(2011) introduced Gesasha, and the ascospore dimensions and shape are quite

similar to L. conchicola. However, Gesasha diagnostic characters include a

periphysate neck, asci with an apical thickening, and pore features that exclude

L. conchicola (Abdel-Wahab & Nagahama 2011). The two currently accepted

Nais species differ from L. conchicola in ascospore dimensions and deliquescing

asci (Pang et al. 2003a, Jones et al. 2009).

Acknowledgements

The authors are grateful to Dr. Eric H.C. McKenzie (Landcare Research, Auckland,

New Zealand) and Dr. Rui-Lin Zhao (Southwest Forestry University, Yunnan, China)

for reviewing and improving the manuscript. We thank the Mushroom Research

Centre (Mae Taeng District, Chiang Mai Province, Thailand) for providing funding

and facilities for this study. The BIOTEC Central Research Unit (National Center for

Genetic Engineering and Biotechnology, Thailand Science Park, Bangkok, Thailand) is

thanked for providing facilities for the southern Thailand collecting trip. The Global

Research Network for Fungal Biology and King Saud University are thanked for

supporting this work. The authors also thank Anupong Klaysuban (BIOTEC), Jariya

Sakayaroj (BIOTEC), Satinee Suetrong (BIOTEC) and Sita Preedanon (BIOTEC)

for their assistance with fieldwork. Dr. Ying Zhang (University of Hong Kong, Hong

Kong SAR, P.R. China) and Phongeun Sysouphan Khong (Mae Fah Luang University,

Thailand) are thanked for their helpful suggestions. Kanjana Niraphai (MFU) is thanked

for herbarium assistance.

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Abdel-Wahab MA, Nagahama T. 2011. Gesasha (Halosphaeriales, Ascomycota), a new genus with

three new species from the Gesashi mangroves in Japan. Nova Hedwigia 92: 497-512.

http://dx.doi.org/10.1127/0029-5035/2011/0092-0497

Crane J, Shearer C. 1986. Nais glitra, an ascomycete from red mangrove in Everglades National

Park, Florida. Transactions of the British Mycological Society 86: 509-512.

Cribb A, Cribb JW. 1956. Marine fungi from Queensland. II. Papers of the Department of Botany,

University of Queensland 3: 97-105.

Hoéhnk W. 1955. Studien zur Brack- und Seewassermykologie. V. Hohere Pilze des submersen

Holzes. Veréffentlichungen des Institutes fir Meeresforschung in Bremerhaven 3: 199-227.

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Series 12.

MYCOTAXON

http://dx.doi.org/10.5248/117.351

Volume 117, pp. 351-358 July-September 2011

Craspedodidymum and Corynespora spp. nov. and

a new anamorph recorded from southern China

Li-Guo Ma, JIAN Ma, YI-DONG ZHANG & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Two new species are described and illustrated: Craspedodidymum fujianense

from dead branches of Acacia confuse and Corynespora fujianensis from dead branches of

Myrioneuron faberi. Craspedodidymum proliferans is a new record for China. These three

species were collected from tropical and subtropical forests in southern China.

KEY worDs — microfungi, taxonomy

Introduction

China is considered an important world reservoir of biodiversity. Many

wood-inhabiting fungi collected in China have recently been published (Dai

et al. 2009, Dai & Li 2010, Ma et al. 2010). As part of studies on the fungal

diversity of southern China, numerous collections of anamorphic fungi on

dead wood were made from tropical and subtropical forests. Among these,

two undescribed species and a new record were found. These three species

are described, illustrated and compared with similar species. The 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).

Taxonomy

Craspedodidymum fujianense L.G. Ma & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 561700

CoLoNnlI4_E in substrato naturali effusae, atrobrunneae. Mycelium partim superficiale vel

immersum. CONIDIOPHORA Mmacronematosa, mononematosa, solitaria, simplicia, erecta,

recta vel leniter flexuosa, laevia, crassitunicata, septata, brunnea vel atrobrunnea, ad

apicem pallida, 240-320 ym longa, 6.5-8.5 um lata. CELLULAE CONIDIOGENAE integratae,

terminales, monophialidicae, cylindricae, brunneae, 21.5-25 x 11-12 um, cum collaretto

cupulato, 6.5-7.5 um longo, ad apicem 6.5-9.5 um lato, ad basim 4.5-5.5 um diamm

352 ... Ma & al.

= s

2 3

20um 20um

Fic. 1. Craspedodidymum fujianense. a, b. Conidiophores, conidiogenous cells

and conidia. c. Conidiogenous cells with collarettes. d. Conidia.

praeditae. Conip1A 13-15.5 x 7.5-10 um, crassitunicata, laevia, brunnea, oblonga, 0-

septata.

Type: China, Fujian Province: the National Forest Park of Wuyishan, on dead branches

of Acacia confusa Merr. (Mimosaceae), 15 Aug. 2009, L.G. Ma (holotype, HSAUP

H1010-2; isotype HMAS 146089).

EryMoLoey: in reference to the province where the type was found.

Craspedodidymum & Corynespora spp. nov. (China) ... 353

Co.onligs on the natural substratum effuse, dark brown. Mycelium superficial

and immersed. CONIDIOPHORES macronematous, mononematous, solitary,

simple, erect, straight or slightly flexuous, smooth, thick-walled, septate,

brown to dark brown, paler towards the apex, 240-320 um long, 6.5-8.5 um

wide. CONIDIOGENOUS CELLS integrated, terminal, monophialidic, cylindrical,

brown, swollen at the subapical region, 21.5-25 x 11-12 um, with a collarette

at the apex. Collarette funnel-shaped, 6.5-7.5 um high, 6.5-9.5 um wide at

the apex, 4.5-5.5 um diam. at the base. Conrp1a 13-15.5 x 7.5-10 um, thick-

walled, smooth, brown, oblong, 0-septate.

Holubova-Jechova (1972) established Craspedodidymum for C. elatum

Hol.-Jech., the type species. The genus is characterized by macronematous,

mononematous, simple or branched conidiophores bearing integrated, terminal,

enteroblastic, monophialidic, cylindrical, apically swollen conidiogenous cells

with a large and distinct funnel-shaped terminal collarette (Holubova-Jechova

1972, Ellis 1976, Yanna et al. 2000). Craspedodidymum pulneyense Subram. &

Bhat has been placed in synonymy with C. proliferans (Bhat & Kendrick 1993).

Eleven species are currently recognized in this genus, none of which were

described from China. Craspedodidymum is reported for the first time from

China.

Craspedodidymum fujianense is unique in this genus in producing oblong

Q-septate conidia without a papilla. It most closely resembles C. siamense

Pinruan, which also produces 0-septate conidia that are rounded at the base

(Pinruan et al. 2004). However, C. fujianense differs by its oblong, shorter and

wider conidia.

Craspedodidymum proliferans V. Rao & de Hoog, Stud. Mycol. 28: 64, 1986.

Fic. 2

Cotonizs on the natural substratum effuse, dark brown. Mycelium

partly superficial and partly immersed. CONIDIOPHORES macronematous,

mononematous, solitary, erect, simple, straight or slightly flexuous, percurrently

proliferating, smooth, thick-walled, septate, brown to dark brown, unbranched,

paler towards the apex, up to 350 um long, 7.5-8.5 um wide, slightly swollen

at the base. CONIDIOGENOUS CELLS integrated, terminal, monophialidic,

cylindrical, brown, swollen at the subapical region, 20.5-24 x 9.5-10 um,

with a collarette at the apex. Collarette funnel-shaped, 4.5-6.5 um high, 7-7.5

um diam. at the opening, narrowing to 4-4.5 um diam at the base. ConrDIA

9.5-13 x 8.5-12 um, thick-walled, smooth, dark brown, obovoid, subglobose to

trapezoid, truncate at the base, 0-septate.

SPECIMEN EXAMINED: China, Fujian Province: the National Forest Park of Wuyishan,

on dead branches of Magnolia paenetalauma Dandy (Magnoliaceae), 15 Aug. 2009, L.G.

Ma, HSAUP H1031 (duplicate HMAS 146090).

354 ... Ma & al.

20um

20um 20um

Fic. 2. Craspedodidymum proliferans. A. Conidiophores, conidio-

genous cells and conidia. B. Conidiophore with percurrent proli-

feration. C. Conidia.

Craspedodidymum proliferans is reported for the first time from China.

Compared with the morphological characters of the species as described by

Rao & de Hoog (1986), both of the collections have obovoid, subglobose to

trapezoid, 0-septate conidia with truncate base, with almost identical conidial

size (9.5-13 x 8.5-12 um vs. 10-14 x 8-11 um). We believe they are the same

Craspedodidymum & Corynespora spp. nov. (China) ... 355

species. C. proliferans is most similar to C. cubense J. Mena & Mercado (Mercado

& Mena 1992) in conidial shape, septation and base, but differs from C. cubense

by its much narrower conidia.

Corynespora fujianensis L.G. Ma & X.G. Zhang, sp. nov. FIG. 3

MycoBank MB 561701

COLONI4E in substrato naturali fuscae, effusae. Mycelium semper superficiale, ex hyphis

ramosis, septatis, subhyalinis vel pallide brunneis, laevibus, 2-5 am crassis compositum.

ConrpioPHoRA singula vel fasciculata, erecta, interdum ramosa, recta, cylindrica, septata,

verruculosa, pallide brunnea vel brunnea, per usque ad 7 proliferationes percurrentes

successivas cylindricae elongascentia, 700-1300 um longa, 4-5.5 um crassa. CELLULAE

CONIDIOGENAE monotreticae, in conidiophoris incorporatae, crassitunicata, terminales,

cylindricae, 9.5-14 um longa, 6.5-9 wm crassa, brunnea. Conip1A recta vel leviter

curvata, singula, acrogena, obclavata, longer attenuata, laevia, brunnea, crassitunicata,

4-10-distoseptata, 31-90 x 6.5-10 um.

Type: China, Fujian Province: the National Forest Park of Wuyishan, on dead branches

of Myrioneuron faberi Hemsl. ex F.B. Forbes & Hemsl. (Rubiaceae), 15 Aug. 2009, L.G.

Ma (holotype, HSAUP H1006-2; isotype, HMAS 146094).

EryMoLoey: in reference to the province where the type was found.

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

superficial, composed of branched, septate, subhyaline to pale brown, smooth-

walled hyphae, 2-5 um thick. CONIDIOPHORES arising singly or in groups, erect,

sometimes branched, straight, cylindrical, septate, verruculose, pale brown to

brown, with up to 7 successive percurrent cylindrical proliferations, 700-1300

um long, 4-5.5 um thick. CONIDIOGENOUS CELLS monotretic, integrated,

terminal, thick-walled, cylindrical, 9.5-14 um long, 6.5-9 um wide, brown.

Conip1a straight or slightly curved, formed singly, acrogenous, obclavate,

tapering to the apex, smooth, brown, thick-walled, becoming gradually paler

towards the apex, 4-10-distoseptate, 31-90 x 6.5-10 um.

The genus Corynespora was erected by Giissow (1906). Wei (1950) emended

the diagnosis of the genus and clarified the conidiogenesis of C. cassiicola (Berk.

& M.A. Curtis) C.T. Wei [= C. mazei Giissow, the type species]. Corynespora

is characterized by macronematous, mononematous, simple, or branched

conidiophores with monotretic, determinate, or percurrent conidiogenous

cells, and obclavate to cylindrical, distoseptate, solitary, or catenate conidia.

More than 100 species have been validly described under Corynespora, 24 of

which were described from China, 4 parasitic on plant leaves (Guo 1984), and

20 saprobic on deciduous stems or wood (Zhang & Ji 2005, Zhang & Shi 2005,

Zhang & Xu 2005, Shang & Zhang 2007, Wang & Zhang 2007, Zhang & Zhang

2007, Ma & Zhang 2007, Ma et al. 2008, Zhang et al. 2008, 2009).

Among described Corynespora species, C. jasminicola Meenu et al. (Meenu et

al. 1998) and C. combreti M.B. Ellis (Ellis 1963) are most similar to C. fujianensis

356 ... Ma & al.

50um

b .

50um 50um

d ~~ @

+ |

Fic. 3. Corynespora fujianensis. a. Conidiophores and conidia. b. Unbranched conidiophores

and conidiogenous cells. c-d. Branched conidiophores and conidiogenous cells.

Gmiit= <= —

GI <heucK™

e. Conidia.

in having obclavate conidia and branched conidiophores. The conidia of

C. fujianensis are smaller with fewer septa than those of C. jasminicola, and

Craspedodidymum e& Corynespora spp. nov. (China) ... 357

are not rostrate while those of C. combreti are sometimes rostrate. In addition,

C. fujianensis differs by possessing verruculose and more slender conidiophores.

Acknowledgments

The authors are grateful 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).

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79-81.

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358 ... Ma & al.

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431-436.

MY COTAXON

http://dx.doi.org/10.5248/117.359

Volume 117, pp. 359-363 July-September 2011

A new species of Lepiota (Agaricaceae) from southwestern China

Jun FE LIANG” & ZHu L. YANG”

‘Research Institute of Tropical Forestry, Chinese Academy of Forestry,

Guangzhou 510520, P. R. China

*Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany,

Chinese Academy of Sciences, Kunming 650204, P. R. China

“CORRESPONDENCE TO: jfliang2000@163.com

ABSTRACT — A new species, Lepiota pallidiochracea, is described. It is characterized by

the ochraceous-red squamules on pileus, large ellipsoid to oblong spores, polymorphic

cheilocystidia, and a trichodermium of elongate pileus covering, which is apically attenuate.

Kew worps — Agaricales, lepiotaceous fungi, taxonomy

Introduction

The eastern Himalaya is one of the twenty-five hotspots for biodiversity

(Myers et al 2000), and macrofungi are very rich in the area (Yang 2005). Many

new agarics and aphyllophoroid fungi have recently been described from there

(Yang et al. 2004; Dai et al. 2007; Yuan & Dai 2008; Li et al. 2009; Dai 2010,

2011; Li et al. 2011). This paper is another contribution on the lepiotaceous

fungi occurring in the region.

The genus Lepiota (Pers.) Gray contains more than 400 described species

(Kirk et al 2008), and it is reasonable to expect many representatives of this

cosmopolitan genus in China. However, rather few taxa have been originally

described from China (Chiu 1948, Bi et al 1986, Yang 1994, Tolgor & Li 2004,

Wang & Yang 2005a,b). In our research on Lepiota in China, in addition to

several recently published new species (Liang et al 2009, 2010, 2011, Liang &

Yang 2011), several interesting and undescribed taxa still await documentation.

The present paper describes one new Lepiota species from eastern Himalaya.

Materials & methods

Macro-morphological features were recorded in the field. Material was dried using

an electric drier and deposited in the Herbarium of Cryptogams, Kunming Institute

of Botany, Chinese Academy of Sciences (HKAS) and Research Institute of Tropical

360 ... Liang & Yang

Forestry, Chinese Academy of Forestry (RITF). Terminology for descriptive terms

follows Vellinga & Noordeloos (2001). Color designations are from Kornerup and

Wanscher (1981).

For microscopic observations, sections of fruitbodies were made by hand and

mounted in 5% KOH, Congo red, and Melzer’s reagent. Basidiospores were mounted

in cresyl blue to test for a metachromatic reaction (Singer 1986). Size ranges were

determined for basidia, basidiospores, cheilocystidia, and elements of the pileipellis,

based on ocular micrometer measurements of at least 20 elements of each character.

The abbreviation [n/m/p] indicates that measurements were made on n basidiospores

in m basidiomata from p collections. Dimensions of basidiospores are given using a

notation of the form (a—)b-c(-d). The range b-c contains a minimum of 90% of the

measured values. Extreme values are given in parentheses. The following abbreviations

are used: Q refers to the length/breadth ratio of basidiospores; Q refers to the average

Q of all basidiospores + sample standard deviation.

Taxonomy

Lepiota pallidiochracea J.F. Liang & Zhu L. Yang, sp. nov. Fic. 1-4

MyYCOBANK 561241

Pileus sordide albus, squamulis densibus, rubro-ochraceis, rubro-brunneis vel brunneis.

Stipes subcylindricus, squamulis brunneis. Basidiosporae 9.0-12.0 x 6.0-9.0 um,

ellipsoideae vel lato fusiformes. Cheilocystidia diversa, clavata, fusiformia vel hamata.

Pleurocystidia nulla. Squamulae pilei ex trichodermiis attenuatis terminalibus compositae.

Fibulae praesentes.

Type: China, Tibet, Changdu Co., near Qiongka Bridge, 27 July 2004, Zhu L. Yang 4194

(HKAS 45579 holotype).

Erymo.oey: pallidiochracea (Lat.), referring to pale ochraceous squamules on the

pileus.

Basidiomata (Fic. 1) small-sized. Pileus 1.5-4 cm diam., campanulate with

small umbo when young, plano-convex with obtuse umbo at maturity, cream to

whitish, at centre brownish red (10D6-8) to dark brown (9E4-8), around centre

densely covered with concentric squamules; squamules more or less uplifted,

minute, ochraceous-red (8C7-8), brown (7D8) to reddish brown (9C7-8).

Lamellae free, subventricose, moderately crowded with lamellulae, dirty white

to cream. Stipe 2-4 x 0.2-0.3 cm, subcylindrical, hollow; surface glabrous and

white at the apex, light ochraceous (7C6-7) with scattered ochraceous-red

(8C7-8), brown (7D8) to reddish brown (9C7-8) squamules at the low part.

Annulus whitish, membranous, evanescent. Odor none. Taste not recorded.

Basidiospores (Fic. 2) [63/3/2] 9-12(-14) x 6-8(-9) um [Q = 1.29-1.85,

Q = 1.56 + 0.18], ellipsoid to oblong, sometimes ovoid with small hilar

appendage in side view, with neither suprahilar depression nor germ pore,

ellipsoid in frontal view; hyaline, smooth, slightly thick-walled, dextrinoid,

weakly congophilic, not metachromatic in Cresyl Blue. Basidia 25-37 x 9-14

um, clavate, 4-spored, rarely 2-spored. Lamella edge sterile. Cheilocystidia

Lepiota pallidiochracea sp. nov. (China) ... 361

Fic. 1-4 Lepiota pallidiochracea (All from holotype, HKAS 45579).

1. Basidiomata. 2. Basidiospores. 3. Cheilocystidia. 4. Pileus covering.

(Frc. 3) 15-40 x 5-10 um, variable in shape, subcylindrical to narrowly clavate,

or fusiform, lageniform to utriform; walls smooth, thin, hyaline in KOH,

congophilic. Pleurocystidia absent. Pileus covering (Fic. 4) a trichodermium of

elongate, apically attenuate terminal elements 60-256 x 6.5-11 um, often with

some short narrowly clavate elements interspersed, with pale yellowish brown

parietal and intracellular pigment in upper part; fine incrustation sometimes in

basal part of elements. Clamp connections common.

ADDITIONAL SPECIMEN EXAMINED: CHINA: YUNNAN PROV., SHANGRI-LA COUNTY,

Daxiagu, alt. 3060 m, 20 July 2008, Jun FE. Liang 799 (RITF 660).

HABITAT & DISTRIBUTION: in small groups, gregarious, saprotrophic and terrestrial

on slope, in summer. Known only from southwestern China.

Discussion: Lepiota pallidiochracea is characterized by its ochraceous-red

squamules on the pileus, which is covered with a trichodermium of long,

elongate, apically attenuate and short narrowly clavate elements, large ellipsoid

to oblong spores, and polymorphic cheilocystidia.

Based on the trichodermium type of the pileus squamules and the ellipsoid

basidiospores, L. pallidiochracea may tentatively be placed in L. sect. Ovisporae

362 ... Liang & Yang

(J.E. Lange) Kithner (Singer 1986, Vellinga 2001), although species in the

section usually have spores smaller than 10 um (Singer 1986). Recent molecular

phylogenetic studies indicate that section Ovisporae is not monophyletic, so

that re-evaluation of this section is needed (Vellinga 2003, Liang et al 2011).

Phylogenetically, the new species may be referred to section Lepiota or clade 1

of Lepiota s.l. (Vellinga 2003) until a new taxonomic system is proposed.

Several Lepiota species have similarly shaped spores. The polymorphic

cheilocystidia coupled with the larger spores help distinguish L. pallidiochracea

from L. brunneolilacea Bon & Boiffard (also a member of L. sect. Ovisporae),

L. helveola Bres. and L. ochraceoaurantiaca Dennis (Bon 1996, Candusso &

Lanzoni 1990, Dennis 1952). The larger spores are reminiscent of L. oreadiformis

Velen. (in sect. Lepiota), which, however, has much larger, fusiform to

amygdaliform spores and narrowly clavate or utriform cheilocystidia (Vellinga

2001).

Acknowledgments

We are grateful to Dr. Xiang-Hua Wang and Dr. Yan-Chun Li for offering advice,

suggestions, and analytical assistance. We are greatly indebted to Dr. Yu-Cheng Dai and

Dr. Ping Zhang for their critical reviewing the manuscript. This study is supported by

the National Natural Science Foundation of China (No. 31070014), the Joint Fund of the

National Natural Science Foundation of China and Yunnan Province (No. U0836604),

the National Basic Research Program of China (No. 2009CB522300), the Foundation of

RITF (RITFKYYW 2010-10), and the Commonweal Industry-specific Foundation of

State Forestry Administration (No. 201104057).

Literature cited

Bi ZS, Li TH, Zheng GY. 1986. New and rare species of Agaricales. Acta Mycol Sinica Suppl. 1:

288-296.

Bon M. 1996. Die Grofspilzflora von Europa 3 Lepiotaceae. IHW- Verlag: Eching (Germany). 141 p.

Candusso M, Lanzoni G. 1990. Fungi Europaei 4. Lepiota s.1. Giovanna Biella: Saronno. 743 p.

Chiu WF. 1948. The Amanitaceae of Yunnan. Sci Rept Natl Tsing Hua Univ Ser B, Biol, Psychol Sci

3(3): 165-178.

Dai YC, 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 131-343.

Dai YC, 2011. A revised checklist of corticioid and hydnoid fungi in China for 2010. Mycoscience

521:69=79:

Dai YC, Yu CJ, Wang HC, 2007. Polypores from eastern Xizang (Tibet), western China. Ann Bot

Fenn 44: 135-145.

Dennis RW. 1952. Lepiota and allied genera in Trinidad, British West Indies. Kew Bull 7: 459-499.

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

10th Edition. CABI Publishing, Wallingford.

Kornerup A, Wanscher JH. 1981. Taschenlexikon der Farben. 3. Aufl. Muster-Schmidt Verlag,

Gottingen.

Li YC, Yang ZL, Tolgor B, 2009. Phylogenetic and biogeographic relationships of Chroogomphus

species as inferred from molecular and morphological data. Fungal Divers 38: 85-104.

Lepiota pallidiochracea sp. nov. (China) ... 363

Li YC, Feng B, Yang ZL, 2011. Zangia, a new genus of Boletaceae supported by molecular and

morphological evidence. Fungal Divers 49: 125-143.

http://dx.doi.org/10.1007/s13225-011-0096-y.

Liang JF, Yang ZL. 2011. Two taxa close to Lepiota cristata from China. Mycotaxon 116: 387-394.

http://dx.doi.org/10.5248/116.387.

Liang JE, Xu J, Yang ZL. 2009. Divergence, dispersal and recombination in Lepiota cristata from

China. Fungal Divers 38: 105-124.

Liang JF, Yang ZL, Xu J, Ge ZW. 2010. Two new unusual Leucoagaricus species (Agaricaceae) from

tropical China with blue-green staining reactions. Mycologia 102: 1141-1152.

http://dx.doi.org/10.3852/09-021.

Liang JF, Yang ZL, Xu DP. 2011. A new species of Lepiota from China. Mycologia 103: 820-830.

http://dx.doi.org/10.3852/10-216.

Myers N, Mittermeier RA, Mittermeier CG, da Fonseca GAB, Kent J. 2000. Biodiversity hotspots

for conservation priorities. Nature 403: 853-858.

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

Tolgor B, Li Y. 2004. Lepiota squamulosa, a new species from China. J Fung Res 2 (3): 49-50.

Vellinga EC. 2001. Lepiota (Pers.: Fr.) S.R. Gray. 109-151, in: ME Noordeloos et al. (eds). Flora

Agaricina Neerlandica, Vol. 5. A.A. Balkema, Rotterdam.

Vellinga EC. 2003. Phylogeny of Lepiota (Agaricaceae) - evidence from nrITS and nrLSU sequences.

Mycol Prog 2: 305-322.

Vellinga EC, Noordeloos ME. 2001. Glossary. 6-11, in: ME Noordeloos et al. (eds). Flora Agaricina

Neerlandica, Vol. 5. A.A. Balkema, Rotterdam.

Wang HC, Yang ZL. 2005a. Notes on Lepiota shixingensis and an allied new species (Basidiomycetes).

Nova Hedwigia 81(3-4): 463-469.

Wang HC, Yang ZL. 2005b. A new species of Lepiota (Agaricaceae, Basidiomycetes) from China.

Mycotaxon 91: 51-54,

Yang ZL. 1994. Clarkeinda, Lepiota, Leucoagaricus, Leucocoprinus and Macrolepiota. 122-131, in:

Ying JZ, Zang M (eds.). Economic macrofungi of southwestern China. Science Press: Beijing.

Yang ZL. 2005. Diversity and biogeography of higher fungi in China. 35-62, in: Xu J (ed).

Evolutionary genetics of fungi. Norfolk (UK): Horizon Bioscience.

Yang ZL, Weif M, Oberwinkler F. 2004. New species of Amanita from the eastern Himalaya and

adjacent regions. Mycologia 96: 636-646.

Yuan HS, Dai YC, 2008. Polypores from northern and central Yunnan Province, southwestern

China. Sydowia 60: 147-159.

MY COTAXON

http://dx.doi.org/10.5248/117.365

Volume 117, pp. 365-366 July-September 2011

Passalora wangii comb. nov. from the genus Tandonella

FENG- YAN ZHAI * YING-LAN GUO ” YING-JIE Liu? & Yu Li?

"Henan Institute of Science and Technology, Xinxiang 453003, China

*Institute of Microbiology, Chinese Academy of Science, Beijing 100101, China

3Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,

Jilin Agricultural University, ChangChun 130118, China

*CORRESPONDENCE TO: yuli966@126.com

ABSTRACT —The anamorphic fungus Tandonella wangii is recombined as Passalora wangii.

The species was originally collected on leaves of Ligularia sp. during a taxonomic survey

carried out in Motianling, Inner Mongolia, China.

KEY worps — hyphomycete, imperfect fungi, taxonomy

Tandonella S.S. Prasad & R.A.B. Verma (Prasad & Verma 1970) was one of the

anamorph genera for Mycosphaerella, usually plant pathogenic, symptomless or

almost so but also often causing leaf lesions. Differentiating between Passalora

and Tandonella, which comprised taxa with superficial secondary mycelium,

synnematous conidiophores, and catenate conidia, was difficult.

When Crous & Braun (2003: 19-22) emended the circumscription of

Passalora, they reduced Tandonella to a synonym of Passalora, which otherwise

differed from Tandonella by the formation of solitary conidia and the absence

of superficial mycelium and synnematous conidiophores. The emended

Passalora embraces a wide morphological variation with secondary mycelium

that is absent or well developed, external, and superficial, conidiophores that

are solitary, fasciculate, or in sporodochial to synnematous conidiomata, and

conidia that range from solitary to catenate and in simple or branched chains.

Because Tandonella is no longer tenable as a separate genus, we here transfer

the previously reported species, T: wangii (Zhai et al. 2006), to Passalora.

Passalora wangii (EY. Zhai, Y.L. Guo & Yu Li) FY. Zhai, Y.L. Guo & Yu Li, comb.

nov.

MycoBank: MB519619

= Tandonella wangii EY. Zhai, Y.L. Guo & Yu Li, Mycosystema 25(3): 374. 2006.

366 ... Zhai & al.

DESCRIPTION AND ILLUSTRATION: Zhai et al (2006: 375, Fig.1).

HABITAT AND DISTRIBUTION: Known only from the type collection, on leaves

of Ligularia sp. (Asteraceae), from Inner Mongolia.

Acknowledgements

We express our deep appreciation to Prof. Shu-Yan Liu, Jilin Agricultural University,

and Prof. Shoji OHGA, Kyushu University, for their valuable suggestions, kind help, and

earnest assistance in the course of the submission of this manuscript.

Literature cited

Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs: 1. Names published in Cercospora

and Passalora. CBS Biodiversity Series No. 1: 19-22.

Prasad SS, Verma RAB. 1970. A new genus of Moniliales from India. Indian Phytopathology 23:

111-113.

Zhai FY, Guo YL, Li Y. 2006. A new species of Tandonella on Compositae. Mycosystema 25(3):

374-375.

MY COTAXON

http://dx.doi.org/10.5248/117.367

Volume 117, pp. 367-371 July-September 2011

A new species of Terriera (Rhytismatales, Ascomycota) from China

ZHONG-ZHOU YANG! YING-REN LIN" & CHENG-LIN Hou”

' 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 *: *lyr@ahau.edu.cn & houchenglincn@yahoo.com

ABSTRACT —A new Terriera species, T: huangshanensis on leaves of Eurya muricata var. huiana,

is described. The species is placed in Terriera based on the presence of strongly carbonized

extensions adjacent to the ascoma opening, somewhat thin-walled cells at the marginal parts

of the ascoma composed of colorless to light brown textura angularis-prismatica, and the

absence of lip cells. The new species is similar to T: minor, which is distinguished by smaller

ascomata with rounded ends, textura prismatica in the corner between the covering and basal

stroma, paraphyses branching 2-3 times in the apical 30-40um, sequentially ripening asci,

and ascospores tapering towards the ends. The type specimen is deposited in the Herbarium

of Forest Fungi of Anhui Agricultural University, China (AAUF).

KEY worpDs —taxonomy, Rhytismataceae, Theaceae

Introduction

Terriera B. Erikss. is a member of Rhytismataceae (Rhytismatales,

Leotiomycetes, Ascomycota) (Kirk et al. 2008). Since Eriksson (1970) established

Terriera as anew genus for T: cladophila (Lév.) B. Erikss., 17 species and 2 varieties

have been reported. Terriera species are distributed worldwide and associate

with both angiosperms and gymnosperms. In the past, members of this genus

were placed in Lophodermium Chevall., Hypoderma De Not., Clithris (Fr.)

Bonord., and Dermascia Tehon (Johnston, 2001). Although the classifications

of Hodhnel (1917) and Tehon (1935), based heavily on developmental

characteristics of the ascomatal primordium, have been criticized and rejected

by later researchers, they appear highly significant for recognizing Terriera as

phylogenetically distinct from the highly heterogeneous Lophodermium s. lat.

(Lantz et al. 2011). Ortiz-Garcia et al. (2003) noted that ‘Terriera minor differs

from Lophodermium in structure of the ascomatal primordium? Johnston

(2001) emphasized the importance of characters such as ascus and ascospore

size, apical shape of the ascus and paraphysis, and features of the conidiomata

368 ... Yang & al.

in his taxonomy of Terriera. Therefore in Johnston (2001) and Ortiz-Garcia et

al. (2003), he accepted into the genus several species with a typically Terriera-

like ascoma such as Clithris arundinacea Penz. & Sacc., Hypoderma fuegianum

(Speg.) Kuntze, Lophodermium javanicum Penz. & Sacc., and L. sacchari Lyon.

Ortiz-Garcia et al. (2003) attempted to explain phylogenetic relationships

within Lophodermium by rDNA-ITS sequence analyses of Lophodermium

species and representatives of other rhytismataceous genera. The results

showed that Lophodermium from pine hosts formed an independent clade

with Meloderma Darker and Elytroderma Darker but were distally related to

Terriera. A more comprehensive survey by Lantz et al. (2011) indicated that

Terriera is monophyletic, while Lophodermium as currently circumscribed

contains as many as 10 independently evolved species clusters.

For China, only one species of Terriera, T. brevis (Berk.) P.R. Johnst., was

reported in Hong Kong (Frohlich & Hyde 2000). In the present paper we

describe a new Terriera species from the Huangshan Mountains in Anhui

Province, China.

Materials & methods

Mature fruit bodies were selected from the collected specimen. External shapes,

size, color and opening ways of ascomata and conidiomata as well as zone line were

observed under the dissecting microscope. Materials were then rehydrated in water for

15min, after which 10-15 um thick fruitbody sections were cut by a freezing microtome

and mounted in lactic acid or cotton blue with water pretreatment for viewing outlines

of ascomata and conidiomata in vertical section. Gelatinous sheaths surrounding

ascospores and paraphyses were examined in water or 0.1% (w/v) cotton blue in lactic

acid. Color of diversified structures and ascospore contents were observed in water.

Measurements and drawings were made using materials mounted in 5% KOH and

from 30 asci, ascospores, and paraphyses for each specimen. Line and point integrated

illustrations of internal structures of fruit bodies were drawn using the microscopic

painting device (Panasoianic XSJ-2). The type specimen is deposited in the Herbarium

of Forest Fungi of Anhui Agricultural University, China (AAUF).

Taxonomy

Terriera huangshanensis Z.Z. Yang, Y.R. Lin & C.L. Hou, sp. nov. FIGURES 1-7

MyYCoBANK 512114

Ascomata amplitudine admodum variabilia, 550-1900 x 200-550 ym, elliptica,

subfusiformia vel rimis ternis aperientia. Paraphyses filiformes, ad apicem plerumque

gradatim tumidae vel irregulatim semel aut iterum ramosae. Asci in simul maturescentes,

8-spori. Ascosporae 58-90 x 1.5-2 um, filiformes, hyalinae, aseptatae, vagina gelatinosa

1-1.5 wm crassa indutae.

TyPeE: On leaves of Eurya muricata var. huiana (Kobuski) Hu & L.K. Ling (Theaceae),

China. Anhui, Mt Huangshan, Wenquan. alt. ca 700 m, 12 June 2006, Y. R. Lin et al.

L2217 (HOLOTYPE AAUF68325).

ErymMo.oey: huangshanensis, referring to the place where the specimen was collected.

Terriera huangshanensis sp. nov. (China) ... 369

22 BB

RAF

N25

eee

we,

“\

ORE

HOC FAR THR

PIR ya

ee ES WIE?

20 um

Figs 1-7. Terriera huangshanensis on fallen leaves of Eurya muricata var. huiana. 1. Habit

on leaf. 2. Detail of ascomata and conidiomata. 3. Portion of ascoma in median vertical

section. 4. Paraphyses and asci. 5. Discharged ascospores. 6. Conidioma in vertical section.

7. Conidiogenous cells and conidia.

AscomatTa developing on both sides of fallen leaves, principally on the upper

side of the leaf, scattered to clustered, sometimes confluent in groups of two or

three, in drab or gray-white bleached areas 17-25 mm diam. In surface view,

ascomata varied in dimensions, 550-1900 x200-550 um, elliptical, fusiform

370 ... Yang & al.

or subfusiform, straight or curved (lunate), sometimes 3-lobed or triangular,

ends rounded to subacute, strongly raised above the surface of the substrate

at maturity, opening by a single longitudinal split which is branched in the

triangular ascomata. Lips absent, split extending almost the whole length of

the ascoma. Entirety of ascomata black, matt or slightly glossy, the edge well

defined. In median vertical section, ascomata subepidermal with epidermal

cells becoming filled with fungal tissue as ascomata develops, 175-205 um

deep. COVERING STROMA 18-22 um thick near the centre of the ascoma, slightly

thicker towards the edges, extending to the basal stroma, consisting of dark

brown to light black, thick-walled textura angularis and textura globulosa with

cells 3.5-5.5 um diam. Along the edge of the ascoma opening there is a 12-20

um thick extension adjacent to the covering stroma which covers the hymenium,

and which is comprised of strongly carbonized tissue with no obvious cellular

structure. ExCIPULUM very poorly developed, closely adhering to sides of the

extension and the covering stroma. BASAL STROMA 10-18 um thick, dark

brown, consisting of 2-4 rows of 4-7 um diam., angular, thick-walled cells.

SUBHYMENIUM 15-25 um thick, composed of hyaline textura angularis and

textura porrecta, with a colorless to gray-brown, 12 -30 um thick, of textura

angularis mixed with textura prismatica at the edge of the ascoma. HYMENIUM

often extending beyond the top of the extension when ripening. PARAPHYSES

120-140 x 1-1.2 um, filiform, thin-walled, hyaline, branching 1-2 times and

slightly swollen at the apex, with a ca 1 um thick gelatinous matrix, forming

a distinct epithecium above the asci. Asci ripening synchronously, 100-120 x

5-7 um, narrow-cylindrical, thin-walled and equal, apex rounded or subacute,

not bluing in iodine, discharging spores through a small apical hole, 8-spored,

stalk 15-28 um in length. Ascospores borne in a fascicle, sometimes helically

arranged, 58-90 x 1.5—2 um, filiform, slightly tapered towards the base, hyaline,

aseptate, thin-walled, covered by a 1-1.5 um thick gelatinous sheath.

Conip1oMaTA on both sides of leaves, predominantly on the upper side,

scattered to crowded, sometimes coalescing. In surface view, conidiomata

75-130 x 70-90 um, round to elliptical, raising the leaf surface, lightly brown

but dark brown, grey-brown or dark brown in the regions of the edge and

the surrounding of apical ostiole after discharging spores. In vertical section,

conidiomata subcuticular, somewhat lenticular in outline, 50-65 um deep.

UPPER WALL poorly developed, only present in surrounding of the ostiole. BASAL

WALL well-developed, 6-8.5 um, dark brown, consisting of 2-3 rows of 2-3.5

um diam., angular, slightly thick-walled cells. SUBCONIDIOGENOUS LAYER ca 8

um thick, composed of very light, thin-walled, angular cells. CONIDIOGENOUS

CELLS 8-12 x 2-3 um, cylindrical, slightly tapered towards the apex, hyaline,

proliferating sympodially. Conrp1a 4.5-6 x ca 1 um, cylindrical, hyaline,

aseptate.

Terriera huangshanensis sp. nov. (China) ... 371

ComMENTs —Terriera huangshanensis is distinctive within the genus because

of the synchronously ripening asci and the hymenium that often overtops the

extension of the covering stroma when mature. The most widely distributed

species, T. minor (Tehon) P.R. Johnst., is similar but differs in many aspects: its

ascomata have rounded ends and are not associated with conidiomata and zone

lines, the textura prismatica between the covering and basal stroma is poorly

developed, asci ripen sequentially, and the ascospores are 0-1 septate and

taper slightly towards both ends (Johnston 1988, 1989a, b). The type species

T. cladophila is distinguished from T. huangshanensis by subcuticular, circular

to elliptical, non-curved ascomata that are associated with brown diffuse zone

lines; the textura prismatica between the covering and basal stroma composed

of vertically oriented cells; sequentially ripening asci; and a subhymenium

consisting of textura angularis and textura intricata (Minter 1996).

Acknowledgments

We are grateful to Dr. PF. Cannon and Dr. W.-Y. Zhuang for critically reading the

manuscript, to Dr. P.R. Johnston and Dr. D.W. Minter for providing related literature, and

to Dr. S.-J. Wang, Dr. L. Chen, and Dr. X.-M. Gao for field investigations and micrographs.

The 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), the Natural Science Foundation of

Anhui Province of China (No. 070411005), and PHR (KZ201110028036).

Literature cited

Eriksson B. 1970. On Ascomycetes on Diapensales and Ericales in Fennoscandia. Symb. Bot. Upsal.

19: 1-71.

Frohlich J, Hyde KD. 2000. Palm microfungi. Fungal Diversity Press. Hong Kong. 364 p.

Hohnel F. von. 1917. System der Phacidiales v. H. Ber. Deutsch. Bot. Ges. 35: 416-422.

Johnston PR. 1988. An undescribed pattern of ascocarp development in some non-coniferous

Lophodermium species. Mycotaxon 31: 383-394.

Johnston PR. 1989a. Lophodermium (Rhytismataceae) on Clusia. Sydowia 41: 170-179.

Johnston PR. 1989b. Rhytismataceae in New Zealand 2. The genus Lophodermium on indigenous

plants. New Zealand J. Bot. 27: 243-274.

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.

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

Minter DW. 1996. Terriera cladophila. IMI Descr. Fungi & Bact. no. 1296.

Ortiz-Garcia S, Gernandt DS, Stone JK, Johnston PR, Chapela IH, Salas-Lizana R, Alvarez-

Buylla ER. 2003. Phylogenetics of Lophodermium from pines. Mycologia 95: 846-859.

http://dx.doi.org/10.2307/3762013

Tehon LR. 1935. A monographic rearrangement of Lophodermium. Illin. Biol. Monogr. 13: 1-151.

MY COTAXON

http://dx.doi.org/10.5248/117.373

Volume 117, pp. 373-380 July-September 2011

New Bulgarian records of fungi associated

with glacial relict plants

CVETOMIR M. DENCHEV , DIMITAR Y. STOYKOV,

EKATERINA F. SAMEVA & BorRIs ASSYOV

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

2 Gagarin St., 1113 Sofia, Bulgaria

CORRESPONDENCE TO *: cmdenchev@yahoo.co.uk

ABSTRACT — The paper contributes to knowledge of Bulgarian fungal diversity on glacial

relict vascular plants. Eleven new country records of ascomycetes and anamorphic fungi are

presented with descriptions, namely Asteromella silvarum, Cainiella johansonii, Gnomoniella

vagans, Hendersonia culmicola, Leptosphaerulina dryadis, Melasmia mougeotii, Phaeosphaeria

nardi, Pseudomassaria islandica, Septoria macropoda, S. tenella, and Sydowiella dryadis.

Microbotryum silenes-acaulis is also an addition to the list reflecting the recent changes in

M. violaceum s. lat. New data are included about the distribution of Leptosphaeria doliolum,

Nodulosphaeria modesta, Ophiognomonia gei-montani, and Phaeosphaeria juncina, all

previously recorded in Bulgaria. In addition, a brief review is presented of previously

encountered fungal species on glacial relicts in the country.

KEY worDs — taxonomy

Introduction

The study of fungi on relict hosts is of considerable interest as it may

contribute to understanding fungal biogeography and fungus-host interactions,

given that many parasitic and saprotrophic fungi demonstrate preferences for

particular host plants. This research also contributes to better understanding

of the need to establish appropriate practices for simultaneous conservation

of specialized parasitic and saprotrophic fungi and their hosts (Denchev &

Bakalova 2002, Denchev 2005). Research into fungal diversity associated with

glacial relict plants has a long history. See Chlebicki (2002) for details and

further references on this topic.

Although fungi associated with glacial relict plants have not previously

been studied in Bulgaria, earlier taxonomic surveys have revealed a number

of ascomycetes, smut, rust, and anamorphic fungi on various Bulgarian relict

plants: on Astragalus alopecurus: Uromyces punctatus J. Schrot.; on Carex atrata:

374 ... Denchev & al.

Puccinia dioicae Magnus; on Carex curvula: Anthracoidea curvulae Vanky &

Kukkonen and Schizonella melanogramma (DC.) J. Schrot.; on Carex rupestris:

Comoclathris deflectens (P. Karst.) Nograsek and Physalospora alpestris Niessl;

on Cerastium eriophorum: Massariosphaeria rubicunda (Niessl) Crivelli; on

Dianthus microlepis: Microbotryum dianthorum (Liro) H. Scholz & I. Scholz; on

Dryas octopetala s. lat.: Isothea rhytismoides (Bab. ex Berk.) Fr. and Wettsteinina

dryadis (Rostr.) Petr.; on Empetrum nigrum: Duplicaria empetri (Fr.) Fuckel; on

Gentiana asclepiadea: Cronartium flaccidum (Alb. & Schwein.) G. Winter; on

Oxyria digyna: Bauhinus vinosus (Tul. & C. Tul.) R.T. Moore (= Microbotryum

vinosum (Tul. & C. Tul.) Denchev) and Puccinia oxyriae Fuckel; on Poa alpina:

Puccinia graminis Pers.; on Polygonum viviparum: Bauhinus bistortarum (DC.)

Denchev (= Microbotryum bistortarum (DC.) Vanky); on Primula deorum:

Uromyces primulae-integrifoliae (DC.) Niessl; on Primula minima: Uromyces

apiosporus Hazsl.; on Rheum rhaponticum: Oidium erysiphoides Fr.; on Salix

herbacea: Rhytisma salicinum (Pers.) Fr. and Melampsora epitea Thiim.; on Salix

lapponum: Hyaloscypha albohyalina (P. Karst.) Boud. and Melampsora lapponum

Lindf.; on Salix reticulata: Melampsora epitea; on Saxifraga stellaris: Puccinia

saxifragae Schltdl; on Vaccinium uliginosum: Naohidemyces vacciniorum

(J. Schrot.) Spooner (Hinkova 1960, 1961; Fakirova 1991; Denchev 1994, 1995,

2001; Dimitrova 1995; Denchev & Negrean 2001; Chlebicki 2002; Zwetko et al.

2004).

The glacial refugia in Bulgaria are of undoubted interest, being among the

southernmost points in Europe for distribution of a significant number of

relict host plants. The authors of this paper aim at further detailed study of the

mycota associated with glacial relict plants in Bulgaria. This initial study has

focused on some parasitic and saprotrophic fungi on relict vascular plants less

sampled during previous mycological studies in this country. The survey has so

far produced some interesting new records that are reported below.

Material & methods

Fungi on Antennaria dioica (L.) Gaertn. (Asteraceae), Bartsia alpina L.

(Scrophulariaceae), Carex atrata L. (Cyperaceae), Dryas octopetala L. (Rosaceae), Festuca

pirinica Horvat ex Markgr.-Dann. (Poaceae), Geum bulgaricum Panci¢ (Rosaceae),

Juncus trifidus L. (Juncaceae), Kobresia myosuroides (Vill.) Fiori (Cyperaceae), Poa alpina

L. (Poaceae), and Silene acaulis (L.) Jacq. (Caryophyllaceae) were examined. Specimens

were collected from three major Bulgarian glacial refugia — Vihren area (Pirin Mts),

Kozyata Stena area (Stara Planina Mts), and the high mountain zone of Rila Mts (Fic. 1).

These were supplemented by examination of older collections of glacial relict plants kept

in the Herbarium of the Institute of Biodiversity and Ecosystem Research, Bulgarian

Academy of Sciences (SOM). All fungal specimens studied have been conserved in

the Mycological Collection of the Institute of Biodiversity and Ecosystem Research

(SOMF).

Fungi associated with glacial relicts (Bulgaria) ... 375

Fic. 1. Collecting areas for fungi on glacial relicts:

1. Kozyata Stena area (Stara Planina Mts), 2. Rila Mts, 3. Vihren area (Pirin Mts).

Microscopic examination was carried out on dried specimens. Slides of ascomata

and conidiomata for LM were prepared in lactophenol, the mounting medium being

gently heated to boiling point and then cooled. The descriptions below are based on

the examined specimens. Primary sources employed for identification of fungi were

the works of Holm (1952, 1979), Miiller (1957), Barr (1965), Saville (1968), Arx (1970),

Vassiljeva (1979, 1987, 1998), Merezhko (1980), Monod (1983), Vanev & van der Aa

(1998), and Chlebicki (2002).

Ascomycetes new for Bulgaria

Cainiella johansonii (Rehm) E. Miill., Sydowia 10: 121, 1957[‘1956’].

PERITHECIA 100-115 x 170-205 um, scattered, immersed, with a protruding

beak. Beak 650 x 60 um, long. Asci 137-141 x 37-41 um, oblong, almost

sessile, with refractive apical ring, I-, 8-spored. AscosporEs (25-)27.0+1.9

(-32) x (11.5-)13.5+1.6(-19) um, I/w (1.5-)2.02+0.17(-2.2) (n = 50), almost

kidney-shaped, hyaline, finally brown, 1-septate (sometimes constricted at the

septum).

SPECIMENS EXAMINED — On petioles of Dryas octopetala: Stara Planina Mts, Kozyata

Stena reserve, 17 Aug 2009, D.Y. Stoykov, B. Assyov & C.M. Denchev (SOMF 29 178);

ditto, near the Boba peak, 42°47'05.5" N, 24°32'45.9" E, 18 Aug 2009, D.Y. Stoykov, B.

Assyov & C.M. Denchev (SOMF 29 173).

376 ... Denchev & al.

CoMMENTS — Cainiella johansonii was described from Germany (Rehm

1904, as Lizonia johansonii) and further reported by Miller (1957) from the

Alps (Italy) on Dryas octopetala. Later it was recorded in Canadian Arctic on

D. drummondii Richardson ex Hook. and D. integrifolia Vahl (Chlebicki 2002).

According to Holm (1979), this fungus is rather common in the Scandes

Mountains (Scandinavia).

Gnomoniella vagans Johanson, Ofvers. Kongl. Svenska Vetensk.-Akad. Férh. 41(9):

163, 1884.

PERITHECIA densely scattered, immersed in leaf tissues. Neck long,

protruding. Asci 45-70 x 10-16 um, clavate, with apical refractive ring, I’, 8-

spored. Ascosporgs (13.5-)16.2+1.3(-18.5) x (5-)7.2+0.9(-8.5) um (n = 25),

ellipsoid to ovoid, hyaline, non septate.

SPECIMEN EXAMINED — On dead leaves of Dryas octopetala: Stara Planina Mts, Ushite

peak, 13 May 2002, leg. B. Assyov, det. D.Y. Stoykov (SOMF 29 172).

ComMENTs — Barr (1959) considers this fungus to be a rare species.

Leptosphaerulina dryadis (Starback) L. Holm, Bot. Not. 132: 86, 1979.

PERITHECIA small, subpyriform, immersed in leaf tissues. Ascr 70-81 x 30-

33 uum, very few, saccate, 8-spored. AscosporEs 27-34 x 10-12 um, hyaline,

about slipper shaped, upper part rounded, cylindrical below, with 5 transverse

septa and one + incomplete longitudinal septum; several additional transverse

and longitudinal septa formed at maturity.

SPECIMEN EXAMINED — On dead leaves of Dryas octopetala: Stara Planina Mts, Kozyata

Stena reserve, near the Boba peak, 42°47'05.5" N, 24°32'45.9" E, 18 Aug 2009, D.Y.

Stoykov, B. Assyov & C.M. Denchev (SOMF 29 174).

ComMMENTs — According to Holm (1979), this fungus is rare. The author

suggested that this species is closely related to Leptosphaerulina pulchra (G.

Winter) M.E. Barr on Potentilla spp., the latter distinguished on the basis of less

pronounced ascospore septation than in L. dryadis.

Phaeosphaeria nardi (Fr.) L. Holm, Symb. Bot. Upsal. 14(3): 124, 1957.

PERITHECIA single, subepidermal, globose or depressed globose. Ascr

62-82 x 9-13 um, cylindric, 8-spored. AscosPorREs (19.5-)21.9+3.1(-27.5) x

(3-)4.5+0.7(-5.5) um (n = 50), yellowish to brownish, 7-8-septate; the third

(fourth) cell from above swollen, of the same length as the lowest cells and

distinctly longer than the second (third) cell; biseriate in the ascus.

SPECIMEN EXAMINED — On dry leaves of Kobresia myosuroides: Pirin Mts, Kazana, on

marble, 1 Oct 1999, leg. P. Vasilev, det. D.Y. Stoykov (SOMF 27 967).

Pseudomassaria islandica (Johanson) M.E. Barr, Mycologia 56: 854, 1965 [‘1964’].

PERITHECIA 190-260 um in diameter, subglobose, immersed. Setae 200-

350 x 17.5-18 um, dark-brownish. Ascr 80-105 x 17.5-22 um, +oblong, with

Fungi associated with glacial relicts (Bulgaria) ... 377

refractive apical ring, I+, 8-spored. AscosporEs (21.5—)23.0+1.6(-26.5) x

(10.5—)10.740.8(-12.5) um (n = 75), ellipsoid, yellowish-hyaline, apiosporous;

irregularly uniseriate in the ascus.

SPECIMENS EXAMINED — On dead leaves of Dryas octopetala: Pirin Mts, along the track

from Vihren peak to Kazana, 2400 m, 41°76'88.7" N, 23°41'25.2" E, 11 Aug 2010, leg.

I. Apostolova & H. Predashenko, det. D.Y. Stoykov (SOMF 27 631); Rila Mts, Sedemte

Ezera, above Babreka lake, 2380 m, 42°12'07.6" N, 23°18'27.2" E, 15 Sep 2009, leg. I.

Apostolova, det. D.Y. Stoykov (SOMF 27 640).

Sydowiella dryadis Lar.N. Vassiljeva, Mikol. Fitopatol. 13(4): 279, 1979.

PERITHECIA ca. 400 um in diameter, superficial, single or in groups, black.

Beak 350-700 x 100-120 um, papillate. Asc 135 x 13 um, cylindric, 8-spored,

with apical ring. Ascospores (14—)16.9+1.7(-19.5) x (6-)7.4+0.62(-9) um,

I/w (1.9-)2.3+0.26(-2.9) (n = 75), ellipsoid, hyaline, two-celled, septum median,

slightly constricted at the septa; uniseriate in the ascus.

SPECIMEN EXAMINED — On dead twigs of Dryas octopetala: Stara Planina Mts, Kozyata

Stena reserve, 17 Aug 2009, D.Y. Stoykov, B. Assyov & C.M. Denchev (SOME 29 177).

COMMENTS — The measurements of asci and ascospores of the Bulgarian

specimen fit well those in the original description of the species (Vassiljeva

1979).

Ascomycetes already known from Bulgaria, found on glacial relict plants

during the study

Leptosphaeria doliolum (Pers.) Ces. & De Not., Comment. Soc. Crittog. Ital. 1(4):

234, 1863.

SPECIMEN EXAMINED — On dry leaves of Poa alpina: Stara Planina Mts, Sveti Plast peak,

1820 m, 16 Jul 1952 (ex SOM), fungus comm. & det. D.Y. Stoykov (SOMF 29 175).

Nodulosphaeria modesta (Desm.) Munk ex L. Holm, Symb. Bot. Upsal. 14(3): 80,

1957.

SPECIMEN EXAMINED — On dead stems and floral parts of Antennaria dioica: Rila Mts,

Sedemte Ezera, 7 Jul 2010, D.Y. Stoykov & C.M. Denchev (SOME 27 629).

ComMENTS — The fungus is so far known in Bulgaria only from Rila Mts on

stems and leaves of Lactuca (Stoykov 2004).

Ophiognomonia gei-montani (Ranoj.) Sogonoy, Stud. Mycol. 62: 58, 2008.

SPECIMENS EXAMINED — On dead leaves of Geum bulgaricum: Rila Mts, Sedemte Ezera,

ca. 2300 m, 21 Jul 1909, leg. B. Davidov (ex SOM), fungus comm. & det. D.Y. Stoykov

(SOME 29 171); Rila Mts, along the track from Smrudlivoto lake to Kirilova Polyana

locality, 21 Jul 1998, D.Y. Stoykov (SOMF 22 511); Rila Mts, Yakoroudski circus, on the

slopes of Mt Kovatch, above Murtvoto Ezero, ca. 2200 m, 30 Aug 2008, leg. B. Assyov &

R. Vassilev, det. D.Y. Stoykov (SOME 29 179).

ComMENts — A description of this species based on Bulgarian specimens on

Geum bulgaricum appears in Stoikov (2000).

378 ... Denchev & al.

Phaeosphaeria juncina (Auersw.) L. Holm, Symb. Bot. Upsal. 14(3): 127, 1957.

SPECIMEN EXAMINED — On leaves and floral parts of Juncus trifidus: Rila Mts, Sedemte

Ezera, in a rock crack, 42°12'97.2" N, 23°19'21.0" E, 7 Jul 2010, D.Y. Stoykov & C.M.

Denchev (SOME 27 625).

ComMMENTS — In Bulgaria so far recorded only from Vitosha Mt, on Juncus

conglomeratus L.

Anamorphic fungi new for Bulgaria

Asteromella silvarum Petr., Ann. Mycol. 23: 112, 1925.

Pycnipi epiphyllous, separated, arranged in rows between the veins of the

leaves, subepidermal, then erumpent, globose, thick-walled, dark brown, 70-

100 um in diameter. Ostioles central, circular, 8-12 um in diameter. CONIDIA

rod-shaped, cylindrical, with rounded ends, straight, unicellular, hyaline, 2.5-4

x 1-1.5 um.

SPECIMEN EXAMINED — On leaves of Carex atrata: Rila Mts, 2550 m, 24 Sep 1955, leg. I.

Bondev (ex SOM), fungus comm. & det. E. Sameva (SOME 29 164).

Hendersonia culmicola Sacc., Michelia 1(2): 201, 1878.

PYCNIDIA gregarious between the veins of the leaves, immersed, then

erumpent, globose or subglobose, pale brown or cinnamon-brown, 75-115

um in diameter. Ostioles rounded, 10-15 um in diameter, surrounded by dark

brown cells. Conip1A cylindrical, narrow clavate, straight or slightly curved,

the base blunt or subtruncate, the apex rounded or pointed, with 3-6 distinct

septa, some constricted at the septa, pale olivaceous, in mass brownish, 25-35

x (2.5-) 3-4 um.

SPECIMEN EXAMINED — On dried parts of leaves of Festuca pirinica: Pirin Mts, below

Vihren peak (2900 m), 9 Aug 1938, leg. B. Achtarov (ex SOM), fungus comm. & det. E.

Sameva (SOMF 29 165).

Melasmia mougeotii (Desm.) Arx, Verh. Kon. Ned. Akad. Wetensch., Afd. Natuurk.,

2de Reeks 51(3): 106, 1957.

LEAF spots black and spread out on both surfaces of leaves. STROMATA

amphigenous, mostly hypophyllous, crustaceous, flat, subepidermal, black,

gregarious, discrete, sometimes confluent in clusters, multilocular; circular,

1-2 mm in diameter, or oblong and irregular, 3-5 x 1-2 mm. LocuL! rounded,

125-250 um in diameter. Conrp1opHorEs filiform, straight, hyaline, 20-30 x

1 um. Conrp1A numerous, cylindrical with rounded ends, straight or slightly

curved, unicellular, hyaline, (3.5-)4-6(-7) x 1.5-2(-2.5) um.

SPECIMENS EXAMINED — On leaves of Bartsia alpina: Pirin Mts, below Vihren peak,

2009, leg. V. Vladimirov, det. E. Sameva (SOMF 29 168); ditto, 11 Aug 2010, leg. H.

Pedashenko & I. Apostolova, det. E. Sameva (SOMF 29 169).

Septoria macropoda Pass., Fungi Parm. Septor.: no. 141, 1879.

PycNnIDIA mainly epiphyllous, scattered or arranged in irregular lines,

globose or slightly flattened, semi-imersed, thin-walled, brown, 55-125 um in

Fungi associated with glacial relicts (Bulgaria) ... 379

diameter. Ostioles rounded or somewhat ellipsoidal, 15-25 um in diameter.

ConipiA filiform, straight or curved, mostly with obtuse ends, some slightly

tapered at the apex, unicellular or with 2-3 indistinct septa, hyaline, 20-40 x

JIS pm,

SPECIMEN EXAMINED — On leaves of Poa alpina: Rila Mts, Skakavitsa, 20 Aug 1956, leg.

S. Petrov (ex SOM), fungus comm. & det. E. Sameva (SOMF 29 166).

Septoria tenella Cooke & Ellis, Grevillea 8(45): 11, 1879.

PycnipiA spread over both leaf surfaces, gregarious or forming indistinct

rows, globose or subglobose, immersed, thick-walled, dark brown, 120-220

um in diameter. Ostioles indistinct. Conip1a filiform, straight, curved to arch-

shaped or slightly flexuous, gradually tapered towards the both ends, subacute

at the apex, unicellular, hyaline, 22-100 (-109) x 1-1.5 (-2) um.

SPECIMEN EXAMINED — On leaves of Festuca pirinica: Pirin Mts, below Vihren peak,

2900 m, 9 Aug 1938, leg. B. Achtarov (ex SOM), fungus comm. & det. E. Sameva (SOMF

29 165).

A smut fungus new for Bulgaria

Microbotryum silenes-acaulis M. Lutz et al., Mycol. Res. 112: 1289, 2008.

An anthericolous smut fungus on Silene acaulis has been reported from

Bulgaria by Klika (1926) and Denchev (2001), as a species of Ustilago or

Microbotryum, respectively. In 2008, a new cryptic species of Microbotryum on

Silene acaulis was described as M. silenes-acaulis. Most probably, the following

records refer to this cryptic species and therefore, it is considered here as a new

species for Bulgaria.

LITERATURE RECORDS — On Silene acaulis: Rila Mts, below Mussala peak, ca. 2900 m, J.

Klika (Klika 1926, as Ustilago violacea; Denchev 2001, as Microbotryum violaceum s. lat.).

Acknowledgements

We gratefully acknowledge Dr David W. Minter (CABI Europe, Egham, UK) and

Dr Vadim A. Melnik (V.L. Komarov Botanical Institute, St. Petersburg, Russia) for

serving as pre-submission reviewers. The study of fungi associated with relict plants was

financed by project BG0034-GAE-00100-E-V1 - EEA FM, Conservation of biodiversity

in hot-spots of glacial relict plants in Bulgaria.

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MY COTAXON

http://dx.doi.org/10.5248/117.381

Volume 117, pp. 381-404 July-September 2011

Two new Ceratocystis species associated with

mango disease in Brazil

MARELIZE VAN WYK’, BRENDA D. WINGFIELD’, ALI O. AL-ADAWI?,

CARLOS J. ROSSETTO?, MARGARIDA FUMIKO ITO?* & MICHAEL J. WINGFIELD’

"Department of Genetics, Forestry and Agricultural Biotechnology Institute (FAB),

University of Pretoria, Pretoria 0002, South Africa

*Ghadafan Agriculture Research Station, Ministry of Agriculture, Sohar, 311, Sultanate of Oman

*Instituto Agrondmico-IAC, Campinas-SP, 13001-970, Brazil

‘Cnpq Researcher, Centro de Pesquisa e Desenvolvimento de Fitossanidade-IAC, Brazil

*CORRESPONDENCE TO: marelizevw@nicd.ac.za

ABSTRACT —Mangifera indica, a disease known as mango blight, murcha or seca da

mangueira in Brazil, is caused by the canker wilt pathogen Ceratocystis fimbriata sensu

lato. It is also closely associated with infestation by the non-native wood-boring beetle

Hypocryphalus mangiferae (Coleoptera: Scolytinae). The aim of this study was to characterize

Ceratocystis isolates obtained from diseased mango trees in Brazil. Identification was based

on sequence data from ITS1+5.8S+ITS2 rDNA, part of the Beta-tubulin 1 gene, and part of

the Transcription Elongation Factor 1-alpha gene. The Brazilian isolates grouped in two well

defined and unique clades within C. fimbriata s.1. These were also distinct from C. manginecans,

which causes a similar disease associated with H. mangiferae in Oman and Pakistan. Based

on sequence comparisons and morphological characteristics, isolates representing the two

phylogenetic clades are described as C. mangicola sp. nov. and C. mangivora sp. nov.

KEY worps — agricultural crop, bark beetles

Introduction

A disease typified by wilting of the leaves, flowers, and stems of mango trees

(Mangifera indica L. (Anacardiaceae) mango) was first reported from Brazil in

the 1930's (Viégas 1960, Ploetz 2003). The disease, commonly referred to as

“mango blight’, “seca” or “murcha da mangueira’, represents one of the most

important constraints to mango production in Brazil (Ploetz 2003). The causal

agent of this disease was identified as Ceratocystis fimbriata Ellis & Halst. sensu

lato (s.1.) (Viégas 1960, Piza 1966, Ribeiro 1980).

Ceratocystis fimbriata s.l. was first recognized as possibly encompassing more

than one taxon by Webster & Butler (1967a, b), who showed host specificity

382 ... Van Wyk & al.

amongst isolates of the fungus. Isolates of C. fimbriata s.l. are morphologically

similar, but many can be differentiated through DNA sequence analyses. During

the past decade, numerous new and cryptic species in the C. fimbriata complex

have been described. Examples include the African fungus C. albifundus M.].

Wingf. et al. (Wingfield et al. 1996, Barnes et al. 2005), C. larium M. van Wyk

& M.J. Wingf. (Van Wyk et al. 2009a), C. cacaofunesta Engelbr. & T.C. Harr.

(Engelbrecht & Harrington 2005), C. fimbriatomima M. van Wyk & M.J. Wingf.

(Van Wyk et al. 2009b), C. curvata M. van Wyk & M.J. Wingf., C. ecuadoriana

M. van Wyk & M.J. Wingf., and C. diversiconidia M. van Wyk & M.J. Wingf. (Van

Wyk et al. 2011). In the strict sense, C. fimbriata is restricted to those isolates

related to the sweet potato black-rot pathogen, first described by Halsted (1890)

from diseased Ipomoea batatas (L.) Lam. (Convolvulaceae) (sweet potato)

tubers in the USA (Engelbrecht & Harrington 2005). An alternative view is

that phylogenetically different C. fimbriata s.l. isolates from various Brazilian

hosts might represent populations of C. fimbriata s.s. rather than discrete taxa

(Ferreira et al. 2010).

Ceratocystis species require wounds to infect trees (De Vay et al. 1963, Kile

1993). In Brazil, mango blight is closely associated with the wood-boring beetle

Hypocryphalus mangiferae Stebbing (Coleoptera: Scolytinae) that is native to

southern Asia (Wood 1982, Butani 1993, Atkinson & Peck 1994). It has been

hypothesized that this insect aids in the dissemination of the fungus in Brazil

(Ribeiro 1980, Yamashiro & Myazaki 1985, Ploetz 2003). Interestingly, the

same beetle is associated with Ceratocystis manginecans M. van Wyk et al. that

causes a serious disease of Mango trees in Oman and Pakistan (Al Adawi et al.

2006, Van Wyk et al. 2005; 2007a) and that has the same symptoms as mango

blight in Brazil.

When Van Wyk et al. (2007a) described C. manginecans, only two

C. fimbriata s.l. isolates from diseased mango in Brazil were included. These

isolates differed phylogenetically from C. manginecans but were not treated as

novel due to the small number of isolates available. Recently, a larger collection

of C. fimbriata s.l. isolates associated with mango blight in Brazil has become

available for study. The aim of this investigation was to compare these isolates

with C. manginecans and thus to determine their identity.

Materials and methods

Isolates

A total of 15 isolates (TABLE 1) from diseased Mango trees obtained in Sao Paulo

State in Brazil were transferred to 2% Malt Extract Agar (MEA) (20 g/L) (Biolab,

Midrand, South Africa) and maintained at room temperature (~25°C). All cultures

used are maintained in the culture collection (CMW) of the Forestry and Agricultural

Biotechnology Institute (FABI, University of Pretoria, South Africa). Representative

Ceratocystis spp. nov. (Brazil) ... 383

isolates have also been deposited with the Centraalbureau voor Schimmelcultures (CBS,

Utrecht, The Netherlands). Cultures of representative isolates bearing fruiting structures

of the fungi were dried on 30% glycerol and have been deposited with the National

Collection of Fungi (PREM), South Africa.

TABLE 1. Ceratocystis spp. for which isolates or sequences were used in this study.

SPECIES ISOLATE . GENBANK ACC. # Host ORIGIN

C. acaciivora CMW22563 EU588656, Acacia mangium Indonesia

EU588636,

EU588646

CMW22564 EU588657, A. mangium Indonesia

EU588637,

EU588647

C. albifundus CMW4068 DQ520638 A. mearnsii RSA

EF070429

EF070400

CMW5329 AF388947 A. mearnsii Uganda

DQ371649

EF070401

C. atrox CMW 19383 EF070414 Eucalyptus grandis _ Australia

CBS120517 EF070430

EF070402

CMW 19385 EF070415 E. grandis Australia

CBS120518 EF070431

EF070403

C. cacaofunesta CMW15051 DQ520636 Theobroma cacao Costa Rica

CBS152.62 EF070427

EF070398

CMW 14809 DQ520637 T. cacao Ecuador

CBS115169 EF070428

EF070399

C. colombiana CMW9565 AY233864 soil Colombia

CBS121790 AY233870

EU241487

CMW5751 AY 177233 Coffea arabica Colombia

CBS121792 AY177225

EU241493

CMW9572 AY 233863 Citrus reticulata Colombia

AY233871

EU241488

C. caryae CMW 14793 EF070424 Carya cordiformis USA

CBS114716 EF070439

EF070412

CMW 14808 EF070423 C. ovata USA

CBS115168 EF070440

EFO070411

C. curvata CMW22442 FJ151436 Eucalyptus Colombia

CBS122603 FJ151448 deglupta

FJ151470

CMW 22435 FJ151437 E. deglupta Colombia

CBS122604 FJ151449

FJ151471

C. diversiconidia CMW 22445 FJ151440 Terminalia Colombia

CBS123013 FJ151452 ivorensis

FJ151474

CMW 22446 FJ151443 T. ivorensis Colombia

FJ151455

FJ151477

384 ... Van Wyk & al.

TABLE 1, continued.

SPECIES ISOLATE No. GENBANK ACC. # Host ORIGIN

C. ecuadoriana CMW 22092 FJ151432 E. deglupta Colombia

CBS124020 FJ151444

FJ151466

CMW 22093 FJ151433 E. deglupta Colombia

CBS124021 FJ151445

FJ151467

C. fimbriata s.s. CMW 15049 DQ520629 Ipomaea batatas USA

CBS141.37 EF070442

EF070394

CMW1547 AF264904 I. batatas Papua New

EF070443 Guinea

EF070395

C. fimbriata s.1. C1345 AY157966 Eucalyptus sp. Brazil

C1987 AY585344 Eucalyptus sp. Brazil

C2041 AY585345 Acacia mearnsii Brazil

CMW 14811 AY526288 Colocasia esculenta _ Brazil

CBS115171

C1905

CMW 14791 AY526286 C. esculenta Brazil

CBS114713

C1865

C1900 AY526287 C. esculenta Brazil

C2032 AY526289 C. esculenta Brazil

C925 AY157967 Gmelina arborea Brazil

CMW 14806 AY526292 Ficus carica Brazil

CBS115166

C1782

CMW 14796 AY526307 Colocasia esculenta USA, Hawaii

CBS114720

C1715

CMW 14804 AY526306 C. esculenta USA, Hawaii

CBS115164

C1714

BPI596162 AY526305 C. esculenta China

C1558 AY157965 Mangifera indica Brazil

C. fimbriatomima CMW 24174 EF190963 Eucalyptus sp. Venezuela

CBS121786 EF190951

EF190957

CMW24176 EF190964 Eucalyptus sp. Venezuela

CBS121787 EF190952

EF190958

C. larium CMW25434 EU881906 Styrax benzoin Indonesia

CBS122512 EU881894

EU881900

CMW25435 EU881907 S. benzoin Indonesia

CBS122606 EU881895

EU881901

C. mangicola CMW 14797 AY953382 Mangifera indica Brazil

CBS114721 EF433307

C1688 EF433316

CMW27306 FJ200256 M. indica Brazil

FJ200269

FJ200282

CMW28907 FJ200257 M. indica Brazil

FJ200270

FJ200283

CMW28908 FJ200258 M. indica Brazil

FJ200271

FJ200284

TABLE 1, continued.

SPECIES

(C. mangicola)

C. manginecans

C. mangivora

C. neglecta

C. obpyriformis

C. papillata

ISOLATE No.

CMW28913

CMW28914

CMW13851

CBS121659

CMW13852

CBS121660

CMW23634

CMW23628

CMW15052

CBS600.70

C74

CMW27304

CBS127204

CMW27305

CBS128340

CMW27307

CMW28909

CMW28910

CMW28911

CMW28912

CMW28916

CMW17808

CBS121789

CMW 18194

CBS121017

CMW23807

CBS122608

CMW23808

CBS122511

CMW8857

CMW8856

CBS121793

GENBANK ACC. #

FJ200259

FJ200272

FJ200285

FJ200260

FJ200273

FJ 200286

AY953383

EF433308

EF433317

AY953384

EF433309

EF433318

EF433302

EF433303

EF433298

EF433306

EF433315

FJ200261

FJ200274

FJ200287

FJ 200262

FJ200275

FJ200288

FJ 200263

FJ200276

FJ200289

FJ200264

FJ200277

FJ200290

FJ200265

FJ200278

FJ200291

FJ200266

FJ200279

FJ200292

FJ 200267

FJ200280

FJ 200293

FJ200260

FJ200281

FJ200294

EF127990

EU881898

EU881904

EF127991

EU881899

EU881905

EU245004

EU244976

EU244936

EU245003

EU244975

EU244935

AY233868

AY233878

EU241483

AY233867

AY233874

EU241484

Ceratocystis spp. nov. (Brazil) ... 385

Host

M. indica

Hypocryphalus

mangiferae

M. indica

Eucalyptus sp.

Acacia mearnsii

A. mearnsii

Annona muricata

Citrus limon

ORIGIN

Brazil

Oman

Pakistan

Brazil

Colombia

South Africa

Colombia

386 ... Van Wyk & al.

TABLE 1, continued.

SPECIES

(C. papillata)

C. pirilliformis

C. platani

C. polychroma

C. polyconidia

C. populicola

C. smalleyi

C. tanganyicensis

C. tsitsikammensis

C. variospora

ISOLATE No.

CMW 10844

CMW6569

CMW6579

CBS118128

CMW 14802

CBS115162

CMW23918

CMW 11424

CBS115778

CMW11436

CBS115777

CMW23809

CBS122289

CMW23818

CBS122290

CMW14789

CBS119.78

CMW 14819

CBS114725

CMW 14800

CBS114724

CMW26383

CBS114724

CMW15991

CBS122295

CMW15999

CBS122294

CMW 14276

CBS121018

CMW 14278

CBS121019

CMW20935

CBS114715

CMW20936

CBS114714

GENBANK ACC. #

AY 177238

AY177229

EU241481

AF427104

DQ371652

AY528982

AF427105

DQ371653

AY528983

DQ520630

EF070425

EF070396

EF070426

EF070397

EU426554

AY528970

AY528966

AY528978

AY528971

AY528967

AY528979

EU245006

EU244978

EU244938

EU245007

EU244979

EU244939

EF070418

EF070434

EF070406

EF070419

EF070435

EF070407

EF070420

EF070436

EF070408

EU426553

EU426555

EU426556

EU244997

EU244969

EU244929

EU244998

EU244970

EU244939

EF408555

EF408569

EF408576

EF408556

EF408570

EF408577

EF070421

EF070437

EF070409

EF070422

EF070438

EF070410

Host

Coffea arabica

Eucalyptus nitens

E. nitens

Platanus

occidentalis

Platanus sp.

Syzygium

aromaticum

S. aromaticum

Acacia mearnsii

A. mearnsii

Populus sp.

Carya cordiformis

C. cordiformis

A. mearnsii

Rapanea

melanophloeos

R.. melanophloeos

Quercus alba

Q. robur

ORIGIN

Colombia

Australia

USA

Greece

Indonesia

South Africa

Poland

USA

Tanzania

South Africa

USA

Ceratocystis spp. nov. (Brazil) ... 387

TABLE 1, concluded.

SPECIES ISOLATE No. GENBANK ACC. # Host ORIGIN

C. virescens CMW 11164 DQ520639 Fagus americana USA

EF070441

EF070413

CMW3276 AY528984 Q. robur USA

AY528990

AY529011

C. zombamontana CMW 15235 EU245002 Eucalyptus sp. Malawi

EU244974

EU244934

CMW15236 EU245000 Eucalyptus sp. Malawi

EU244972

EU244932

*CMW numbers are in the Culture collection of the Forestry and Agricultural Biotechnology Institute

(FABI), University of Pretoria, South Africa; CBS numbers are in the Centraalbureau voor

Schimmelcultures (CBS), Utrecht, The Netherlands; C numbers are in the T. Harrington collection

Iowa State University, USA; BPI numbers are in the US National Fungus collection.

Phylogenetic comparisons

DNA was extracted from the isolates obtained from mango in Brazil according to

Van Wyk et al. (2006). Three sets of analyses were run on DNA sequence data obtained

from these isolates. The first dataset comprising the Internal Transcribed Spacer region

1 and 2 including the 5.8S rRNA operon (ITS) included sequences for all species in the

C. fimbriata s.1. complex as well as most C. fimbriata sequences available in GenBank

(http://ncbi.nlm.nih.gov) and from a variety of plants in Brazil. For the second dataset,

three gene regions were targeted for PCR including the ITS, part of the Beta-tubulin 1

(Bt) gene, and part of the Transcription Elongation Factor 1-alpha gene (EF1-a). Data

for these three gene regions were combined. The third dataset consisted of only the

isolates from Brazil and from mango with each gene region (ITS, Bt, EFl-a) treated

separately.

DNA amplification was achieved with the primer sets ITS1 and ITS4 (White et al.

1990), Btla and Bt1b (Glass & Donaldson 1995), and EF1F and EF1R (Jacobs et al. 2004),

following the protocols described by Van Wyk et al. (2006). Amplification was assessed

with the aid of gel electrophoresis in the presence of ethidium bromide. PCR amplicons

were purified using 6% Sephadex G-50 columns (Steinheim, Germany) and sequenced

in both directions using the ABI PRISM™ Big DYE Terminator Cycle Sequencing Ready

Reaction Kit (Applied BioSystems, Foster City, California), with the same primers as

those used for DNA amplification. Sequencing reactions were run on an ABI PRISM™

3100 Autosequencer (Applied BioSystems, Foster City, California, USA).

Sequences were analysed using the software programme Chromas Lite 2.01

(http://www.technelysium.com.au). Sequence data obtained in this study for Brazilian

isolates from mango were compared with those residing in the C. fimbriata s.l. clade

for Ceratocystis obtained from GenBank or those previously published (Van Wyk

et al. 2005, 2007a,b, 2009a,b, 2011). These sequences were aligned using MAFFT

(http://timpani.genome.ad.jp/%7emaftt/server/) (Katoh et al. 2002) and confirmed

manually. Thereafter, the C. fimbriata s.1. dataset was analyzed using PAUP version

4.0b10* (Swofford 2002). Sequences for the three gene regions were analyzed separately

and a partition homogeneity test (Swofford 2002) was used to determine whether the

388 ... Van Wyk & al.

three datasets (ITS, Bt and EFl-a) could be combined. The combined analyses were

run as described in Van Wyk et al. (2009b). Sequences derived from this study were

deposited in GenBank (TABLE 1) and the accompanying datasets and trees are deposited

in TreeBase (http://purl.org/phylo/treebase/phylows/study/TB2:S11630).

A modeltest (MrModeltest2) was run on each gene region to determine nucleotide

substitution rates (Nylander 2004) for incorporation into Bayesian analyses (MrBayes

version 3.1.1) to determine whether nodes obtained with PAUP had statistical support

(Ronquist & Huelsenbeck 2003). One million trees were generated using the Markov

Chain Monte Carlo (MCMC) procedure. Four chains, two hot and two cold, were

utilized to obtain the results. Trees were sampled every 100" generation and printed.

Tree likelihood scores were assessed to determine the number of trees that had formed

before stabilization to avoid including trees that had formed before convergence. Trees

outside the point of convergence were discarded by means of the burn-in procedure

(Ronquist & Huelsenbeck 2003).

Molecular Evolutionary Genetics Analysis (MEGA) 4 (Tamura et al. 2007) was used

to determine the level of variation between the isolates from a wide range of hosts in

Brazil for the ITS region only. In addition, this approach was applied for the ITS, Bt, and

EF 1-a for the isolates obtained from mango trees in Brazil and including C. manginecans

previously described from Oman and Pakistan. Sequences for each of the three gene

regions were inspected to determine the number of fixed alleles between them.

An allele network was drawn using the software TCS (Clement et al. 2000). The

dataset consisted of the combined gene regions (ITS, Bt and EF1-a) of all the isolates

obtained from mango in Brazil and including C. manginecans and C. fimbriatomima.

Culture characteristics and morphology

Based on the phylogenetic comparisons, two groups (Bl and B2) of isolates

emerged. Three representatives from each of the two groups (CMW 14797, CMW27306,

CMW28907 and CMW15052, CMW27304, CMW27305) were randomly selected for

growth studies in culture at different temperatures. The isolates were grown for 14 days

on 2% MEA, after which 5mm diam. plugs were transferred to the centers of 90mm

Petri plates containing 2% MEA. These plates were incubated at temperatures between

5 and 35°C at five degree intervals. Five plates were used for each isolate at each of the

test temperatures and the entire experiment was repeated once. The colony colours for

isolates were assigned using the colour charts of Rayner (1970).

For microscope studies, the same six isolates, representing the two groups (Bl

and B2) that were used to compare culture characteristics were selected. These

cultures were grown for 10 days on 2% MEA plates. Fungal structures were selected

and mounted in lactic acid on glass slides. Photographic images were captured with a

Carl Zeiss compound microscope and using a Zeiss Axio Vision camera system. For

isolates CMW14797 and CMW28305, 50 measurements were made for taxonomically

relevant morphological characteristics, while 10 measurements were taken for isolates

CMW27306, CMW28907, CMW15052 and CMW27304. The averages and standard

deviations (stdv) were computed for all the measurements that are presented in the

descriptions as (minimum-—) mean minus stdv - mean plus stdv (-maximum). Where

the minimum value was the same as the mean minus the stdv, a parenthetical minimum

was not included.

Ceratocystis spp. nov. (Brazil) ... 389

Results

Phylogenetic comparisons

ITS sequences for species in C. fimbriata s.1. —including unidentified isolates

from Colocasia (Araceae) (taro), Mangifera (Anacardiaceae) (mango), Gmelina

(Lamiaceae) (yemane) and Ficus (Moraceae) (fig) in Brazil— gave a 614 bp

dataset for 83 isolates. This dataset consisted of 234 constant, 11 parsimony

uninformative, and 369 parsimony informative characters. Of the five trees

obtained in these analyses, one was selected for presentation (Fic. 1). The tree

had the following characteristics; tree length = 1279, Consistency Index = 0.6,

Rescaled Index = 0.5, Retention Index = 0.9.

MrModeltest2 selected the GTR+I+G model for the ITS gene region. These

settings were included in the Bayesian analyses and four thousand trees were

discarded because they were obtained outside the point of convergence. The

Bayesian probabilities obtained in MrBayes were included in the phylogram

(Fic. 1) obtained in PAUP. The probabilities obtained in the Bayesian analyses

were similar to the support values obtained in PAUP.

The isolates from Brazil grouped into several polyphyletic clades (Fic. 1).

These included a well-supported clade (Bootstrap 86%) represented by two

isolates, one from Acacia (Mimosaceae) the other from Eucalyptus (Myrtaceae)

A second clade included only isolates from taro (Bootstrap 85%). Isolates

from mango and a Eucalyptus and Gmelina isolate resided in a discrete clade

(Bayesian 91%), and C. manginecans isolates grouped in a clade sister to that

clade (Bootstrap 87%). A group of isolates from mango resided in a clade with

strong support (Bootstrap 99%, Bayesian 86%), and an isolate from fig was

sister to that clade.

Amplicons of ~500 bp (ITS and £-tubulin) and ~800 bp (EFl-a) were

obtained for the Brazilian isolates of C. fimbriata s.l. from mango (TaBLE 1).

The PHT resulted in a low P-value (P=0.01), possibly due to the small amount

of variation in the Bt gene region. Although the P-value was low, this value

remained acceptable (Sullivan 1996, Cunningham 1997) to support combination

of the data for the three gene regions. The combined dataset for the three gene

regions consisted of a total of 1971 characters, 1066 of which were constant, 57

were parsimony-uninformative, and 848 were parsimony informative. Twenty-

two most parsimonious trees were obtained, one of which (Fic. 2) was selected

for presentation (Tree length = 2361, Consistency Index = 0.6, Rescaled Index

= 0.5, Retention Index = 0.9).

MrModeltest2 selected the GIR+I+G model for the ITS gene region, the

GTR+G model for the Bt gene region and the HKY+I+G for the EFl-a gene

region. These settings were included in the Bayesian analyses. Two thousand

trees were discarded as they were outside of the point of convergence. The

posterior probabilities for the branch nodes were included in the tree obtained

390 ... Van Wyk & al.

with PAUP (Fic. 2). The posterior probabilities supported the bootstrap values

obtained using PAUP.

The isolates from mango in Brazil grouped in two distinct clades (B1 and

B2), with high bootstrap support (100% and 100%, respectively). These two

phylogenetic groups were sister to C. manginecans, the species most closely

related to them. All other species considered in this study, formed well-

supported and distinct clades, confirming their unique nature.

The single ITS gene tree (Fic. 3) had a structure similar to the tree based

on combined sequences for the three gene regions. The two groups of isolates

from mango (B1 and B2) in Brazil grouped apart from C. fimbriatomima and

C. manginecans with high bootstrap support. The single gene trees for the Bt and

EF1-a gene region did not distinguish between the two groups of isolates from

mango in Brazil but they did distinguish C. fimbriatomima and C. manginecans

from these two groups.

The number of fixed alleles between the four groups (three from mango

and one from Eucalyptus), C. manginecans, the two groups of isolates obtained

from Mango in Brazil (Group B1 and Group B2) and C. fimbriatomima varied

within and between groups (TABLE 2). Analysis of the combined dataset for the

three gene regions using TCS resulted in two allele trees (Fic. 4). Ceratocystis

fimbriatomima was represented on its own while all three taxa from diseased

mango including C. manginecans and the two groups identified in this study,

resided in a single allele tree.

TABLE 2. Comparison of differing sequences and number of fixed alleles in Ceratocystis

spp. from mango and the closely related species C. fimbriatomima. Shaded cells

indicate variations within each species.

| C.mangicola | ACT tt S| TCE

| C.manginecans | iT —CiE t—“(i*dL

emanate

[eomangineamns

[ecnbratoina

[eC mangeola

[mangers PTT

[eC Fimbratomina PO

Ceratocystis spp. nov. (Brazil) ... 391

97 CMWI1S049 C. fmbriata s.s.

CMW154? C firdriata s.s.

B7G00)- CMWI1S0S1 C cacanfinssta

93 CMW14809 C. cacagfimesta

{0% CMW22562 C acaciivera

| CMW2253 C acaciivera

| orl CMWS761 C colanikona

LOOPED’ CMEWS?51 C. colombiana

| AY157966 Babpus Brazil

AY 585345 Acacia saree Brazil

1002100) CMW 4802 C phatani

CMW23918 C plates

PS; CM'W14811 Taro Brazil

95 —bh] ~ CMW14791 Tuo Brazil

AYS25287 Tao Brazil

AYS25229 Taro Brazil

AYRE Eucahytus Brazil

CMW28907 Margo Brazil Bl

en CMW28913 Mangp Brazil

CAIW28914 Mangp Brazil

AYI57967 Gmeline arborea Brazil

CM W13851 C muangnecans

CM W13852 C nunginecans

CMW2363 C nunginecans

CMW23623 C munginecans

CMW24174 C findratomima

| *' CM W24176 C findbriatomima

CMW27307 Mangp Brazil B2

CMW1486 Fig Brazil

POLO) CAIW1 0844 C paprilata

CMW5746 C papillae

208% JCMW17808 C. welecta

~ CMW181% C oweglecta

oats CM W22092 C. ecuadmiaw

100@00)_ — CMW22442 C. curvata

CMW22432 C ewvate

1000) 6 CMW22445 C dierszonda

CMW22446 C diversicadia

100G.0C CMW14276 C tsitstkanmesis

CMW14278 C ssitsikanenmsis

10100 CMWI15S991 Crangarpicensis

CMW15992 Ctangayicensis

£290) CMW14795 Taro US A

&5@3) CMW1 4804 Taro US A

CD! $y AYS26005 Tao China

aan CMW19383 C atrox

CMW19385 C arrox

CMWES8 C piniliifornys

saw CMWES79 C. pinilixfona's

93(100) > ICMW23808 C. cbyyriforna's

mace nt CMW2380? C. obyyryfornas

cog CMW2B8? C. polconidia

’ CMW23818 C. poiconidia

a CMW S236 C. sonbanaztana

10000)’ CMI WI 5242 C sombanctana

CMW11424 C polyelronn

100 1OOCUO)'CMIW1435 C. polychromn

1000.00 CM W4058 C absfioxus

9809) CMWSG29 C abbjfioxtus

1000.0G CMW24% C lariuun

CMW2543? C briven

9900! CMW14789 C popuiicola

£099) CMW14819 € pepulicoia

10000) - CMIW20935 C varios pora

2.495) CMW20836 C varios pora

CMW14800 C soaailey?

1000.00) CMW25383 C. smalleyt

CMW1488 C carpae

SOD CMW14793 C cayas

CMW111&4 C wrescens

CMW3276 C virescens

—

FIGURE 1. Phylogenetic tree based of the ITS gene region for Ceratocystis mangicola (B1),

C. mangivora (B2), and other species in the C. fimbriata s.1. complex including isolates from Brazil

obtained from various hosts. Bootstrap values are indicated at the branches, with Bayesian support

in brackets.

392 ... Van Wyk & al.

100100) CM W15049 C frdniate s.s.

@9 CMW? C firbriatas.s.

CMWIS051 C cacaofimesta

100 FLOO)- cary 4809 C cacacfimesta

(C4IW15052 Mangp Brazil

100000) ChIW27304 Mango Brazil

CIW27305 Mango Brazil

aooy| | 4827307 Mango Brazil |,

|_| CAf28909 Mango Brazil

100) CHIW289 10 Mango Brazil

$7000) CMW28911 Mango Brazil

CMIW289 12 Margo Brazil

| CHIW289 16 Mango Brazil

9800) CMW13851 C. mmgirecans

CMW13852 C mamgnrecans

CHIW14797 Mangp Brazil

rT CMIW27306 Mangp Brazil

CMW28907 Mango Brazil Bl

CIW28908 Mangp Brazil

(C2IW28913 Mang Brazil

100(100), CMW28914 Mangp Brazil

I CMW24174 C fintriawnim

92000)

(100)

CMW24176 C. ferdriatonom

100000) Im CMW22562 C acaaivra

90(78) CM W22583 C acaniwre

100100) CMW22442 C curvata

CMW22432 C cuvate

CMW 4802 C platens

CMW23918 C plantar

CMW10844 C popiilata

CMW5746 C papillata

CM W5S761 C colombiana

00100) ‘CM WS751 C colombiara

9799 P89) CMW 7808 C negkcta

CMW18194 C negkcta

OB) F cMWw22092 C ecradonina

- WOO) 0441729003 C aciadorions

1007100) _fCMW22445 C dive sicomidia

CMW22446 C diversiconmdia

1000100) CM WI 5991 C tanganyicensis

28100 CMWI1 5992 € tangayrcensis

000100) CM W14276 C tsitstkamesnsis

100) CMW14298 C tsitsitkerremnsis

89— CM W569 C. pinillyfor mis

96:99) CMWE579 C. pirilifirms

"ree 3 Bk ate sects Neate

obzyrforns

balance 000 85|__ ICM W23809 C. pobconidia

100000)’ cn w23818 C pobvonidia

CMW152% C sombanrtana

100(100)' cpu 5242 C sombamontara

96100) 99100); CMW11424 & podcirom

271.00 CMW1 1436 C popelvonn

CMW19383 C atvox

1000100)" cnpw 9385 & avon

00000) fF CMW4058 C altyfindus

iSalontd 10002) CMW5329 C albyfindus

000100 CM W254%5 & lariuem

CMW25437 C. laium

1000100) ¥

984100)

98000

1000.00)-— CMW 4789 & popultcola

CMW14819 € populicola

1000.00) | CMW20035 C variospora

CMW20936 C wriospora

100000)) CATWA 4800 C sali

CMW26383 C sri

| CMW14808 C cargve

CMW14793 C canue

100000)

7000

CMW11164 C wrescens

CM W2278 C wires cons

cei

FIGURE 2. Phylogenetic tree based on the combined regions of the ITS, 6-tubulin and EF1-a for

Ceratocystis mangicola (B1), C. mangivora (B2), and other species in the C. fimbriata s.l. species

complex. Bootstrap values are indicated at the branches, with Bayesian support in brackets.

Aa eke ae cre

CMV 4811 Taro Graal

Ceratocystis spp. nov. (Brazil) ...

#5 a1 hep eee Taro Brazil

&Y526287 Taro Brazil

AVB3B358 ta18 ae

ell 4¢8 Hs pret ahs,

28907 Mango Braal

cae margs Beat

aiSbeer eh sis eos ae

chy eed

anao Bra

AY585344 EucalyptusBrazil

. MANGMECANS

FAANGMECINS

B1 (C. mangicob)

TY_le Masog & manenecans

ac CM 33867

canta aneus

MANGMECINS

ash ais e260 Bes srlh

4 ero Brazil

36 const geaie.me ang Baal azil

AYS26292 Ficus cance Braal

b. CMW24174 C. finbriatonima

CMW24176 C. finbriaomma

CMW229 16 Mango Brazil

CMW1 5052 Margo Brazil

CMW27204 Margo Brazil

——!

CMW24174 C findriatomma

CMW241% C finbdbriatomima

66. CMWI13851 C nunginecans

CMW13852 C »uamginecans

CMW14797 Mango Brazil

CMW27305 Mango Brazil

CMW2730? Mango Brazil

CM W2290? Mango Brazil

CMW229 10 Mango Brazil

CMW28911 Margo Brazil

CM W229 12 Mango Brazil

B2 (C. mangivore)

393

FIGURE 3. Three separate unrooted phylogenetic trees representing three gene regions. Isolates

representing the two groups from Brazil mango as well as Ceratocystis manginecans and

C. fimbriatomima were included. a. ITS. b. Bt. c. EFl-a.

394 ... Van Wyk & al.

FiGurE 4. An allele network of the two groups of isolates from Brazil as well

as a closely related species C. manginecans also isolated from mango trees and

Ceratocystis fimbriatomima. The numbers represent CMW numbers (TABLE 1).

Culture characteristics and morphology

Isolates representing Group B1 were morphologically similar to other species

in C. fimbriata s.|. They produced a banana odour, typical of fungi in this group.

After 2 weeks on 2% MEA, the colonies had a dark brown (snuff brown, 15”k)

colour (Rayner 1970) with large numbers of perithecia visible on the surface of

the cultures. At 5°C and 35°C, no growth was observed after 7 days. At 10°C

(8 mm), 15°C (22 mm), 20°C (36 mm) and 30°C (20 mm) diminished growth

was observed after 7 days while the optimum temperature for growth of these

isolates was 25°C (44 mm).

Ceratocystis spp. nov. (Brazil) ... 395

Isolates residing in Group B2 were similar to those in Group B1 producing a

banana odour and they had a similar morphology. After 2 weeks on 2% MEA,

the colonies also had a dark brown (snuff brown, 15”k) colour (Rayner 1970)

with many perithecia produced on the culture surface. As with the isolates in

Group B1, after seven days, there was no growth at either 5 or 35°C. Some

growth was observed at the other temperatures tested 10°C (7 mm) and 15°C

(22 mm), 20°C (41 mm), 30°C (36 mm) and 25°C (45 mm) represented the

optimum temperature for growth.

Taxonomy

Based on DNA sequence comparisons and (to a lesser extent) morphology,

isolates from mango in Brazil could be separated into two distinct groups. These

groups represent previously unknown species that are described as follows:

Ceratocystis mangicola M. van Wyk & M.J. Wingf., sp. nov. FIGURE 5

MycosBank MB511886

Hyphae ostiolares hyalinae divergentes convergentesque, (47-)57-73(-79) um longa.

Conidiophorae biformes; primariae phialidicae, lageniformes, hyalinae; secondariae

copiosae, tubiformes, apicibus expansis, hyalinae.

Tye: Brazil, Sao Paulo State, from diseased Mangifera indica trees, isolated C.J. Baker

C1688, 2000. (holotype PREM60182 (culture dried on 30% glycerol); culture ex-type

CMW 14797 = CBS114721).

Erymo ocy: The epithet refers to the fact that the fungus occurs on mango.

Colonies brown (15”k) on 2% MEA. Odour banana. Hyphae smooth and

segmented. Ascomatal bases globose to sub-globose, dark-brown to black,

(125-)139-199(-230) um wide, (115—-)136-192(-236) um long. Ascomatal

necks brown becoming lighter towards apices (541—)766-980(-1103) um long,

(21-)26-36(-46) um wide at base, (15-)19-27(-33) um wide at tip. Ostiolar

hyphae of two types; hyaline, divergent and convergent, (47—)57-73(-79) um

long. Asci evanescent, not seen. Ascospores hyaline, hat-shaped, 3-4 um long,

3-4 um wide excluding sheath, 5-6 um wide including sheath.

Thielaviopsis ANAMORPH: Conidiophores of two morphological forms.

Primary conidiophores phialidic, lageniform, hyaline, (59-)71-119(-140)

um long, (3-)4-6(-7) um wide at base, 5-7(-8) um wide at broadest point,

3-5(-8) um wide at tips. Secondary conidiophores, abundant, tube-like, flaring

at apices, hyaline, (53-)72-114(-148) um long, 4-6(-7) um wide at bases

and 6-8(-9) um wide at tips. Conidia of two types. Primary conidia, hyaline,

cylindrical, (15-)18-24(-29) um long, (3-)4-6 um wide. Secondary conidia,

abundant, hyaline, barrel to sub-globose shaped, (6-)7-9(-11) um long, 6-8

um wide. Chlamydospores rare, brown, thick-walled, globose to sub-globose,

(12-)14-16(-17) um long by (9-)11-13(-14) um wide.

396 ... Van Wyk & al.

FiGurE 5. Morphological characteristics of Ceratocystis mangicola (from holotype): a. Globose

ascomata. b. Divergent ostiolar hyphae. c. Convergent ostiolar hyphae. d. Primary phialidic

conidiophore. e. Secondary conidiophore with emerging chain of barrel-shaped conidia. f. Dark

colored chlamydospores. g. Hat-shaped ascospores. h. Cylindrical conidia. i. Chain of barrel-

shaped conidia. Scale bars: a = 100 um, b-c, e-f, h-i = 10 um, d = 20 um, g = 5 um.

Ceratocystis spp. nov. (Brazil) ... 397

HABITAT & DISTRIBUTION: Isolated from Mangifera indica trees and

associated with the wood-boring scolytine Hypocryphalus mangiferae. Known

from Sao Paulo State, North West Brazil.

ADDITIONAL SPECIMENS EXAMINED: BRAZIL. SAO PAULO STATE, VOTUPORANGA,

from diseased Mangifera indica trees, isolated C.J. Rossetto 13750-1, 2007, PREM60183,

living culture CMW27306; isolated C.J. Rossetto 13959, 2008, PREM60184, living

culture CMW28907; isolated C.J. Rossetto 13911, 2008, PREM60186, living culture

CMW28913; SAO PAULO STATE, SANTA BARBARA D°OESTE, from diseased Mangifera

indica trees isolated C.J. Rossetto 13977, 2008, PREM60185, living culture CMW28908;

SAO PAULO STATE, PINDORAMA, from diseased Mangifera indica trees isolated C.J.

Rossetto 13966, 2008, PREM60187, living culture CMW28914.

NoTEs: Ceratocystis mangicola (B1; CMW14797) is distinguished from all other

species in the C. fimbriata s.l. complex based primarily on phylogenetic

inference. However, it also has ostiolar hyphae that are both divergent and

convergent as opposed to being only divergent in most species of this genus.

Ceratocystis mangivora M. van Wyk & M.J. Wingf., sp. nov. FIGURE 6

MycosBank MB512368

Bases ascomatum globosae vel obpyriformes. Colla ascomatum brunnea, apicem versus

pallescentia, apice in duo vel plura ramose. Conidiophorae biformes; primariae phialidicae

lageniformes hyalinae; secondariae abundantes tubiformes apice expansae hyalinae.

Chlamydosporae absunt.

TyPE: Brazil. Sao Paulo State, Campinas, from diseased Mangifera indica trees, isolated

C.J. Rossetto 12132, 2001. (holotype PREM60570 (culture dried on 30% glycerol);

culture ex-type CMW27305 = CBS128340).

Erymo.oey: The epithet refers to the fact that the fungus causes a disease on mango.

Colonies brown (15”k) on 2% MEA. Odour banana. Hyphae smooth and

segmented. Ascomatal bases globose to obpyriform, dark-brown to black,

(171-)188-244(-295) um wide, (174—-)192-256(-310) um long. Ascomatal

necks brown becoming lighter towards apices, branching at apices into two

or more necks, (394—)437-575(-654) um long, (21-)26-34(-40) um wide at

base, (16-)19-29(-35) um wide at tip. Ostiolar hyphae hyaline, divergent and

convergent, (60-—)75-91(-96) um long. Asci evanescent, not seen. Ascospores

hyaline, hat-shaped, 3-5 um in length, 4-6 um wide excluding sheath, 5-8 um

wide including sheath.

Thielaviopsis ANAMORPH: Conidiophores of two morphological forms.

Primary conidiophores phialidic, lageniform, hyaline, (70—)78-106(-124) um

long, (3-)5-7 um wide at base, 5-7(-8) um wide at broadest point, 3-5 um

wide at tips. Secondary conidiophores, abundant, tube-like, flaring at apices,

hyaline, (42—)62-100(-118) um long, (3-)4-6 um wide at bases and (4-)6-8(-

9) um wide at tips. Conidia of two types: Primary conidia, hyaline, cylindrical,

(12-)16-24(-31) um long, 2-5 um wide. Secondary conidia, abundant, hyaline,

loa)

Ceratocystis spp. nov. (Brazil) ... 399

barrel-shaped, (8—)9-13(-15) um long, (5-)6-8(-9) um wide. Chlamydospores

absent.

HABITAT & DISTRIBUTION: Isolated from Mangifera indica trees. Associated

with the wood-boring scolytid Hypocryphalus mangiferae. Known from Sao

Paulo State, Central East Brazil.

ADDITIONAL SPECIMENS EXAMINED: BRAZIL, SAo Pauto State, from diseased

Mangifera indica trees, isolated M. Barreto Figueiredo, 1970, PREM60188, living culture

CMW15052 = CBS600.70; BRAZIL, SAo PauLo STATE, CAMPINAS, from diseased

Mangifera indica trees, isolated C.J. Rossetto 12093, 2001, PREM601839, living culture

CMW27304 = CBS127204; isolated C.J. Rossetto 12036, 2001, PREM60190, living

culture CMW27307; BRAZIL, SAo PAULO STATE, SANTA BARBARA D?OESTE, from

diseased Mangifera indica trees, isolated C.J. Rossetto 13988, 2008, PREM60191, living

culture CMW28909; isolated C.J. Rossetto 13987, 2001, PREM60192, living culture

CMW28910. BRAZIL, SAo PAULO STATE, TUPI (NEAR PIRACICABA), from diseased

Mangifera indica trees, isolated C.J. Rossetto 13989, 2008, living culture CMW28911;

isolated C.J. Rossetto 13989-1, 2008, living culture CMW28912; BRAZIL, SAo PAULO

STATE, VALINHOS, from diseased Mangifera indica trees, isolated C.J. Rossetto 13986,

2008, living culture CMW28916.

Notes: Isolates of C. mangivora (B2; CMW27305) can have ascomatal necks that

branch dichotomously at the apices with ostiolar hyphae being either divergent

or convergent. Isolates of this species also did not produce chlamydospores in

culture.

Discussion

Results of this study showed that a relatively large collection of Ceratocystis

isolates from mango trees suffering from Mango blight in Brazil reside in two

distinct phylogenetic clades. These groups are, furthermore, distinct from

C. manginecans, which causes a similar disease of mango in Oman and Pakistan

(Al Adawi et al. 2006, Van Wyk et al. 2005, 2007a). The isolates residing in

these two groups are consequently treated as distinct taxa and the names

C. mangicola and C. mangivora have been provided for them.

The mango tree blight in Brazil, known for almost a century, was previously

ascribed to C. fimbriata s.l., which we now recognize represents a relatively

large number of cryptic taxa. These species are morphologically very similar,

and although individual species can be distinguished from their closest

relatives, recognition based solely on morphological characteristics would

FiGurE 6. Morphological characteristics of Ceratocystis mangivora (from holotype): a. Globose

to obpyriform ascomata. b. Ascomatal neck branching into two apices with both convergent and

divergent ostiolar hyphae. c. Convergent ostiolar hyphae. d. Hat-shaped ascospores. e. Primary

conidiophore. f. Secondary conidiophore with emerging chain of barrel-shaped conidia. g. Chain

of cylindrical conidia. h. Chain of barrel-shaped conidia. Scale bars: a = 100 um, b-c, e-g = 10 um,

d,h=5 um.

400 ... Van Wyk & al.

be very difficult. This situation exists for many groups of fungi, for example

Fusarium species in the Gibberella fujikuroi species complex (Leslie et al. 1992,

O'Donnell et al. 2000). Ceratocystis mangicola is phylogenetically most closely

related to C. manginecans, a known pathogen of mango and other crops in

Oman and Pakistan. Ceratocystis mangivora (also described in this study) has

no well-defined sister group but is also closely related to the other two mango

pathogens. Isolates of these species were not only distinct from each other but

also phylogenetically distinct from the mango pathogen, C. manginecans. The

species phylogenetically most closely related to these mango pathogens from

Brazil is C. fimbriatomima, which was first isolated from Eucalyptus trees in

Venezuela (Van Wyk et al. 2009b).

Phylogenetic analyses of sequences for the ITS gene region strongly

supported separation of C. mangicola and C. mangivora. In contrast, the Bt

and EF1-a gene regions for the single gene trees showed little or no variation

between C. mangicola and C. mangivora. This is not uncommon for species in

the C. fimbriata complex (Van Wyk et al. 2010). Similarly the allele trees for four

species; C. fimbriatomima and the three mango pathogens, C. manginecans,

C. mangicola, and C. mangivora showed that the three species from mango

were most closely related to each other. This suggests a common ancestor for

the three mango pathogens and the fact that they have probably undergone

speciation relatively recently.

Ceratocystis mangicola and C. mangivora are morphologically very similar,

both producing dark brown cultures with a banana odour that is characteristic

of many species of Ceratocystis. However, isolates representing the two species

could be distinguished from each other based on various micro-morphological

characteristics. Thus, C. mangicola isolates have both divergent and convergent

ostiolar hyphae, a characteristic not noted for other C. fimbriata s.l. species

except C. mangivora described here. Isolates of C. mangivora consistently

display branched ascomatal necks that give rise to either convergent or divergent

ostiolar hyphae. Furthermore, similar to some species in C. fimbriata s.L.,

C. mangivora did not produce chlamydospores in culture while these structures

are very obvious in cultures of C. mangicola. Ceratocystis mangicola isolates

also have globose to sub-globose ascomatal bases compared to the globose to

obpyriform bases in C. mangivora.

A previous study on C. fimbriata s.l. from different hosts (including mango

in Brazil) treated the isolates as a genetically diverse population representing

a single taxon (Ferreira et al. 2010). It would have been interesting to include

data from that study’s isolates in our own research, which might have provided

a more robust species delimitation for C. mangicola and C. mangivora. The

unavailability of the isolates for study and the absence of their sequence data

from GenBank preclude comparisons at that level.

Ceratocystis spp. nov. (Brazil) ... 401

An interesting aspect of the mango blight disease in Brazil and in Oman is

the fact that the pathogens are associated with the same wood-boring insect

(H. mangiferae) in both areas of the world. Hypocryphalus mangiferae is a

monophagous bark beetle found only on Mangifera species (Schedl 1961).

Its source area is likely the same as mango trees in tropical Asia (Wood 1982,

Butani 1993, Kostermans & Bompard 1993). Both tree and beetle have been

introduced into Brazil (Wood 1982, Butani 1993, Kostermans & Bompard

1993).

Both C. mangicola and C. mangivora are suspected to be native to Brazil.

As with most Ceratocystis species, a wound is required for C. mangicola and

C. mangivora to infect mango trees (Silva et al. 1959). Intensive studies of

diseased mango trees in Brazil have shown that H. mangiferae is the only

insect present during the early disease stages. Xyleborus species (Coleoptera:

Curculionidae) typically appear when the disease spreads down towards the

larger branches and Cerambycidae only appear when the disease reaches

the trunk regions (Silva et al. 1959, Castro 1960, Medeiros & Rossetto 1966,

Rossetto et al. 1980). Studies have also shown that H. mangiferae is the primary

vector of C. mangicola and C. mangivora in Brazil (Ribeiro & Rossetto 1971).

A similar vector relationship has also been shown for C. manginecans in Oman

(Al Adawi et al. 2006, Van Wyk et al. 2007).

The fact that H. mangiferae has become associated with three cryptic species

of Ceratocystis is not surprising. Species in this group of fungi easily establish

relationships with insects (Kile 1993, Roux & Wingfield 2009), probably

facilitated by the fruity aromas that they produce. The association between

C. mangicola, C. mangivora, and C. manginecans and H. mangiferae is very

similar to emerging new associations between ambrosia beetles and tree

pathogens such as those found in Laurel Wilt Disease in the USA (Mayfield et

al. 2008

In this study we chose to provide names to species reflected by phylogenetic

lineages (C. mangicola, C. mangivora, and C. manginecans) rather than to treat

them as population components of the single species C. fimbriata. In doing

so we could easily define distinctly different phylogenetic groupings and

provide a mechanism to distinguish their differences. Defined clades showing

such differences are undoubtedly valuable in studying important diseases,

including aspects of host pathogen interaction and resistance. Furthermore,

providing separate names for C. mangicola, C. mangivora, and C. manginecans

facilitates quarantine procedures and efforts to curb the global movement of

tree pathogens (Wingfield et al. 2001, Slippers et al. 2005).

Acknowledgements

We thank the National Research Foundation (NRF), members of the Tree Protection

Co-operative Programme (TPCP), the THRIP initiative of the Department of Trade

402 ... Van Wyk & al.

and Industry, and the Department of Science and Technology (DST)/NRF Centre of

Excellence in Tree Health Biotechnology (CTHB) for funding. We also thank Dr. Hugh

Glen for providing the Latin descriptions and for suggesting the names for the new

species. Peer reviews of the manuscript were provided by Prof. W.RO Marasas and

Dr. E. Roets, for which we are most grateful.

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MY COTAXON

http://dx.doi.org/10.5248/117.405

Volume 117, pp. 405-422 July-September 2011

The genus Cladonia (lichenized Ascomycota, Cladoniaceae)

in South Korea

XIN YU WANG’, YOGESH JOSHI’? & JAE-SEOUN HuR’™*

'Korean Lichen Research Institute, Sunchon National University, Sunchon 540-742, Korea

*Department of Botany. S.S.J. Campus, Almora 263601, Uttarakhand, India

* CORRESPONDENCE TO: jshur1@sunchon.ac.kr

AsBsTRACT — During a comprehensive study of Cladonia from South Korea, 41 taxa were

recognized, including C. dehiscens, C. floerkeana, C. macroptera, and C. rappii var. exilior,

new to South Korea. Brief information and discussion are provided for each taxon, together

with a key to the taxa recorded from South Korea.

Key worps — lichen-forming fungus, taxonomy, new records

Introduction

Cladonia is one of most common lichen genera, widely distributed on all

continents and including more than 400 species worldwide (Ahti 2000). Park

(1990) was the first to conduct an expert study of lichens in South Korea.

Her ‘Macrolichen flora of South Korea; however, included only 22 Cladonia

species, and the omission of many species we have since collected has made

identification difficult. Thus, we began comprehensive research on the genus

in South Korea with extensive field surveys from 2003 to 2010 and found

several Cladonia species previously not known in this country. Here we provide

information for 41 Cladonia taxa and a key to the South Korean species. Here

we follow the taxonomic system set forth by Ahti (2000), separating South

Korean Cladonia into five sections based on the secondary metabolites as

identified by High Performance Liquid Chromatography (HPLC): Cladonia,

Cocciferae, Helopodium, Perviae, and Unciales. Cladina is treated as a subgenus

of Cladonia.

Material and methods

We examined 496 Cladonia specimens collected during 2003-2010 in South Korea,

and deposited in the Korean Lichen Research Institute (KoLRI), Sunchon National

University. External morphological descriptions were based on air-dried material, and

406 ... Wang, Joshi & Hur

all specimens were observed under a Nikon SMZ645 stereomicroscope. The chemical

characters were determined by color reaction according to Yoshimura (1974): K (10%

aqueous KOH solution), C (saturated aqueous NaOCl solution), KC (10% KOH

followed by C solution), P (5% alcoholic solution of para-phenylenediamine). Chemical

compounds were identified using Thin Layer Chromatography (Culberson 1972; Elix et

al. 1987; Orange et al. 2001; White & James 1985).

Taxonomy

Key to the sections of Cladonia

1. Podetia ecorticate, surface dull and webby ...................... A. Subg. Cladina

Li BOGS tT a-CORMCAC tamale cancels teavenals sents eh ete na sid cel taht ROR ALR Ces Rothe 2

2 SAPOLMCCIAANG PHC TATE... 5.4 eee tsb dae bta waa Iadra tenet lotta aoahlatts Waatish « B. Sect. Cocciferae

2. Apothecia.and pycmidia DFOWN.. «54.13 5 dence sane t Re niet mene ee mei? betlenend bAletee tek tens 3

3. Podetia yellowish and smooth, containing usnic acid............. C. Sect. Unciales

3. Podetia green, gray or brown, not yellowish, without usnic acid (except in

CG ECHL OPUS obi ce tia ples ttle ae # eggs 2 tied ie 4 eae FS alt gle AE ae Oe es 4

4. Basal squamules well developed and large, podetia small or sometimes

evanescent, withoutcups. .. 6.76. [ses esteoe teal staal os D. Sect. Helopodium

4. Basal squamules usually persistent, podetia usually with well developed cups,

axils-and-cup center closed: i. 3 s/he +34: og dihece a a Aner ganze 38h: E Sect. Cladonia

4. Basal squamules evanescent, podetia branched, without cups, axils open

and thecup-certer perforate, g 4s. 24.0034 nee 34 neerde nerd 4 neat E. Sect. Perviae

Key to the Cladonia taxa recorded from South Korea

A. Cladonia subg. Cladina

li. Podetia sray, KC lackinio<wsinie acide «cious 36.6 vy wie x ely oe bee nee nak 2

1. Podetia yellowish, KC+ yellow, containing usnic acid

Sera PRES Netley tag Seria kx ilaaete 5 Death aetiaal 2. C. arbuscula subsp. beringiana

2. Ultimate branches pointing in one direction, polychotomously branched, final

branchlets: 2= 4) ou Ao setathe Sembee atl bent 32. C. rangiferina subsp. rangiferina

2. Podetia small and slim, dichotomously branched, final branchlets 2

ip ene Rell ice SEAR ce ee ng Rene A a, 33. C. rangiferina subsp. grisea

B. Cladonia sect. Cocciferae

LePodetia usually topped with cups .1o. cued tse a dec gestae Asc teban des debs piscd apace eocnahssa debate 2

1 -PoCetiasWARROULCUDS 3 whi. 8. Wut, Sots eR Wk hey wees lod bey lek! ets ihe cate eh arto 5

2 POG EU AS OLEAN ALC rg Fo. lit ce Mesto elarhcn Rar at Ri et me 29. C. pleurota

2. Podetia esorediate, with granules or squamules.............. 0. eee e ee eee eee 3

3. Podetia usually covered with granules, containing squamatic acid.. 19. C. granulans

3, Podetia-covered with squaimulés.or scaly-plates...... « ca.c0 o. vias ge siete co aide eete Se dates 4

Cladonia in South Korea... 407

4. Podetia covered with squamules, containing didymic acid..... 24. C. metacorallifera

4. Podetia covered with scaly plates, containing zeorin................ 6. C. coccifera

5. Podeétia-distinctly- corticate;sorediate. oo ax. 6 nj Selene ht eine ee we 14. C. floerkeana

5, (Pocetigm OSthyecOettca te, wee. Fo ihe hoo Rs beta Aes oR bl Neate erage salah 6

6. Farinose soredia throughout the podetia, corticate only at the basal parts,

Gisuially without SGUATIIUTES: 8. tins Sa ehive ty ie a el NE eta whale 22. C. macilenta

6. Podetia entirely decorticated, granulose soredia mixed with squamules

Rh ate Mic Ped cl NN hl dle Waa de PO tricle Pred te Ped BD PE ele Peed 11. C. didyma

C. Cladonia sect. Unciales

Podetia bifurcating or forming small cups at apices, containing barbatic acid

ddascka bly dshekon-st yicenieg dtsekg hcl vekes cringe doled zela dhe g chen od syplag cin zploo 1. C. amaurocraea

Podetia pointed and richly branched, containing squamatic acid....... 41. C. uncialis

D. Cladonia sect. Helopodium

Lf Pear younaliisspersist, podetiae ares +a. ee a PG tS | oe 2

1. Podetia usually present, primary thallus small or evanescent .................... 3

2. Primary thallus grayish green, rather large and thick, K+ yellow

Ft NAN Le DL Oe Le, eee RL ee LAR, Ee 39. C. symphycarpia

. Primary thallus yellowish green, small, K-....................088 4. C. caespiticia

3. Podetia with squamules, P+ yellow, containing psoromic acid ...... 10. C. dehiscens

3. Podetia verruculose on surface, P+ red, containing fumarprotocetraric acid

STE Re Mote sPaticle TPs sracecas trae la orate Partey ees ents pees oe 27. C. peziziformis

E. Cladonia sect. Perviae

1. Podetia coarsely sorediate on the tips ...................0000. 36. C. scabriuscula

| MOUS a CSOECCIALG ah Pre notary aceate -cormtel er amulet RT he wt way 4 Sees Pe Ie 2

2. Podetia K+ yellow, containing atranorin, primary thallus abundant 40. C. turgida

2-Podetia:K—, prtvary thallusevanescent. b6-0-4 state oce ww eects weal Bore ae a 3

3. Podetia P+ red, containing fumarprotocetraric acid.............. cece eee ee eee 4

3. Podetia P= contaiiine-squarni ale ACiCe rast. sceetey oor eee 2 etd Oe AL al 6

4. Podetia up to 10 cm long, with abundant squamules ............ 23. C. macroptera

4, Podetia shorter and slim, with few squamules ............... cee eee eee eee 5

5. Podetia simply branched, usually without apothecia...... 15. C. furcata var. furcata

5. Podetia with cymose branches, usually topped with apothecia

So eng GA Sins ood att eng Sher og dbo ong dra setng dh eet dt eee Ge 16. C. furcata var. racemosa

6. Funnel-like cups present, with large perforation in the center, podetial surface

smooth and lackinge-squamules. ... 0 se ea ei etree need veered ees 9. C. crispata

6. Squamules abundant on the podetia, narrow cups present or not... 37. C. squamosa

E. Cladonia sect. Cladonia

1. Podetia yellowish green, containing usnic acid ................. 3. C. bacilliformis

1. Podetia not yellowish, without usnic acid... 1.0.00... . cece e eee e ee Z

408 ... Wang, Joshi & Hur

NN DD Vo Ff FP WW NW LY

POdEtia eSOre di abe ax.2 Sale. Fp Sriale Py Seiala es se lelyye SNe Raye Seay ancy lyse eh dlecey Ad 3

PPOCET AOL eI ALC A CIewy Acree aiki ea ithe) Suey all es ay ins Les dP doy Brady Bs eH Ras ade 12

“Podetiansually topped withteups 4.0% Ait 28 fowt. ott Whee! eee EE TR ae OE eh 4

. Podetia without cups or with shallow cups........ 0... cece eect ees 9

. Podetia proliferating from the center of the cups........... 34. C. rappii var. exilior

“Proliferation absent.or-on-the marein of thescups ct ors sotars cede cdi ee ete see 5

PPodetiaswithidistinct dee prCUp se, saceios x nrie tes mays H moringe ¥ eniong ¥ woven Hdtame tewone Houde 6

Cups shallow, Corticate thsidé OF Lhe Cups.2: 4.055% lahat YiahOal ule B al Wat heh EOS LEY 8

. Podetia P+ yellow, containing psoromic acid................... 38. C. subconistea

. Podetia P+ red-brown, containing fumarprotocetraric acid...................0.. vi

. Cups small, K+ yellow, containing atranorin ................... 21. C. kurokawae

. Cups large, goblet shaped, scaly plated inside of the cups, K-, without atranorin

aeatty Sorte ing Sota veer Pagan aucatn auastieo eee arena ateeee at 30. C. pyxidata

. Cups regular, apothecia large, base of the podetia not blackened

MAS, Cer OR ere 8 Bere ee ARC eee eA 18. C. gracilis subsp. turbinata

8. Cups irregular, base of the podetia blackened, with white areolae . . .28. C. phyllophora

9. Podetia with smooth and continuous cortex, with areolae..................00. 10

9. Podetia lacking continuous cortex, largely decorticated..................0000. 11

10. Podetia with abundant squamules, usually pointed at apices...... 12. C. fenestralis

10. Podetia with few squamules, topped with small shallow cups

haveaghijare G4 isin e dQ ihm ed 4 Raed baerd 4 nip daiheer a4 Oe 17. C. gracilis subsp. gracilis

11. Podetia pointed at apices, rarely branched, with few squamules, sometimes small

ald oeariitl elk ey: Face eBags ioe age ee Page Pas Pape gs | eee 32. C. ramulosa

11. Podetia with many squamules on the lower parts, small cups present on the tips,

ApOtheCiailar Ge A eG ha Dace Poed ce ce ooP a Pes [Me ode ioc 26. C. mongolica

12.-Podetia-pointed or with tiny cups at ApIees on. Pen el ia Min wkd insecure Mike 13

IZ APOCetia-WIthODVIGUS CUS JP Al cares ties: Settee a sable otieatlin: anpaaltaata inate seat y ae 15

13. Podetia corticate only at the base, with farinose soredia throughout

Bt rll Nl ah vokeet tec alata gna sl sl Lis eputat Aes omtadl. Ley oad i Seg ahah dae 7. C. coniocraea

13, Podetia corticate, with patches oFsoralia nn”... 48 wet ek kW we ably ot 14

14. Primary thallus esorediate, podetia up to 4 cm tall, with soralia on upper

14. Primary thallus sorediate, podetia short, with patches of farinose soredia

Berd as ary iG Sanne vary earache aed gah ta Frise leek ane shal LO aa 26. C. ochrochlora

15. Cups shallow, margin irregular and asymmetrical....................0. 34. C. rei

LS Cups deep, Mar Sie Ntire ari C MOND mec. o yikes Ola Mo de gla og tas Me od eset so acer Pte 16

16. Soredia granulose, cups large and broad....................6. 5. C. chlorophaea

WGsSOPEATA TAPIA OSE jeainres oat. Movacte a ianastha Manatee Mawar ne earn cee hans cunank ae eeare cet nace 17

17. Podetia short, cups small, K+ yellow, containing atranorin......... 20. C. humilis

17. Podetia covered with farinose soredia, K-, without atranorin...... 13. C. fimbriata

Cladonia in South Korea ... 409

Figure 1: A. Cladonia dehiscens; B. C. floerkeana; C. C. macroptera; D. C. rappii var. exilior.

Scale bars: A, B, D = 1 cm; C =2 cm.

1. Cladonia amaurocraea (Flérke) Schaer.

REMARKS — This species is characterized by its yellowish podetia, frequently

tipped with small cups. Cladonia uncialis resembles this species, but it does not

have cups, and it contains squamatic acid but not barbatic acid. C. arbuscula is

also yellowish with usnic acid, but its podetia lack a cortex.

This species is reported from Japan, North America and Europe (Thomson

1984; Yoshimura 1974). In South Korea, common, usually on rock, mostly

found in the northern part and over the altitude of 800 m.

REPRESENTATIVE SPECIMENS (OF 15) EXAMINED — SOUTH KOREA. GANGWON

Prov., Taebaek city, Mt. Naejang, 35°29'47.2"N 126°53'44.5"E, alt. 1080 m, Hur 030419;

Mt. Sorak, 38°07'14.5"N 128°23'03.5"E, alt. 1355 m, Hur 041490; GYEONGBUK PRov.,

Mungyeong city, Mt. Juhul, 36°46'58.5"N 128°05'21.3"E, Hur 040160; GYEONGNAM

Prov., Mt. Gaya, 35°49'11.3"N 128°07'18.2"E, alt. 1440 m, Hur 040215.

2. Cladonia arbuscula subsp. beringiana Ahti

REMARKS — ‘The subspecies is resembles C. rangiferina but with yellowish

podetia due to the presence of usnic acid. A worldwide complex with many

subspecies and chemotypes, C. arbuscula is usually represented by C. arbuscula

subsp. beringiana (Ahti 2000) in eastern Asia and (in Japan) subsp. mitis

(Yoshimura 1974, Harada et al. 2004).

410 ... Wang, Joshi & Hur

This subspecies is reported from North America, Europe, Japan and China

(Harada et al. 2004; Thomson 1984; Wei 1991). In South Korea, only one

specimen was found on soil during our study.

SPECIMEN EXAMINED — SOUTH KOREA. GYEONGBUK PRov., Mt. Juwang,

36°23'47.7"N 129°08'50.5"E, alt. 270 m, Hur 050584.

3. Cladonia bacilliformis (Nyl.) Vain.

REMARKS — Characterized by having soredia all over the podetia, this

species might be confused with the sorediate species C. ochrochlora and

C. coniocraea but differs in having yellowish podetia and containing usnic acid

as the main compound.

Cladonia bacilliformis is also known from North America and China

(Thomson 1984; Wei 1991). In South Korea, very rare, found only in some low

altitude mountains growing on soil or rock.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PRov., Mt. Ilrim, 34°40'43.2"N

127°00'41.8"E, alt. 360 m, Hur 050375. Mt. Cheongwan, 34°32'39.8"N 126°56'51.5"E,

alt. 200 m, Hur 050527. GYEONGGI PRov., Mt. Gakhul, 38°06'55.1" N 127°22'49.6" E,

alt. alt. 826 m, Hur 080336.

4. Cladonia caespiticia (Pers.) Flérke

REMARKS — Diagnosed by very short (< 0.5 cm) podetia and pycnidia on

the primary thallus, C. caespiticia might be confused with C. symphycarpia but

lacks norstictic acid and has a smaller thinner primary thallus.

Also known from North America, Europe, South Africa, and Asia (Park

1990); not common or widespread in South Korea, where it grows on soil or

rock at altitudes below 900 m.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PRrov., Mt. Baekun, 35°04'17.4""N

127°39'27.1"E, alt. 840 m, Hur 041268. GANGWON PRov., Mt. Sorak, 38°10'46.7"N

128°21'54.7"E, alt. 430 m, Hur 041536. CHUNGNAM PRov., Mt. Gaeryeong, 36°21'30.7"N

127°12'44.5"E, alt. 750 m, Hur 041620.

5. Cladonia chlorophaea (Flérke ex Sommerf.) Spreng.

REMARKS — Characterized by wide-open cups and granulose soredia,

C. chlorphaea could be confused with C. grayi, which differs in producing

grayanic acid, and C. pyxidata, which has granular squamules but no soredia.

Widely distributed from the temperate zone to the Antarctic/Arctic (Ahti

2000), C. chlorophaea is rather common in South Korea, where it grows on

exposed soil (or sometimes rock or moss) at altitudes above 1000 m.

REPRESENTATIVE SPECIMENS (OF 13) EXAMINED — SOUTH KOREA. JEju Prov., Mt.

Halla, 33°05'48.8"N 126°55'11.6"E, alt. 1325 m, Hur 030371. GANGWON PRov., Mt.

Hambaek, 37°11'48.6"N 128°55'06.5"E, alt. 1194 m, Hur 030689. Mt. Odae, 37°47'55.3"N

128°32'53.3"E, alt. 1490 m, Hur 040472. JEONBUK PRov., Mt. Palgong, 35°36'01.8"N

127°27'57.4"E, alt. 688 m, Hur 050400.

Cladonia in South Korea... 411

6. Cladonia coccifera (L.) Willd.

REMARKS — Characterized by large, red apothecia on the proliferation,

surface squamules, and usnic acid and zeorin as the secondary metabolites,

C. coccifera resembles C. pleurota but lacks soredia while C. pleurota has

granulose soredia.

Circumpolar in the northern hemisphere south to the Himalayas (Stenroos

1989), C. coccifera is uncommon in South Korea where it is found on rock or

soil at 15-1000 m or above.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PRov., Gohong county, Sorok

Island, 35°48'11.2"N 129°18'47.3"E, alt. 15 m, Hur 030063-1. GANGWON PRov., Mt.

Gariwang, 37°24'05.0"N 128°32'39.5"E, alt. 937 m, Hur 080058, Hur 080065. Mt. Tuda,

37°26'36.8"N 128°59'35.3"E, alt. 761 m, Hur 080110. GYEONGBUK PRov., Mt. Joryeong,

37°48'27"N 128°03'32"E, alt. 500 m, Hur 080303.

7. Cladonia coniocraea (Flérke) Spreng.

REMARKS — ‘This species may be confused with C. ochrochlora but has

extensively spreading farinose soredia over podetia rather than round soralia.

It is mostly decorticated, while C. ochrochlora has a well-developed cortex all

over the podetia.

Known from the northern hemisphere and Australia (Park 1990),

C. conciocraea is uncommon in South Korea where it is found on rock or moss.

SPECIMENS EXAMINED — SOUTH KOREA. GANGwon PRov., Mt. Naejang, 35°29'47.2"N

126°53'44.5"E, alt. 1080 m, Hur 030423. Mt. Sorak, 38°06'40.4"N 128°24'26.8"E, alt.

1335 m, Hur 041452. Jeyu Prov., Jeju Island, 33°24'26.6"N 126°29'46.0"E, alt. 600 m,

Hur 040634-1.

8. Cladonia cornuta (L.) Hoftm. subsp. cornuta

REMARKS — Very close to C. coniocraea, which is corticate only at the base,

this subspecies has a more extensive cortex and patches of soralia. Cladonia

ochrochlora is also similar, but it is short and stout, its podetia are usually topped

with small cups, and the primary thallus has soredia, while C. cornuta subsp.

cornuta lacks cups, is slim and tall, and has an esorediate primary thallus.

Circumpolar, boreal, arctic, and in the southern hemisphere (Thomson

1984), the subspecies is rather common throughout South Korea, usually

growing on soil, sometimes on wood or rock.

REPRESENTATIVE SPECIMENS (OF 22) EXAMINED — SOUTH KOREA. JEONNAM PRov.,

Mt. Jiri, 35°18'39.4"N 127°35'54.3"E, alt. 1620 m, Hur 040294. Jeyu Prov., Jeju Island,

33°21'30.4"N 126°31'19.3"E, alt. 1710 m, Hur 040740. JEONBUK PRov., Mt. Naejang,

35°29'46.1"N 126°53'56.7"E, alt. 600 m, Hur 050021. GANGWON PrRov., Mt. Sorak,

38°08'53.5"N 128°20'05.3"E, alt. 1080 m, Hur 050279.

9. Cladoonia crispata (Ach.) Flot.

REMARKS — The funnel-like cups make this species quite unique. Cladonia

crispata occurs on soil and may be confused with C. furcata, which, however,

412 ... Wang, Joshi & Hur

does not form cups and the tips and contains fumarprotocetraric acid but not

squamatic acid.

Circumpolar, arctic, temperate, it is also found in South America (Thomson

1984), C. crispata is very rare in South Korea, where it usually grows on soil or

thin soil over rock at altitudes c. 1000 m.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PRrRov., Mt. Baekun, 35°10' N

127°60' E, alt. 1045 m, Hur 030291. GANGWON PRov., Mt. Taebaek, 37°06'15.5" N

128°56'06.2" E, alt. 1000 m, Hur 030391, Hur 030394.

10. Cladonia dehiscens Vain. Fic. 1A

REMARKS — New to South Korea, C. dehiscens may be confused with

C. peziziformis, but it contains psoromic acid only, the podetia surfaces are

partly decorticated and squamulose; C. peziziformis contains atranorin, and the

surface is verruculose without squamules.

Previously known only from China (Wei 1991), the species is rather rare

in South Korea, where it was found only found at c. 250 m on Mt. Gumsan

(Gyeongnam province), growing on rock.

SPECIMENS EXAMINED — SOUTH KOREA. GYEONGNAM PRov., Namhae county, Mt.

Gumsan, 34°45'38.5"N 128°02'55.5"E, alt. 249 m, Hur 070944, Hur 070945.

11. Cladonia didyma (Fée) Vain.

REMARKS — In having red apothecia and soredia, C. didyma is resembles

C. macilenta. The latter species, however, has thamnolic acid as the main

chemical compound and usually farinose soredia all over the podetia, while

C. didyma has didymic acid and the granulose soredia mix with squamules.

Pantropical, extending to warm temperate areas (Ahti 2000). In South Korea,

only one specimen was found on bark from the Jeju Island.

SPECIMEN EXAMINED — SOUTH KOREA. Jrju Prov., Jeju Island, 33°26'04.4"N

126°34'01.7"E, alt. 545 m, Hur 040878.

12. Cladonia fenestralis Nuno

REMARKS — ‘This species might be confused with C. gracilis, but

C. fenestralis has many more squamules on the podetia (which are usually

taller) and inconspicuous cups. Cladonia gracilis has regular shallow cups on

the tips of the podetia.

This species has been reported from Southeast Asia (Sipman 1993) and

South Korea (Hur et al. 2005). During our study, only two specimens were

found growing on rocks from Jeonbuk and Gangwon provinces.

SPECIMENS EXAMINED — SOUTH KOREA. JEONBUK PRov., Mt. Seonun, 35°29'46.9"N

126°53'40.7"E, alt. 300 m, Hur 030240. GANGWoN PRov., Mt. Naejang, 35°29'47.2"N

126°53'44.5"E, alt. 1080 m, Hur 030421.

Cladonia in South Korea... 413

13. Cladonia fimbriata (L.) Fr.

REMARKS — Characterized by slim podetia, narrow cups, and farinose

soredia, C. fimbriata is closely resembles C. chlorophaea, which differs in wider

cups and granulose soredia.

A common temperate to boreal and arctic-antarctic species (Ahti 2000),

C. fimbriata is very rare in South Korea, found only in Chungbuk province,

growing on rock.

SPECIMEN EXAMINED — SOUTH KOREA. CHUNGBUK PRov., Mt. Sobaek, 36°57'18"N

128°28'52.3"E, alt. 1389 m, Hur 030769.

14. Cladonia floerkeana (Fr.) Florke Fic. 1B

REMARKS — New to South Korea, C. floerkeana might be confused with

another sorediate species C. macilenta, but the latter species has farinose

soredia and the whole podetia are almost entirely decorticated, while

C. floerkeana is corticate over most of the podetia, its soredia are granulose, and

it has squamules on the podetia.

Also reported from North America, Europe, Australasia (Ahti 2000), and

East Asia (Wei 1991), C. floerkeana is very rare in South Korea, where it was

found growing on bark at c. 300 m on Mt. Juwang.

SPECIMEN EXAMINED — SOUTH KOREA. GYEONGBUK PROv., Mt. Juwang,

36°23'47.7"N 129°08'50.5"E, alt. 270 m, Hur 050587.

15. Cladonia furcata (Huds.) Schrad. var. furcata

REMARKS — ‘This variety is easily recognizable by having a well-developed

cortex, simple branches, and a few squamules. It has many variations and forms;

some Korean specimens, which are rather long and stout (< 8 x 2 mm), would

have been referred to C. furcata var. pinnata (Fink 1904), but this has recently

been synonymised with C. furcata var. furcata (Hur et al. 2005).

A widespread warm to cold temperate species (Ahti 2000), C. furcata is the

most common species in South Korea, widely distributed from low to high

altitudes, usually growing on humus or soil over rock, under the shady forest.

REPRESENTATIVE SPECIMENS (OF 53) EXAMINED — SOUTH KOREA. JEONNAM

Prov., Mt. Baekun, 35°10' N 127°60' E, alt. 1050 m, Hur 030289. GANGWON PRov.,

Mt. Taebaek, 37°06'25.9"N 128°56'47.8"E, alt. 963 m, Hur 030633. CHUNGBUK PROv.,

Mt. Sobaek, 36°57'17.6"N 128°28'47.1"E, alt. 1394 m, Hur 030778. GYEONGBUK PROv.,

Mt. Juhul, 36°46'58.5"N 128°05'21.3"E, alt. 675 m, Hur 040157. CHUNGNAM PRov.,, Mt.

Gaeryong, 36°21'25.6"N 127°12'35.3"E, alt. 770 m, Hur 041628.

16. C. furcata var. racemosa (Hoffm.) Florke

REMARKS — One additional variety of C. furcata has been found in Korean

specimens, which differs from other species by the subradiate branching

414 ... Wang, Joshi & Hur

towards the apex, the microsquamules on the surface, and the apothecia

commonly present on the cymose branch tips. Thallus P + red, containing

fumarprotocetraric acid.

SPECIMEN EXAMINED — SOUTH KOREA. GANGWON PRov., Mt. Odae, 37°47'43.3"N

128°34'22.0"E, alt. 1240 m, Hur 040481.

17. Cladonia gracilis (L.) Willd. subsp. gracilis

REMARKS — Similar to C. furcata, C. gracilis subsp. gracilis is much more

slender and has fewer squamules and podetia tips usually topped with cups. The

similar C. maxima lacks a primary thallus and has much longer, unbranched

podetia. The subspecies also differs from C. cornuta by lacking soredia.

Distributed from circumpolar regions to South America (Thomson 1984),

this subspecies is rare in South Korea, usually found growing on rockat altitudes

above 900 m.

SPECIMENS EXAMINED — SOUTH KOREA. JEONBUK PRov., Mt. Naejang, 35°29'46.1"N

126°53'56.7"E, alt. 600 m, Hur 050020. Jeyu Prov., Mt. Halla, 33°21'53.8"N

126°30'40.3"E, alt. 1695 m, Hur 090207. GANGwon PRov., Mt. Jobong, 37°56'10.7"N

128°33'74.7"E, alt. 980 m, Hur 090256.

18. Cladonia gracilis subsp. turbinata (Ach.) Ahti

REMARKS — This subspecies differs from subsp. gracilis mainly by having

large, wide cups and being covered with squamules. The similar C. rappii

differs in proliferating from the center of the podetial cups. The also similar

C. phyllophora differs in having regular cups and a smooth cortex.

Cosmopolitan (Ahti 1980), C. gracilis subsp. turbinata is very common and

widespread in South Korea on soil or mosses over rock at altitudes above 1000

REPRESENTATIVE SPECIMENS (OF 24) EXAMINED — SOUTH KOREA. JEju Prov., Mt.

Halla, 33°05'48.8"N 126°55'11.6"E, alt. 1655 m, Hur 030361; alt. 800 m, Hur 030368.

JEONNAM PRov., Mt. Jiri, 35°19'09.4"N 127°41'15.9"E, alt. 1600 m, Hur 040348.

GANGWON PRov., Mt. Taebaek, 37°06'10.4"N 128°57'16.1" E, alt. 1120 m, Hur 041021.

19. Cladonia granulans Vain.

REMARKS — Characterized by slender podetia covered by squamules or

granules on the podetia, C. granulans is quite similar to C. metacorallifera but

lacks didymic acid and has no proliferation on the cup margins.

Originally described from Japan and also known from North America

(Thomson 1984), this species is rare in South Korea and usually found at

altitudes above 1000 m, growing on bark or soil.

SPECIMENS EXAMINED — SOUTH KOREA. GANGWON PRov., Mt. Naejang,

35°29'47.2"N 126°53'44.5"E, alt. 1050 m, Hur 030418. Jeyu Prov., Jeju Island,

33°21'42.4"N 126°32'56.9"E, alt. 1670 m, Hur 040819. CHUNGBUK PRov., Mt. Sobaek,

36°55'47.5" N 128°27'22.9" E, alt. 1301 m, Hur 070338.

Cladonia in South Korea... 415

20. Cladonia humilis (With.) J.R. Laundon

REMARKS — Cladonia humilis, which contains atranorin, is difficult

to differentiate from C. chlorophaea, which lacks atranorin and has larger

podetia.

A widespread temperate species (Ahti 2000), C. humilis is uncommon in

South Korea, where it is found on soil over rock at low altitudes.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PRov., Sorok Island, 35°48'11.2""N

129°18'47.3"E, alt. 60 m, Hur 030056. GYEONGBUK PRov., Mt. Juhul, 36°46'58.5"N

128°05'21.3"E, alt. 657 m, Hur 040156; 36°46'46.0"N 128°06'02.1"E, alt. 845 m, Hur

040151. CHUNGBUK PRov., Mt. Joryeong, 37°01'33.3"N 128°11'59.2"E, alt. 285 m, Hur

061110.

21. Cladonia kurokawae Ahti & S. Stenroos

REMARKS — Characterized by small granules rather than real soredia,

somewhat small podetia and cups, fumarprotocetraric acid, and atranorin,

C. kurokawae resembles C. subconistea, which lacks fumarprotocetraric acid

and contains psoromic acid as the main compound.

Originally described from East Asia (Lai 2000), C. kurokawae is rather

common in South Korea where it is found mostly on soil or mossy rock in low

altitude mountains.

REPRESENTATIVE SPECIMENS (OF 17) EXAMINED — SOUTH KOREA. JEONNAM PRov.,

Mt. Baekun, 36°57'14.5"N 128°29'18.8"E, alt. 1000 m, Hur 0300182-2. JEONBUK PROV.,

Mt. Naejang, 35°48'11.2"N 126°18'47.3"E, alt. 600 m, Hur 030453. GYEONGNAM PROv.,

Mt. Gumsan, 34°45'44.2"N 127°59'29.6"E, alt. 290 m, Hur 040031. GYEONGGI PRov.,

Mt. Myeongji, 37°55'57.7"N 127°28'53.3"E, alt. 223 m, Hur 080669.

22. Cladonia macilenta Hoftm.

REMARKS — ‘This species is characterized by having red apothecia and

farinose soredia all over the podetia. TLC detected two chemical races in

Korean materials: (1) didymic and thamnolic acids; (2) didymic, barbatic, and

usnic acids. The horizontal primary thallus generally contains only thamnolic

acid (Asahina 1970).

Widespread worldwide (Ahti 2000), C. macilenta is also common and

widespread in South Korea, where it usually grows on rotting wood, bark,

humus, or soil at altitudes above 500 m.

REPRESENTATIVE SPECIMENS (OF 14) EXAMINED — SOUTH KOREA. GANGWON

Prov., Mt. Hambaek, 37°11'48.6"N 128°55'06.5"E, alt. 1194 m, Hur 030685. Mt.

Seokbyeong, 37°34'29.9"N 128°51'21.8"E, alt. 686 m, Hur 080188. Jeyu PRov., Jeju

Island, 33°24'26.6"N 126°29'46.0"E, alt. 600 m, Hur 040634. JEONNAM PRov., Wando

Island, 34°21'10.3"N 126°41'10.9"E, alt. 535 m, Hur 050142.

23. Cladonia macroptera Rasanen Fig. 1C

REMARKS — Characterized by long (< 10 cm) podetia, C. macroptera differs

from C. furcata by tall thick podetia with rare branches and densely squamulose

416 ... Wang, Joshi & Hur

and an areolate cortex that differs from the more or less continuous cortex in

C. furcata.

Previously known only from China and Canada (Brodo et al. 1987, Wei

1991), C. macroptera is new to South Korea where two specimens were found

growing on moss over rock.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PROv., Mt. Duryon, 37°06'27.6"N

128°56'48.8"E, alt. 200 m, Hur 030314. GANGWON PRov., Mt. Odae, 37°47'36.2"N

128°34'37.0"E, alt. 1195 m, Hur 040483.

24. Cladonia metacorallifera Asahina

REMARKS — ‘This species is characterized by having cups topped with red

apothecia, without soredia but squamulose and contains squamatic acid. It may

be confused with C. coccifera, but the latter species usually has farinose granules

resembling soredia inside of the cups and contains zeorin.

Also known from North America, Japan, and China (Thomson 1984; Wei

1991), C. metacorallifera is uncommon in South Korea where it usually grows

on soil over rock at 1000-1700 m altitudes.

SPECIMENS EXAMINED — SOUTH KOREA. GANGwWON PROv., Mt. Naejang, 35°29'47.2"N

126°53'44.5"E, alt. 1080 m, Hur 030420. Mt. Sorak, 38°06'42.8"N 128°24'21.8"E, alt.

1360 m, Hur 041466. Jeyu Prov., Jeju Island, 33°21'32.9"N 126°31'54.9"E, alt. 1920 m,

Hur 040758. JEONNAM PRov., Mt. Jiri, 35°19'40.9"N 127°44'15.0"E, alt. 1564 m, Hur

060686; 35°18'59.5"N 127°42'26.1"E, alt. 1659 m, Hur 060955.

25. Cladonia mongolica Ahti

REMARKS — Characterized by dense squamules on the lower podetia and

usually with large apothecia, C. mongolica resembles C. scabriuscula but lacks

soredia near the podetia tips.

Also reported from China (Wei 1991), the species is rather rare in South

Korea and grows on soil or rock. Kwang-Hee Moon reported C. mongolica from

Mt. Sorak (Moon 1999), while we have found more specimens from different

localities in the country.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PRoOv., Mt. Duryon, 37°06'27.6"N

128°56'48.8"E, alt. 200 m, Hur 030318. GANGWON PRov., Mt. Hambaek, 37°11'48.6"N

128°55'06.5"E, alt. 1194 m, Hur 030686. CHUNGBUK PRov., Mt. Sobaek, 36°57'27.0"N

128°26'40.7"E, alt. 618 m, Hur 030711.

26. Cladonia ochrochlora Flérke

REMARKS — Characterized by farinose soredia, this species might be

confused with C. coniocraea, which differs by being sorediate all over the

podetia while C. ochrochlora has continuous cortex. It is also similar to the

yellowish C. bacilliformis but lacks usnic acid so the thallus color is greener.

Cladonia in South Korea... 417

Mainly distributed in temperate and boreal areas (Ahti 2000), C. ochrochlora

is not very common in South Korea, where it occurs mostly in the northern

part at altitudes above 1000 m.

SPECIMENS EXAMINED — SOUTH KOREA. JEONNAM PRov., Mt. Jiri, 35°18'07.4"N

127°34'00.1"E, alt. 1440 m, Hur 060266. JEoNBUK PRov., Mt. Deogyu, 35°51'05.3"N

127°44'55.4"E, alt. 1576 m, Hur 060506. GANGWON PrRov., Mt. Taebaek, 37°05'46.1"N

128°55'05.6"E, alt. 1572 m, Hur 070633. Mt. Gariwang, 37°24'05.0"N 128°32'39.5"E, alt.

937 m, Hur 080026, Hur 080029.

27. Cladonia peziziformis (With.) J.R. Laundon

REMARKS — Reported previously from South Korea (Wang et al. 2008),

C. peziziformis is characterized by short verruculose podetia and roundish

primary squamules. It is very similar to C. cariosa, which, however, contains

atranorin. It also resembles C. subcariosa, which has significantly elongated

primary squamules.

Primarily a temperate species that prefers mineral soil, C. peziziformis is

common in the eastern United States, Europe, Japan, and China (Ahti 2000). In

South Korea, only one specimen from Jeonnam province was found, growing

on soil.

SPECIMEN EXAMINED — SOUTH KOREA. JEONNAM PRov., Mt. Baeka, 35°09'08.3"N

127°09'45.2"E, alt. 550 m, Bae 080032.

28. Cladonia phyllophora Ehrh. ex Hoffm.

REMARKS — Characterized by whitish spots near the basal parts and quite

similar to C. gracilis subsp. turbinata, C. phyllophora has more irregular cups, a

decorticated area under the cups, and a base that usually blackens.

Distributed in circumpolar, boreal, and low arctic regions (Thomson 1984),

C. phyllophora is common in South Korea, usually on soil over rock at altitudes

above 1000 m,.

REPRESENTATIVE SPECIMENS (OF 10) EXAMINED — SOUTH KOREA. JEONNAM

Prov., Mt. Jiri, 35°20'05.0"N 127°42'49.4"E, alt. 1525 m, Hur 040394. GANGWON

Prov., Mt. Hambaek, 37°11'48.6"N 128°55'06.5"E, alt. 1194 m, Hur 030687. Mt. Sorak,

38°09'17.4"N, E128°26'25.6"E, alt. 1255 m, Hur 050300.

29. Cladonia pleurota (Flérke) Schaer.

REMARKS — Characterized by cups bearing granulose soredia, C. pleurota

may be mistaken for C. carneola when the pycnidia are brownish but differs in

having zeorin and isousnic acid.

Circumpolar in the arctic and the boreal forest (Thomson 1984), C. pleurota

is not common in South Korea and is restricted to the northeastern part, usually

on rock, soil, and at the base of the trees.

418 ... Wang, Joshi & Hur

SPECIMENS EXAMINED — SOUTH KOREA. GANGwon PRov., Mt. Gariwang,

37°24'05.0"N 128°32'39.5"E, alt. 937 m, Hur 080057. Mt. Taebaek, 37°12'35.7"N

128°55'13.0"E, alt. 1400 m, Hur 080297. GYEONGBUK PRov., Mt. Joryeong 37°48'27"N

128°03'32"E, alt. 500 m, Hur 080304.

30. Cladonia pyxidata (L.) Hoffm.

REMARKS — Characterized by bell-shaped cups covered inside and out by

round squamules, C. pyxidata it may be confused with C. chlorophaea, which

differs in usually wider and goblet shaped cups (that are sometimes narrow

when young) that have granulose soredia inside the cups.

This is a cosmopolitan species (Thomson 1984). In South Korea, it is not

widespread, growing on rock or soil, at the elevation lower than 1000 m.

SPECIMENS EXAMINED — SOUTH KOREA. CHUNGBUK PRov., Mt. Worak, 36°51'36.6"N

128°05'27.4"E, alt. 245 m, Hur 041158. GANGWoN PRov., Mt. Sorak, 38°08'53.5"N

128°20'05.3"E, alt. 1000 m, Hur 050275. JEONNAM PRov., Mt. Dalma, 34°22'45.2"N

126°35'11.6"E, alt. 456 m, Hur 050344.

31. Cladonia ramulosa (With.) J.R. Laundon

REMARKS — ‘The species is characterized by squamules on the lower part of

the podetia and partly decorticated. Morphologically, C. rei is very similar, but

C. ramulosa lacks powdery soredia on the upper podetia and has translucent

decorticated areas on the podetia. Cladonia scabriuscula, which might be

confused with C. ramulosa, lacks homosekikaic acid.

A widespread temperate to tropical species (Ahti 2000), it is rather common in South

Korea, usually on soil or rock and found at altitudes from sea level to above 1000 m.

REPRESENTATIVE SPECIMENS (OF 26) EXAMINED — SOUTH KOREA. JEONNAM PRov.,

Sorok Island, 35°48'11.2"N 129°18'47.3"E, alt. 60 m, Hur 030058. GANGWON PRov., Mt.

Taebaek, 37°06'15.5"N 128°56'06.2"E, alt. 1000 m, Hur 030392. Mt. Odae, 37°47'43.3"N

128°34'22.0"E, alt. 1240 m, Hur 040480. Jeyu Prov., Jeju Island, 33°34'00.1"N

126°45'44.4"E, alt. 10 m, Hur 090031.

32. Cladonia rangiferina (L.) Weber ex EH. Wigg. subsp. rangiferina

REMARKS — ‘The subspecies is easily recognizable by having grayish and

webby podetial surface, with branches curved and facing a same direction. It

is very similar to C. rangiferina subsp. grisea, but the branchlets of the latter

subspecies are mainly isotomic dichotomously branched, and the podetium is

thinner and smaller.

Circumpolar arctic to temperate in distribution (Thomson 1984), it is rather

common over rock or soil in South Korea at altitudes above 1000 m.

REPRESENTATIVE SPECIMENS (OF 30) EXAMINED — SOUTH KOREA. Jgju Prov.,

Mt. Halla, 33°57'32.6"N 126°29'17.5"E, alt. 1980 m, Hur 030357. GYEONGBUK PROV.,

Cheongryangsa Temple, 36°47'25.2"N 128°55'25.4"E, alt. 885 m, Hur 040102. JEONNAM

Prov., Mt. Jiri, 35°19'03.9"N 127°41'22.7"E, alt. 1660 m, Hur 040352. Gangwon Prov.,

Mt. Gariwang, 37°24'05.0"N 128°32'39.5"E, alt. 937 m, Hur 080059.

Cladonia in South Korea... 419

33. Cladonia rangiferina subsp. grisea Ahti

REMARKS — See under C. rangiferina subsp. rangiferina.

The subspecies is fairly common on Japanese mountains and also known

from Korea, Taiwan and China (Ahti 1961). In South Korea, common and

usually growing on rock under the open canopy.

REPRESENTATIVE SPECIMENS (OF 8) EXAMINED — SOUTH KOREA. JEONNAM PRov.,

Mt. Wolchul, 34°45'29.5"N 126°40'46.5"E, alt. 400 m, Hur 030115. Jeyu Prov., Jeju

Island, 33°21'37.7"N 126°32'40.3"E, alt. 1755 m, Hur 040838. GANGWON PrRov., Mt.

Sorak, 38°06'40.1"N 128°24'33.9"E, alt. 1335 m, Hur 041437. GYEONGBUK PRov., Mt.

Juwang, 36°23'47.7"N 129°08'50.5"E, alt. 270 m, Hur 050590.

34. Cladonia rappii var. exilior(Abbayes) Ahti Fic. 1 D

REMARKS — A new record for South Korea, this variety is characterized by

proliferating podetia and podetia cups that are usually very small. It is similar

to C. cervicornis, which has 4-6 proliferation tiers compared to the 2-3 tiers

usual for this variety.

Cladonia rappii is distributed in North America, Asia, Africa, and Melanesia

(Ahti 2000). In South Korea, only this variety is found and mainly occurs in the

southwestern part of Korea, usually growing on rocks.

SPECIMENS EXAMINED — SOUTH KOREA. CHUNGBUK PROv., Mt. Sobaek, 36°57'05.5"N

128°29'27.4"E, alt. 1375 m, Hur 030758. Mt. Joryeong, 36°49'00.9"N 128°02'53.7"E, alt.

784 m, Hur 061048. JEONNAM PRov., Mt. Jiri, 35°15'59.2"N 127°34'40.7"E, alt. 530 m,

Hur 040231; 35°19'54.5"N 127°43'04.7"E, alt. 1657 m, Hur 060701. GYEONGNAM PRov.,

Mt. Gaya, 35°48'36.9"N 128°08'20.9"E, alt. 746 m, Hur 060103.

35. Cladonia rei Schaer.

REMARKS — Characterized by largely decorticated podetia with small cups

and coarse soredia, C. rei resembles C. gracilis subsp. gracilis, except for the

presence of the soredia, and homosekikaic acid. It could be confused with

C. scabriuscula, which lacks cups on the apex and homosekikaic acid and has

squamulose podetia.

This is a cosmopolitan species (Galloway 2007). In South Korea, it is rather

rare and grows on soil or rock at altitudes below 1000 m.

SPECIMENS EXAMINED — SOUTH KOREA. GANGWoN PRov., Mt. Taebaek,

37°06'28.4"N 128°56'47.8"E, alt. 944 m, Hur 030626. JEONNAM PRov., Mt. Baeka,

34°32'33.1"N 126°55'46.7"E, alt. 334 m, Hur 050556.

36. Cladonia scabriuscula (Delise) Leight.

REMARKS — Characterized by dichotomously branched podetia and having

granulose soredia at the apex, C. scabriuscula may be confused with C. squamosa,

except for the different chemical compounds and absence of squamatic acid.

It also resembles C. furcata, which differs in having a continuous cortex and

lacking soredia on the podetia.

420 ... Wang, Joshi & Hur

Widespread worldwide in primarily temperate, oceanic regions (Ahti 2000),

C. scabriuscula is rather common and widespread throughout South Korea,

usually on rock or soil over rock.

REPRESENTATIVE SPECIMENS (OF 33) EXAMINED — SOUTH KOREA. GANGwon PRov.,

Mt. Odae, 37°47'43.3"N 128°34'22.0"E, alt. 1240 m, Hur 040482. JeonBuK PRov.,

Mt. Deogyu, 35°52'13.5"N 127°46'42.3"E, alt. 830 m, Hur 050061. JEONNAM PRov.,

Mt. Hulseok, 34°41'21.4"N 126°40'51.4"E, alt. 203 m, Hur 050476.

37. Cladonia squamosa (Scop.) Hoftm.

REMARKS — ‘This species is hard to identify when the squamules are greatly

reduced and appear to be granular or even soredia; key characters are possession

of squamatic acid and the lack of real soredia.

Widespread in temperate and colder regions in both northern and southern

hemispheres (Ahti 2000), C. squamosa is common but not widespread in South

Korea on soil or rock, sometimes at the base of a tree.

REPRESENTATIVE SPECIMENS (OF 9) EXAMINED — SOUTH KOREA. JEONNAM PRov.,

Mt. Wolchul, 36°56'50.5"N 128°29'51.2"E, alt. 400 m, Hur 030137. CHUNGBUK PROv.,

Mt. Sobaek, 36°57'11.2"N 128°28'37.2"E, alt. 1404 m, Hur 030792. GANGWON PRov.,

Mt. Sorak, 38°06'43.0"N 128°24'21.3"E, alt. 1375 m, Hur 041470.

38. Cladonia subconistea Asahina

REMARKS — Diagnosed by psoromic acid, C. subconistea resembles C. humilis

in having small podetia and cups but lacks the soredia and fumarprotocetraric

acid found in the latter.

Known only from east Asia (Harada et al. 2004, Hur et al. 2005, Wei 1991),

C. subconistea is very rare in South Korea, where it is found growing on rock,

usually at altitudes below 500 m.

SPECIMENS EXAMINED — SOUTH KOREA. GANGwon PRov., Mt. Chiak, 37°17'43.6"N

128°01'20.1"E, alt. 515 m, Hur 040562. GYEONGNAM PRov., Mt. Cheondae, 36°09'16.0"N

127°37'19.1"E, alt. 196 m, Hur 061145.

39. Cladonia symphycarpia (Florke) Fr.

REMARKS — Characterized by its short podetia (< 1 cm) and containing

norstictic acid, C. symphycarpia (frequently misspelled as “symphycarpa,’ see

Ahti 2000) usually lacks podetia, so that it must be recognized based on the

primary thallus alone.

Widespread in Eurasia and northern North America (Ahti 2000), the species is

rather rare in South Korea and is restricted to the Sorak Mountains, growing on soil

over rock.

SPECIMENS EXAMINED — SOUTH KOREA. GANGwoON PRov., Mt. Sorak, 38°07'37.6"N

128°27'48.5"E, alt. 1300 m, Hur 041581; 38°09'37.7"N 128°19'26.5"E, alt. 660 m, Hur

050256, Hur 050260.

Cladonia in South Korea... 421

40. Cladonia turgida Hoftm.

REMARKS — The species is characterized by having atranorin, a large primary

thallus, and distorted podetia with a well-developed cortex.

The species is circumpolar, arctic to north temperate (Thomson 1984). In

South Korea, not widespread, usually occurs in the southern part, on soil over

rock.

REPRESENTATIVE SPECIMENS (OF 11) EXAMINED — SOUTH KOREA. JEONNAM

Prov., Mt. Jiri, 35°15'59.2"N_ 127°34'40.7"E, alt. 530 m, Hur 040227; 35°19'40.9"N

127°44'15.0"E, alt. 1564 m, Hur 060687. Mt. Dalma, 34°22'45.4"N 126°35'13.6"E, allt.

445 m, Hur 050355. Jeyu Prov., Mt. Halla, 33°21'18.8"N 126°30'00.4"E, alt. 1492 m,

Hur 080778.

41. Cladonia uncialis (L.) FH. Wigg.

REMARKS — Some specimens from Japan are reported to contain

hypothamnolic acid (Yoshimura 1974) and squamatic acid is reported in

material from America (Thomson 1984), but neither chemotype is found in

Korean specimens. C. uncialis is unique in having yellowish and dichotomously

branched podetia and forming dense tufts. Quite similar to C. amaurocraea,

C. uncialis differs in having usually smaller podetia and usnic acid and lacking

cups and no barbatic acid.

The species is widespread, circumpolar, and arctic to temperate (Ahti 2000).

Rare in South Korea, only one specimen has been found from Gyeongnam

province, growing on soil.

SPECIMEN EXAMINED — SOUTH KOREA. GYEONGNAM PrRov., Mt. Gaya, 35°49'11.3"N

128°07'18.2"E, alt. 1440 m, Hur 040220.

Acknowledgments

The authors are grateful to Dr Laszl6 L6k6s (Botanical Department of the Hungarian

Natural History Museum) and Dr. Hiroshi Harada (Natural History Museum & Institute,

Chiba, Japan) for reviewing the manuscript and providing valuable comments. This work

was supported by a grant from The Korea National Research Resource Center Program

(Grant 2010-0000660) and Korean Forest Service Program (KNA 2010) through Korea

National Arboretum.

Literature cited

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Zool. Bot. Fenn. “Vanamo” 32(1): 1-160.

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Ahti T. 2000. Cladoniaceae. Flora Neotropica Monograph 78. New York, New York Botanical

Garden Press.

Asahina Y. 1970. Lichenologische Notizen (231-234). J. Jap. Bot. 45: 99-100.

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British Columbia, Canada. Mycotaxon 28: 99-110.

422 ... Wang, Joshi & Hur

Culberson CE. 1972. Improved conditions and new data for the identification of lichen products

by a standardized thin-layer chromatographic method. J. Chromatography 72: 113-125.

http://dx.doi.org/10.1016/0021-9673(72)80013-X

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University, Canberra. 103 p.

Fink B. 1904. Further notes on Cladonias III. Cladonia furcata and Cladonia crispata. Bryologist

4: 52-58.

Galloway DJ. 2007. Cladonia. Flora of New Zealand - lichens, 2"¢ ed., vol. 1, pp. 344-418. Manaaki

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Harada H, Okamoto T, Yoshimura I. 2004. A checklist of lichens and lichen-allies of Japan.

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Hur JS, Koh YJ, Harada H. 2005. A checklist of Korean lichens. Lichenology 4: 65-95.

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Laundon JR. 1984. The typification of Withering’s neglected lichens. Lichenologist 16: 211-239.

http://dx.doi.org/10.1017/S002428298400044X

Moon KH. 1999. Lichens of Mt. Sorak in Korea. J. Hattori Bot. Lab. 86: 187-220.

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http://dx.doi.org/10.2307/3243619

Sipman HJM. 1993. Lichens from Mount Kinabalu. Tropical Bryology 8: 285.

Stenroos S. 1989. Taxonomy of the Cladonia coccifera group. 1. Ann. Bot. Fenn. 26: 157-168.

Thomson JW. 1984. Cladonia. American arctic lichens 1. The macrolichens. pp 98-175. Columbia

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Wang XY, Hur H, Lee YM, Bae FE, Koh YJ, Hur JS. 2008. Cladonia peziziformis (lichenized

Ascomycota, Cladoniaceae) new to Korea. Mycobiology 36(3): 193-194. http://dx.doi.

org/10.4489/MYCO.2008.36.3.193

Wei JC. 1991. Cladina and Cladonia. An enumeration of lichens in China. pp. 62-82. International

Academic Publishers, Beijing, China.

White FJ, James PW. 1985. A revised guide to the microchemical techniques for the identification

of lichen substances. British Lichen Society Bulletin 57(Supplement): 1-41.

Yoshimura I. 1974. Cladoniaceae. Lichen flora of Japan in color. pp. 124-159. Hoikusha Publishing

Co., Ltd., Osaka, Japan.

MY COTAXON

http://dx.doi.org/10.5248/117.423

Volume 117, pp. 423-427 July-September 2011

First record of Resupinatus poriaeformis (Agaricomycetes)

from South America

GEORGEA SANTOS NOGUEIRA-MELO™, LEIF RYVARDEN?”

& TATIANA BAPTISTA GIBERTONI-

* Universidade Federal de Pernambuco, Departamento de Micologia,

Av. Nelson Chaves s/n, CEP 50760-420, Recife, PE, Brazil

*University of Oslo, Department of Botany, P. O. Box 1045, Blindern, N-0316, Oslo, Norway

*CORRESPONDENCE TO: georgeacomea@hotmail.com

ABSTRACT — Resupinatus poriaeformis is reported as new to South America. Description

of the species and a key to the accepted species of the genus reported to South America are

provided.

Key worps — Tricholomataceae, diversity

Introduction

Resupinatus was validated by Gray in 1821 and comprises about 15-20 valid

species (www.indexfungorum.org; http://www.cbs.knaw.nl; Kirk et al. 2008).

It is a cosmopolitan genus with small, resupinate, or pendent basidiomata, a

partially gelatinized context, and clamped hyphae (Singer 1962; Thorn 1986;

Pegler 1986). Recently Thorn et al. (2005) placed some cyphelloid fungi or

“reduced agarics” with cup-shaped basidiomata into Resupinatus based on

molecular and morphological studies.

Although some species of Agaricales have been registered in mangroves,

there are no reports of Resupinatus from this ecosystem (Hibbett & Binder

2001; Baltazar et al. 2009). However there are five reports of the genus from

South America: R. alboniger (Pat.) Singer 1978, R. applicatus (Batsch) Gray

1821, R. dealbatus (Berk.) Singer 1973, R. graminum (Singer) Singer 1973, and

R. hyalinus (Singer) Thorn et al. 2005. Of these, R. applicatus, R. dealbatus,

and R. hyalinus occur in Brazil (Singer 1943; 1973; 1989; Pegler 1997; Thorn &

Barron 1986). We present a description of a new occurrence and a key to the

Resupinatus species in South America.

424 ... Nogueira-Melo, Ryvarden & Gibertoni

Material & methods

Brazilian mangroves extend from 4°30'N to 28°30'S and despite the limited floristic

diversity vary significantly in plant growth form, species distribution patterns, and stand

structure (Schaeffer-Novelli et al. 1990). From March 2009 to March 2010, 36 field trips

were undertaken in four mangroves in Pernambuco state in northeast Brazil: Maria

Farinha(07°51'24.8"S34°50'32.7"W), Itamaraca (07°46'52.6"S34°52'53.3"W), Maracaipe

(08°32'22.8"S 35°00'29.1"W), and Rio Formoso (08°41'20.8"S 35°06'06.6" W).

After collection, the specimen was analyzed macro- (shape, color, hymenial surface)

and micromorphologically (hyphal system, presence/absence and measurements of

sterile structures and basidiospores). Microscopical observations were made from

slide preparations with 5% KOH, stained with 1% of aqueous phloxine, and Melzer’s

reagent (Ryvarden 1991). Color designation followed Watling (1969). The material was

incorporated to URM.

Scanning electron microscopy (SEM) was conducted at the Laboratdério de

Microscopia Eletrénica (DF/UFPE). Sections were removed from dried basidiomata

and mounted directly on aluminum stubs using carbon adhesive tabs. The fragments

were coated with 8-13 nm of gold using a Baltec SCD050 sputter coater and examined

with a JEOL JSM-5900 scanning electron microscope.

Taxonomy

Resupinatus poriaeformis (Pers.) Thorn, Moncalvo & Redhead,

Mycologia 97(5): 1148 (2006) FIGURE 1-2

= Peziza anomala var. poriaeformis Pers., Syn. meth. fung. 2: 656 (1801)

BASIDIOMATA annual, resupinate, cup-shaped, up to 2-3mm, gregarious,

subiculum tomentose to cottony, up to 0.1 mm, grayish (smoke grey 34 to

mouse grey 35), without reaction in KOH. HyMENIAL suRFACE smooth, dark

grey (drab 33 to mouse grey 35). HYPHAL SYSTEM monomitic; generative hyphae

hyaline, clamped, thin-walled. Basrp1a hyaline, clavate, guttulate, 25-30 x 5-7

um, 4-sterigmata. CysTIDIA absent. DENDROHYPHIDIA present. BASIDIOSPORES

globose, hyaline, thin-walled, smooth, inamyloid, acyanophilous, 4.5-5 um in

diam.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Mangrove of Maria Farinha, on dead

Rhizophora mangle L. (Rhizophoraceae), 28.V1.2009, G.S. Nogueira-Melo et al. NM006

(URM 82257).

REMARKS: Resupinatus poriaeformis is characterized by small grey cup-shaped

basidiomata and globose basidiospores (4.5-5 um diam., subglobose and < 6

ium in the literature). The species resembles R. hyalinus based on the basidioma

size and smooth hymenial surface. However, the basidiospores of R. hyalinus

are elliptical (6-6.5 x 3-3.4 um). The other species reported from South

America have larger basidiomata (0.5-4 cm) and lamellate hymenial surface.

Additionally, the basidiospores are ellipsoid (5.8-7.8 x 2.7-3.6 um) in R.

alboniger, cylindrical (5-8 x 2.5-3 um) in R. dealbatus, and subglobose (4.4-6

Resupinatus poriaeformis new to South America ... 425

> Aa

Fic. 1-2. Resupinatus poriaeformis. 1. Basidiospores, 2. Basidiomata. Scale bar = 2 mm.

x 4—4.8 um) in R. applicatus (Corner 1981; Ryvarden & Gilbertson 1993; Singer

1943, 1962; Thorn et al. 1986).

DISTRIBUTION — Ryvarden & Gilbertson (1993) and Bernicchia (2005)

report this species as cosmopolitan. Records are known from Honduras, Italy,

New Zealand, Puerto Rico, South Africa and Sweden (Cunningham 1963;

426 ... Nogueira-Melo, Ryvarden & Gibertoni

Ryvarden 1988; Ryvarden & Gilbertson 1993; Pennycook & Galloway 2004;

Thorn et al. 1986, 2006; Bernicchia 2005, O database).

Key to the Resupinatus species recorded from South America

ka; Hymenial:surface lainellates 4ih..-24ih- <a dinars dihaceg dined dh pee 2 Hheckw Al Ratt neeea'd 2

WoW skigencsonel ecithd cletecyns\e\ci Wor emir s ane Morin» Winmren® Wireren) Mirren} Wimertn ) erran® Wirar an Wien b

2a. Basidiomata reniform to flabelliform up to 2.5-4cm................ R. dealbatus

2b. Basidiomata cupulate to flabelliform up tol cm ........ eee eee ee eee eee 3

Sa ey sticiab elements taGkenGst.29 Aid50 Bite et WE UE et OEE Wh EE ta 8 R. graminum

5b, Gystidialelenients present, sdis.+adieegd need add nee dd beerdd barew dibeenaid ange A 4

4a. Basidiospores subglobose, 4-4.5X5 UM ........ cece eee eee eee R. applicatus

4a. Basidiospores ellipsoid to cylindrical, slightly curved 5.8-7.8 x 2.7-3.6 um

borie hia pak ones atteh we Mansy ituard cei aaea tine tearm howe ceittiers ucheark uelteaws R. alboniger

5a. Basidiospores globose to subglobose, 4.5 x 6 um diam. ........... R. poriaeformis

5b. Basidiospores ellipsoid, 6-6.5 x 3.3-4 UM ........ eee eee eee eee R. hyalinus

Acknowledgments

We would like to thank Dr. A. Bernicchia and Dr. R.G. Thorn for critically reviewing

the manuscript, Sérgio Santos for assistance with the SEM, and Leandro Agra for taking

the photo. Further, we acknowledge the CNPq for the master scholarship of GSNM, the

FACEPE (APQ 0444-2.03/08) and the Pés-Graduacao em Biologia de Fungos (UFPE)

for financial support.

Literature cited

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basidiomycetes (Basidiomycota) in mangroves. Mycotaxon 107: 221-224.

Bernicchia A. 2005. Polyporaceae s.l. Edizione Candusso, Alassio. 808 p.

Corner EJH. 1981. The agaric genus Lentinus, Panus and Pleurotus with particular reference to

Malaysian species. J. Cramer, Germany. 169 p.

Cunningham GH. 1963. The Thelephoraceae of Australia and New Zealand. New Zealand

Department of Scientific and Industrial Research, Bulletin 145. 359 p.

Hibbett DS, Binder M. 2001. Evolution of marine mushrooms. Biological Bulletin 201: 319-322.

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

Publishing, Wallingford. 771 p.

Pegler DN. 1986. Agaric flora of Sri Lanka. Kew Bulletin Additional Series 12.519 p.

Pegler DN. 1997. The agarics of Sao Paulo, Brazil. Royal Botanic Garden, Kew. 68 p.

Pennycook SR, Galloway DJ. 2004. Checklist of New Zealand “fungi”. 401-488, in: Introduction to

fungi of New Zealand. Fungi of New Zealand/Nga Harore 0 Aotearoa, vol. 1. Fungal Diversity

Research Series 14.

Ryvarden L. 1988. Type studies in the Polyporaceae 19. Species described by M.C. Cooke. Mycotaxon

31: 45-58.

Ryvarden L. 1991. Genera of polypores - nomenclature and taxonomy. Synopsis Fungorum 5:

1-363.

Ryvarden L, Gilbertson RL. 1993. European polypores, vol. 2. Fungiflora, Oslo. pp. 394-743.

Resupinatus poriaeformis new to South America ... 427

Schaeffer-Novelli Y, Citrén-Molero G, Adaime RR. 1990. Variability of mangrove ecosystems along

the Brazilian coast. Estuaries 13(2): 204-218.

Singer R. 1943. Type studies on Basidiomycetes II. Mycologia 32(2): 142-163.

Singer R. 1962. The Agaricales in modern taxonomy. 2™ ed. Germany J: Cramer. 915 p.

Singer R. 1973. Diagnoses fungorum novorum agaricalium III. Beihefte zur Sydowia 7. 106 p.

Singer R. 1989. New taxa and new combinations of Agaricales (Diagnoses fungorum novorum

agaricalium IV). Fieldiana Botany 21: 1-133.

Thorn RG, Barron GL. 1986. Nematoctonus and the tribe Resupinateae in Ontario, Canada.

Mycotaxon 25(2): 321-453.

Thorn RG, Moncalvo JM, Redhead SA, Lodge DJ, Martin MP. 2005. A new poroid species of

Resupinatus from Puerto Rico, with a reassessment of the cyphelloid genus Stigmatolemma.

Mycologia 97(5): 1140-1151. http://dx.doi.org/10.3852/mycologia.97.5.1140

Watling R. 1969. Colour identification chart. Her Majesty’s Stationary Office, Edinburgh.

MY COTAXON

http://dx.doi.org/10.5248/117.429

Volume 117, pp. 429-434 July-September 2011

New recombinations in Glomeromycota

FRITZ OEHL', GLADSTONE ALVES DA SILVA?,

BRUNO TOMIO GOTO? & EWALD SIEVERDING*4

'Federal Research Institute Agroscope Reckenholz-Tanikon ART, Organic Farming Systems,

Reckenholzstrasse 191, CH-8046 Ziirich, Switzerland

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

Av. Prof. Nelson Chaves s/n, Cidade Universitaria, 50670-420, Recife, PE, Brazil

*Departamento de Botanica, Ecologia e Zoologia, CB, Universidade Federal do Rio Grande

do Norte, Campus Universitario, 59072-970, Natal, RN, Brazil

‘Institute for Plant Production and Agroecology in the Tropics and Subtropics,

University of Hohenheim, Garbenstrasse 13, D-70599 Stuttgart, Germany

*CORRESPONDENCE TO: fritz.oehl@art.admin.ch

ABSTRACT — Combined morphological and molecular biological analyses have achieved

major advances in the taxonomy of fungal species in phylum Glomeromycota. In this study,

we analyzed which species might not yet be attributed to their correct genus, focusing on

Acaulospora myriocarpa, A. undulata, A. nicolsonii, and Scutellospora nodosa. Based on spore

wall structure and phylogenetic support, we recombined the taxa as Archaeospora myriocarpa,

Ar. undulata, Ambispora nicolsonii, and Cetraspora nodosa.

Key worps — molecular phylogeny, rDNA

Introduction

Combined morphological and molecular phylogenetic analyses have

thoroughly changed the classification of the phylum Glomeromycota (e.g.

Morton & Redecker 2001, Oehl et al. 2008, 2011a). Today, there are three

glomeromycotan classes, five orders, 14 families, and 29 genera (Oehl et al.

2011a,d). When species were attributed to these newly established higher taxa,

it was known that some species were incorrectly placed due to insufficient

available morphological or phylogenetical information (Oehl et al. 2008,

2011a,b,e). In this paper, we consider four of those species —Acaulospora

myriocarpa, Ac. undulata, Ac. nicolsonii, and Scutellospora nodosa. As a result

of new available information, we transfer these species to their proper genera

based on their morphological features.

430 ... Oehl & al.

Material & methods

Specimens

Holotype or isotype materials of Ac. myriocarpa (OSC, herbarium of Oregon State

University, Corvallis, U.S.A.), Ac. undulata (OSC), and Sc. nodosa (DPP, the Plant

Pathology herbarium, Academy of Agriculture, Szczecin, Poland) were analyzed. Type

material of Ac. nicolsonii (Walker et al. 1984) was either never deposited at OSC or lost

during shipping (Richard Halse & Joey Spatafora, curators of OSC, pers. communication);

therefore here we rely on the reinterpretation by Walker et al. (1984) of the original

species description and illustrations and on comparison of Ac. nicolsonii with the type

materials of Ambispora gerdemannii and Am. fennica in Walker et al. (2007).

Morphological analyses

Morphological descriptions of spores (including sporiferous saccules, sporogenous

cells, germination shields, and subcellular structures) are based on observations of

specimens mounted in polyvinyl alcohol-lactic acid-glycerol (PVLG) and in a mixture

of PVLG+Melzer’s reagent (Brundrett et al. 1994). Spore structure terminology follows

Goto & Maia (2006), Sieverding & Oehl (2006), Oehl et al. (2006), Spain et al. (2006),

Palenzuela et al. (2008, 2010, 2011), Goto et al. (2008) and Oehl et al. (201 1a-—c) for

acaulosporoid, ambisporoid, archaeosporoid, entrophosporoid, and gigasporoid spore

forming species.

Results

Acaulospora myriocarpa and Acaulospora undulata

Both Ac. myriocarpa and Ac. undulata form bi-walled spores laterally on

the neck of a sporiferous saccule. The outer wall is bi-layered and the inner

wall triple-layered. The staining reaction of the mycorrhizal structures is faint

to absent, and vesicle formation has been observed only infrequently in pure

cultures of the two species (Schenck et al. 1986) and then possibly due to

misinterpretation (Sieverding own observations). These features attribute both

species unequivocally to the genus Archaeospora J.B. Morton & D. Redecker

emend. Spain (Spain 2003).

Archaeospora myriocarpa (Spain, Sieverd. & N.C. Schenck) Oehl, G.A. Silva, B.T.

Goto & Sieverd., comb. nov.

MycoBank MB 561657

= Acaulospora myriocarpa Spain, Sieverd. & N.C. Schenck, Mycotaxon 25: 112. 1986.

Archaeospora undulata (Sieverd.) Sieverd., G.A. Silva, B.T. Goto & Oehl, comb. nov.

MycoBank MB 561662

= Acaulospora undulata Sieverd. Angew. Bot. 62: 373. 1988.

Acaulospora nicolsonii

Acaulospora nicolsonii regularly forms spores on the lateral branch (ice.

pedicel) of a sporiferous saccule. Its spores have three walls: an outer wall,

which during degradation shows cracking fissures known only for Ambispora

New recombinations in Glomeromycota ... 431

species, a middle wall, and an inner wall. Since the spore morphology exactly

matches the features of typical acaulosporoid spores in Ambispora C. Walker et

al. (= ‘acaulo-ambisporoid’ spores), we transfer the species to this genus (Spain

et al. 2006, Walker et al. 2007, 2008).

Ambispora nicolsonii (C. Walker, L.E. Reed & FE. Sanders) Oehl, G.A. Silva,

B.T. Goto & Sieverd., comb. nov.

MycoBank MB 561663

= Acaulospora nicolsonii C. Walker, L.E. Reed & FE. Sanders,

Trans Br. Mycol. Soc. 83: 360. 1984.

Scutellospora nodosa

New observations on the germination shield of the type material of Sc.

nodosa reveal that the shields are hyaline and multiply lobed, which is typical

for Racocetraceae species (e.g. Oehl et al. 2010). Since the species has triple-

walled spores (Blaszkowski 1991, Oehl et al. 2008), we transfer it to the genus

Cetraspora Oehl et al. (Racocetraceae).

Cetraspora nodosa (Blaszk.) Oehl, G.A. Silva, B.T. Goto & Sieverd., comb. nov.

MycoBank MB 561664

= Scutellospora nodosa Btaszk., Mycologia 83: 537. 1991.

Discussion

Archaeospora has now three species— two with smooth spore walls (Ar.

trappei, Ar. myriocarpa) and one with an ornamented (undulating) spore

surface (Ar. undulata).

Ambispora has now three species (Am. callosa, Am. fecundispora, Am.

leptoticha) for which only glomoid spore formation is known and two species

(Am. jimgerdemannii, Am. nicolsonii) for which only acaulosporoid spore

formation is known. All other Ambispora species have spore bimorphism (Am.

appendicula, Am. brasiliensis, Am. fennica, Am. gerdemannii, Am. granatensis)

(Spain et al. 2006, Walker 2008, Goto et al. 2008, Palenzuela et al. 2011).

Three major characteristics strongly support transfer of Ac. nicolsonii to

Ambispora: i) the presence of three walls (outer, middle, inner), ii) the clear

pedicel on the sporiferous saccule neck, referred to as a ‘stalk’ in the protologue

(Walker et al. 1984), and iii) outer spore walls that have the typical ‘cracking

fissures as known for outer walls of almost all known Ambispora species, e.g.

Am. appendicula, Am. gerdemannii, Am. fennica, and Am. brasiliensis (see

illustrations in Walker et al. 1984; Spain et al. 2006, Goto et al. 2008). The

absence of fracturing (i.e., ‘plate-like splitting’) of the middle wall layers in Am.

nicolsonii (Walker et al. 2007) might be the most important morphological

difference between it and Am. gerdemannii and Am. fennica, while the outer

walls in both Am. nicolsonii and Am. gerdemannii do not stain in Melzer’s

reagent but do in Am. fennica (Walker et al. 2007; Oehl, pers. observation).

432 ... Oehl & al.

The recent transfer of Am. brasiliensis to Acaulospora by Kriger et al. (2011)

cannot be accepted; the spore morphology of the Scottish material described

and illustrated as Acaulospora brasiliensis by Kriger et al. (2011) does not match

the morphology of Am. brasiliensis, and their sequenced material apparently

represents a true Acaulospora species rather than authentic Am. brasiliensis.

Kriger et al. (2011) unfortunately ignored the major characters elaborated by

Spain et al. (2006) for Ambispora species and detailed by Goto et al. (2008) for

Am. brasiliensis.

There have been doubts about the shield morphology of Cetraspora nodosa

(Scutellospora nodosa in Oehl et al. 2008), and sequences of the fungus deposited

in public databases during more than 10 years were apparently misidentified.

New combined morphological and molecular phylogenetic analyses, based

on identified sequences recently placed in the public databases, now show a

congruency between morphological and molecular data supporting C. nodosa

as closely related to C. helvetica, C. pellucida, and C. gilmorei (Oehl et al. 2010,

2011b) within the monophyletic genus Cetraspora.

Acknowledgments

We acknowledge the valuable comments and revisions of Felipe Wartchow

(Universidade Federal de Paraiba, Joao Pessoa, Brazil), Eduardo Furrazola (Instituto

de Ecologia y Sistematica, La Habana, Cuba), and Javier Palenzuela (EEZ, CSIC,

Granada, Spain) on the manuscript and appreciate the corrections by Shaun Pennycook,

Nomenclature Editor, and suggestions by Lorelei L. Norvell, Editor-in-Chief. This study

was supported by the Swiss National Science Foundation (SNSF, Project 315230_

130764/1), and by the Fundacao de Amparo a Ciéncia e Tecnologia do Estado de

Pernambuco (FACEPE) and the Universidade Federal de Pernambuco (UFPE) which

provided grants to EF. Oehl as ‘Visiting Professor’

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Guelph, Mycologue Publications, Guelph, Ontario.

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group molecularly distinct from Zygomycota. Mycotaxon 96: 129-132.

Goto BT, Maia LC, Oehl FE. 2008. Ambispora brasiliensis, a new ornamented species in the arbuscular

mycorrhiza forming Glomeromycetes. Mycotaxon 105: 11-18.

Kriiger M, Walker C, SchiiSler A. 2011. Acaulospora brasiliensis comb. nov. and Acaulospora alpina

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morphological characters. Mycologia 93: 181-195.

New recombinations in Glomeromycota ... 433

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MY COTAXON

http://dx.doi.org/10.5248/117.435

Volume 117, pp. 435-483 July-September 2011

Pouzarella (Agaricales, Entolomataceae) species from

New South Wales (Barrington Tops National Park)

and northeastern Queensland, Australia

Davin L. LARGENT**, SARAH E. BERGEMANN?’, GRIFFIN A. CUMMINGS’,

KATHRYN L. RYAN’, SANDRA E. ABELL-DAVIS? & SKYE MOORE‘

"Biological Sciences, Humboldt State University, 1 Harpst St, Arcata CA 95521 United States

Biology Department, Middle Tennessee State University,

PO Box 60, Murfreesboro TN 37132 United States

3School of Marine and Tropical Biology, Australian Tropical Herbarium,

James Cook University, PO Box 6811, Cairns QLD 4870 Australia

+Hunter Central Rivers Catchment Management Authority, Paterson NSW 2421 Australia

*CORRESPONDENCE TO: mrp@humboldtl.com

ABSTRACT —Ten Pouzarella species are described from New South Wales and northern

Queensland, Australia, of which eight (P albostrigosa, P. farinosa, P. fusca, P. lageniformis,

P. pamiae, PB. parvula, P. pilocystidiata and P. setiformis) are new to science and two

(P. debilis and P. lasia) are reported for the first time for Australia. Phylogenetic analyses

of the mitochondrial small subunit rRNA (mtSSU), two variable domains (D1, D2) of the

25-288 large subunit rDNA (LSU), and a portion of the second largest subunit of the RNA

polymerase (RPB2) gene support separation of these species into three clades. Morphological

characters common to these clades are described, and a key to species based on the characters

that are distinct between clades is included.

Key worps — Basidiomycota, phylogeny, mtSSU, LSU, RPB2

Introduction

The genus Pouzarella Mazzer (Entolomataceae, Agaricales) was first

monographed on a worldwide basis by Mazzer (1976). He proposed replacing

Pouzaromyces Pilat with Pouzarella because the type species of Pouzaromyces,

Agaricus fumosellus G. Winter, had no extant type material and the protologue

description had been interpreted as probably referring to a coprinaceous

species (Mazzer 1976, Noordeloos 1984, 1992; see Baroni et al. 2008 regarding

nomenclature). The name Pouzarella has since been accepted by many

researchers (Largent 1994, Karstedt et al. 2007, Baroni & Ortiz 2002, Baroni et

al. 2008, Horak 2008).

436 ... Largent & al.

Pouzarella combines the following features: typically mycenoid basidiomata

(often resembling a smallish Inocybe (Fr.) Fr.); strongly fibrillose, squamulose, or

hispid squamulose pilei; often radially strigose stipe bases; tramal hyphae with

externally incrusting pigments; pileipellis and stipitipellis hyphae with parietal

and/or externally incrusting pigments; a pileipellis (at least on the pileus center)

with erect to semi-erect hyphae composed of chains of cells; nodulose-angular,

heterodiametric basidiospores with 6-9 facets; and a hymenium frequently, but

not always, with dark brown contents or aborted basidia.

Because there have been no Pouzarella cited in Australian publications

on Entolomataceae Kotl. & Pouzar (Gates & Noordeloos 2007, Gates et al.

2009, Noordeloos & Gates 2009, May & Wood 1997, Grgurinovic 1997), we

provide the first reports and descriptions of Pouzarella in Australia. Of the ten

Pouzarella species covered, four are newly described from the Barrington Tops

National Park in central New South Wales, while four new species and two new

records are cited for the wet tropical rainforests of northeastern Queensland.

We also present a phylogeny based on a supermatrix of 1985 characters from

three partial gene regions: mitochondrial small subunit (mtSSU), 25-288

nuclear rDNA large subunit (LSU), and a portion of the gene encoding the

second largest subunit of RNA polymerase gene (RPB2). The sequence analyses

strongly support separation of the ten Australian taxa into three clades, each of

which can be diagnosed by micromorphological characters.

Materials & methods

Macromorphological and micromorphological features

Specimens were collected during February—April in 2009 and 2010 from different

sites within the Wet Tropics Bioregion in northeastern Queensland and in April 2010 in

the Barrington Tops National Park of central New South Wales. Basidiomata collected

in the field were stored in plastic containers for transport to the laboratory. Macroscopic

features were described from recently collected fresh materials. Field GPS coordinates

for each collection were taken using a Garmin GPSmap 60CSx. Colors were described

subjectively and coded according to Kornerup & Wanscher (1978), with color plate

designations noted in parentheses. Color plate abbreviations itemize page number,

column, and row (e.g., 8D-F5-6 = p. 8, columns D-F, rows 5-6.)

Dried specimens were examined microscopically with a trinocular research-grade

Nikon Labophot compound microscope fitted with bright field optics according to

Largent (1994: 1-3); basidiospore measurements follow Baroni & Lodge (1998: 681).

Digital microphotographs were made using a Nikon Coolpix 990 camera focused

through the trinocular head of the compound microscope.

All microscopic measurements were obtained using a GTCO Corporation Graphic

Digitizer, Model DP5A-111A, connected to a laptop computer. The software utilized,

Measure Me 101 v 1.0, was modified for use on a contemporary computer from the

BASIC program, Metrics5 (David Malloch, University of Toronto). The mathematical

factors determined using this program include: arithmetic mean (x,,) of basidiospore

Pouzarella (Australia) ... 437

length and width + standard deviation for n objects measured; quotient of basidiospore

length by width (E) indicated as a range for n objects measured; mean of E values (Q)

+ standard deviations. The sample size (n) equals the quotient (n = x/y) of the total

number of microscopic structures measured (x) and the number of basidiomata studied

(y).

All collections for New South Wales cited in ‘Additional collections examined’ are

deposited in The Plant Pathology Herbarium, Orange Agricultural Institute (DAR); the

Queensland collections were split with duplicates deposited in the Australian Tropical

Herbarium (CNS) and The Queensland Herbarium (BRI). All holotype and isotype

collections are deposited in the herbaria designated in the Latin descriptions using

acronyms from Holmgren et al. (1990).

DNA sequences and phylogenetic analyses

Lyophilized basidiomata were pulverized (4 mps for 20 sec) with glass beads (FastPrep

FP120 homogenizer, QBiogene, Carlsbad CA USA). DNA was extracted with 2 x

cetyl trimethylammonium bromide (CTAB) buffer, purified with phenol-chloroform-

isoamyl alcohol (25:24:1) bound to silica columns (GeneClean Turbo Columns, MP

Biomedicals, Solon OH USA), and washed with 70% ethanol (Baumgartner et al. 2010).

Portions of three loci were amplified using Polymerase Chain Reaction (PCR). PCR

primers used were MS1 and MS2 for mtSSU (White et al. 1990); Ctb6 and Tw13 for

nuclear rDNA LSU (variable domains D1 and D2; White et al. 1990); and for the RPB2

portion either pb2i6f and rpb2i7r (Co-David et al. 2009) or our Pouzarella-specific

primers, rpb2-PouF (5° to 3’ - GAA GGT CAA GCT TGT GGT C) and rpb2-PouR (5’ to 3’

- CAT GCT YGG ATG GAT TTC).

The mtSSU and LSU genes were amplified in 25 uL reactions containing 1x

GoTaq Flexi Buffer (Promega, Madison WI USA), 2 mM MgCl, 0.2 mM dNTPs, 500

nM forward and reverse primers, 0.2 mg/mL bovine serum albumin, 0.2 M betaine,

0.025 U Taq polymerase, and 1-2 uL of diluted template DNA. RPB2 was optimally

amplified according to the primer pair used. The rpb2-PouF and rpb2-PouR primer-

based amplifications utilized 25 uL reactions using the above protocol, except that

betaine was increased to 1 M. The Co-David et al. (2009) based RPB2 amplifications

increased the concentrations of primers rpb2-i6f and rpb2-i7r to 1 uM and the betaine

concentration to 1 M. PCR cycling conditions were as follows: mtSSU and LSU: 94°C

for 1 min, followed by 40 cycles of 94°C for 1 min, 50°C for 1 min, and 72°C for 1 min,

followed by a final extension of 72°C for 7 min. RPB2: EITHER the touchdown protocol

using primers rpb2-i6f and rpb2-i7r (Co-David et al. 2009) or [Pouzarella primers

rpb2-PouF, rpb2-PouR] 95°C for 5 min, followed by 40 cycles of 95°C for 30 sec, 50°C

or 52°C for 1 min 30 sec, and 72°C for 1 min, followed by a final extension of 72°C for

7 min. Unresolved sequences with overlapping chromatograms were resolved by sub-

cloning PCR products with the TOPO TA cloning kit (Invitrogen, Carlsbad CA USA)

according to Bergemann et al. (2009).

PCR products were cleaned using 1 uL of ExoSAP-IT (GE Healthcare, Pittsburgh,

PA USA). Sequencing reactions were carried out in 10 uL reactions containing 1 uL of

Applied Biosystems BigDye ver. 3.1 (Applied Biosystems, Carlsbad CA USA), 0.4 um

primer, 0.875 x Sequencing Buffer (Applied Biosystems, Carlsbad CA USA) and 1 uL

PCR water. Cycling parameters followed: 39 cycles of 96°C for 10 sec, 50°C for 5 sec,

438 ... Largent & al.

60°C for 4 min with an infinite hold at 15°C. Reactions were purified with 3 M NaOAc,

250 mM EDTA (pH 8) and ethanol by centrifugation at 2500 G. Reactions were washed

once with 70% EtOH and centrifuged at 2500 G for 15 min. Precipitated products were

stored at -20°C.

Forward and reverse sequences for each locus were generated on an Applied

Biosystems 3130xl Genetic Analyzer at Middle Tennessee State University. Sequence

contigs were edited in Sequencher 4.8 (GeneCodes Corp., Ann Arbor, MI USA) and

manually aligned in SE-Al (Rambaut 2002). Prior to tree estimation, the model selection

feature in TOPALi v. 2.5 (Milne et al. 2004) was used to test for the best-fit model of

nucleotide substitution using three selection criteria (AIC1 = Akaike information

criterion, AIC2 = second order correction and BIC = Bayesian information criterion).

Each criterion selected general time-reversible (GTR) as the best fitting substitution

model. All phylogenetic analyses were conducted on the combined dataset (mtSSU +

LSU + RPB2) using Lyophyllum decastes (Fr.) Singer, L. leucophaeatum (P. Karst.) P.

Karst., and Calocybe carnea (Bull.) Donk as outgroups and including two additional taxa

(Entoloma violaceovillosum Manim. & Noordel. and E. araneosum (Quél.) M.M. Moser)

known to have typical Pouzarella features (Co-David et al. 2009). Maximum likelihood

(ML) analyses implemented in TOPALi v. 2.5 (Milne et al. 2004) used a heuristic search

algorithm based on a rapid branch swapping operation (PhyML; Guindon & Gascuel

2003). The level of clade support was assessed using 100 bootstrap (BS) replicates.

Bayesian inferences (BI) were implemented in MrBayes ver. 3.1.2 (Ronquist &

Huelsenbeck 2003) using the GTR set to two runs of one million generations sampling

every 100 generations. After convergence, Bayesian posterior probabilities (BPP) were

determined after calculating a 50% majority rule consensus tree.

GenBank accession numbers for each sequence are listed at the end of the type

description for each species along with the habitat data and GPS co-ordinates of the

holotype collection. Sequences for all other collections are listed after the collection

number in the ‘Other collections examined’ section. GenBank accession numbers for

other species included in the phylogenetic analyses are as follows (mtSSU, LSU, and

RPB2, respectively): Entoloma violaceovillosum (GQ289345, GQ289205, GQ289273),

E. araneosum (GQ289293, GQ289153, GQ289225), Calocybe carnea (AF357097,

AF223178, DQ367432), Lyophyllum leucophaeatum (AF357101, AF223202, DQ367434),

and L. decastes (AF357136, AF042583, DQ367433).

Results

We obtained sequences for all three loci (mtSSU, LSU, RPB2) for 22

Pouzarella collections except that we omitted one RPB2 sequence (for P

parvula DL Largent 9901) after several amplification attempts failed to produce

PCR amplicons that could be sequenced. The dataset comprised 686 (mtSSU),

661 (LSU), and 638 (RPB2) aligned characters assembled into a supermatrix

consisting of 1985 characters. Of these, 122 mtSSU, 61 LSU, and 104 RPB2

characters were excluded at the 5’ or 3’ ends due to heterogeneity in the lengths

of the sequences obtained for different samples.

We conducted ML and BI analyses on the combined dataset. As the two tree

topologies were fully congruent, we present only the ML tree with corresponding

Pouzarella (Australia) ...439

BPP values included for comparison. Most branches are strongly supported

by BS values greater than 70% and BPP values greater than 0.95, although in

some cases, high levels of BPP support (0.99-1.00) were found when BS values

were less than 70%. The Australian collections formed three clades (PLATE 21):

Clade I formed a well-supported (BS >70, BPP = 1.00) group comprising five

species (Pouzarella lasia, P. albostrigosa, P. fusca, P. debilis, P. setiformis). Clade

II included two species (P. pamiae, P. farinosa) with high BPP (0.99) but low BS

(<70%) support. Clade III formed a well-supported group (BS >70, BPP = 1.00)

of three species (P. pilocystidiata, P. parvula, P. lageniformis). The clades can

be distinguished using micromorphological features (cheilocystidia, aborted

basidia, and pileocystidia; see Key). The Australian taxa are not resolved as

monophyletic in relation to the two European taxa included (E. araneosum and

E. violaceovillosum), though the grouping of the Australian clade III with these

species received low BS (but high BPP) support.

Taxonomy

1. Pouzarella pilocystidiata Largent & Skye Moore, sp. nov. PLATES 1-2

MycoBank MB 519567

Habitu Pouzarellae fulvostrigosae similis, sed centro pilei hispido-squamuloso, atrofusco,

cheilocystidiis versiformibus, incrustationibus externis in hyphis tramalibus distinctis

valdis, caulocystidiis copiosis, pseudocystidiis nullis atque superficie pilei argentea differt.

Type — Australia, New South Wales, central Hunter District, Barrington Tops National

Park, end of Blue Gum Loop Track, 32°08'31.7"S 151°30'56.4"E, 366.4 m, 22 April 2010,

DL Largent 9932 (holotype DAR), sequences: HQ876543 (mtSSU), HQ876521 (LSU),

HQ876500 (RBP2).

EryMoLocy — Derived from the Latin pilus (= hair) + cystidium, referring to the long

pileocystidia.

PitEus 8-19 mm broad, 2-7 mm high, convex or conic-campanulate and

suggestively umbonate becoming broadly convex and non-umbonate with

expansion, densely hispid-squamulose on the disc, hispid-squamulose to

squamulose near the disc, matted appressed squamulose to appressed fibrillose

elsewhere, with age the marginal area becomes orange-white to orange-gray (5-

6A-B2) as the trama is exposed through separation of the cuticle; squamules and

fibrils dark brown (5-7E-F5-6) on dark blond to brown (5E-F4) background, dull,

opaque and not hygrophanous; margin decurved at all times, not translucent,

even. TRAMA less than 1 mm thick, dark brown. TasTE farinaceous. ODOR

mild or somewhat pungent. LAMELLAE 3-8 mm long, 1-5 mm high, narrow

when young, moderately broad, broad or ventricose when mature, at first light

brownish-orange (6C4 camel) then orange-white to orange-gray or reddish-

white to orangish-gray (5-7A-B2-3), with age developing brown, irregularly

arranged spots, adnate to short decurrent or with a distinct decurrent tooth,

440 ... Largent & al.

close then subdistant; margin whitish and + fimbriate because of cystidia,

becoming eroded with age, bruising brownish with handling. LAMELLULAE 3

(2 short, 1 medium to medium-long) between lamellae. StrpE 23-45 mm long,

0.5-2 mm broad, equal, dark brown (5-7E-F4-6), darkens upon handling to

dark brownish-gray to grayish-brown (7F2-3), at first squamulose-hispid with

erect squamulose tips at the apex, elsewhere entirely covered with entangled

squamules and fibrils, with age, rain or handling the squamules and fibrils get

plastered on the surface and thus, the stipe appears fibrillose; stipe base with

orange cinnamon strigose hairs. BRUISING REACTION absent.

BASIDIOSPORES nodulose angular with 6-8 angles, 9.2-14.6(-15.1) x 5.4-8.7

(-9.7) um (x, = 11.6 + 1.1 x 7.2 + 0.7 um; E = 1.22-2.08; Q = 1.63 + 0.16;

n = 118/4). Basip1A 4-sterigmate, cylindro-clavate to clavate, moderately long

to long, 32.5-50.1 x 6.3-12.7 um (x, = 40.8 + 4.4 x 10.2 + 1.6 um; E = 2.90-6.27;

Q = 4.12 + 0.81; n = 34/4). CHEILOCYSTIDIA abundant, colorless, thin-walled,

versiform (clavate, acuminate, aciculate, ventricose-rostrate), most common

near the pileal margin and on the lamellulae, 39.5-82.8 x 4.3-24.0 um

(x = 57.8 + 11.4 x 13.5 + 4.3 um; E = 2.27-12.34; Q = 4.90 + 2.45; n = 24/4).

PLEUROCYSTIDIA absent. PSEUDOCYSTIDIA absent. ABORTED BasIpIA absent.

HYPHAE OF LAMELLAR TRAMA very long, similar in size to Inocephalus

(Noordel.) P.D. Orton, 164.1-748.0 x 3.8-26.8 um. PILEIPELLIS 45-400 um

deep, composed of entangled strands of thick-walled hyphae in chains of up to

10 cells, entirely erect and trichodermial when young, upon pileal expansion

remaining so in the center, semi-erect near the disc and repent at the margin.

PILEOCYSTIDIA cylindric, cylindro-clavate, narrowly obclavate, 28.1-171.3 x

6.8-20.1 um (x, = 79.7 + 25.7 x 11.3 + 3.6 um; E = 3.56-10.81; Q = 7.16 +

2.40; n = 19/3). HYPHAE OF THE PILEAL TRAMA Similar in length and width to

the lamellar trama. STIPITIPELLIS loosely entangled layer of hyphae, 45-205

uum deep. CauLocystip1A cylindric to cylindro-clavate 53.2-125.4 x 4.5-9.4

um. OLEIFEROUS HYPHAE absent. Liporp BODIES absent. PIGMENTATION with

strong, external incrustations in the pileal, lamellar and stipe tramas; parietal as

internal incrustations in the pileipellis and stipitipellis.

ECOLOGY AND DISTRIBUTION: Solitary or scattered in soil either amongst

leaf litter or under a log in amongst rocks, subtropical and temperate gallery

rainforests; central Hunter district, New South Wales.

ADDITIONAL COLLECTIONS EXAMINED —AUSTRALIA. New SouTH WALES, central

Hunter District, Barrington Tops National Park, Williams Day Use Area, end of blue

gum track, 32°09'03.0"S 151°31'28.2"E, 366.4 m, 8 April 2010, DL Largent 9848 (mtSSU

HQ876542, LSU HQ876520, RPB2 HQ876499); Pool of Reflections track, 32°08'29.5"S

151°30'55.0"E, 366.4 m, 20 April 2010, DL Largent 9921; Lion’s Rock Track, 32°09'10.2"S

151°3'38.6"E, 366.4 m, 26 April 2010, DL Largent 9949 (mtSSU HQ876544, LSU

HQ876522, RPB2 HQ876501).

CoMMENTS— Pouzarella pilocystidiata is diagnosed by a pileus with a

squamulose-hispid disc and elsewhere appressed squamulose to appressed

Pouzarella (Australia) ... 441

PLATE 1 - Pouzarella pilocystidiata. A: Basidiomata (DLL 9932, HOLOTYPE); B: Lamellae (DLL 9932);

C: Pileus surface (DLL 9932); D: Strigose stipe base, stipe surface (DLL 9949).

fibrillose, a tomentulose to densely hirsute stipe apex, a dark brown stipe that

bruises dark grayish-brown with a strigose base with light-colored to orange

cinnamon hairs, a farinaceous taste, basidiospores generally smaller than 12.0

442 ... Largent & al.

PLaTE 2 - Pouzarella pilocystidiata. A: Basidiospores (1000x) (DLL 9949); B: Basidia (400x)

(DLL 9959); C: Stipitipellis (100x) (DLL 9949); D: Cheilocystidia (400x) (DLL 9932, HOLOTYPE);

E: Pileipellis on and near disc (100x) (DLL 9921); F: Pileocystidia (400x) (DLL 9949).

Pouzarella (Australia) ... 443

x 7.5 um, basidia narrower than 11.0 um, pileipellis and stipitipellis <400 um

thick, and <175 um long cylindro-clavate pileocystidia and caulocystidia.

Pouzarella pilocystidiata morphologically resembles P. fulvostrigosa (Berk. &

Broome) Mazzer, which is distinguished by a densely silvery fibrillose mouse

gray pileus, lageniform cheilocystidia, minutely incrusted tramal hyphae,

scattered pseudocystidia, and caulocystidia (Noordeloos 1979).

Australian taxa similar to Pouzarella pilocystidiata

Pouzarella pilocystidiata shares with P. lageniformis and P. parvula nodulose

angular basidiospores (x, <13.5 um long), externally incrusted tramal

hyphae, abundant, thin-walled, versiform (rostrate, ventricose, lageniform)

cheilocystidia, long cylindro-clavate pileocystidia, and the absence of aborted

basidia. Morphological features unique to Pouzarella pilocystidiata include

the small hispid-squamulose dark brown/dark blond pileus, densely hirsute

stipe surface, farinaceous taste, basidiospore size, rare acuminate pileocystidia,

abundant cylindro-clavate caulocystidia, and lack of setiform pileocystidia.

Pouzarella lageniformis is differentiated by a 11 x 2 mm hispid-squamulose

orange-white/grayish orange pileus, entirely pruinose stipe surface, mild taste,

slightly larger basidiospores, acuminate pileocystidia, and lack of setiform

and aculeate thick-walled caulocystidia. Pouzarella parvula differs by its

smaller matted squamulose pileus, adnexed lamellae, glabrous stipe, larger

basidiospores, versiform (some rostrate ventricose) cheilocystidia, entangled

pileipellis, and lack of aborted basidia and caulocystidia.

2. Pouzarella lageniformis Largent & Skye Moore, sp. nov. PLATES 3-4

MycoBaAnk MB 519568

A speciebus congenericis omnibus ceteris pileo juventute omnino hispido-squamuloso,

superficie pilei omnino pallide griseo-brunnea squamulis aurantiaco-albis ornata,

apice stipitis pruinoso, alibi stipite fasciculis fibrillarum rarenter praedito, basidiosporis

10.4-13.8 x 6.6-8.7 um, mediane 12.06 x 7.55 um, basidiis mediane 36.9 x 11.9 um,

pileocystidiis clavatis acuminatis vel aculeatis, cheilocystidiis versiformibus aliquibus

rostrato-ventricosis, caulocystidiis aculeatis differt.

Type — Australia, New South Wales, central Hunter District, Barrington Tops National

Park, Williams Day Use Area, end of Blue Gum Track, 32°09'01.9"S 151°31'28.7"E, 365.5

m, 18 April 2010, DL Largent 9895 (holotype DAR), sequences: HQ876545 (mtSSU),

HQ876523 (LSU), HQ876502 (RBP2).

EryMoLocy — Derived from the Latin lagena (= flask) + forma (= shaped), referring to

the flask-shaped or rostrate ventricose cheilocystidia.

PitEus 11 mm broad, 2 mm high, broadly convex, when young entirely erect

hispid-squamulose with the squamules somewhat scattered, when mature

remaining so on and near the center, semi-erect near the center, repent

elsewhere, squamules orange-white (5-6A1-2), background light grayish-brown

(6C-D3), overall grayish-orange (6B-C2), dull, dry, opaque, not hygrophanous,

444 ... Largent & al.

PLATE 3 - Pouzarella lageniformis (DLL 9895, HOLOTYPE)

A: Basidiomata; B: Lamellae; C: Pileus surface; D: Stipe surface.

Pouzarella (Australia) ... 445

striate with striae colored as the background. Marain decurved, fringed in

places. TasTE and Opor indistinct. LAMELLAE 5 mm long, 1.5 mm high, broad

and ventricose, short-decurrent, orange-gray (5A2), light-colored when dried,

subdistant, margin suggestively and minutely serrulate, whitish. LAMELLULAE

3 (2 short, 1 medium to medium long) between lamellae. Stipe 33 mm long,

1 mm broad at apex, 0.75 mm broad at base, slightly tapered, entirely pruinose

with the pruinae scattered and orange-white to orange-gray on a grayish-

brown (6D3) background, dull, hollow; stipe base densely pruinose. BRUISING

REACTION absent.

BASIDIOSPORES angular-nodulose with 6-8 angles, heterodiametric in

profile view, 6-angled and isodiametric in polar view, 10.4-13.8 x 6.6-8.7 um

(x = 12.14 1.0 x 7.6 + 0.6 um; E = 1.38-1.87; Q = 1.6 + 0.14 (heterodiametric);

n = 29/1). Basip1a 4-sterigmate, sterigmata 0.7-3.5 um long, base of basidia

3.3-5.8 um wide, relatively small, full of small droplets, 29.3-43.6 x 9.9-14.2

ym (x_ = 36.9 + 3.3 x 11.9 + 1.4 um; E = 2.56-4.32; Q = 3.13 + 0.48; n = 13/1).

CHEILOCYSTIDIA abundant, scattered, versiform (long ventricose-rostrate,

acuminate), 47.5-133.6 x 8.7-28.8 um (x, = 85.6 + 22.4 x 17.9 + 6.0 um;;

B.°=%3.16-10:75;" 0 = 5,24. +92282-n =-10/1). PLEUROGCYSTIDIA” absent:

PSEUDOCYSTIDIA absent. ABORTED BASIDIA absent. HYPHAE OF LAMELLAR

trama relatively long 230.2-418.7 x 11.3-31.4 um. PILEIPELLIS 117-333 um

deep, composed of loosely entangled, long strands of hyphae with chains of

3-5 cells, individual cells relatively long, entirely erect to semi-erect and

trichodermial in the center when young, upon pileal expansion remaining

so in the center, becoming semi-erect near the center and repent elsewhere.

PILEOCysTIDIA very long, clavate, aculeate, or acuminate, 105.4-192.7 x

10.7-20.2 um. HYPHAE OF THE PILEAL TRAMA 194.6-451.3 x 9.4-17.1 um.

STIPITIPELLIS 30-150 um deep, composed of scattered strands of thick-walled

hyphae. CauLocystip1A aculeate, thick-walled, 20.5-144.4 x 5.0-9.2 um.

PIGMENTATION externally incrusted on the tramal hyphae of the pileus, stipe,

lamellae, parietal with internal incrustations in the pileipellis and stipitipellis.

ECOLOGY AND DISTRIBUTION: Solitary amongst leaf litter, subtropical gallery

rainforest; known only from type locality.

ComMMENTS— Pouzarella lageniformis is diagnosed by an entirely hispid

squamulose young pileus with orange-white squamules on a light grayish

brown background, stipe with a pruinose apex and rare fibrillose bundles

elsewhere, basidiospores averaging 12.1 x 7.6 um, basidia generally >11.0 um

long, clavate, acuminate, or aculeate pileocystidia, versiform (some rostrate

ventricose) cheilocystidia, and aculeate caulocystidia.

See P. pilocystidiata for comparisons with Australian and other taxa related

to P. lageniformis.

446 ... Largent & al.

B- . “=

PiatE 4 - Pouzarella lageniformis (DLL 9895, HOLOTYPE). A: Basidiospores (1000x); B: Basidia

and narrow rostrate-ventricose cheilocystidia (400x); C: Pileocystidia and pileipellis (180x);

D: Cheilocystidia (100x); E: Stipitipellis (100x); F: Thick-walled caulocystidium (400x).

Pouzarella (Australia) ... 447

3. Pouzarella parvula Largent & Skye Moore, sp. nov. PLATES 5-6

MycoBank MB 519569

A speciebus congenericis omnibus ceteris pileo parvo, 6 mm lato, 2 mm alto, implexo,

squamuloso, lamellis adnexis, stipite glabro, basidiosporis mediane 13.23 x 7.7

um, cheilocystidiis versiformibus aliquibus rostrato-ventricosis, pileipelle implexa,

pseudocystidiis caulocystidiisque nullis differt.

Type — Australia, New South Wales, Central Hunter District, Barrington Tops National

Park, Williams River Day Use Area, end of Blue Gum Track, 32°09'13.6"S 151°31'39.7"E,

355.1 m, 18 April 2010, DL Largent 9901(holotype DAR), sequences: HQ876546

(mtSSU), HQ876524 (LSU), HQ876543 (RBP2).

Erymotocy — Derived from the Latin parvulus (= very little), referring to the

very small pileus.

PILEUS 6 mm broad and 2 mm high, convex, entirely densely matted squamulose,

overall orange-gray to grayish-orange (5B2-3) dull, even, dry, opaque and not

hygrophanous; margin decurved, fringed, eroded and + crenulate. TASTE AND

Opor indistinct. LAMELLAE 2 mm long, 1 mm deep, narrow, adnexed, orange-

gray (5B2), distant; margin smooth but lighter in color; 1 lamellula between

lamellae. Stipe 20 mm long, <1 mm broad, equal, dull, pruinose at the apex,

glabrous elsewhere, orange-gray to grayish-orange (5B2), hollow and very

fragile; stipe base not pruinose. BRUISING REACTION absent.

BASIDIOSPORES nodulose angular with 6-8(-9) angles, heterodiametric in

profile view, 6-angled and isodiametric in polar view, 10.5-15.0 x 6.3-8.6 um,

(x, = 13.2 + 1.0 x 7.7 + 0.6 um; E = 1.35-2.04; Q = 1.72 + 0.14; n = 29/1).

Basip1a 4-sterigmate, cylindro-clavate to clavate 32.5-40.4 x 10.8-14.8

um, (x, = 36.3 + 2.6 x 12.8 + 1.5 um; E = 2.47-3.37; Q = 2.87 + 0.29; n =

9/1). CHEILOCYSTIDIA solitary to scattered, versiform (aciculate, acuminate,

obclavate, rostrate-ventricose), 36.0-88.6 x 14.8-21.3 um. PLEUROCYSTIDIA

absent. PsEUDOCysTIDIA absent. ABORTED Basrip1a absent. PILEIPELLIS

100-200 um deep, an entangled layer of hyphae composed of chains of thick

walled cells, individual cells long and slender. PiteocystTip1< cylindro-clavate,

aciculate, aculeate to distinctly setiform 57.6-106.0 x 6.1-13.2 um, (x, = 74.8

+ 16.8 x 8.6 + 2.2 um; E = 7.01-16.00; Q = 9.04 + 2.6; n = 11/1). STIPITIPELLIS

a cutis from near the apex to the base of the stipe; apex overlooked because

of stipe size. CAULOCYSTIDIA absent where studied. PIGMENTATION distinctly

externally incrusted on the hyphae of the pileal stipe and lamellar trama;

parietal and in the form of internal incrustations in the pileipellis.

ECOLOGY AND DISTRIBUTION: Solitary in soil amongst mosses on a bank cut

in a wet sclerophyll subtropical gallery rainforest; known only from the type

locality.

ComMENTS— Pouzarella parvula is differentiated by a small (6 x 2 mm) matted

squamulose orange-gray pileus, adnexed lamellae, stipe pruinose at the apex

and glabrous elsewhere, basidiospores averaging 13.2 x 7.7 um, versiform

448 ... Largent & al.

PLATE 5 — Pouzarella parvula (DLL 9901, HOLOTYPE). A: Field photo of basidiomata;

B: Lamellae and lamellulae; C: Pileal surface and eroded pileal margin.

Pouzarella (Australia) ... 449

: a e's |

PLATE 6 - Pouzarella parvula (DLL 9901, HOLOTYPE). A: Basidiospores (1000x); B: Basidia

and cheilocystidium (400x); C: Pileipellis near pileus margin (100x); D: Cheilocystidia (400x);

E: Setiform pileocystidia (400x); F: Setiform pileocystidium (400x).

450 ... Largent & al.

(some rostrate ventricose) cheilocystidia, entangled pileipellis with setiform

pileocystidia, and lack of aborted basidia and caulocystidia.

Pouzarella minuta (E. Horak) E. Horak from New Zealand (Horak 2008),

Pouzaromyces napaliensis Desjardin & T.J. Baroni from the Hawaiian Islands,

and Entoloma testaceostrigosum Manim. & Noordel. from India have nearly

the same size pileus as P parvula. Pouzarella minuta differs by a smaller

(2-4 x 1 mm) stipe densely covered with brownish or white fibrils and much

larger basidiospores (16-20 x 10-12 um) (Horak 1980, 2008). Entoloma

testaceostrigosum and Pouzaromyces napaliensis both possess setiform

pileocystidia and caulocystidia and lack cheilocystidia (Desjardin & Baroni

1991, Manimohan et al. 2006).

See P. pilocystidiata for comparisons with Australian species with a

resemblance to P. parvula.

4, Pouzarella pamiae Largent, sp. nov. PLATES 7-8

MycoBank MB 519570

Habitu Pouzarellae squamifoliae similis, sed coloribus basidiomatum fulvis, pileo latiore

usque ad 3-8 mm, stipite minore, 12-25 x 0.5-0.75 mm atque contusis atro-fuscis differt.

TypE — AUSTRALIA, Queensland, Cook Region, Mossman Gorge National Park,

16°28'17.6"S 145°19'51.7"E, 84.4 m, 18 March 2010, DL Largent 9794 (holotype BRI,

isotype CNS), sequences: HQ876539 (mtSSU), HQ876517 (LSU), HQ876496 (RBP2).

Erymo.ocy —in honor of Pamela Largent, the collector and inspiration for the study of

entolomatoid fungi in northeastern Queensland.

PILEus 3-8 mm broad, 2-4.5 mm high, at first conic to conic-campanulate and

densely erect squamulose-hispid over the entire surface expanding to convex

and eventually convex-campanulate, remaining erect squamulose hispid on

the center but becoming appressed squamulose from the margin first and

then to near the center, at first entirely reddish-golden to brownish-orange

(6-7C-D5-7) darkening to medium brown (6-7E-F5-6 teak brown to burnt

umber) then dark brown (6-7E-F6-7 burnt umber to dark brown), with

maturity remaining darker on the disc but lightening to yellowish-white to

orangish-white (4-5A2-3), squamule tips lose color and separate showing

the whitish-yellow background, not umbonate, opaque, dry and dull; margin

decurved and fringed with squamule tips then some eroded as the tips collapse

with age. TASTE indistinct. OpDor indistinct to pungent. LAMELLAE 3-4 mm

long, 1.25-1.5 mm high, pale orange (6A3) then brownish (6E5 sunburn) with

basidiospore maturity, narrow and then moderately broad, subdistant to nearly

distant; no bruising reaction observed in the margin, fimbriate to serrulate and

light-colored when lamellulae are mature. LAMELLULAE nearly non-existent

with at most one lamellula between lamellae. Stipe 12-25 mm long, 0.5 mm

broad at the apex, 0.75 mm broad at base, equal to slightly clavate, at first covered

by a yellowish-white to orangish-white (4-6A2-3) dense layer of entangled

Pouzarella (Australia) ... 451

PLaTE 7 - Pouzarella pamiae (DLL 9794, HOLOTYPE). A: Basidiomata, note densely hispid

squamulose pileal surface and stipe base colored like pileus; B: Pileus with fringed pileal margin

(left), lamellae (center) and young pileus surface (right).

452 ... Largent & al.

fibrils, with maturity the fibrils darken to colors similar to those of the pileus,

at times the consistency and color of the fibrils at the base are identical to those

on the pileal surface, eventually the stipe surface becomes slightly roughened as

the fibrils collapse onto the surface and then the stipe darkens to medium dark

brown (6E6); the surface discolors to dark brown (near 6F6) when bruised;

stipe base strigose or matted. BRUISING REACTION absent.

BASIDIOSPORES 7-9-angled, nodulose-angular, subisodiametric to hetero-

diametric in profile view, 5-6-angled and isodiametric in polar view, 10.0-15.9

(-17.3) x 7.1-11.2 um (x, = 13.07 + 1.3 x 8.5 + 0.8 um; E = 1.18-1.85; Q = 1.53

+ 0.16; n = 82/2). Basrp1A 4-sterigmate, clavate, tapered, filled with a droplet,

31.8-48.3 x 11.5-15.3 um (x, = 42.3 + 5.6 x 13.6 + 1.2 um; E = 2.17-3.78;

Q=3.11 +0.4; n= 11/1). CHErLocystip1a abundant, broadly clavate to broadly

acuminate, with a faint brownish cytoplasmic pigment, + thick-walled, 47.1-77.6

x 10.1-26.2 um and located as terminal cells arising from tramal hyphae with at

least one subterminal cell inflated. PLEUROCysTID1A absent. PSEUDOCYSTIDIA

absent. ABORTED BASIDIA inconspicuous or rare. HYPHAE OF THE LAMELLAR

TRAMA 79.3-240.3 x 5.9-15.3 um. PILEIPELLIS 111-551 um deep, an entangled

layer of long, rather slender hyphae with cells + in chains, in young specimens

entirely erect and trichodermoid, in older specimens trichodermoid only in

the center, semi-erect towards the margin and repent at the margin; distinctly

thick-walled in the apical 1-5 cells. PiLEocystip1a cylindric, cylindro-clavate,

broadly clavate, or broadly obclavate, thick-walled and with parietal pigment,

26.0-78.7 x 6.6-13.0 um (x, = 46.8 x 9.8 um; E = 2.87-7.52; Q = 4.8; n = 9/1).

HYPHAE OF THE PILEAL TRAMA Same size and pigmentation as lamellar trama.

STIPITIPELLIS similar to the pileipellis except with the addition of clusters of

hymenial elements at the apex; CAuLocystTrp1A similar to the pileocystidia,

36.1-79.1 x 8.4-19.9 um. PIGMENTATION externally incrusted in the trama,

parietal and faintly externally incrusted in the pileipellis and stipitipellis.

ECOLOGY AND DISTRIBUTION: scattered in tan-colored soil beneath and

protected by an overhanging boulder in a complex mesophyll vine forest on

well-drained alluvium (Mossman Gorge National Park) or solitary in hard-

packed soil (Danbulla National Park, Lake Euramoo).

ADDITIONAL COLLECTIONS EXAMINED —AUSTRALIA. QUEENSLAND, Cook Region,

Mossman Gorge National Park, 16°28'17.2"S 145°19'50.2"E, 86.6 m, 26 March 2010,

DL Largent 9834 (mtSSU HQ876541, LSU HQ876519, RPB2 HQ876498); Danbulla

National Park, Lake Euramoo Track, 17°09'42.3"S 145°37'45.6"E, 762.6 m, 21 March

2010, DL Largent 9808 (mtSSU HQ876540, LSU HQ876518, RPB2 HQ876497).

ComMMENTS— Pouzarella pamiae is diagnosed by its 3-8 mm broad densely

squamulose-hispid pileus with brownish-orange squamules, long (12-25 mm)

narrow stipe that bruises dark brown and has a base covered with squamules

identical to those on the pileus, thick-walled acuminate to broadly clavate

cheilocystidia, and basidiospores averaging 13.1 x 8.6 um.

Pouzarella (Australia) ... 453

PiaTE 8 — Pouzarella pamiae. A: Basidiospores (1000x) (DLL 9808); B: Basidia (400x) (DLL

9794, HOLOTYPE); C: Pileocystidia (100x) (DLL 9794); D: Thick-walled cheilocystidia with

brownish pigment (400x) (DLL 9794); E: Cylindro-clavate pileocystidia (180x) (DLL 9794);

F: Stipitipellis and chains of hyphae with terminal caulocystidia (100x) (DLL 9794).

454 ... Largent & al.

Other species with a brownish-orange, densely squamulose-hispid pileus

and stipe include P fulvolanata (Berk. & Broome) Mazzer and P. myoderma

(Berk. & Broome) TJ. Baroni (both from Ceylon), P squamifolia (Murrill)

Mazzer from Jamaica, P. sepiaceobasalis (E. Horak) T.J. Baroni from Argentina,

and P ferreri T.J. Baroni et al. from the Dominican Republic, Brazil, and Costa

Rica.

Mazzer (1976) described P. fulvolanata with yellowish to tawny colors and

basidiospores in the same size range (12-16 x 7-8 um) as P. pamiae, but it

can be differentiated by subpyriform cheilocystidia and larger pileus (13 mm

broad) and stipe (32 mm x 2 mm). Noting their similar stature, coloration,

and pileus and stipe surfaces, Mazzer (1976) differentiated P fulvolanata and

P. squamifolia based on distribution and stipe color: P squamifolia, described

from Jamaica, has a pallid stipe with ferruginous hairs whereas P._ fulvolanata,

described from Ceylon, has tawny hairs on the stipe.

The basidiospores, basidia, cheilocystidia, pileipellis, pileocystidia,

stipitipellis and caulocystidia of P pamiae are identical to those described for the

Jamaican holotype of P. squamifolia (Mazzer 1976, Baroni et al. 2008). However,

P. squamifolia is differentiated by ferruginous colors (tawny in P pamiae), wider

(10 mm broad) pileus, larger (40 x 1.5 mm) stipe, and the absence of a bruising

reaction.

Pouzarella sepiaceobasalis is differentiated from P pamiae by the longer

basidiospores (15-19 um) and longer stipe (60 mm) with a base that gradually

becomes blue-black to black (Baroni et al. 2008).

Pouzarella ferreri is separated by a more reddish coloration, longer (<60 mm)

stipe, larger (~16.2 x 10.7 um) basidiospores, globose to sphaeropedunculate

cheilocystidia, and black staining reactions (Baroni et al. 2008).

On the basis of type studies, Pegler (1977, 1986) synonymised A. fulvolanatus

Berk. & Broome under A. myodermus Berk. & Broome, for which he reported

basidiospore measurements of 9-13(-15) x 5.5-7.0 um (average 10.5 + 1.0 x

6.5 + 0.34 um). He could not verify the presence of cheilocystidia, commenting,

“Lamella-edge not revived.” Based on further type studies, Horak (1980)

synonymised both A. myodermus and A. fulvolanatus under Agaricus lasius, for

which he reported basidiospores measuring 10.5-13.5 x 6-8 um, cheilocystidia

absent, and robust basidiomata with <20 mm broad pilei and 15-50 x 1-3

mm stipes. Therefore, P pamiae is differentiated from both Pouzaromyces

myodermus sensu Pegler (1977) and Pouzaromyces lasius sensu Horak (1980)

by larger basidiospores, abundant clavate to acuminate cheilocystidia, and a

smaller stature.

Australian taxa related to Pouzarella pamiae

The ML phylogeny weakly (BS <70) supports the clade including P. pamiae

and P. farinosa. Both species possess + thick-walled pigmented napiform to

Pouzarella (Australia) ...455

clavate cheilocystidia and versiform pileocystidia. The two species are easily

distinguished by basidiospore size (~15.5 x 10.0 um in P. farinosa) and coloration

(orange-gray squamules on a grayish-brown background in P farinosa).

5. Pouzarella farinosa Largent & Skye Moore, sp. nov. PLATES 9-10

MycoBank MB 519571

Habitu Entolomate dysthali (subg. Pouzarellae) similis, sed sapore farinaceo, squamulis

pilei aurantiaco-griseis, cheilocystidiis latioribus, 21.1-51.1 x 16.7-30.9 um differt.

Type — Australia, New South Wales, Cook Region, Central Hunter District, Barrington

Tops National Park, Pool of Reflections Track, 32°08'15.8"S 151°30'38.2"E, 727.6 m,

22 April 2010, DL Largent 9934 (holotype DAR), sequences: HQ876538 (mtSSU),

HQ876516 (LSU), HQ876495 (RBP2).

Erymo.ocy — derived from the Latin farinosus (= mealy), referring to the mealy or

farinaceous taste.

Piteus 8-14 mm broad, 2-6 mm high, convex to broadly convex, in some

with a slight, broad umbo, at first entirely squamulose-hispid with densely

packed upright tufts of orange-gray (5B2) squamules on a grayish-brown (5E3)

background, the overall color light grayish-brown (5C-D3), upon expansion

remaining so in or near the center, becoming appressed squamulose and then

appressed fibrillose towards the margin as the squamules get pressed onto the

surface, dry, dull, opaque, not hygrophanous but becoming striate with age;

margin decurved then slightly uplifted, striate with age, the striae grayish-brown

(5D-E3). Taste slightly to distinctly farinaceous. ODorR indistinct. LAMELLAE

3-6 mm long, 1-2 mm high, narrow, adnate with a distinct subdecurrent tooth,

subdistant to distant, at first brownish-gray (5C3) then orange-white (6A2)

with maturation of the basidiospores; margin whitish, smooth when very

young eventually becoming minutely serrulate to fimbriate because of cystidia.

STIPE 30-54 mm long, 1-1.25 mm broad, equal, fragile, dull, at first entirely

orange-gray (5B2) and covered with dense visible tangled masses of fibrils and

squamules that are more abundant towards the basal part of the stipe, with

maturity and handling, the overall color grayish-brown (6C3-4) with areas

darkening to dark brown (5F6) with age as the fibrils and squamules become

appressed to the stipe surface. BRUISING REACTION absent.

BASIDIOSPORES 6-8-angled and nodulose angular in profile view, 6-angled in

polar view, 12.0-19.8 x 7.9-11.9 um (x, =15.5+1.8 x 10.0 £0.84 um; E= 1.2-1.8;

Q = 1.56 + 0.14 (heterodiametric); n = 63/2). Bastp1a 4-sterigmate, broadly

clavate to clavate, tapered, 4-sterigmate with the sterigma long, slender and

nearly needle-like with large droplets, 40.6-53.1 x 13.6-20.3 um (x, = 47.2 +

3.5 x 16.3 + 1.7 um; E = 2.29-3.49; Q = 2.29 + 0.34; n = 18/2). CHEILOCYSTIDIA

abundant and forming a sterile layer mostly near the pileal margin, nearly

globose, vesiculate, or nearly napiform, + thick-walled, 25.1-51.7 x 16.7-30.9

456 ... Largent & al.

PLaTE 9 — Pouzarella farinosa.

A: Basidiomata and stipe surface (DLL 9934, HOLOTYPE); B: Pileus surface (DLL 9900).

Pouzarella (Australia) ...457

uum, (x_ = 37.3 + 8.2 x 23.0 + 4.3 um; E = 1.14-2.61; Q = 1.65 + 0.40; n = 14/1).

PLEUROCYSTIDIA absent. PsEuDOCyYsTIDIA absent. ABORTED BAsIpIA absent.

HYPHAE OF THE LAMELLAR TRAMA Subparallel with relatively short cells,

narrow and slender but more broad than those in the pileal trama, 4.4-87.4 x

3.6-10.7 um. PILEIPELLIS a densely entangled layer of hyphae in chains of 4-6

cells, + palisade and erect in the center and near the center, semi-erect near

the center and repent near the margin. PILEOCysTIDIA relatively short, clavate,

broadly clavate or nearly napiform on or near the center, 21.4-54.1 x 8.6-20.4

um (x, = 42.4 411.5 x 14.2 + 4.3 um; E = 1.35-4.87; Q = 3.24 + 1.24;n =9/1).

HYPHAE OF THE PILEUS TRAMA Similar to but more narrow than those in the

lamellar trama. STIPITIPELLIS 76-233 um, similar to the pileipellis, composed

of abundant clusters of hyphae in 5-6-celled chains. CAuLocystTb1A clavate,

cylindro-clavate, narrow to broadly aculeate, or even acuminate, longer than

the pileocystidia, 32.4—76.6 x 9.1-20.6 um, (x, = 55.8 + 14.4 x 14.3 + 3.0 um;

E = 1.57-8.13; Q = 4.17 + 1.74; n = 14/2). OLEIFEROUS HYPHAE absent. LIPOID

BODIES absent in the trama; large globules present in the basidia. PIGMENTATION

strongly incrusted on the pileal, lamellar and stipe trama; parietal and internally

incrusted in the stipitipellis and pileipellis.

ECOLOGY AND DISTRIBUTION: Solitary to gregarious in soil, often amongst

rocks and ferns in warm temperate and subtropical gallery rainforests, Central

Hunter District, New South Wales, Australia.

ADDITIONAL COLLECTIONS EXAMINED — AUSTRALIA. New SoutTH WALEs, Cook

Region, Barrington Tops National Park, Williams Day Use Area, end of Blue Gum Loop

Track, 32°09'03.3"S 151°31'28.2"E, 366.9 m, 18 April 2010, DL Largent 9900 (mtSSU

HQ876537, LSU HQ876515, RPB2 HQ876494).

ComMMENTS— Pouzarella farinosa is diagnosed by an entirely squamulose-

hispid pileus covered by densely packed upright tufts of orange -gray squamules,

basidiospores measuring 12.0-19.8 x 7.9-11.9 um, nearly globose, vesiculate,

or nearly napiform cheilocystidia, and the lack of aborted basidia.

The new species resembles Entoloma dysthales (Peck) Sacc. f. dysthales in

basidiospore size, cheilocystidia, and overall stature and coloration. However,

E. dysthales lacks the orange-gray colors of the P farinosa squamules and

possesses a mild (not farinaceous) taste and odor, abundant aborted basidia,

and longer, narrower (21-82 x 8-21 um) cheilocystidia. Our collections are

similar to Gates 2265, the proposed holotype for a new species to be described

in a forthcoming book on the entolomatoid fungi in Tasmania (Gates pers.

comm.), but differs in the absence of aborted basidia, cheilocystidial shape

(clavate to broadly clavate or obovoid in Gates 2265), and farinaceous taste.

See P pamiae for comparisons with taxa closely related to P farinosa.

458 ... Largent & al.

PLATE 10 - Pouzarella farinosa. A: Basidiospores (1000x) (DLL 9900); B: Pileipellis (400) (DLL

9934, HOLOTYPE); C: Basidia (400x) (DLL 9900); D: Thick-walled cheilocystidia with brownish

pigment (400x) (DLL 9934); E: Pileocystidia (400x) (DLL 9900); F: Acuminate caulocystidia

(400x) (DLL 9934).

Pouzarella (Australia) ... 459

6. Pouzarella setiformis Largent & Abell-Davis, sp. nov. PLATES 11-12

MycoBank MB 519586

Habitu Pouzaromyceti transito similis, sed pileo juventute hispido squamuloso,

squamulis primo pallido-coloratis tum cito atro-griseo-brunneo, postremo aetate

pallide vel mediocriter brunneolo-griseo decolorantibus, peileocystidiis caulocystidiisque

multicellularibus setiformibus differt.

Type — Australia, Queensland, Cook Region, Danbulla National Park, Lake Euramoo

Track, 17°09'41.1"S 145°37'46.9"E, 714.5 m, 21 March 2010, DL Largent 9809 (holotype

BRI, isotype CNS), sequences: HQ876547 (mtSSU), HQ876525 (LSU), HQ876503

(RBP2).

EryMoLocy — derived from the Latin seta (= bristle) + forma (= shape), for setiform

pileocystidia and caulocystidia.

Piteus 8.5-12.5 mm broad, 3-5.5 mm high, convex then broadly convex,

scurfy with + erect light colored (6C3) brownish-orange + hispid squamules

on the disc, tomentulose to tomentose elsewhere, overall dark grayish-brown

(6-7E-F2-3) fading to light or medium brownish-gray (6D3) with age, dull,

opaque, dry, even when young becoming + sulcate and striate with age, not

hygrophanous; margin decurved then plane, minutely hispid. Taste and ODoR

indistinct. LAMELLAE 3-5.5 mm long, 1-2 mmhigh, narrowto moderately broad,

adnexed to adnate, subdistant to distant, dark brownish-gray to dark grayish-

brown at first (7-8E-F2-3); margin smooth and concolorous. LAMELLULAE 1-3

(2 short, 1 medium long) between lamellae. StrpE 15-25 mm long, 0.75-1.0

mm broad, equal, stiff but hollow, fragile and breaks easily, with abundant stiff,

light brownish-gray (6D3 or 7C3) + tomentulose pointed hairs and squamules

at the apex, pruinose to fibrillose dark brownish-gray (6-7E-F3); elsewhere,

dull, suggestively strigose at the base. BRUISING REACTION absent.

BASIDIOSPORES nodulose-angular with 6-8 angles, heterodiametric in profile

view, isodiametric and 6-angled in polar view, 8.2-13.9 x 4.8-7.9 um (x, = 10.7

+ 1.0 x 6.3 + 0.7 um; E = 1.4-2.5; Q = 1.73 + 0.24 (heterodiametric); n = 56/2).

BASIDIA 25.7-32.9 x 8.5-12.2 um; E = 2.11-3.85; Q = 2.95. CHEILOCYSTIDIA

absent. PsEUDOCysTIDIA absent. ABORTED Basip1A often with dark brown

contents in 3% KOH, aborted sterigma and/or aborted basidiospores often

present, 26.2-41.0 x 6.8-10.7 um. Subhymenium composed of thin and

short hyphae. HyPHAE OF LAMELLAR TRAMA long and broad, 58.3-253.7

x 3.1-26.6 um. PILEIPELLIS a dense compactly entangled chain of 3-5 cells,

55-144 um deep in the center and towards the margin, at the margin with tufts

of hyphae, 93-150 um deep. Prteocystip1A cylindro-clavate in the center to

near the margin, 23.9-64.9 x 8.8-17.4 um (x, = 40.5 x 12.8 um; E = 2.20-4.07;

Q = 3.13; n = 9/1), at the margin thick-walled and setiform, 42.4-69.9 x

4.5-5.7 um. STIPITIPELLIS at the apex with tufts of heavily incrusted hyphae,

74-190 um deep, entangled hyphae between the tufts. CAULOCYSTIDIA

aculeate to aciculate, clear and + setiform when in abundant tufts, 47.0-121.0 x

460 ... Largent & al.

PLATE 11 - Pouzarella setiformis (DLL 9809, HOLOTYPE). A: Basidiomata;

B: Pileipellis (100x); C: Basidiospores (1000x); D: Basidia and pseudocystidia (400x).

Pouzarella (Australia) ... 461

¥ io »*

PLaTE 12 - Pouzarella setiformis. A: Pileocystidia with parietal pigmentation (400x) (DLL 9809,

HOLOTYPE); B: Multicellular setiform pileocystidia (400x) (DLL 9809); C: External incrustations

on tramal hyphae of pileus (400x) (DLL 9810); D: Multicellular setiforma caulocystidia (100x)

(DLL 9809).

4.8-7.9 um; E = 8.44-17.50; Q = 10.89; clear and acuminate when not in tufts,

17.4-61.3 x 5.5-11.6 um; E = 1.93-6.15; Q = 3.13. PIGMENTATION heavily

incrusted on the outer walls of the tramal hyphae of the stipe, pileus and

lamellae; parietal and heavily incrusted on inner and outer cell walls of the

pileipellis and stipitipellis excluding the pileocystidia and caulocystidia; thick-

walled, not incrusted, + suggestively parietal but mostly clear in the setiform

pileocystidia and caulocystidia.

ECOLOGY AND DISTRIBUTION: Solitary or scattered in hard packed soil in a

complex mesophyll vine forest; northeastern Queensland.

ADDITIONAL COLLECTIONS EXAMINED — AUSTRALIA. QUEENSLAND, Cook Region,

Danbulla National Park, Lake Euramoo track, 17°09'41.6"S 145°37'40.7"E, 714.5 m, 21

March 2010, DL Largent 9810 (mtSSU HQ876548, LSU HQ876526, RPB2 HQ876504).

462 ... Largent & al.

CoMMENTS— Pouzarella setiformis is diagnosed by a pileus that is hispid-

squamulose on disc and elsewhere tomentulose to tomentose with hairs that

are initially light colored but quickly become dark gray brown before fading

to light or medium brownish-gray with age and a young even margin that

becomes + sulcate and striate with age, nodulose-angular, heterodiametric

basidiospores averaging 10.7 x 6.3 um, the absence of cheilocystidia, and the

presence of aborted basidia, setiform pileocystidia and caulocystidia, and

encrusted pileipellis and stipitipellis hyphae.

Pouzaromyces napaliensis from the Hawaiian Islands, P. transitus E. Horak

from Papua New Guinea, and Entoloma testaceostrigosum from India have

nodulose-angular heterodiametric basidiospores and setiform caulocystidia and

lack cheilocystidia, features common to P. setiformis. Pouzaromyces napaliensis

is distinguished by its slightly larger (11.2-15.4 x 7-9.6 um) basidiospores,

uniformly beige pileus and lamellae, smaller size (2-5 mm broad pileus, 3-4

mm long stipe), and weakly incrusted tramal hyphae (Desjardin & Baroni

1991). Pouzaromyces transitus differs in its black to soot-brown basidiomata and

pileus hairs and single-celled setiform caulocystidia (Horak 1980). Entoloma

testaceostrigosum is separated by its larger (12.5-17.0 x 7-10 um) basidiospores

and absence of encrusted pileipellis and stipitipellis hyphae (Manimohan et al.

2006).

Australian taxa related to Pouzarella setiformis

Pouzarella setiformis, P. albostrigosa, P. fusca, P. lasia and P. debilis form

a monophyletic group that share a brown squamulose hispid pileus, stipe

surface composed of a layer of entangled brownish fibrils, pileipellis composed

of a chain of cells with clavate to broadly clavate pileocystidia, more or less

stipitipellis with occasional entirely clavate to broadly cylindro-clavate

caulocystidia, basidiospores averaging 10.0-12.0 um long, aborted basidia that

become reddish-brown in 3% KOH, and absence of cheilocystidia.

Within this group P setiformis differs from P debilis, P. fusca, P. albostrigosa,

and P lasia by a stipe surface with abundant stiff, + tomentulose pointed hairs

that is pruinose to fibrillose except for the apex squamules and a stipitipellis and

pileipellis with scattered to abundant setiform caulocystidia and pileocystidia

(most evident at the margin). The other taxa in this clade (P. debilis, P. fusca,

P. albostrigosa and P. lasia) possess instead a stipe surface that is densely wooly

or covered with abundant loose fibrils to nearly glabrous and non-setiform

cystidia.

Pouzarella debilis, P. fusca, and P. albostrigosa share a convex to broadly

convex pileus, a relatively shorter (typically <20 mm) stipe, aborted basidia

typically imbedded in the hymenium, and caulocystidia, pileocystidia, and

many subterminal cells with strong parietal pigmentation comprising coarse

bands and obvious internal incrustations. Pouzarella fusca produces a young

Pouzarella (Australia) ... 463

pileus that is opaque and black to dark gray brown, basidiospores averaging

10.0 x 7.3 um, and caulocystidia of 2 types (some thick-walled and pigmented).

Pouzarella albostrigosa has light colored squamules when young, basidiospores

averaging 11.5 x 7.7 um, and caulocystidia of 2 types (some thick-walled and

pigmented). Pouzarella debilis combines light-colored squamules when young,

basidiospores averaging 10.4 x 6.3 um, and thin-walled + colorless caulocystidia,

similar in shape to the pileocystidia.

Lastly, P lasia produces a conic to campanulate pileus (at least when young),

a stouter <50 mm long stipe, aborted basidia that distinctly project beyond

the hymenium, caulocystidia and pileocystidia with obscure bands and minute

internal incrustations, and absence of setiform pileocystidia and caulocystidia.

7. Pouzarella debilis (Corner & E. Horak) Largent & Abell-Davis, comb. nov.

MycoBank MB 519572 PLATES 13-14

= Pouzaromyces debilis Corner & E. Horak, Beih. Nova Hedwigia 65: 42 (1980).

PiLEus 5 mm broad, 4 mm high, convex, at first entirely opaque and hispid-

squamulose with the squamules off-white (5A-B1-2) and erect in the center,

suberect elsewhere, base color of the center grayish-brown (6F3-4) and

elsewhere slightly lighter grayish-brown (6E-F3-4) when faded remaining so

on the disc but becoming appressed squamulose and striate to the margin,

elsewhere with the base color light grayish-brown (near 6C3-4) and the striae

darker grayish-brown (6E3-4), dry, dull, not hygrophanous, even; margin

decurved, minutely fringed and crenulate at the very edge of the margin.

TASTE and Opor unknown. LAMELLAE 7.5 mm long, uncinate with a short

decurrent tooth, close to subdistant, narrow (1 mm deep); margin smooth and

concolorous. LAMELLULAE very short, commonly 1, infrequently 3 (all short)

between 2 lamellae. StrpE 15 mm long, 0.5 mm broad, equal, on the surface

with loosely entangled fibrils or scattered pruina, obvious at the apex and base,

inconspicuous elsewhere, surface fibrils similarly colored to those of the pileus,

base color grayish-orange (6B-C3-4) and upon handling the base bruises a

grayish-brown (6E3-4), hollow and fragile because of size. BRUISING REACTION

absent.

Basip1ospores nodulose-angular with 6-8 angles, heterodiametric in

profile view, isodiametric and 6-angled in polar view, 8.7-12.0 x 5.0-7.4 um,

(x = 10.44 + 0.8 x 6.3 + 0.6 um; E = 1.36-2.27; Q = 1.67 + 0.22; n = 29/1).

BasIDIA 4-sterigmate, clavate, tapered, 27.6-36.5 x 8.9-12.8 um, (x, = 31.8 +

2.7 x 11.1 + 1.2 um; E = 2.33-3.49; Q = 2.88 + 0; n = 10/1). CHEILOCYSTIDIA

absent. PSEUDOCYSTIDIA absent. ABORTED BASIDIA abundant on the gill edge

and face, colored reddish-brown in 3% KOH, often with 1-2 sterigma and

aborted basidiospores, slightly projected beyond the hymenium, 33.6-46.8

x 7.4-11.6 um. HYPHAE OF LAMELLAR TRAMA Subparallel, moderately long,

464 ... Largent & al.

PLATE 13 — Pouzarella debilis (DLL 9784). A-B: Basidiomata.

narrow to fairly wide, 184.9396.8 x 6.2-30.8 um. PILEIPELLIS with scattered,

entangled hyphae composed of chains of 3-5 cells, subpellis without inflated

cells, hyphae similar to pileal trama. PitEocystip14 broadly cylindro-clavate

35.1-58.4 x 9.7-20.0 um (E = 2.49-4.10; Q = 3.2). STIPITIPELLIs with scattered

areas of clustered hyphae similar to those of the pileipellis, 29-103 um long, a

cutis between the clusters. CAULOCYSTIDIA similar to pileocystidia, 20.7-54.4 x

7.0-16.5 um (x, = 34.9 + 10.6 x 11.6 + 2.7 um; E = 1.82-6.41; Q = 3.14 + 1.31;

n = 16/1). PIGMENTATION moderately externally incrusted in the trama of the

pileus, stipe and lamellae; parietal in the form of minute to moderate internal

incrustations in the hyphae of the stipitipellis and pileipellis.

ECOLOGY AND DISTRIBUTION: Solitary in dark crumbly soil on an open

hillside in complex mesophyll vine forest of cloudy to moist highlands on

granite; Mt. Hypipamee National Park. Previously reported from Papua New

Guinea and the Solomon Islands.

Pouzarella (Australia) ... 465

PiaTE 14 — Pouzarella debilis (DLL 9784). A: Basidiospores (1000x); B: Basidium (black arrow),

pseudocystidium (white arrow) (400x); C: Caulocystidia (400x); D: Pileipellis hyphae with clavate

to broadly cylindro-clavate pileocystidia (400x).

COLLECTIONS EXAMINED — AUSTRALIA. QUEENSLAND, Cook Region Mt. Hypipamee

National Park, Falls Loop Track, 17°25'36.9"S 145°29'07.9"E, 714.5 m, 16 March 2010,

DL Largent 9784 (holotype BRI, isotype CNS; mtSSU HQ876550, LSU HQ876528,

RPB2 HQ876506).

ComMENTS— Pouzarella debilis is diagnosed by small fragile basidiomata

(pileus 5 mm broad, stipe 15 x 0.5 mm), a hispid-squamulose pileus at first

with off-white squamules that are erect on the disc and elsewhere suberect over

a medium to dark grayish-brown base color and in age becoming striate from

the margin inwards and fading to light grayish-brown, aborted basidia that

project slightly beyond the hymenium, absence of cheilocystidia, caulocystidia

resembling the pileocystidia, and basidiospores averaging 10.4 x 6.3 um,

(Q = 1.67). The macro- and micro-morphological features of the Australian

466 ... Largent & al.

collection P debilis are nearly identical with those described for Pouzaromyces

debilis from Papua New Guinea and the Solomon Islands (Horak 1980).

Pouzarella domingensis T.J. Baroni from the Dominican Republic and

P. dunstervillei (Dennis) Mazzer have moderate-sized basidiospores and lack

cheilocystidia, features also noted for P debilis. Pouzarella domingensis differs

in the lack of light-colored squamules in young pilei, inflated subpellis hyphae,

and larger basidiospores averaging 11.0 x 7.6 um (Baroni et al. 2008), while

P. dunstervillei (which also lacks light-colored squamules in young pilei) has

a nearly glabrous pileus and stipe (Mazzer 1976). Pouzarella pulverea (Rea)

Mazzer from Great Britain and the Netherlands, Entoloma romagnesii Noordel.

from France, and E. lomapadum Manim. et al. from India also have similarly

sized basidiospores, but all three lack light-colored squamules in the young

pilei and possess thin walled cheilocystidia (Mazzer 1976, Manimohan et al.

1995, Noordeloos 1979, 2004).

See P. setiformis for comparisons with other Australian entolomatoid fungi

similar to P. debilis.

8. Pouzarella fusca Largent & Abell-Davis, sp. nov. PLATES 15-16

MycoBank MB 519573

Habitu Entolomate romagnesii (subg. Pouzarellae) similis, sed pileo infuscate atro-

brunneo, pigmento extrorsum in hyphis ad partes pileipellis stipitipellisque pertinentibus

incrustato, pigmentatione perietali pileocystidiorum cauocystidiorumque, cheilocystidiis

nullis differt.

Type — Australia. Queensland, Cook Region, Dinden National Park, 17°02'14"S

145°36'4"E, 727.6 m, 24 February 2009, DL Largent 9623 (holotype BRI, isotype CNS),

sequences: HQ876549 (mtSSU), HQ876527 (LSU), HQ876505) (RBP2).

EryMoLocy — derived from Latin fuscus (= dark brown), referring to the dark blackish-

brown coloration.

Piteus 3.5-10 mm broad, 1.5-3.0 mm high, consistently convex, minutely

but entirely squamulose, erect in the center, imbricate squamulose elsewhere,

becoming appressed squamulose at the very margin, black to a very dark gray

brown (not matching any color to Kornerup & Wanscher (1978), fading to dark

gray-brown (6F3) except a medium gray-brown (6E-F3) on the margin, dull,

opaque, not hygrophanous, not striate; trama dark gray to black, less than 0.5

mm thick above the stipe; margin slightly incurved to decurved, entire. TASTE

mild. ODOR somewhat pungent. LAMELLAE 2.5-3.5 mm long, 0.5-1.0 mm high,

concolorous with the pileus, adnate, subdistant and narrow; margin smooth

and concolorous. LAMELLULAE 3 (2 short, 1 medium) between lamellae. STIPE

8-17 mm long, 0.8-2.0 mm broad, equal, dark gray (6F1), minutely and entirely

squamulose to covered with loose fibrils; basal tomentum scarce, strigose base

present. BRUISING REACTION absent.

Pouzarella (Australia) ... 467

“

PiaTE 15 - Pouzarella fusca (DLL 9623, HOLOTYPE).

A: Basidiomata; B: Basidiospores (1000x); C: Basidia with basidiospores (400x).

468 ... Largent & al.

Basipiospores distinctly angular to slightly nodulose, in profile view

typically 6-angled, often 7-8-angled, nearly or definitely heterodiametric,

8.6-11.5 x 6.1-8.7 um (x, = 10.0 + 0.8 x 7.3 + 6 um; E = 1.22-1.54; Q = 1.38

+ 0.09 (heterodiametric); n = 28/1). Basip1a 4-sterigmate, sterigma rather

long, 4.2-5.5 um; clavate, origin unusual for entolomatoid fungi, originating

from a basal cell in the subhymenium, 33.9-47.2 x 8.9-12.6 um (E = 2.8-4.7;

Q = 3.83); base 3.0-6.5 um. PLEUROCYSTIDIA absent. PSEUDOCYSTIDIA absent.

ABORTED BASIDIA abundant, but scattered on the lamellae, clavate, often

with aborted sterigma, not projecting beyond the hymenium and filled with

opaque contents that stain reddish-brown in 3% KOH, 35.0-44.6 x 8.7-13 um

(x_ = 40.9 x 11.0 um; E = 3.1-4.6; Q = 3.78; n = 7/1). HYPHAE OF THE LAMELLAR

TRAMA rather short, 29.6-179.0 x 4.7-11.0 um. PILEIPELLIS 240-400 um deep,

composed of chains of cells, erect or suberect, on or near the disc and towards

the margin, becoming repent at the margin. PILEOCysTip1A clavate to broadly

cylindric, 36.5-118.9 x 5.2-21.4 um (E = 2.6-6.1; Q = 5.04). HYPHAE OF THE

PILEAL TRAMA rather short, 45.5-105.4 x 2.1-8.0 um. STIPITIPELLIs similar

to the pileipellis and composed of solitary caulocystidia. CAULOCYSTIDIA

of two types; cylindro-clavate and the terminal cells comprised of a loosely

entangled hyphae, 20.8-73.2 x 4.7-10.9 um (E = 3.6-8.1; Q = 5.53), and clavate

to broadly napiform and the terminal cells comprised of solitary hyphal walls

and in clusters with the hyphae originating from a common base, 22.3-57.7 x

9.1-14.3 um (E = 1.7-5.4; Q = 2.71). HYPHAE OF THE STIPE TRAMA Parallel to

subparallel, 60.5-336.3 x 4.5-12.5 um. OLEIFEROUS HYPHAE rare in the trama

of the stipe, pileus and lamellae. Lipoip BoptrEs present in the basidia, absent

in the aborted basidia and tramal hyphae of the pileus, lamellae and stipe.

PIGMENTATION externally incrusting the hyphae of the basal portion of the

pileipellis and stipitipellis and distinct and strong on the tramal hyphae of the

lamellae, pileus and stipe; suggestively externally incrusting the outer hyphal

and more obviously parietal on the inside the hyphal walls of the terminal

2-3 cells of the pileipellis and stipitipellis, including the pileocystidia and the

caulocystidia.

ECOLOGY AND DISTRIBUTION: Scattered or connate in dark soil on high

banks along road just above bridge at end of Davies Creek Road in a simple to

complex notophyll vine forest, Dinden National Park.

ComMMENTS— Pouzarella fusca is diagnosed by the dark gray to dark blackish-

brown to black coloration when young, a pileus that is initially squamulose

on the disc and imbricate squamulose near the margin, only slightly nodulose

PLATE 16 - Pouzarella fusca (DLL 9623, HOLOTYPE). A: Pseudocystidium, origin from incrusted

hyphae in subhymenium, basidium origin from non-incrusted hyphae (400x); B: Externally

incrusted pigment on walls of lamellar trama (400x); C: bullet-shaped caulocystidia (400x);

Pouzarella (Australia) ... 469

D: cylindro-clavate caulocystidia (400x); E: Pileipellis near pileus center (100x); F: Cylindro-

clavate to clavate pileocystidia (400x).

470 ... Largent & al.

6-8 angled basidiospores averaging 10.0 x 7.3 um, caulocystidia of two types

(cylindro-clavate or napiform to clavate), and abundant aborted basidia.

The distinctive features of P fusca are not found in any other Pouzarella

species. Entoloma romagnesii described from France has similarly shaped and

sized basidiospores but has a dark yellow-brown pileus and stipe, broadly

ellipsoid to clavate cheilocystidia, and non-encrusted pileus and stipe hairs

(Noordeloos 1979).

See P. setiformis for comparisons with other Australian entolomatoid fungi

similar to P. fusca,.

9. Pouzarella albostrigosa Largent & Abell-Davis, sp. nov. PLATES 17-18

MycoBank MB 519587

Habitu Pouzarellae lasiae similis, sed pileo convexo vel late convexo, base stipitis alba

distincte strigosa, stipite minus robusto usque ad 22 mm longo, pseudocystidiis in hymenio

inclusis, caulocystidiis pileocystidiisque pigmentatione parietali in parietibus internis e

fasciis tumoribusque composita differt.

Type — Australia, Queensland, Cook Region, Mt. Hypipamee National Park, Kauri

Creek Track, 17°25'35.7"S 145°29'11.4"E, 964.7 m, 10 March 2009, DL Largent 9641

(holotype BRI, isotype CNS), sequences: HQ876557(mtSSU), HQ876535(LSU),

HQ876513 (RBP2).

Erymo.Locy — derived from the Latin albus (= white) + strigosus (= bristly), referring

to the white stipe base.

PiLEus 3-10 mm broad, 0.8-2.5 mm high, convex to broadly convex, at first

opaque, entirely hispid-squamulose with light colored squamules (between

white and orange-white or reddish-white, 5-7A-B1-2) on a dark smoky brown

to blackish-brown background (6-7F2-3), upon expanding and at maturity

remaining hispid-squamulose on the disc, appressed squamulose to appressed

fibrillose toward the margin and appressed fibrillose at the margin with an

overall color of blackish-brown on the disc (close to but darker than 7F2) and

slightly lighter elsewhere (close to but darker than 6-7F2-3) and becoming

striate up to the center, eventually fading overall to dark grayish-brown

(5F3) but remaining striate, dull, not hygrophanous and even; margin entire,

decurved and opaque then striate. TasTE indistinct to suggestively farinaceous.

Opor indistinct. LAMELLAE 2.5-5.5 mm long, adnate to uncinate, subdistant

to distant, narrow to moderately broad (1.0-1.8 mm high), dark gray to dark

brownish-gray (7F2), and then appearing dark grayish-brown (5E-F4) and

eventually light grayish-brown (5C-D3) after basidiospore maturation; margin

smooth, somewhat lighter than the surface. StrpE 6-22 mm long, 0.8-1.0 mm

broad at the apex, equal, covered with a dense layer of light colored fibrils

or squamules on a dark brown (5-7E-F3-4) faintly fibrillose surface, solid at

first, then hollow, basal tomentum absent to scarce; basal area pruinose just

above white, distinctly strigose base. BRUISING REACTION stipe bruising dark

Pouzarella (Australia) ... 471

PLATE 17 - Pouzarella albostrigosa (DLL 9641, HOLOTYPE).

A: Basidiomata stature; B: Pileal surface; C: Lamellae and lamellulae.

gray brown when surface fibrils become appressed to the surface. BRUISING

REACTION absent.

BASIDIOSPORES moderately nodulose-angular with 7-9 angles,

heterodiametric in profile and side views, isodiametric and 6-angled in polar

view, 9.1-15.6 x 6.7-9.2 um (x, = 11.5 + 1.4 x 7.7 + 0.6 um; E = 1.27-2.16; Q=

1.49 + 1.6;n=57/3). BAsipIA 2-4 sterigmate, clavate and tapered, colorless in 3%

KOH, 21.0-69.4 x 7.1-13.2 um (x, = 38.8 + 14.0 x 9.7 + 1.6 um; E = 2.06-6.71;

Q = 4.07 + 1.50; n = 24/2). PLEUROCYSTIDIA absent. PSEUDOCYSTIDIA absent.

ABORTED BASIDIA very abundant, nearly always with 2-4 sterigma and a few

with aborted basidiospores, clavate and tapered, equal to but more often % to “4

more narrow at the base than the basidia, embedded in the hymenium and not

projecting beyond the basidia, often but not always, originating from the outer

portions of the lamellar trama, dark reddish-brown in 3% KOH, 28.6-41.9 x

472 ... Largent & al.

PLaTE 18 - Pouzarella albostrigosa. A: Basidiospores (1000x) (DLL 9641: HOLOTYPE); B:

Pseudocystidia (black arrows) (400x) (DLL 9641); C: Pileipellis in disc area (100x) (DLL 9663);

Pouzarella (Australia) ... 473

7.1-12.2 um (E = 2.69-4.33; Q = 3.71 + 0.48; n = 11/1). LAMELLAR TRAMAL

HYPHAE did not revive well and was not measured. PILEIPELLIS up to 100-150

um deep, composed of clusters (i.e. the squamules) of 5-10 laterally attached

hyphae, each hypha with 5-10 cells; the clusters tapered with 5-10 hyphae

laterally agglutinated at the base and 2-4 hyphae laterally agglutinated at the

apex, erect on the disc, semi-erect towards the margin and nearly repent at the

margin. PILEOCYSTIDIA typically narrowly cylindro-clavate, at times broadly

cylindro-clavate, rarely clavate, 22.5-63.6 x 5.7-21.6 um (x, = 41.2 +9.1 x 11.1

+ 3.8 um; E = 1.42-8.40; Q = 4.20 + 1.80; n = 36/3) PILEAL TRAMAL HYPHAE did

not revive well and was not measured. STIPITIPELLIS a cutis of pigmented hyphae

between abundant scattered clusters of pigmented hyphae with clavate, broadly

obclavate or broadly cylindro-clavate thick-walled; pigmented caulocystidia

and scattered clusters of loosely entangled hyphae, 5-10 cells long, with

cylindro-clavate to clavate, colorless caulocystidia. CAULOCYSTIDIA 21.0-69.4

x 7.9-13.2 um (n = 24/3). STIPE TRAMAL HYPHAE parallel, 86-330 x 6-29 um

(n = 17/1). OLEIFEROUS HYPHAE rare to nearly absent, reddish-brown in 3%

KOH. PIGMENTATION distinct, coarsely and externally incrusting the tramal

hyphae of the stipe and most probably the pileus and lamellae; parietal and with

bands and bumps on the inner walls of the pileipellis, stipitipellis, pileocystidia

and some caulocystidia, externally incrusting the outer walls of the stipitipellis;

uniformly cytoplasmic in some pileocystidia and caulocystidia.

ECOLOGY AND DISTRIBUTION: Solitary in dark crumbly soil on hillside with

water pipe above a trail or in hard packed soil within a complex mesophyll vine

forest of high rainfall; northeastern Queensland.

ADDITIONAL COLLECTIONS EXAMINED — AUSTRALIA. QUEENSLAND, Cook Region,

Mt. Hypipamee National Park, 17°25'35.7"S 145°29'11.4"E, 964.7 m, 18 March 2009, DL

Largent 9666; Danbulla National Park, Lake Euramoo Track, 17°09'42.3"S 145°37'45.6"E,

762.6 m, 18 March 2009, DL Largent 9663 (mtSSU HQ876558, LSU HQ876536, RPB2

HQ876514).

ComMMENTS— Pouzarella albostrigosa typically has a convex pileus that is hispid-

squamulose on the disc and imbricate squamulose elsewhere and initially

covered with white to orange-gray or reddish-gray tipped squamules that

become dark brownish-gray with age and exposure, a <22 long stipe covered

by light-colored layer of entangled fibrils overlying longitudinally appressed

dark brown fibrils and with distinct white strigose fibrils at the base, nodulose-

angular basidiospores averaging 11.5 x 7.7 um, absence of cheilocystidia, and

the presence of aborted basidia embedded in the hymenium, pileocystidia and

caulocystidia with internal parietal pigmentation in the form of bands and

D: Pileocystidia (400x) (DLL 9641); E: Stipitipellis squash mount showing chains of cells and

caulocystidia as terminal cells (400x) (DLL 9641); F: Stipitipellis periclinal section showing chains

of cells with caulocystidia as terminal cells (400x) (DLL 9641).

A7A ... Largent & al.

bumps (some uniformly pigmented) and both thick-walled pigmented and

thin-walled colorless caulocystidia.

See Pouzarella setiformis for comparisons with Australian taxa related to

P. albostrigosa.

10. Pouzarella lasia (Berk. & Broome) Largent & Abell-Davis, comb. nov.

MycoBAnk MB 519574 PLATES 19-20

= Agaricus lasius Berk. & Broome, J. Linn. Soc., Bot. London. 11: 539 (1871).

PitEus 4-10 mm broad, 2.5-7 mm high, conic to campanulate, at first hispid-

squamulose on the disc and imbricate squamulose elsewhere, remaining so

on the disc when mature but becoming appressed squamulose towards the

margin and appressed fibrillose on the margin, the squamules entirely pallid

to brownish-orange (5C3) at first, with age darkening from the base towards

the tip, dark brownish-gray (5E-F3), eventually entirely dark brownish-

gray (5-7F2-3) on the disc and a lighter brownish-gray elsewhere (5-7E2-3);

marginal fibrils concolorous with nearby squamules dull, opaque and even

at first, becoming striate when faded; margin decurved, entire. TasTE and

Opor indistinct. LAMELLAE 2-6 mm long, rarely adnexed to more commonly

uncinate, subdistant, narrow to moderately broad (1-2 mm deep) when 2-6

mm long, when young entirely brownish-gray (5E-F3-4) and then appearing

brownish-orange (5C3) with a lighter-colored margin with basidiospore

maturation, margin smooth. LAMELLULAE typically 3 between lamellae and in

2 tiers (2 short and 1 medium long). Stipe 11-40(-50) x 0.5-1.3 mm, equal,

with a superficial, orange-gray (5B2-3) to brownish-orange (5C3) layer of

entangled fibrils overlying longitudinally appressed, grayish-brown (5D-E3-4)

fibrils, solid becoming hollow with age; basal tomentum scarce to absent; stipe

base pallid and obscurely strigose. BRUISING REACTION observed in the stipe

(7F2-3) but not the strigose base, darkening upwards.

BasipDIospoREs nodulose-angular with 7-9 angles in profile and side

views, heterodiametric, 8.9-14.5 x 5.1-8.7 um (x, = 11.5 + 1.2 x 7.2 + 0.6 um;

E = 1.29-1.99; Q = 1.61 + 0.14; n = 99/4). Basip1A 2-4 sterigmate, clavate

but hardly tapered, colorless in 3% KOH, 34.6-42.7 x 7.7-11.8 (widest point)

x 5.2-7.9(base) um (E = 2-96-5.28; Q = 4.02). PLEUROCyYSTIDIA absent.

PsEUDOCYSTIDIA absent. ABORTED BASIDIA abundant, nearly always with

2-4 sterigma and a few with aborted basidiospores, clavate and tapered to

a narrow end, often embedded in the hymenium but a few projecting up to

17 um beyond the basidia, often but not always, originating from the outer

portions of the lamellar trama, dark reddish-brown in 3% KOH, 36.2-59.0 x

7.1-11.2(widest point) x 2.8-5.2(base) um (E = 4.76-5.58; Q = 5.02). LAMELLAR

TRAMAL HYPHAE parallel to subparallel, 22.8-215.2 x 4.0-25.7 um; narrow

hyphae abundant, broad hyphae rare to uncommon; subhymenium indistinct.

Pouzarella (Australia) ...475

PLATE 19 — Pouzarella lasia (DLL 9662).

A: Basidiomata stature; B: Pileus surface; C: stipe surface.

476 ... Largent & al.

PILEAL TRAMAL HYPHAE 56.0-162.3 x 3.0-13.5 um. PILEIPELLIS up to 240 um

deep, composed of clusters (squamules) of 5-10 laterally attached hyphae, each

hypha with 10-20 cells that have parietal pigmentation; the clusters tapered with

5-10 hyphae at the base and 2-4 hyphae at the apex, erect on the disc, semi-erect

towards the margin and nearly repent at the margin. PILEOCysTIDIA typically

narrowly to broadly cylindro-clavate, at times clavate to nearly napiform,

24,4-66.1 x 6.8-19.0 um (x, = 41.7 x 11.6 um; E = 2.0-6.9; Q = 3.89; n = 13/1).

STIPITIPELLIS a cutis between abundant, densely scattered clusters of colorless

1-3 celled hyphae with clavate, broadly obclavate or broadly cylindro-clavate

caulocystidia and scattered clusters of loosely entangled hyphae, 5-10 cells

long, with cylindro-clavate to clavate, colorless caulocystidia. CAULOCYSTIDIA

16.5-66.4 x 7.1-13.9 um. STIPE TRAMAL HYPHAE parallel, 101.4-355.5 x

5.4-13.4 um. OLEIFEROUS HYPHAE rare in the pileal and stipe trama, absent in

the gill trama, reddish-brown in 3% KOH. PIGMENTATION coarsely externally

incrusted on the tramal hyphae of the lamellae, pileus and stipe and in the

basal cells of the pileipellis; obscurely internally parietal in the hyphae of the

stipitipellis and in the outermost 1-3 cells of the hyphae of the pileipellis.

ECOLOGY AND DISTRIBUTION: Scattered to gregarious in hard packed soil

in complex mesophyll vine forest in areas with high rainfall; Lake Euramoo,

Danbulla National Park, Ceylon and Papua New Guinea.

COLLECTIONS EXAMINED — AUSTRALIA. QUEENSLAND, Cook Region, Danbulla

National Park, Lake Euramoo Track, 17°09'43.0"S 145°37'44.7"E, 763.2 m, 18 March

2009, DL Largent 9662 (mtSSU HQ876551, LSU HQ876529, RPB2 HQ876507); 19

March 2009, DL Largent 9670 (mtSSU HQ876552, LSU HQ876530, RPB2 HQ876508),

DL Largent 9671; 21 March 2010, DL Largent 9807 (mtSSU HQ876555, LSU HQ876533,

RPB2 HQ876511), DL Largent 9811 (mtSSU HQ876556, LSU HQ876534, RPB2

HQ876512), DL Largent 9812; 24 March 2010, DL Largent 9818, 9827; Cook Region,

Danbulla National Park, Kauri Creek Track, 17°07'49.1"S 145°35'58.2"E, 725.1 m,

21 February 2010, DL Largent 9729 (mtSSU HQ876553, LSU HQ876531, RPB2

HQ876509); 13 March 2010, DL Largent 9778 (mtSSU HQ876554, LSU HQ876532,

RPB2 HQ876510).

ComMMENTS— Pouzarella lasia is diagnosed by a typically conic to campanulate

to parabolic pileus that is hispid-squamulose on the disc and imbricate

squamulose elsewhere and covered when young with pallid to brownish-orange

tipped squamules that become dark brownish-gray with age and exposure;

a <50 mm long stipe covered by an orange-gray to brownish-orange layer of

entangled fibrils overlying longitudinally appressed grayish-brown fibrils and

with an obscurely strigose base, nodulose-angular basidiospores averaging

11.5 x 7.2 um, absence of cheilocystidia, presence of aborted basidia that

project beyond the hymenium, internally parietal pigmented pileocystidia and

caulocystidia, + thick-walled strongly encrusted basal pileipellis hyphae, and a

nondistinctive taste and odor.

Pouzarella (Australia) ... 477

PLATE 20 - Pouzarella lasia (DLL 9662). A: Basidiospores (1000x); B: Basidia (colorless) and

pseudocystidia (dark) (400x); C: Pileipellis on disc (100x); D: Pileocystidia (400x); E: Stipitipellis

(100x); F: Caulocystidia (400x).

478 ... Largent & al.

In Australia, we observed 30-40 P. lasia basidiomata in the same habitat

and locality twice over a 2-4 day period in mid-March 2009 and mid- to late-

March 2010. Most pilei from basidiomata collected during the beginning days

possessed light-colored squamules over the entire surface but the squamule

base of the remaining basidiomata began to develop dark gray brown tints.

Pileal surfaces from basidiomata collected at the end of the 2-4 day period had

only dark gray brown squamules and were otherwise morphologically identical

to Pouzaromyces lasius as described and illustrated by Horak (1980). With

the exception of the light colored squamules, the basidioma features from the

beginning of the time period also matched those of Pouzaromyces lasius. Horak

apparently either overlooked the light colored squamules in young specimens

or examined only mature specimens.

Horak (1980) and Pegler (1977) studied the holotype of Agaricus lasius.

Referring to his type study in his Pouzaromyces lasius description, Horak

noted basidiospores measuring 10.5-13.5 x 6-8 um, cheilocystidia absent,

and the terminal (1-3 cell thick) hairs on stipe [and pileus?] as thin-walled

and remaining cells as thick-walled. For the holotype, Pegler (1977) reported

basidiospores averaging 14 x 8.5 um (11.5-15.5 x 8-9.5 um) and a 4-um

thickness for the cell walls of the pileal hairs. It is rather perplexing to have two

investigators study the same specimens and report such distinct differences.

Pouzarella lasia, P domingensis from the Dominican Republic, Pouzaromyces

napaliensis from the Hawaiian Islands, and Entoloma testaceostrigosum from

the state of Kerala in India lack cheilocystidia and have similarly shaped and

sized basidiospores. Pouzarella domingensis differs in the presence of inflated

cells in the pileal subpellis (Baroni et al. 2008), while Pouzaromyces napaliensis

has aciculate to setiform caulocystidia and pileocystidia, smaller basidiomata

(2.5 mm broad pileus, 3-4 x <1 mm stipe), and a pale yellowish-brown pileus

(Desjardin & Baroni 1991). Entoloma testaceostrigosum is separated by its larger

(12.5-17.0 x 7-10 um) basidiospores and absence of encrusted pileipellis and

stipitipellis hyphae (Manimohan et al. 2006).

Pouzarella lasia was found fruiting abundantly in two locales. All these

collections were made in a similarly complex mesophyll vine rainforest

in Danbulla National Park in northeastern Queensland. Four collections

—DL Largent 9662 (18 March 2009), 9670 (19 March 2009), 9807 and 9811

(21 March 2010)— came from populations in identical habitats on the Lake

Euramoo Track, while two —DL Largent 9729 (21 February 2010), 9778

(13 March 2010)— were from the same population and identical habitat on

the Kauri Track. We found no evidence of genetic isolation among populations

separated by 4 km and the level of allelic variation within each locus is

consistent with the variation expected in interbreeding populations with a

large effective population size. Future studies of these and other populations

Pouzarella (Australia) ... 479

of P. lasia may provide insight into the extent of gene flow or genetic isolation

among populations.

See P. setiformis for comparisons with Australian taxa related to P lasia,.

Discussion

In the past decade, efforts to gain insight into the morphological features that

unite mushroom-forming fungi with their sequestrate relatives (Basidiomycota,

Agaricales) have pinpointed some unique characters that can form the basis of

natural classifications (e.g. Moncalvo et al. 2004, Matheny et al. 2006). However,

defining these boundaries for large and diverse families such the Entolomataceae

with over 1500 mushroom-forming species and few sequestrate taxa (Largent

1994, Moncalvo et al. 2004, Co-David et al. 2009) can be challenging. The

basis for the early classification of all members in the Entolomataceae is the

pinkish basidiospore deposit and angular basidiospores observed in polar

view, features that may form the basis of a natural classification based on the

observed monophyletic support using DNA sequence data (Moncalvo et al.

2004, Matheny et al. 2006, Co-David et al. 2009).

Despite the support for the monophyly of the Entolomataceae, defining

generic boundaries within the family may be challenging. For example,

several of the proposed generic concepts based on morphological features

have been questioned based on the lack of observed monophyletic support

in phylogenetic analyses (Moncalvo et al. 2002, 2004, Co-David et al. 2009).

Large genera that exemplify these problems include Entoloma (Fr.) P. Kumm.,

recovered as either polyphyletic (Moncalvo et al. 2004) or paraphyletic (Co-

David et al. 2009). A similar lack of support has been observed for Claudopus

Gillet, Leptonia (Fr.) P. Kumm., Rhodocybe Maire, and Nolanea (Fr.) P. Kumm.

(Moncalvo et al. 2002, Co-David et al. 2009). Although it is unquestionably

an under-sampled genus, monophyly of Pouzarella is supported by molecular

data (from only two collections, the European Entoloma araneosum and Indian

E. violaceovillosum; Co-David etal. 2009). In this paper, we includea phylogenetic

analysis of two European (Co-David et al. 2009) and ten Australian Pouzarella

species; given the exclusion of other entolomataceous genera, we understand

that our phylogeny neither refutes nor supports the monophyly of Pouzarella.

Nonetheless, our sequence analyses support delineation of the ten Australian

species based on micromorphological features, and features common to

these clades are highlighted as characters that may form the basis of a natural

classification for other genera.

One interesting pattern apparent in the phylogeny is the strong support

for the monophyletic clade containing P lageniformis, P. parvula, and P.

pilocystidiata. All species possess long cylindro-clavate pileocystidia, abundant

thin-walled lageniform to ventricose-rostrate cheilocystidia, and lack aborted

480 ... Largent & al.

DL Largent 9807 Pouzarella lasia

DL Largent 9662 Pouzarella lasia

0.1 substitutions/site

1.00] ' DL Largent 9729 Pouzarella lasia I

DL Largent 9641 Pouzarella albostrigosa

DL Largent 9623 Pouzarella fusca

DL Largent 9784 Pouzarella debilis

DL Largent 9810 Pouzarella setiformis

DL Largent 9809 Pouzarella setiformis

DL Largent 9794 Pouzarella pamiae

1.00 # DL Largent 9834 Pouzarella pamiae

DL Largent 9808 Pouzarella pamiae Il

DL Largent 9934 Pouzarella farinosa

DL Largent 9900 Pouzarella farinosa

DL Largent 9949 Pouzarella pilocystidiata

1.00] DL Largent 9932 Pouzarella pilocystidiata

DL Largent 9848 Pouzarella pilocystidiata Il

DL Largent 9901 Pouzarella parvula

DL Largent 9895 Pouzarella lageniformis

Entoloma ‘‘Pouzarella”’ violaceovillosum

Entoloma “Pouzarella” araneosum

Calocybe carnea

Lyophyllum leucophaeatum

Lyophyllum decastes

PLATE 21 - Phylogenetic relationships of Pouzarella based on Maximum Likelihood analysis of

mtSSU, LSU, and RPB2 of 10 Australian, one European, and one Indian species. Tree topologies

based on Maximum Likelihood and Bayesian estimation were congruent. Thick branches represent

nodes that are supported by bootstrap values of 70% or greater and values above each branch are

the Bayesian posterior probability values computed from the 50% majority rule consensus tree.

Shaded areas represent the three clades recovered. Taxa are labeled with the collection identification

number and name of taxon.

basidia (unusual in Pouzarella). Strong support was also observed for another

clade comprising P. albostrigosa, P. debilis, P fusca, P. lasia, and P. setiformis.

One unique morphological feature common to this clade is the conspicuous

absence of cheilocystidia, a feature shared by other tropical species including

Pouzaromyces minutus (Horak 1980), Pouzaromyces napaliensis (Desjardin

& Baroni 1991), and Entoloma testaceostrigosum (Manimohan et al. 2006).

Pouzarella pamiae and P. farinosa formed a monophyletic group, although with

low ML BS support. Both possess a squamulose-hispid stipe surface and + thick

walled cheilocystidia — features also encountered in the tropical P ferreri and

temperate P. dysthales (Peck) Mazzer (Baroni et al. 2008, Noordeloos 2004).

Future studies that include these and other representative Entolomataceae

taxa are needed to better assess phylogenetic relationships and diagnostic

Pouzarella (Australia) ... 481

morphological characters that could form the basis of a natural classification

of Pouzarella.

Acknowledgments

Fieldwork in Australia was supported by the Largent family trust and we are

particularly grateful for the support of Pamela Largent. Fieldwork and logistical support

was provided by the Australian Tropical Herbarium and the School of Marine and

Tropical Biology, James Cook University. The DNA sequences generated in this study are

based upon work supported by the National Science Foundation under Grant No. DRI

0922922 awarded to SE Bergemann. Ms. Patricia Eckel provided the Latin diagnoses.

Comments by the two reviewers, Dr. Timothy Baroni and Dr. Andrew Methven, and

by the nomenclature editor Dr. Shaun Pennycook, were also helpful. We wish to thank

Dr. Genevieve Gates for sharing unpublished illustrations and descriptions. We also

acknowledge Dr. Todd Osmundson for a critique of an earlier draft of this paper.

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Noordeloos ME. 1992. Entoloma s.1. in Fungi Europaei vol. 5. Ed. Candusso: Alassio, Italy.

Noordeloos ME. 2004. Entoloma s.l. in Fungi Europaei vol. 5A. Ed. Candusso: Alassio, Italy.

Noordeloos ME, Gates MA. 2009. Preliminary studies in the genus Entoloma in Tasmania II.

Cryptogamie, Mycologie 30(2): 107-140.

Pegler DN. 1977. A revision of the Entolomataceae (Agaricales) from India and Sri Lanka. Kew

Bulletin 32: 189-220. http://dx.doi.org/10.2307/4117266

Pegler DN. 1986. Agaric flora of Sri Lanka. Kew Bulletin Additional Series 12: 1-519.

Rambaut A. 2002. Se-Al: Sequence alignment editor. Available at: http://evolve.zoo.ox.ac.uk/

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

Saccardo PA. 1887. Sylloge Hymenomycetum, Vol. I. Agaricineae. Sylloge Fungorum 5:1-1146.

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, DH Gelfand, JJ Sninsky, TJ White

(eds). PCR protocols: a guide to methods and applications. Academic Press, Inc.: San Diego,

California.

Pouzarella (Australia) ... 483

Key to Pouzarella species from central New South Wales (N)

—

Qo Oo

and northeastern Queensland (Q)

. Cheilocystidia present; aborted basidia absent or if present, rare and colorless

. Cheilocystidia absent; aborted basidia present and typically abundant and

staining dark reddish-brown in 3% KOH ...............0... 0 00a 6 (Clade I)

. Cheilocystidia acicular, aculeate, narrowly acuminate, finally rostrate ventricose;

pileocystidia cylindro-clavate, with Q = 7.0 or more ............. 3 (Clade III)

. Cheilocystidia globose, napiform, vesiculose pyriform or clavate to broadly

acuminate; pileocystidia versiform, withaQ<5.0 .............. 5 (Clade II)

. Pileus orange white to grayish-orange; stipe apex pruinose; taste mild;........... 4

. Pileus squamules and fibrils dark brown on dark blond to brown background;

stipe apex tomentulose to densely hirsute or with abundant bundles of

TAHA SyctAStE TALIM ACCOUS e§ t-cetng debe oto} dy sctad days ed da feted deh; 1. P. pilocystidiata (N)

. Pileus hispid-squamulose; stipe entirely pruinose; pileocystidia at most

AGUTMIN ALC we anew ls tga swle snus pede nailed a yulans he 2. B. lageniformis (N)

. Pileus entirely densely matted squamulose; stipe pruinose at the apex,

glabrous elsewhere; at least some pileocystidia setiform ....... 3. P. parvula (N)

. Pileus at first entirely reddish-golden to brownish-orange, then darkening

to medium teak brown to burnt umber; cheilocystidia broadly clavate to

broadly. acuminate -:4.. 2.2 2 aie epee ek Mina na eh ae le 4. P. pamiae (Q)

. Pileus with densely packed upright tufts of orange gray squamules on a

grayish-brown background, overall light grayish-brown; cheilocystidia

at first nearly globose then napiform, vesiculose, or pyriform.. 5. P. farinosa (N)

. Stipitipellis with scattered to abundant setiform caulocystidia; pileipellis

with setiform pileocystidia, most evident at the margin ..... 6. B. setiformis (Q)

. Stipitipellis with cylindric to clavate to obclavate caulocystidia; setiform

catilocystidia-and ipileoeystidia abSent, hve, ait... weet. wieclan Wheeden olemadn-esathrabey yah Es

. Pileus broadly convex to convex; stipe up to 22 mm long; aborted basidia

embedded in the hymenium or rarely extending beyond the hymenium

AA MAL ee SCE ees SC het Let Ment C ne ee Le eee eee 8

. Pileus conic, campanulate or parabolic; stipe <50 mm long; aborted basidia

extending beyond the hymenium by <17 um ................. 10. P. lasia (Q)

. Caulocystidia of one type, similar to the pileocystidia ............. 7. P. debilis (Q)

. Caulocystidia of two types: 1) cylindro-clavate, the end cells of loosely entangled

hyphae, similar to the pileocystidia; 2) clavate, obclavate, or broadly napiform

and typically solitary and not in loose hyphal clusters ....................0. 9

. Pileus minutely but entirely squamulose, black to a very dark gray-brown,

without off-white squamules when young; pileus opaque when mature;

stipe base pallid and obscurely strigose ...................0004 8. P. fusca (Q)

. Pileus entirely hispid-squamulose and with off-white (5A-B1-2) squamules

when young, base color dark grayish-brown; pileus striate when mature;

stipe base white and distinctly strigose .................. 9. P. albostrigosa (Q)

MY COTAXON

http://dx.doi.org/10.5248/117.485

Volume 117, pp. 485-497 July-September 2011

Amanita vernicoccora sp. nov.

—the vernal fruiting ‘coccora’ from California

DIMITAR BOJANTCHEV’ , SHAUN R. PENNYCOOK? & R. MICHAEL Davis?

'MushroomHobby.com, 345 Shipwatch Lane, Hercules, CA 94547, USA

?Manaaki Whenua Landcare Research, Private Bag 92 170, Auckland, New Zealand

*Department of Plant Pathology, University of California, Davis, CA 95616, USA

* CORRESPONDENCE TO: dimitar@pontix.com

ABSTRACT — We describe Amanita vernicoccora, a vernal fruiting species known as the

“spring coccora’ in California. Sequence analyses of four DNA regions and phenotypic

traits demonstrate that A. vernicoccora, long considered a pale-colored form of the autumnal

fruiting A. calyptroderma (the ‘fall coccora’), is a unique species. Morphological and genetic

data support both species in section Caesareae. We also address some nomenclatural and

taxonomic intricacies surrounding application of the proper name for the fall-fruiting

coccora.

Key Worps — Amanitaceae, fungal taxonomy, nrLSU, nrITS data

Introduction

The popular name —‘coccora or ‘coccoli’— was coined by Italian- Americans

in California for two valuable edible mushrooms of genus Amanita Pers.

collected during the fall and spring. The autumnal fruiting A. calyptroderma

(referenced here as the ‘fall coccora, Fic. la) and the vernal fruiting

A. vernicoccora (referenced here as the ‘spring coccora, Fic. 1b) are closely

related and were for many years considered conspecific seasonal color forms.

Both taxa represent subg. Amanita sect. Caesareae Singer and are close relatives

of A. caesarea (Scop.) Pers. —the European ‘Caesar's amanita— a popular

edible in Europe, especially in Italy. The coccora has a notable thick cottony veil

that leaves a distinctive monolithic velar remnant on the pileus. The association

with a thick-walled cocoon enveloping the Amanita egg is the etymological

origin of the popular name.

Due to the perceived conspecificity of the two coccora, the autumnal name

was used for both species, currently referenced as either A. calyptroderma or

A. lanei. Both names are still used in various reference materials and field

486 ... Bojantchev, Pennycook & Davis

Fic. 1. a) The fall coccora Amanita calyptroderma. b) The spring coccora Amanita vernicoccora.

guides, and there is general confusion as to which is preferable. The key details

of the rich and intricate nomenclatural and taxonomic history surrounding the

choice of the proper scientific name for the fall coccora are discussed here.

Nomenclatural and taxonomic history of the fall coccora

Amanita calyptroderma G.F. Atk. & V.G. Ballen, Science, n.s. 29: 944. 1909.

?= Amanita calyptrata Peck, Bull. Torrey Bot. Club 27:

14. 1900 [nom. illegit., non Lam. 1783].

= Venenarius calyptratus Murrill, Mycologia 4: 241. 1912 [nom. nov.].

= Venenarius lanei Murrill, N. Amer. Fl. 10: 75. 1914 [nom. nov. superfl.].

= Amanita lanei Sacc. & Trotter, Syll. Fung. 23: 5. 1925 [nom. nov.].

The fall coccora was first validly named as Amanita calyptrata (Peck 1900) based

on autumnal collections from fir woods in Oregon and notes by Dr. H. Lane.

When first published, this name was legitimate, but under the retrospective

application of the current International Code of Botanical Nomenclature

(ICBN Art. 53.1; McNeill et al. 2006), it is an illegitimate later homonym of

A. calyptrata Lam. Murrill (1912) “recombined” the name as Venenarius

calyptratus —under ICBN Art. 58.1, this must be interpreted as a legitimate

nom. nov., with priority dating from 1912. Two years later, Murrill (1914) had

become aware of the later homonymy of Peck’s binomial (illegitimate under the

1907 American Code of Botanical Nomenclature) and published V. lanei as a

nom. nov. based on Peck’s description and typification. However, under ICBN

Art. 52.1, the name V. lanei was a superfluous (ie., illegitimate) synonym of

V. calyptratus, and consequently the “recombination” Amanita lanei published

by Saccardo & Trotter (Trotter 1925) must also be interpreted as a legitimate

nom. nov. under ICBN Art. 58.1.

In 1909 George Atkinson described A. calyptroderma (Atkinson 1909a,b)

based on autumnal collections, several diagnostic photographs of fresh

material, and supplemental notes from Virginia Ballen from Brookdale,

Santa Cruz Co., California. According to the Atkinson papers in the CUP

Herbarium, the collections were sent from California in fresh condition and

Amanita vernicoccora sp. nov. (California, USA) ... 487

arrived at Cornell University on Dec 7, 1908 with some basidiomata partly

decayed. The major elements of the collection notes match the A. calyptroderma

protologue (Atkinson 1909a,b). The date of the collections, the description, and

the photographs published in Atkinson (1909c) leave no doubt that the name

A. calyptroderma was applied to the familiar fall coccora (Fic.1a, Fic. 10).

Apparently, Atkinson (1909c) was aware of Peck’s earlier species and makes

a single reference to A. calyptrata in the footnote of his description —“closely

related to Amanita calyptrata Peck, Bull. Torr. Bot. Club 27:14.1909 [sic, =

1900], but differs in color and other characters.’ Contrary to Atkinson's view,

most resident California taxonomists consider A. calyptroderma and A. lanei

(= A. calyptrata) conspecific. Taxonomically, we believe that A. calyptroderma

faithfully represents the fall coccora. Moreover, if we accept conspecificity with

A. lanei/V. calyptratus (dating from 1912), A. calyptroderma (dating from 1909)

has nomenclatural priority. [It should be noted that under the ICBN, until 1981

the correct name for this conspecific taxon was Amanita calyptrata, because it

remained legitimate and had priority over A. calyptroderma. This situation was

reversed when the 1981 International Botanical Congress (Sydney) moved the

starting point date for agaric nomenclature back from 1821 to 1753, making

A. calyptrata Peck a nom. illegit.]

Nevertheless, there are lingering doubts about the exact identity of

A. lanei, as the original description by Peck notes green tinges on the pileus.

In our experience, the fall coccora frequently develops olive-brown tints in age

or when exposed to colder weather but we would not describe the color as

green (Fic. 10c). Olive-brown tints, for example, are commonly observed on

specimens from the Sierra Nevada foothills. It is also possible that the stronger

color qualification of “green” was used instead of “olive,” a common style in

many early descriptions. Nonetheless, the green color has been stressed in

numerous taxonomic treatments as an important diagnostic character. Zeller

(1931) states: “These two species, A. calyptrata Peck and A. calyptroderma

Atkinson and Ballen, are very similar and difficult to distinguish. The main

distinction is the greenish tinting of the pileus and gills in A. calyptrata...”

Hotson (1936) supported this view that the green tints were a key diagnostic

character in his treatment of the amanitas of Washington. Although it is

unclear whether these authors personally collected green- tinted specimens,

we cannot discount the possibility that A. lanei refers to a rarely collected

unique species from the more northern areas of the Pacific Northwest. This

prospect is additionally corroborated by the suggested association of A. lanei

with fir per its original description. In contrast, the fall coccora is primarily

associated with broadleaved trees in California. Therefore, we believe that

on both nomenclatural and taxonomic grounds, the correct name for the fall

coccora is A. calyptroderma.

488 ... Bojantchev, Pennycook & Davis

Murrill (1912, 1914) and Zeller (1931) synonymized Amanita calyptroderma

and A. calyptratoides Peck, although it appears that Murrill never collected or

examined either species. Current researchers do not accept this synonymy,

and we also consider it to be clearly an error. The macromorphological traits

of A. calyptratoides —such as the relatively smaller size (4-8 cm), gray-brown

to plumbeous pileal colors, and particularly the evanescent to inconspicuous

annulus— contrast sharply with the features of A. calyptroderma as supported

by the very diagnostic original photographs (Atkinson 1909c). The current

interpretation of A. calyptratoides refers to a small species, common under oaks,

with primary distribution of southern California and Mexico (Tulloss 2011).

The conspecificity of fall vs. spring coccora

The conspecificity of the fall and spring fruiting coccora was assumed at

the time of the first descriptions and that view generally has not changed.

While Peck and Murrill do not seem to have been aware of the existence of a

spring fruiting species, Atkinson was and discussed it at length. The original

description of A. calyptroderma (Atkinson 1909a,b) clearly referenced the fall

species based on the time of collection and the colors he specified — “pileus

maize yellow to pale chrome yellow.’ Very soon after that, based on the input

of his experienced California correspondent, Virginia Ballen, he offered a more

elaborate description of the pileus colors (Atkinson 1909c), including the spring

form, “The pileus is a maize-yellow in its bright-colored forms and varies to a

pale straw color or Naples yellow (R.) in the vernal forms’

Regarding the vernal fruiting habit of the coccora, Atkinson (1909c)

developed an elaborate hypothesis:

“It occurs in the high Sierras and in the Coast Range. Probably the entire

summer season is needed for the growth and extension of the mycelium

in the forest mold, so that the huge fruit bodies are developed in late

autumn and early spring. While we have as yet no information bearing

on the time of origin of the fundament of the fruit bodies, it is likely that

all of them are formed during the summer and late autumn, and that the

second crop, which appears early in the spring, is composed of plants

which have lived through the winter in a partially developed condition.

The autumn crop ceases about the last of December, while the spring

crop begins about the middle of March.”

Contemporary authors of influential field guides in California (Arora 1986,

Wood & Stevens 2009) continue to reflect the notion of the conspecificity and

seasonal color differences between the fall and spring coccora. At the same

time, the awareness that they are likely different species has grown in recent

years. Smith et al. (2007) first reported that the fall and spring coccora differ

significantly in their ITS sequences.

Amanita vernicoccora sp. nov. (California, USA) ... 489

Materials & methods

Methods for morphological studies and DNA extraction, PCR conditions and

primers, and amplicon clean up and sequencing follow Bojantchev & Davis (2011).

Collections are stored in the Bojantchev private herbarium or at the University of

California herbarium in Berkeley (UC) as noted.

All Amanita nrITS and nrLSU sequences from the public databases GenBank (http://

www.ncbi.nlm.nih.gov) and UNITE (http://unite.ut.ee/) were downloaded, reviewed,

and selected for quality and identification congruity. Sequence alignments were

generated with MAFFT v6.821b (Katoh et al. 2002) with the G-INS-i global alignment

iterative refinement strategy. No gap opening and extension penalties were set for better

resolution of the variable sectors within nrITS, while the default higher penalties were

left for the more conserved nrLSU region. The alignments were visually inspected and

corrected where needed.

The evolutionary history of 248 Amanita nrLSU sequences was inferred using the

Maximum Likelihood method based on the Tamura-Nei model as implemented by

MEGAS (Tamura et al. 2007).

Nine Amanita sect. Caesareae nrITS sequences, representing four taxa, were selected

for higher resolution phylogenetic analysis (Fic. 3). Amanita muscaria (L.) Lam.

was selected as outgroup. Six sequences representing four taxa —the Asian Amanita

hemibapha (Berk. & Broome) Sacc., A. caesarea, A. calyptroderma, A. vernicoccora—

were sourced from GenBank. Three sequences of A. calyptroderma and A. vernicoccora

(including the holotype) are from the authors’ collections. Two nrITS sequences of

Amanita jacksonii Pomerl. (GenBank: AY436461, HQ539807) were excluded due

to quality and coverage concerns. The evolutionary history was inferred using the

Maximum Parsimony method as implemented by MEGAS5 (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 nine most parsimonious trees (length = 187), which differed only in the topology of

the terminal nodes.

Phylogenetic results

Four gene regions —nrITS, nrLSU, RPB2, 6-tubulin— were sequenced

from A. calyptroderma (RMD07036, not photographed) and A. vernicoccora

(RMD07020, Fic. 9B). Direct comparisons of these four gene regions support

TABLE 1. The four DNA regions sequenced from collections of Amanita calyptroderma

and A. vernicoccora.

DNA regions A. calyptroderma A. vernicoccora BLAST maximum

[coll. RMD07036] i [coll. RMD07020] : identity index difference

nrITS GQ250400 GQ250401 4%

sep GR a a Sn a eae as beans tes Al seit 7 a

Sa aera oe abi Mo Beate OP baeacucte ans tar ats pe pa

B-tubulin GQ401356 GQ401357 4%

490 ... Bojantchev, Pennycook & Davis

subgenus Lepidella

section Vaginatae

-- Amanita jacksonii (AF097376) North Carolina, USA

4] Amanita jacksonii (HQ539703) Massachusetts, USA

Amanita hemibapha var. ochracea (AF024458) Yunnan, China

| Amanita calyptroderma (GQ250415) California, USA

'— Amanita vernicoccora (GQ250416) California, USA

L Amanita caesarea (AF024443.) Spain

/Torrendia clade

aeasesaey) "99S

eylueulpy snuebgns

section Amanita

Fic. 2. Phylogenetic tree inferred by maximum likelihood analysis of 248 Amanita nrLSU

sequences. The tree shows the position of sect. Caesareae relative to the collapsed subtrees of the

other Amanita subgenera and sections. GenBank accession numbers are enclosed in brackets.

the phenotypic and ecological determination that taxonomically they are

different species and have been on separate evolutionary paths (TABLE 1).

The Maximum Likelihood tree (Fic. 2) shows the position of sect. Caesareae

within genus Amanita with the major subgenera and sections compressed. Our

research shows strong phylogenetic support for sect. Caesareae and generally

agrees with Drehmel et al. (1999) and Justo et al. (2010). One most parsimonious

MP tree is given in Fic. 3.

90 Amanita vernicoccora (JN133297) black oak, California, USA*

Amanita vernicoccora (GQ250401) black oak, California, USA

91]5! Amanita vernicoccora (EF559283) live oak, California, USA

99, Amanita calyptroderma (GQ250400) tanoak, California, USA

g | Amanita calyptroderma (DQ974693) live oak, California, USA

Amanita caesarea (AY486237) Spain

Amanita hemibapha (FJ441038) Yunnan, China

Amanita hemibapha (AB015699) Yunnan, China

10 | Amanita hemibapha (FJ375334) Yunnan, China

Amanita muscaria (DQ060908) Alaska, USA

BeDIeSIVED *JN9S

Fic. 3. Phylogenetic tree inferred by maximum parsimony analysis of nine Amanita sect.

Caesareae nrITS sequences with A. muscaria as the outgroup. The tree shows the position of

A. vernicoccora relative to its closest neighbors in sect. Caesareae. 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. The type collection is marked with *.

Amanita vernicoccora sp. nov. (California, USA) ... 491

Fic. 4. Amanita vernicoccora DBBO00201.

Taxonomy

Amanita vernicoccora Bojantchev & R.M. Davis, sp. nov. Fics 4-8

MycoBank MB 561705

Pileo 60-180 mm lato, hemispherico, dein planoconvexo, glutinoso, flavo, margine striato,

cum fragmento volvae magno persistento albo. Lamellis confertis, liberis, albis. Stipite 50-

140 mm longo, cylindrico, albo. Annulo superiore, striato, albo. Volva ampla, membranosa,

crassa. Carne albido. Sapore miti. Species vernalis. Sporis 9-12 x 6-7 um, late ellipsoideis,

hyalinis. Basidiis 42-64 x 11-14 um, tetrasporigeris, fibulis praesentibus.

Type: USA. California: El Dorado County, Georgetown, Dru Barner Campground,

under Quercus kelloggii, 21 May 2011, Bojantchev DBB43538 (Holotype UC 1860906;

Genbank nrITS JN133297).

ErymMo.oey: from the Latin vernus = vernal and coccora = the local name for this

mushroom.

P1LEus 60-180 mm diam., hemispherical to convex when young, plano-convex

to plano-concave with age; margin straight, short striate; color uniform yellow

to pale yellow; with a white, monolithic, central velar remnant, cottony thick

at first, thinning with age. LAMELLAE crowded, 10-18 mm broad, white to pale

cream, even, seceding to free, lamellulae common. STIPE 50-140 mm long,

15-30 mm wide, cylindrical, white with yellow tints at age, context hollow or

stuffed with a cottony or jelly-like substance in buttons. ANNULUS superior,

membranous, pendant, upper surface striate, cottony-thick at first, thinning and

collapsing with age, white to pale yellow with age. VoLva ample, thick, friable,

free, sac-like at first, thinning and collapsing with age, white. CONTEXT white

to pale yellow. Opor mild at first, pungent with age, frequently interpreted as

fishy. TasTE mild to pungent. MACROCHEMICAL REACTIONS 5% KOH negative,

492 ... Bojantchev, Pennycook & Davis

wi i

Fic. 5. Amanita vernicoccora (UC 1860906, holotype): a) Basidiospores. b) Hymenial layer

composed of basidia, basidioles, and subhymenial layer formed of densely packed irregular to

pyriform cells; clamped basidia (see arrow). Tissues are stained in Congo Red.

i |

Lib Ean. . Gee eee

Fic. 6. Amanita vernicoccora (UC 1860906, holotype): a) Filamentous and vesiculose cells from the

outer surface of the universal veil. b) Inflated cells from the upper surface of the annulus.

10% NH,OH negative, 3% phenol slowly pinkish-lilac on all surfaces, Guaiac

negative, 10% FeSO, negative. Spore Deposit white.

BASIDIOSPORES (8.5—)9.2-11.8(-12.5) x (5.7-)6.2-7.1(-7.5) um (mean 10.5

x 6.5 um), Q = 1.43-1.77, Q. = 1.61 (N = 183, 5 basidiomata, 4 collections),

broadly ellipsoid, with a prominent lateral apiculus, hyaline, inamyloid. BAsip1a

42-64 x 11-14 um, 4-spored, clavate, clamped, sterigmata 4-6 um long.

SUBHYMENIAL LAYER composed of several layers of irregular to pyriform cells

10-30 x 8-22 um, frequently clamped LAMELLAR TRAMA divergent, composed

of filamentous to swollen hyphae 8-24 um wide, occasionally clamped. CysT1p1a

not observed. PILEIPELLIS an ixocutis to ixotrichoderm, densely interwoven

within a gelatinous matrix, 200-320 um thick, hyphae 2-7 um wide, branched,

clamped, intracellular pigment. ANNULUS composed of filamentous cells, 2-6

um wide with clusters of inflated clavate, pyriform or mucronate cells 25-50

x 16-24 um, mainly on the upper surface. UNIVERSAL VEIL formed of dense

filamentous hyphae 1.5-8 um wide, clamped, interspersed with vesiculose cells

Amanita vernicoccora sp. nov. (California, USA) ... 493

—on the inner surface: narrow ellipsoid to elongated, 50-120 x 20-50 um; on

the outer surface: broadly ellipsoid to subglobose, 60-180 x 40-150 um, slightly

gelatinized. Stripe TRAMa acrophysalidic, composed of filamentous hyphae 2-8

um wide and inflated hyphae 50-150 x 20-46 um, occasionally clamped.

HABITAT AND DISTRIBUTION — Amanita vernicoccora fruits in the late winter

and spring and is apparently restricted to California. Along the coast and in the

lower elevations of Sierra Nevada (<2000 feet) it fruits in February-March and

is known to associate with evergreen oaks such as live oak (Quercus agrifolia

Née), interior live oak (Quercus wislizenii A. DC.) and blue oak (Quercus

douglasii Hook. & Arn.). Likely associations are also to Pacific madrone

(Arbutus menziesii Pursh) and manzanita (Arctostaphylos manzanita Parry).

Infrequently, A. vernicoccora fruits in the northern coastal areas where it is likely

associated with tanoak (Notholithocarpus densiflorus (Hook. & Arn.) Manos et

al. This species is far more common in the foothills of the Sierra Nevada and

Shasta Cascade range in May-June where the primary association in the lower

elevations (2000-4000 feet) is with the deciduous black oak (Quercus kelloggii

Newb.). At higher elevations A. vernicoccora may switch association to conifers,

but we have not seen it outside of the range of the black oak (<6000 feet).

ADDITIONAL COLLECTIONS EXAMINED: USA. CALIFORNIA: AMADOR CouNnrY, Indian

Grinding Rock State Park, elev. 2400 ft, under black oak, 17 May 2009, Davis RMD07020

(Genbank: nrITS GQ250401, nrLSU GQ250416, RPB1 GQ401359, B-tubulin gene

GQ401357); EL Dorado County, Georgetown, Dru Barner campground, elev. 2540 ft,

under black oak, 26 May 2011, Davis RMD110003; 29 May 2011, Bojantchev DBB43625;

Eldorado National Forest, Crystal basin area, near Union Valley Reservoir, elev. 4870 ft.,

under mixed conifers and black oak, 2 Jun 2006, Bojantchev DBB00118; CALAVERAS

County, Stanislaus National Forest, off Hwy. 120 and Hardin Flat Road exit, elev. 3600

ft., under mixed conifers and black oak, 27 May 2006, Bojantchev DBB00102; Stanislaus

National Forest, off Evergreen Rd, elev. 4100 ft. under mixed conifers and black oak,

22 May 2006, Bojantchev DBB00099; Siskryou County, Dunsmuir, off Hwy 5, near

Dunsmuir airport, elev. 2300 ft. under black oak, 31 May 2008, Bojantchev DBB00201;

Marin County, Marin Watershed, off Bolinas-Fairfax Rd., elev. 800 ft. under live oak,

13 Mar 2008, Bojantchev DBB07809.

Fic. 7. Amanita vernicoccora:

a) UC 1860906, holotype. b) DBB07809 under live oak in the California coastal range.

494 ... Bojantchev, Pennycook & Davis

Fic. 8. Amanita vernicoccora — a study in the development of the basidiocarp: a) Expanding button,

note the jelly like substance in the stipe context, which later turns cottony or leaves it hollow. b)

Note the thick cottony universal veil and the striate upper surface of the annulus. c) Note the

thinning of the universal veil on pileus, friable volva and collapsing annulus.

, & 4 4 ty, a y ‘ | . — d batt = ~ 4 my m

Fic. 10. Amanita calyptroderma, the fall coccora: a) DBB00347. b) DBB00393. c) DBB00491 shows

the frequent olive tinges on the pileus due to exposure to cold weather.

Discussion

Both A. vernicoccoraand A. calyptrodermaare beautiful species, distinguished

by a thick, cottony, white velar remnant on the pileus that remains whole into

maturity. This feature makes them among the easier amanitas to identify.

The edibility of the fall coccora, A. calyptroderma, was extolled by both Peck

(1900) and Atkinson (1909a), who made references to its gastronomical virtue.

Amanita vernicoccora sp. nov. (California, USA) ... 495

While considered conspecific, A. vernicoccora also was presumed edible and is

widely collected for the table. Even though practical experience has shown that

both coccora are relatively safe choices as edibles, confusion with other toxic

Amanita species do occur and the possibility of dangerous misidentification

must be stressed. The standard word of caution is that several reliable taxonomic

factors must be evaluated before an Amanita collection is deemed safe for

consumption.

The greatest danger posed by the pale spring coccora is misidentification

with members of the deadly toxic sect. Phalloideae (Fr.) Quél. —Amanita

ocreata Peck and A. phalloides (Fr.) Link, which also associate with oaks

throughout California. Amanita ocreata is a native species, while A. phalloides

is an import from Europe that has found favorable conditions and spread widely

in California (Pringle et al. 2009). Both can have a large patch of universal veil

on the cap (Fic. 11). In southern California, A. ocreata develops a much more

Fic. 11. Dangerous look-alikes of A. vernicoccora: a) The deadly toxic Amanita ocreata fruits in the

spring under oaks. b) The deadly toxic Amanita phalloides is a native of Eurasia, but a recent import

to California that fruits abundantly under oak in late fall and winter.

Fic. 12. Dangerous look-alikes of A. vernicoccora: a) The toxic Amanita gemmata gr. b) The

toxic Amanita aprica is frequently found with A. vernicoccora in the Sierra Nevada foothills. The

minutely broken velar remnants on the pileus contrast with the monolithic veil remnant of the

spring coccora.

496 ... Bojantchev, Pennycook & Davis

yellowish pileal coloration, which may further confound field identification.

Attention needs to be paid to all features of the basidiocarp, particularly to the

almost complete absence of striations on the pileal margin of the Phalloideae

species.

Other yellow species that can be mistaken for A. vernicoccora are in the

Amanita gemmata (Fr.) Bertill. clade, or gemmatoid amanitas (Fic. 12a), which

fruit in both the fall and spring in California. They also are toxic. In the Sierra

foothills, A. vernicoccora frequently co-occurs with Amanita aprica J.E. Lindgr.

& Tulloss (Fic. 12b). One key feature separating the two species is the manner

in which the veil of gemmatoid amanitas breaks into many small patches and

warts.

An extensive iconography of A. vernicoccora is available at http://www.

mushroomhobby.com.

Acknowledgements

The authors are grateful to Dr. Boris Assyov and Dr. Matthew E. Smith for their

presubmission reviews and comments. Dr. Dennis Desjardin examined the manuscript

and contributed critical analysis. Dr. Kathie Hodge researched the CUP Herbarium

records and revealed important information regarding the A. calyptroderma type

collection. Dr. Boris Assyov reviewed and corrected the Latin diagnosis. We thank

Terry Caudle and Mike Sampson for providing valuable collections. Ron Pastorino and

Debbie Klein contributed research on the current usage of the name “coccora’ in Italy.

Literature cited

Arora D. 1986. Mushrooms demystified: A comprehensive guide to the fleshy fungi. Ten Speed

Press. Berkeley.

Atkinson GE 1909a. A new edible species of Amanita. Science n.s. 29: 944.

http://dx.doi.org/10.1126/science.29.754.944

Atkinson GE. 1909b. Preliminary notes on some new species of Agaricaceae and Clavaria. Ann.

Mycol. 7: 365-376.

Atkinson GE 1909c. A remarkable Amanita. Bot. Gaz. 48: 283-293.

http://dx.doi.org/10.1086/330013

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

Drehmel D, Moncalvo J-M, Vilgalys R. 1999. Molecular phylogeny of Amanita based on large-

subunit ribosomal DNA sequences: implications for taxonomy and character evolution.

Mycologia 91: 610-618. http://dx.doi.org/10.2307/3761246

Hotson JW. 1936. The Amanitae of Washington. Mycologia 28: 63-76.

http://dx.doi.org/10.2307/3754068

Justo A, Morgenstern I, Hallen-Adams HE, Hibbett DS. 2010. Convergent evolution of sequestrate

forms in Amanita under Mediterranean climate conditions. Mycologia 102: 675-688.

http://dx.doi.org/10.3852/09-191

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.

Amanita vernicoccora sp. nov. (California, USA) ... 497

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.

Murrill WA. 1912. The Agaricaceae of the Pacific Coast II. Mycologia 4: 231-262.

Murrill WA. 1914. (Agaricales) Agaricaceae [pars]. North American Flora 10: 1-76.

Peck CH. 1900. New species of fungi. Bull. Torrey Bot. Club 27(6): 14-21.

http://dx.doi.org/10.2307/2477738

Pringle A, Adams RI, Cross HB, Bruns TD. 2009. The ectomycorrhizal fungus Amanita phalloides

was introduced and is expanding its range on the west coast of North America. Mol. Ecol. 18:

817-833. http://dx.doi.org/10.1111/j.1365-294X.2008.04030.x

Smith ME, Douhan GW, Rizzo DM. 2007. Intra-specific and intra-sporocarp ITS variation

of ectomycorrhizal fungi as assessed by rDNA sequencing of sporocarps and pooled

ectomycorrhizal roots from a Quercus woodland. Mycorrhiza 18: 15-22.

http://dx.doi.org/10.1007/s00572-007-0148-z

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

Trotter A. 1925. Supplementum universale, pars X. Basidiomycetae. Sylloge Fungorum 23. 1026 p.

Tulloss RE. 2011. “Studies in the Amanitaceae”.

http://www.amanitaceae.org/? Amanita calyptratoides. Retrieved 05/25/2011

Wood M, Stevens F. 2009. “California Fungi: Amanita lanei”. MykoWeb.

http://www.mykoweb.com/CAF/species/Amanita_lanei.html. Retrieved 05/25/2011

Zeller SM. 1931. Amanita calyptrata and Amanita calyptroderma. Mycologia 23: 225-226.

http://dx.doi.org/10.2307/3753963

MYCOTAXON

http://dx.doi.org/10.5248/117.499

Volume 117, pp. 499-507 July-September 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

The emphasis of this instalment is on tropical mycology, with mycofloristic

accounts of Panama and a National park in Malaysia, a revision of the Boletales

species described by Corner, and cercosporoid taxa from India. The European

Strophariaceae are the focus of a new book in the series FUNGI EuROPAEI, and

the economically important Aspergillus is the topic of a new volume of STUDIES

IN MycoLoey.

This contribution concludes with one new book announcement.

ASCOMYCETES

Taxonomic studies in the genus Aspergillus. Edited by R.A. Samson, J. Varga &

J.C. Frisvad. 2011. SrupiEs In Myco.oey no. 69. CBS-KNAW Fungal Biodiversity

Centre, P.O. Box 85167, 3508 AD Utrecht, The Netherlands. <info@cbs.knaw.nl>.

Pp. iv + 97, illustr. ISBN 978-90-70351-86-1. Price: 40 €.

There have been so many publications on Aspergillus systematics in recent

times, not least “Aspergillus Systematics in the Genomic Era” (STUDIES IN

Myco.oey no. 59, 2007; see Mycotaxon 107: 509-511, 2007), that it might

have been assumed that most species had now been discovered and named.

This collection of six papers, all involving the world’s most respected authorities

on the genus, shows that is far from the case: in even such an exceptionally well

‘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.

500 ... Vellinga, BOOK REVIEW EDITOR

studied genus, we have still a long way to go to approach a complete global

inventory.

In sect. Nigri, four new species are described, taking the total to 26;

additionally A. foetidus is placed as a synonym of A. niger, and molecular data

reveal that two further named species are colour mutants of others. Within the

black species, a separate study shows that growth rate parameters and hydrolase

activities are characters that can assist in species separations in the group.

In addition, an L-arabinose transcriptional activator regulating the pentose

catabolic pathway and involved in enzyme release is described for A. niger; it

appears to be confined to Eurotiales and to have arisen by gene duplication when

the order (or family) split from other filamentous ascomycetes. In sect. Terrei,

several taxa previously treated as subspecies or varieties of A. terreus prove to

be distinct lineages and are recognized as separate species, necessitating the

introduction of three new names; an additional species is newly described, and

the teleomorph-producing Fennellia nivea is found to belong elsewhere. Sect.

Flavi is considered to include 22 species, amongst which are several formerly

placed in other sections, and two new aflatoxin-producers distinguished by

extrolite profiles as well as sequence data; these have cholesterol-reducing

and hepatoxic or neurotoxic extrolites. Finally, 21 species are accepted in sect.

Usti, of which five are described as new. That section includes two species with

teleomorphs —Emericella heterothallica and Fennellia monodii— for which

binominals in Aspergillus (with a new combination for A. monodii) are used

in anticipation of the changes in the CopE (now the INTERNATIONAL CODE

OF NOMENCLATURE FOR ALGAE, FUNGI, AND PLANTS) subsequently effected in

July 2011.

The papers also demonstrate the value of the polyphasic approach, using

several different gene sequences, extrolite, cultural, and other features in

species circumscriptions. It was also gratifying to see such an almost seamless

transition to one name for one fungus adopted here by the key workers on

these organisms, which augurs well for other cases where a monophyletic

group has a particularly familiar anamorph-typified generic name. As has

become the norm in the SrupiEs, all papers are illustrated by exceptionally fine

photographs of conidial heads and colonies, many in colour, and an occasional

SEM micrograph. All struggling to identify Aspergillus species in the sections

treated will need to consult a copy, either by having one to hand or accessing it

free of charge through the CBS website.

Davip L. HAWKSWORTH

Departamento de Biologia vegetal II, Facultad de Farmacia,

Universidad Complutense de Madrid,

Plaza Ramon y Cajal, Madrid 28040, Spain

davidh@farm.ucm.es; d.hawksworth@nhm.ac.uk

MycotTaxon 117 Book Reviews ... 501

Cercosporoid fungi of India. By Kamal, 2010. Bishen Singh Mahendra Pal Singh,

23A New Connaught Place, Dehre Dhun, 248 001 Uttarakhand, India. Pp. vii + 351,

figs 47, pl. 1. ISBN 978-81-211-0753-2. Price: US$ 90.

Kamal, a former professor at Gorakhpur University, has been involved in

the documentation of Indian leaf-inhabiting fungi for over 30 years. In such

speciose groups as the cercosporoid fungi, in which over 3000 species have

been described, periodic regional syntheses are necessary to translate global

monographic work into those accessible at other geographical scales. Following

the landmark monograph of Chupp (1954), which accepted 1419 species, an

account of the Indian representatives was prepared by Vasudeva (1963), who

enumerated 260 species. Now, following on from the critical compilation of

Crous & Braun (2003; see MycoTAxon 89: 518, 2003), Kamal has similarly

provided an update for India. Of the 1815 cercosporoid fungi reported to have

been recorded from India up to 2009, 1050 are accepted, including descriptions

of four new species. These are dispersed between the genera Asperisporium (5

spp.), Cercospora (254), Distocercospora (1), Passalora (143), Prathigada (8),

Pseudocercospora (527), Scolecostigmina (12), Sirosporium (15), Stenellopsis

(1), Stigmina, Verrucisporota (4), and Zasmidium (88). A staggering 129 new

combinations are made, and seven replacement names introduced. It should

be noted that in the case of the new names and some combinations, “Kamal” is

omitted (corrected by an erratum slip pasted in at the front of the book). Also,

where new names are introduced, the authors of the replaced name should not

have been placed in parentheses — for example it should be C. holarrhenigena

“Kamal, nom. nov. and not “(R.K. Srivast., N, Srrivast. & A.K. Srivast.) nom.

nov. (p. 51).

In the arrangement of the entries within the accepted genera, the species

names precede the generic names, an unusual practice following Crous &

Braun (2003). However, here they are not treated as a single alphabetical series

but have been separated into the accepted genera. The core elements of the

nomenclatural information, including types, also seems to have been copied

directly from the 2003 work, with an identical layout and punctuation — and

the author does indicate in the introduction that the work is “based largely on

Crous and Braun (2003)” (p. 5). However, it is stated that type material in the

principle Indian herbaria was examined unless otherwise stated, where decisions

were based on descriptions and illustrations. References to hosts and localities

in India, in some cases Indian specimens, Indian literature reports, and in some

instances notes on the species are added. There are detailed descriptions and

line drawings of the newly described species, while other species are illustrated

to show “various morphotypes in a single genus” (p. 6).

It would have been helpful to provide potential users keys at least to the

genera, although the host list provided may facilitate identification. However,

502 ... Vellinga, BOOK REVIEW EDITOR

with no diagnostic information included (apart from the host plants) in the

species entries, it will be necessary for users to consult original accounts for

details necessary to confirm whether they have that taxon or another on

the same host. Nevertheless, this book represents an incredible amount of

dedicated work and provides the necessary basis for future revisionary studies

using molecular phylogenetic methods — something necessary to resolve

unequivocally species circumscriptions and the extent of host specificities in

the cercosporoid fungi’

Chupp C. 1954. A monograph of the fungus genus Cercospora. Ithaca, NY: C.

Chupp.

Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs. 1. Names

published in Cercospora and Passalora. Utrecht: Centraalbureau voor

Schimmelcultures.

Vasudeva RS. 1963. Indian Cercosporae. New Delhi: Indian Council of Agricultural

Research.

Davip L. HAWKSWwOoRTH

Departamento de Biologia vegetal I, Facultad de Farmacia,

Universidad Complutense de Madrid, Plaza Ramon y Cajal, Madrid 28040, Spain

davidh@farm.ucm.es; d.hawksworth@nhm.ac.uk

BASIDIOMYCETES

Strophariaceae s.l. By M.E. Noordeloos, 2011. Funei Europaer 13. Edizioni

Candusso, Via Ottone Primo 90, 17021 Alassio SV, Italy. <maxcandusso@libero.it>.

ISBN 978-88-905310-0-2. Pp. 648, pl. 377, figs 43. Price 69.00 €.

The Strophariaceae is one of the families in the Agaricales whose concept

has undergone huge changes — thanks to insights derived from DNA-based

phylogenetic analyses. In the morphological sense, it consisted of the brown-

spored Pholiota and the purple-grey dark-spored genera Stropharia, Psilocybe,

and Hypholoma. According to Matheny et al’s (2007) pivotal work, the family

forms one clade with the Hymenogastraceae and together they harbour, in

addition to the traditional genera, Hebeloma, Galerina, Agrocybe, Phaeocollybia,

and Alnicola (Naucoria).

Contrary to what is suggested by the book title, the book deals with the

family as originally described and excluding the modern additional genera: in

other words, Strophariaceae p.p. would have been a better title. However, as

changes in the genus concepts as depicted in recently published phylogenies

(e.g. Matheny et al. 2007) are accepted, the following genera are included:

Deconica (including Melanotus), Flammula, Hemistropharia, Hypholoma,

Kuehneromyces, Leratiomyces, Meottomyces, Phaeonematoloma, Pholiota,

Psilocybe, and Stropharia. This is a very welcome departure from the broad

MycotTaxon 117 Book Reviews ... 503

genus concept as given by the same author in his floristic treatment of this

group for the Dutch agaric flora (Noordeloos 1999). For Pholiota, the recent

European monograph by Holec (2001) is used as a template.

This book follows the general lay-out of the series, with an introduction

providing a history of the family and now current concepts and ideas, a short

introduction to the characters that are of value for the identification of the

species, and the taxonomic part that gives keys to the genera and species,

species descriptions, black-and-white drawings of microscopical characters,

a list of references, and almost 200 pages of colour photos —mainly of the

fruitbodies in their natural habitat, but also with a number of spore photos and

reproductions of important historical illustrations.

The book is bilingual throughout, with the original English text translated

into Italian.

The strength of this publication lies in the combination of descriptions and

illustrations; especially priceless is that multiple photos are given for many

species.

The keys are not easy to use, mainly because the differences between the

species are often subtle and difficult to put into words. I find it very frustrating

that the differences between Stropharia and Leratiomyces are not more clear-

cut, and I keep wondering whether acanthocytes are really restricted to the

genus Stropharia. As with any publication of this size, it is easy to find mistakes

that could have been caught during the publication process.

This book deserves to find a wide use, and not only in Europe. The price is

very reasonable for the amount of information this volume holds.

Holec J. 2001. The genus Pholiota in central and western Europe. Libri botanici

20.

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. 2007

[“2006”] Major clades of Agaricales: a multilocus phylogenetic overview.

Mycologia 98: 982-995.

Noordeloos ME. 1999. Strophariaceae. In Bas et al. (editors) Flora agaricina

neerlandica 4: 27-106. A.A. Balkema, Rotterdam.

Revision of Malaysian species of Boletales s.l. (Basidiomycota) described by

E.J.H. Corner (1972, 1974). By E. Horak, 2011. [MALaysIAN Forest RECORDS

51]. Forest Research Institute Malaysia, 52109 Kepong, Selangor Darul Ehsan,

Malaysia. <FRIM_Publications@frim.gov.my>. Pp. 245, figs. 127. US$ 53.

The 1972 publication of E.J.H. Corner’s BoLETus In Ma aysta (and a 1974

follow-up publication) represents a landmark in the taxonomy of boletoid

504 ... Vellinga, BOOK REVIEW EDITOR

fungi. The importance of this work is twofold. First, it provides the first (and

still largely unsurpassed) extensive exposition of the prodigious biodiversity of

boletes in tropical East Asia. Corner (1972) described nearly 100 new bolete

species; furthermore, he noted that many species could not be described due to

the sheer abundance of collections and environmental conditions that caused

many to decompose before they could be examined, as well as to the abundance

of suitable habitat, stating in regard to the latter: “When I consider the immense

number of places unexplored mycologically in Malaysia, I think 300 species of

boleti may be no exaggeration.” Second, Corner’s book called into question the

tidiness of generic concepts that were constructed around the comparatively

depauperate north temperate mycota. Corner maintained that the defining

characteristics of many bolete genera (such as color of the spore deposit) did

not hold when tested by the tropical mycota, but rather exhibited a continuum

between genera.

Despite the importance of this work, its impact on —and utility for—

subsequent taxonomic studies in tropical Asia and Australasia has been

somewhat diminished by taxonomic out-datedness and the restricted availability

of type collections and other material examined by Corner. In accordance

with his observations on intergradation of characters between genera, Corner

(1972) recognized only four bolete genera — Boletus, Gyroporus, Heimiella (now

Heimioporus), and Strobilomyces— while placing many taxa under subgenera

(e.g., Austroboletus, Leccinum, Tylopilus) that are widely recognized at generic

rank by other workers. Therefore, many taxa are in need of nomenclatural

revision to bring them up to date with current taxonomic concepts. As a result

of high humidity in both field and herbarium storage localities, primitive field

conditions for drying specimens, and storage methods, many of Corner’s

collections were damaged by molds, degraded by storage in alcohol formalin

(or hardened by evaporation of AF during storage), or rendered toxic by

treatment with mercuric chloride; restricted access to collections and restricted

accessibility of important microcharacters have been the result.

Given the importance of Corner’s collections and the difficulties inherent in

their study, the present revision by Egon Horak is a most welcome and important

contribution to the taxonomic study of boletes. Dr. Horak presents revised

descriptions of taxa combining information from Corner’s original protologues

and Latin diagnoses (the latter translated into English to facilitate study), data

gained from additional studies of Corner’s holotypes and other collections,

and data from studies of holotype collections from other collectors (including

Baker, Heim, Hohnel, Hooker, and Ridley, among others) for those species

included but not described by Corner. Horak provides analyses of 160 (124

accepted, 22 doubtful, and 14 rejected, excluded, or invalid) species, including

emended descriptions, line drawings of microscopic features (127 figures),

MycotTaxon 117 Book Reviews ... 505

lists of observations and conclusions from his studies, and nomenclatural

changes necessary for bringing the nomenclature of Corner’s taxa up-to-date

with current concepts. Additional features include taxonomic keys to all of the

treated taxa, a list of additional important taxonomic studies of boletes from

the Far East, a synopsis of all of the treated taxa arranged by currently accepted

genus, and species list indexed by specific epithet. Perhaps most importantly,

Horak brings to this study his own extensive personal experience working in

Malaysia and the Asian tropics, and is therefore able to conceptualize Corner’s

taxa in a broad comparative context.

Any taxonomic revision requires some judgments as to the placement of

species in more inclusive taxa. In the boletes, a longstanding taxonomic issue

involves character weighting of species with pale short-elliptical or phaseoliform

basidiospores; an example of this issue is Rubinoboletus, in which species have

been placed that share this spore morphology but differ significantly in other

characteristics (see Osmundson and Halling 2010). In the present revision,

Horak places a number of species with pale phaseoliform basidiospores in

Gyroporus, and in so doing emphasizes spore morphology over characteristics

(e.g., presence/absence of clamp connections, orientation of hyphae in the stipe)

emphasized by some previous authors. In the case of Gyroporus balloui (Peck)

E. Horak (= Tylopilus balloui (Peck) Singer), this placement is contradicted

by molecular evidence presented after the preparation of Dr. Horak’s volume

(Osmundson and Halling 2010), but other species remain to be evaluated in the

light of DNA sequence data. In the meantime, a provisional placement based

on spore morphology seems reasonable; this example emphasizes that some of

our present taxonomic concepts contain inherent contradictions and may very

well change in the light of additional data and analyses.

In BOLETUS IN MALaysiA, Corner - considering existing taxonomic problems

in the boletes - wrote, “I have no doubt that if the fungus flora of Malaysia

can be explored before the destruction of the main forests or the elimination

of the non-commercial trees, fresh discoveries will resolve these problems.”

Biodiversity discovery, conservation, and the solution to taxonomic problems

share the need for carefully researched scholarly work on the description and

revision of species and higher taxa. Horak states that “the present revision is

only one further step in the direction to unravel the numerous bolete taxa

described from SE-Asia and Australasia;” however, his “one further step” is

a critical one: by providing a detailed re-examination of Corner’s specimens

and by bringing the taxonomy of Corner’s taxa into a modern context, Horak

has made two highly valuable contributions to the study of boletes. In this

outstanding volume, Horak will not only earn the gratitude of boletologists, but

has provided an important tool for assessing diversity, rarity, endemicity and

—unfortunately, but quite likely, given the extent and rate of forest conversion

506 ... Vellinga, BOOK REVIEW EDITOR

in Malaysia— extinction of the morphologically striking and ecologically

important bolete fungi of the East Asian tropics.

Corner EJH. 1972. Boletus in Malaysia. Government Printing Office, Singapore.

263 pp.

Corner EJH. 1974. Boletus and Phylloporus in Malaysia: further notes and

descriptions. Garden's Bull. Singapore 27:1-16.

Osmundson TW, Halling RE. 2010. Tylopilus oradivensis sp. nov.:a newly described

member of the Tylopilus balloui complex from Costa Rica. Mycotaxon 113:

475-483. http://dx.doi.org/10.5248/113.475

Topp W. OSMUNDSON

Department of Environmental Science, Policy & Management

University of California, Berkeley, CA 94720, U.S.A.

toddo@berkeley.edu

MYCOFLORISTICS

A guidebook to the macrofungi of Fraser’s Hill. By B.K. Thi, S.S. Lee, N.

Zanuddin & H.T. Chan, 2011. Strr ALam Dan Rimpa No. 14. Forest Research

Institute Malaysia, 52109 Kepong, Sealngor Darul Ehsan, Malaysia, <FRIM_

Publications@frim.gov.my>. ISBN 978-967-5221-63-7. Pp xi + 93, plates. Price US$

This little guide starts with an introduction to Fraser’s Hill, situated in the state

of Pahang, Malaysia, around 100 km from the capital, Kuala Lumpur. Fraser's

Hill is not only a draw for tourists, it is also well known among mycologists (see

e.g. Tan et al. 2009). The mountainous terrain is accessible along some well-

maintained trails and a road. The world of the fungi is also introduced to the

readers, after which the main part follows with photos and short descriptions

and notes on the most colourful and attractive mushroom-forming fungi of

all taxonomic groups. It is a pleasure to see that the showy purple floccose

Lepiota-like species, recently placed in the new genus Coniolepiota (Vellinga

et al. 2011), is also present in this part of Malaysia, and that another Lepiota-

like taxon in the L. furfuraceipes group (described from China) extends into

Malaysia as well. A nice collection of boletes is illustrated. We hope that this

book will be widely available at the park, opening the eyes of the visitors to the

exciting world of fungi. And that it will serve as an example for other forests in

Malaysia and beyond.

Tan Y-S, Desjardin DE, Perry BA, Vikineswary S, Noorlidah A. 2009. Marasmius

sensu strico in Peninsular Malaysia. Fungal Diversity 37: 9-100.

Vellinga EC, Sysouphanthong P, Hyde KD. 2011. The family Agaricaceae:

phylogenies and two new white-spored genera. Mycologia 103: 494-509.

http://dx.doi.org/10.3852/10-204

MycotTaxon 117 Book Reviews ... 507

Los hongos de Panama. Introduccion a la identificacion de los macroscopicos.

By G. Guzman & M. Piepenbring, 2011. Pp. 386. pl. 798. Price US$ 45.

Panama is a well-known biodiversity hotspot, and yet, little is known about

its fungal diversity. Recently a preliminary checklist of fungi was published

(Piepenbring 2006) in which around 1800 taxa are listed. A new, richly illustrated

book of this Central American state shows the diversity of the macrofungi and

demonstrates that our knowledge of the fungi is still insufiicient with many

species still to be undescribed.

The book is divided into four parts: an introduction to the country, its

habitats, and the fungi; the classification of the fungi; a key to the species treated

in the book; and lastly, but most importantly, the species are introduced. They

are treated alphabetically with colour photos provided for each species and

with line drawings of microscopical details also present. A glossary, index, and

a list of the species in a taxonomic framework complete the work.

A bit unusual, but welcome, for a book like this is that lichens are also

included as well as the slime molds.

The illustrations are often quite large, covering a complete page. In many

cases, several photos are given for one species, showing the mushroom from

different angles or in different stages.

It is really interesting to see that some ‘temperate’ species —such as

Cortinarius violaceus— occur in Panama. But the highlight of a book like this

is that it introduces non-tropical readers to the unfamiliar forms and shapes of

tropical fungi such as Staheliomyces cinctus and Poronia oedipus.

The genus names are in many cases not quite up to date (e.g. Collybia for

what now is called Gymnopus; Coprinus instead of Coprinopsis); photos are not

always sharp, and colours seem to be a little off in several cases.

But the biggest asset of a book like this is that it shows clearly that the times

of fungal discovery and taxonomy are far from over.

Piepenbring, M. 2006. Checklist of fungi in Panama. Preliminary version. Puente

Bioldgica 1: 1-190.

Book ANNOUNCEMENT

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. 414 pp. ill. circa US$ 154.00

MYCOTAXON

http://dx.doi.org/10.5248/117.508

Volume 117, p. 508 July-September 2011

Regional annotated mycobiotas new to www.mycotaxon.com

Mycotaxon is pleased to announce the addition of a new url link on our “web-list” page

at www.mycotaxon.com//resources/weblists.html, which will connect to the checklist PDF

now available for free download from the posted off-site webpage. The content and design

of each list is the sole responsibility of its authors and their three (or more) expert reviewers,

after which those meeting MycotTaxon’s scientific and nomenclatural criteria are accepted

for posting. The author, title, and abstract of our most recently accepted web-list is provided

below.

EURASIA AND THE MID-EAST

Armenia, Azerbaijan, Georgia, Iran, Russia, Turkey

Ghobad-Nejhad, Masoomeh. Updated checklist of corticioid and poroid

basidiomycetes of the Caucasus region. 72 pp.

ApstTrAcT — A first annotated checklist of corticioid and poroid basidiomycetes

of the Caucasus region was published in 2009 (Ghobad-Nejhad et al., Mycologia

Balcanica 6: 123-168, 2009). A new, updated version of the checklist is presented

and is aimed to be regularly updated at an Iranian mycological website. The list

includes 621 accepted species names with their synonym(s) used in the bibliographic

literature. Major updates include: 1) addition of substratum data for the species based

on numerous specimens examined from different herbaria, 2) addition of literature

references, distribution data and species names not present in the earlier version,

and deletion of names confirmed not to occur in the region, and 3) revision of

nomenclature and synonyms. Compared to the 2009 list, 86% of species in the current

checklist are supported by specimens examined. The aim to post the checklist in an

online version is to provide a permanent repository for keeping the list updated with

no unnecessary delay and for making the data available to the mycologists interested

in fungal diversity of the Caucasus biodiversity hotspot. New records for the region

will be published online and the users will be able to learn about the most recent

changes to the checklist via subscription to a feed (RSS). The link to the checklist is:

http://www.myco-lich.com/mycology-of-iran/basidiomycota/caucasus-region/checklists

MYCOTAXON

http://dx.doi.org/10.5248/117.509

Volume 117, pp. 509-515 July-September 2011

Fungal nomenclature.

Changes to publication requirements made at the

XVIII International Botanical Congress in Melbourne

— what does e-publication mean for you?

SANDRA KNaApP’, JOHN MCNEILL? & NICHOLAS J. TURLAND?

' Department of Botany, The Natural History Museum, Cromwell Road, London SW7 5BD, U.K.

? Royal Botanic Garden, Edinburgh, 20A Inverleith Row, Edinburgh EH3 5LR, U.K.

* Missouri Botanical Garden, P.O. Box 299, St. Louis MO 63166-0299, U.S.A.

*CORRESPONDENCE TO: s.knapp@nhm.ac.uk

ABSTRACT — Changes to the INTERNATIONAL CODE OF BOTANICAL NOMENCLATURE are

decided on every six years at Nomenclature Sections associated with International Botanical

Congresses (IBC). The XVIII IBC was held in Melbourne, Australia; the Nomenclature Section

met from 18-22 July 2011 and its decisions were accepted by the Congress at its plenary session

on 30 July. Several important changes were made to the CoDE as a result of this meeting

that will affect publication of new names. Two of these changes will come into effect on

1 January 2012, some months before the Melbourne Copbz is published. Electronic material

published online in Portable Document Format (PDF) with an International Standard Serial

Number (ISSN) or an International Standard Book Number (ISBN) will constitute effective

publication, and the requirement for a Latin description or diagnosis for names of new taxa

will be changed to a requirement for a description or diagnosis in either Latin or English. In

addition, effective from 1 January 2013, new names of organisms treated as fungi must, in

order to be validly published, include in the protologue (everything associated with a name

at its valid publication) the citation of an identifier issued by a recognized repository (such

as MycoBank). Draft text of the new articles dealing with electronic publication is provided

and best practice is outlined.

To encourage dissemination of the changes made to the INTERNATIONAL CODE OF

NOMENCLATURE FOR ALGAE, FUNGI, AND PLANTS, this article will be published in BOTANICAL

JOURNAL OF THE LINNEAN SOCIETY, BMC EVOLUTIONARY BIOLOGY, CLADISTICS,

Mycotaxon, MycoKeys, NEw PHYTOLOGIST, NORTH AMERICAN FUNGI, NOVON, OPUSCULA

PHILOLICHENUM, PHYTOKEYS, PHYTONEURON, PHyTOTAxA, PLOS ONE, SYSTEMATIC

BoTAny, and TAXxon.

Introduction

At the XVIII International Botanical Congress in Melbourne, Australia,

in July 2011, two important changes were made to the INTERNATIONAL

510 ... Knapp, McNeill & Turland

CopE OF BOTANICAL NOMENCLATURE (now the INTERNATIONAL CODE OF

NOMENCLATURE FOR ALGAE, FUNGI, AND PLANTS) that will take effect from

1 January 2012. These changes will affect everyone who publishes names

governed by this Cope. As the Melbourne Cope will not be published until

approximately mid-2012, we felt it would be helpful to outline these changes,

particularly those concerning effective publication in electronic media (in

Articles 29, 30, and 31). For a concise report on all the changes to the CoDE

accepted in Melbourne, see McNeill & al. (2011).

A draft wording of the revised Articles, Notes, and Recommendations on

effective publication is provided to aid editors and publishers in establishing

best practice for implementing this aspect of the Cop. We also outline here

what these changes do NOT mean, in order to guide those wishing to publish

new names and typifications by electronic means. We urge readers to consult

the report of the Special Committee on Electronic Publication accompanying

the changes proposed prior to the Congress (Chapman & al. 2010), wherein the

reasoning for the changes now accepted into the CopE is set out.

Draft wording of revised Articles 29, 30, and 31 and

Recommendations 29A, 30A, and 31A

Here we reproduce the wording of all of the relevant Articles, Notes, and

Recommendations (omitting the Examples), with the changes highlighted in

bold. The wording here is provisional, pending the meeting of the Editorial

Committee in December 2011 to finalize the printed version of the Melbourne

CODE.

Article 29

29.1. Publication is effected, under this Cope, by distribution of printed matter

(through sale, exchange or gift) to the general public or at least to botanical institutions

with libraries accessible to botanists generally. Publication is also effected by electronic

distribution of material in Portable Document Format (PDF; see also Art. 29.3

and Rec. 29A.1) in an online publication with an International Standard Serial

Number (ISSN) or an International Standard Book Number (ISBN). Publication is

not effected by communication of new names at a public meeting, by the placing of

names in collections or gardens open to the public, by the issue of microfilm made from

manuscripts, typescripts or other unpublished material, or by distribution electronically

other than as described above.

29.2. For the purpose of this Article, “online” is defined as accessible electronically

via the World Wide Web.

29.3. Should Portable Document Format (PDF) be succeeded, a successor

international standard format communicated by the General Committee (see Div.

III) is acceptable.

29.4. The content ofa particular electronic publication must not be altered after it is

first issued. Any such alterations are not themselves effectively published. Corrections

or revisions must be issued separately to be effectively published.

Melbourne Code and e-publication ... 511

Recommendation 29A

[Existing Recommendation replaced by the following]

29A.1. Publication electronically in Portable Document Format (PDF) should

comply with the PDF/A archival standard (ISO 19005).

29A.2. Authors should preferably publish in publications that are archived,

satisfying the following criteria as far as is practical (see also Rec. 29A.1):

(a) The material should be placed in multiple trusted online digital repositories,

e.g. an ISO-certified repository;

(b) Digital repositories should be in more than one area of the world and preferably

on different continents;

preferably on different continents is also advisable.

Article 30

30.1. Publication by distribution of electronic material does not constitute effective

publication before 1 January 2012.

30.2. An electronic publication is not effectively published if there is evidence

associated with or within the publication that it is merely a preliminary version that

was, or is to be, replaced by a version that the publisher considers final, in which case

only that final version is effectively published.

30.3. Publication by indelible autograph before 1 January 1953 is effective. Indelible

autograph produced at a later date is not effectively published.

30.4. For the purpose of this Article, indelible autograph is handwritten material

reproduced by some mechanical or graphic process (such as lithography, offset, or

metallic etching).

30.5. Publication on or after 1 January 1953 in trade catalogues or non-scientific

newspapers, and on or after 1 January 1973 in seed-exchange lists, does not constitute

effective publication.

30.6. The distribution on or after 1 January 1953 of printed matter accompanying

exsiccatae does not constitute effective publication.

Note 1. If the printed matter is also distributed independently of the exsiccata, it is

effectively published.

30.7. Publication on or after 1 January 1953 of an independent non-serial work stated

to be a thesis submitted to a university or other institute of education for the purpose

of obtaining a degree is not effectively published unless it includes an explicit statement

(referring to the requirements of the Cope for effective publication) or other internal

evidence that it is regarded as an effective publication by its author or publisher.

Note 2. The presence of an International Standard Book Number (ISBN) or a

statement of the name of the printer, publisher, or distributor in the original printed

version is regarded as internal evidence that the work was intended to be effectively

published.

Recommendation 30A

30A.1. Preliminary and final versions of the same electronic publication should be

clearly indicated as such when they are first issued.

30A.2. It is strongly recommended that authors avoid publishing new names and

descriptions or diagnoses of new taxa (nomenclatural novelties) in ephemeral printed

512 ... Knapp, McNeill & Turland

matter of any kind, in particular printed matter that is multiplied in restricted and

uncertain numbers, in which the permanence of the text may be limited, for which

effective publication in terms of number of copies is not obvious, or that is unlikely

to reach the general public. Authors should also avoid publishing new names and

descriptions or diagnoses in popular periodicals, in abstracting journals, or on

correction slips.

30A.3. To aid availability through time and place, authors publishing nomenclatural

novelties should give preference to periodicals that regularly publish taxonomic articles.

Otherwise, a copy of a publication (whether published as printed or electronic

matter) should be sent to an indexing centre appropriate to the taxonomic group, and

publications that exist only as printed matter should be deposited in at least ten, but

preferably more, botanical or other generally accessible libraries throughout the world.

30A.4. Authors and editors are encouraged to mention nomenclatural novelties in

the summary or abstract, or list them in an index in the publication.

Article 31

31.1. The date of effective publication is the date on which the printed or electronic

matter became available as defined in Art. 29 and 30. In the absence of proof establishing

some other date, the one appearing in the printed or electronic matter must be accepted

as correct.

[Existing Note 1 replaced by the following]

31.2. When a publication is issued in parallel electronic and printed versions,

these must be treated as effectively published on the same date unless the dates of the

versions are different according to Art. 31.1.

31.3. When separates from periodicals or other works placed on sale are issued in

advance, the date on the separate is accepted as the date of effective publication unless

there is evidence that it is erroneous.

Recommendation 31A

31A.1. The date on which the publisher or publisher's agent delivers printed matter

to one of the usual carriers for distribution to the public should be accepted as its date

of effective publication.

Best practice

Authors of new names, editors and publishers will all be interested in

ensuring that the publications including new names are in accordance with

the Melbourne Copg, so that the names therein are effectively published. We

suggest that those publishing in journals or monograph series and books that

have online editions communicate with the editors so that best practice can

be established across the community as quickly as possible. Many publishers

have been carefully addressing the issues involved with the e-publication of

novelties for some time (see Knapp & Wright 2010; guidelines in PLoS ONE,

http://www.plosone.org/static/policies.action#taxon) and considerable interest

in making these new CobDE changes function effectively has been apparent.

Some practices that we feel will help with the initial stages of e-publication

of novelties that are according to the Melbourne CopE are:

Melbourne Code and e-publication ... 513

e Having each article bear the date of publication prominently (as is done

in many journals, for example NEw PHYTOLOGIST or NATURE).

e If an online early version is issued that is not the same as the final

version (and thus not the place of effective publication) stamp each

article with this fact prominently (for example AMERICAN JOURNAL

OF BOTANY).

e Prominent display of the ISSN or ISBN of the publication on each

article will help indexers establish effective publication.

e Publication in journals (or monograph series) that participate in the

CLOCKSS system (see Knapp & Wright, 2010, for a description) or

another international archive and preservation system will ensure

long-term archiving.

e Authors of new names by electronic means should alert the appropriate

indexing center as recommended in Rec. 30.A.3 - this will help

indexers who may otherwise not be aware of electronically published

names.

What these changes do not mean

Although the new Articles and Recommendations use the terms PDF

and PDF/A, this does not mean that publications must be issued only in that

format to be effectively published. For example, some online journals issue

papers in Hypertext Markup Language (HTML) format together with a parallel

PDF version. In such cases, the PDF version will be effectively published.

The stipulation that the General Committee for Botanical Nomenclature will

communicate the acceptability of a new international standard format, should

PDF ever be succeeded, means authors of novelties and the community using

the CopE can remain informed as to advances in the field and that the CopE

will be protected from obsolescence.

Use of the following means of electronic publication will NoT result in

effective publication of novelties under the Melbourne Cope.

¢ Publication on websites or in ephemeral documents available over the

Internet (there are strict criteria for granting of ISSNs — see http://

www.issn.org/).

e Publication in journals without a registered ISSN or e-ISSN.

e Publication in books without a registered ISBN or e-ISBN.

The Recommendation approved to advise the deposition of a hard copy of any

e-publication in a library suggests to botanists an action, but it does not set out

standard practice or a protocol for librarians to follow. Librarians are themselves

in a complex transition zone between publication modalities (Johnson & Luther

514 ... Knapp, McNeill & Turland

2007), and botanists may find them unwilling or unable to accommodate single

hard copy papers as individual accessions should the volume be great.

Two other important changes to the Code relating to

the publication of names

The second change to the CoDE approved in Melbourne to take effect from 1

January 2012 is that the description or diagnosis required for valid publication

of the name of a new taxon of all organisms falling under the CoDE may be in

either English or Latin. This is the current provision for names of plant fossils,

but all new non-fossil taxa have required a Latin description or diagnosis (fungi

and plants from 1 January 1935; algae [including cyanobacteria, if treated

under the Cope] from 1 January 1958). This has no bearing on the form of

scientific names, which continue to be Latin or treated as Latin. Individual

journal requirements for Latin and/or English will, of course, be determined by

the editors of those journals.

A third change to the CopE approved in Melbourne relating to publication

of names, but one not taking effect until 1 January 2013 (not 1 January 2012 as

reported by Miller & al., 2011), is that all new names of organisms treated as

fungi must, as an additional requirement for valid publication, include in the

protologue the citation of an identifier issued by a recognized repository (such

as MycoBank, http://www.mycobank.org/). This will be publicized separately.

The requirement for a unique identifier for new names of fungi on or after 1

January 2013 does Not apply to plants or algae; there is no need for authors of

new names in these groups to request LSIDs (Life Science Identifiers) — or other

identifiers — from indexing centers.

Acknowledgements

SK is supported by the NSF's Planetary Biodiversity Inventory programme (DEB-

0316614, ‘PBI Solanum —- a worldwide treatment). JMcN’s and NJT’s attendance at the

Nomenclature Section of the XVII IBC in Melbourne was supported in part by the

International Association for Plant Taxonomy (IAPT). We thank Katherine Challis

(Kew) for helpful comments.

To encourage dissemination of the changes made to the INTERNATIONAL CODE

oF NOMENCLATURE FOR ALGAE, FUNGI, AND PLANTS, this article will be published

in BOTANICAL JOURNAL OF THE LINNEAN SOCIETY, BMC EVOLUTIONARY BIOLOGY,

CLADISTICSs, MycoTaxon, MycoKeys, NEw PHYTOLOGIST, NORTH AMERICAN FUNGI,

Novon, OPUSCULA PHILOLICHENUM, PHYTOKEYS, PHYTONEURON, PHYTOTAXA, PLOS

ONE, SysTEMATIC BOTANY, and Taxon.

Melbourne Code and e-publication ... 515

Literature cited

Chapman AD, Turland NJ, Watson MF (eds.). 2010. Report of the Special Committee on Electronic

Publication. Taxon 59: 1853-1862.

Johnson RK, Luther J. 2007. The E-only tipping point for journals: what's ahead in the print-to-

electronic transition zone. Association of Research Librarians, Washington DC.

Knapp S, Wright D. 2010. E-publish or perish? Pp. 83-93 in: A. Polaszek (ed.), SysTEMA NATURAE

250 — THE LINNAEAN ARK. Taylor and Francis, London.

McNeill J, Turland NJ, Monro A, Lepschi BJ. 2011. XVIII International Botanical Congress:

preliminary mail vote and report of Congress action on nomenclature proposals. Taxon 60,

in press.

Miller JS, Funk VA, Wagner WL, Barrie F, Hoch PC, Herendeen P. 2011. Outcomes of the 2011

Botanical Nomenclature Section at the XVIII International Botanical Congress. PHyTOKEYs

pe i-3,

MY COTAXON

http://dx.doi.org/10.5248/117.517

Volume 117, pp. 517-519 July-September 2011

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 117

Albahypha Oehl, G.A. Silva, B.T. Goto & Sieverd., p. 308

Albahypha drummondii (Blaszk. & Renker) Sieverd., Oehl, B.T. Goto &

G.A. Silva, p. 308

Albahypha walkeri (Btaszk. & Renker) Sieverd., Oehl, B.T. Goto & G.A. Silva, p. 309

Amanita conara Tulloss & Halling, p. 180

Amanita costaricensis Tulloss, Halling, G.M. Muell. & Singer, p. 189

Amanita garabitoana Tulloss, Halling & G.M. Muell., p. 169

Amanita vernicoccora Bojantchev & R.M. Davis, p. 491

Ambispora nicolsonii (C. Walker, L.E. Reed & EE. Sanders) Oehl, G.A. Silva,

B.T. Goto & Sieverd., p. 431

Anaselenosporella indica Pratibha, Bhat & Raghuk., p. 269

Arachnophora goanensis Pratibha, Bhat & Raghuk., p. 272

Archaeospora myriocarpa (Spain, Sieverd. & N.C. Schenck) Oehl, G.A. Silva,

B.T. Goto & Sieverd., p. 430

Archaeospora undulata (Sieverd.) Sieverd., G.A. Silva, B.T. Goto & Oehl, p. 430

Arthrobotrys latispora H.Y. Su & X.Y. Yang, p. 32

Caloplaca allochroa Y. Joshi, Vondrak & Hur, p. 263

Cephalotrichum acutisporum J.J. Xu & T.Y. Zhang, p. 208

Cephalotrichum castaneum (Y.L. Jiang & T.Y. Zhang) Y.L. Jiang & T.Y. Zhang, p. 224

Cephalotrichum cylindrosporum Y.L. Zhang & T.Y. Zhang, p. 209

Cephalotrichum ellipsoideum H.Q. Pan & T.Y. Zhang, p. 211

Cephalotrichum inflatum Y.L. Jiang & T.Y. Zhang, p. 213

Cephalotrichum longicollum Y.L. Jiang & T.Y. Zhang, p. 213

Cephalotrichum macrosporum Y.L. Jiang & T.Y. Zhang, p. 214

Cephalotrichum oblongum J.J. Xu & T.Y. Zhang, p. 216

Cephalotrichum ovoideum Y.L. Jiang & T.Y. Zhang, p. 217

Cephalotrichum robustum Y.L. Jiang & T.Y. Zhang, p. 218

Cephalotrichum spirale H.M. Liu, H.Q. Pan & TY. Zhang, p. 220

518 ... MycoTAXON 117

Cephalotrichum terricola Y.L. Jiang & T.Y. Zhang, p. 221

Cephalotrichum verrucipes Y.L. Jiang & T.Y. Zhang, p. 223

Cephalotrichum verrucisporum (Y.L. Jiang & T.Y. Zhang) Y.L. Jiang & T.Y. Zhang, p. 224

Ceratocystis mangicola M. van Wyk & M.J. Wingf., p. 395

Ceratocystis mangivora M. van Wyk & MJ. Wingf., p. 397

Cetraspora nodosa (Blaszk.) Oehl, G.A. Silva, B.T. Goto & Sieverd., p. 431

Cortinarius callimorphus Bojantchev & R.M. Davis, p. 3

Corynespora fujianensis L.G. Ma & X.G. Zhang, p. 355

Craspedodidymum fujianense L.G. Ma & X.G. Zhang, p. 351

Daldinia grayana Lar.N. Vassiljeva & S.L. Stephenson, p. 332

Dendrothele latenavicularis Gorjén, p. 102

Diatrypella informis Ellis & Everh. ex Lar.N. Vassiljeva & S.L. Stephenson, p. 335

Ellisembia schimae Jian Ma & X.G. Zhang, p. 247

Endophragmiella clausenae L.G. Ma & X.G. Zhang, p. 280

Endophragmiella nanlingensis S.C. Ren & X.G. Zhang, p. 123

Endophragmiella pentaphylacis L.G. Ma & X.G. Zhang, p. 281

Gymnopus fuscotramus MeSi¢c, Tkaléec & Chun Y. Deng, p. 324

Junghuhnia minor HS. Yuan, p. 256

Lepiota aureofulvella Sysouphanthong, K.D. Hyde, Chukeatirote & Vellinga, p. 62

Lepiota pallidiochracea J.F. Liang & Zhu L. Yang, p. 360

Lepiota papillata Sysouphanthong, K.D. Hyde, Chukeatirote & Vellinga, p. 75

Lignincola conchicola J.K. Liu, E.B.G. Jones & K.D. Hyde, p. 344

Linkosia hibisci Jian Ma & X.G. Zhang, p. 249

Macrolepiota rhodosperma var. velicopia Vizzini & Contu, p. 153

Macrolepiota procera f. fuliginosa (Barla) Vizzini & Contu, p. 158

(lectotypified, epitypified)

Macrolepiota procera f. permixta (Barla) Vizzini & Contu, p. 161

(epitypified)

Minimelanolocus linderae Jian Ma & X.G. Zhang, p. 131

Minimelanolocus mori Jian Ma & X.G. Zhang, p. 133

Moelleriella pumatensis T.T. Nguyen & N.L. Tran, p. 48

Mycena lomavritha Manim., p. 244

= Mycena indica Manim. & Leelav. 1989 (“1988”),

non Mycena indica Sarwal & Rawla 1983

Mycena saloma Aravind. & Manim., p. 240

Nemania pseudoillita (Lar.N. Vassiljeva) Lar.N. Vassiljeva & S.L. Stephenson, p. 339

Passalora barringtoniigena R. Singh, Sham. Kumar & Kamal, p. 138

Passalora wangii (F.Y. Zhai, Y.L. Guo & Yu Li) FY. Zhai, Y.L. Guo & Yu Li, p. 365

Phylloporus fagicola Montoya & Bandala, p. 10

NOMENCLATURAL NOVELTIES & TYPIFICATIONS ..

Pouzarella albostrigosa Largent & Abell-Davis, p. 470

Pouzarella debilis (Corner & E. Horak) Largent & Abell-Davis, p. 463

Pouzarella farinosa Largent & Skye Moore, p. 455

Pouzarella fusca Largent & Abell-Davis, p. 466

Pouzarella lageniformis Largent & Skye Moore, p. 443

Pouzarella lasia (Berk. & Broome) Largent & Abell-Davis, p. 474

Pouzarella pamiae Largent, p. 450

Pouzarella parvula Largent & Skye Moore, p. 447

Pouzarella pilocystidiata Largent & Skye Moore, p. 439

Pouzarella setiformis Largent & Abell-Davis, p. 459

Pseudocercospora kamalii Rajeshkumar, Rahul Sharma & S.K. Singh p. 232

Pseudocercospora miliusae R. Singh, Sham. Kumar & Kamal, p. 140

Rhexodenticula zhengii D.W. Li & Jing Y. Chen, p. 288

Sacculospora Oehl, Sieverd., G.A. Silva, B.T. Goto, I.C. Sanchez & Palenz., p. 311

-519

Sacculospora baltica (Blaszk., Madej & Tadych) Oehl, Palenz., I.C. Sanchez, B.T. Goto,

G.A. Silva & Sieverd, p. 311

Sacculosporaceae Oehl, Sieverd., G.A. Silva, B.T. Goto, ILC. Sanchez & Palenz., p. 310

Septobasidium atalantiae S.Z. Chen & L. Guo, p. 291

Stanjehughesia micheliae Jian Ma & X.G. Zhang, p. 251

Terriera huangshanensis Z.Z. Yang, Y.R. Lin & C.L. Hou, p. 368

Tricispora Oehl, Sieverd., G.A. Silva & Palenz., p. 310

Tricispora nevadensis (Palenz., N. Ferrol, Azcon-Aguilar & Oehl) Oehl, Palenz.,

G.A. Silva & Sieverd., p. 310